OLympus

CLINICAL CHEMISTRY
REAGENT GUIDE
Beckman Coulter, Inc., 250 S. Kraemer Blvd.,Brea, CA 92821, USA.
EC REP Beckman Coulter Biomedical Limited, Lismeehan, O' Callaghan's Mills, Co. Clare, Ireland, Tel: ++353 65 6831100.
EN -i–
This guide is intended for use with:
BECKMAN COULTER AU400
BECKMAN COULTER AU600/600IVD
Version .11
Revision date: 2010-06
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CONTENTS
1. OVERVIEW
2. INSTRUCTIONS
2. 1 Guide Format.
2. 2 Guide Updates.
3. SYMBOLS
4. GENERAL PRECAUTIONS AND WARNINGS
5. NOTES
APPENDIX 1 PRODUCT GROUPINGS
APPENDIX 2 TABLE OF CALIBRATORS AND CONTROLS
APPENDIX 3 GLOSSARY **
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1. OVERVIEW
This guide is designed to provide specific chemistry information for the Beckman Coulter clinical
chemistry systems.
Please note that the latest revision of the Instructions For Use (IFU) and Setting Sheets can be
found on the Beckman Coulter website at; www.beckman coulter.com.
Access to this website requires registration; therefore it is recommended that you register as soon
as possible.
The guide does not include all the requirements governing the safe and effective operation of the
Beckman Coulter clinical chemistry systems.
Refer to the analyser specific User Guide and your specific laboratory procedures governing
other areas, such as corrective actions, QC policies, or hazardous waste disposal that are not
addressed in this guide.
Beckman Coulter logo is a trademark of Beckman Coulter, Inc. and is registered in the USPTO.
2. INSTRUCTIONS
2.1 Guide Format:

This guide contains the Instructions For Use (IFU), which describe how the reagent is to be
used, and Setting Sheets for the Beckman Coulter range of reagent kits, which describe the
required Beckman Coulter Chemistry Analyser settings to use with the specified reagents.
The outer label on the Beckman Coulter reagent kit identifies the Part No. for the reagent and the
applicable revision code for the IFU and the Setting Sheet. Please refer to this each time a
kit is used – see below. Additionally, please ensure the use of the appropriate Setting
Sheet for application with the biological fluid under investigation.
Example: REF: OSR6287
BLOSR6x87.01 Î IFU reference and revision code.

BSOSR6x87.01 Î Setting sheet reference and revision code.
If the indicated revision or a higher revision is not present, then please contact your local
Beckman Coulter support organisation. Alternatively, you may access this document on the
Beckman Coulter website as described in Section 1.
Note: The ‘x’ in the reference code denotes a multiple kit format. For reference code
BLOSR6x87 incorporates OSR6187, OSR6287 and OSR6587.
2.2 Guide Updates:

Updates to IFUs and Setting Sheets will be delivered to you as and when these become
available. These updates should be inserted into the guide and the older version removed
(unless specifically required) so that the latest information is available at all times.
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3. SYMBOLS
The following symbols are used in the labelling of Beckman Coulter System Reagents.
Manufacturer Biological Risk
In Vitro Diagnostic Medical Device Contents
Consult Instructions for Use Protect from Light
Material safety data sheet available to

Temperature Limitation professional users on request.
Catalogue Number Serial Number
Keep Upright Caution, consult accompanying documents
Batch code Product conforms to the IVD directive
Use by:
For IVD performance evaluation only (YYYY-MM-DD or YYYY-MM)
Authorized Representative in the

European Community
4. GENERAL PRECAUTIONS AND WARNINGS

The following precautions and warnings are valid for all tests.
• There are literature reports of very rare cases where gammopathy, especially monoclonal IgM
(Waldenström’s macroglobulinemia), may cause protein precipitation when mixed with reagents giving rise
to turbidity. This may occasionally lead to unreliable results when processing such samples using
photometric methods. Further background information can be obtained from the following paper and its
references: “Berth M, Delanghe J. Protein precipitation as a possible important pitfall in the clinical
chemistry analysis of blood samples containing monoclonal immunoglobulins: 2 case reports and a review
of the literature. Acta Clin Belg. 2004;59:263-73.”
Due to the idiosyncratic nature of this interference it can theoretically occur with any liquid reagent. Where
Beckman Coulter has received reports of this type of interference, or for those reagents considered at
highest risk, a warning statement is included in the Interfering Substances section of the appropriate
reagent.
• Please note that recovery of non-Beckman Coulter controls may vary with reagent lots of immunoassay
products, due to the use of non-human materials in the controls.
• Reagents from different containers should not be intermixed.
• When using serum or plasma collection tubes, follow the tube manufacturer’s processing instructions.
Sample collection devices should be assessed for suitability and regularly monitored. Contact your
collection device supplier for further details.
• Samples containing precipitates must be centrifuged before performing the assay. Patient samples should
be homogeneous. Patient samples should not contain clots or air bubbles.
• Biological materials of human origin contained in these products were tested for Anti-HCV, HbsAg and
Anti-HIV 1/2 on a single donor basis and were found to be non-reactive. As there is no known test method
that can offer complete assurance that products derived from human blood will not transmit infectious
agents, these products should be handled as potentially infectious materials.
• Handle all patient samples as potentially infectious and follow universal precautions as dictated by local or
national regulations (for further details refer to CLSI GP17-A2, ISO15190 or 29CFR1910.1030).
• Handle samples in closed containers to avoid contamination and evaporation.
5. NOTES
* Control Material with values determined by this Beckman Coulter system may be used for this application.
** This is not required for the English Reagent Guide.
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APPENDIX 1 PRODUCT GROUPINGS
IFU Reference Setting Sheet Reference
Enzyme Code Code
OSR6x01 ACP BLOSR6x01.01 BSOSR6x01.01
OSR6x03 ALP BLOSR6x03.01 BSOSR6x03.01
OSR6x04 ALP BLOSR6x04.01 BSOSR6x04.01
OSR6x07 ALT BLOSR6x07.01 BSOSR6x07.02
OSR6x06 α-Amylase BLOSR6x06.01 BSOSR6x06.01
OSR6x82 α-Amylase BLOSR6x82.01 BSOSR6x82.01
OSR6x09 AST BLOSR6x09.01 BSOSR6x09.02
OSR6x14 Cholinesterase BLOSR6x14.01 BSOSR6x14.01
OSR6x79 CK (NAC) BLOSR6x79.01 BSOSR6x79.01
OSR6x155 CK-MB BLOSR6x155.01 BSOSR6x155.01
OSR6x20 GGT BLOSR6x20.01 BSOSR6x20.01
OSR6x29 HBDH BLOSR6x29.01 BSOSR6x29.01
OSR6x26 LDH BLOSR6x26.01 BSOSR6x26.01
OSR6x28 LDH BLOSR6x28.01 BSOSR6x28.01
OSR6x30 Lipase BLOSR6x30.01 BSOSR6x30.01
Metabolite
OSR6x02 Albumin BLOSR6x02.01 BSOSR6x02.01
OSR6x90 Bicarbonate BLOSR6x90.01 BSOSR6x90.01
OSR6x11 Direct Bilirubin BLOSR6x11.01 BSOSR6x11.01
OSR6x12 Total Bilirubin BLOSR6x12.01 BSOSR6x12.01
OSR6x13 Calcium oCPC BLOSR6x13.01 BSOSR6x13.01
OSR6x117 Calcium Arsenazo III BLOSR6x117.02 BSOSR6x117.01
OSR6x16 Cholesterol BLOSR6x16.02 BSOSR6x16.01
OSR6x78 Creatinine BLOSR6x78.02 BSOSR6x78.01
OSR6x204 Creatinine (Enzymatic) BLOSR6x204.01 BSOSR6x204.02
OSR6x21 Glucose BLOSR6x21.02 BSOSR6x21.02
OSR6x40 Glucose - STAT BLOSR6x40.01 BSOSR6x40.01
OSR6x87 HDL-Cholesterol BLOSR6x87.01 BSOSR6x87.01
OSR6x22 Inorganic Phosphorous BLOSR6x22.01 BSOSR6x22.01
OSR6x86 Iron BLOSR6x86.01 BSOSR6x86.01
OSR6x93 Lactate BLOSR6x93.01 BSOSR6x93.01
OSR6x83 LDL-Cholesterol BLOSR6x83.02 BSOSR6x83.01
OSR6x89 Magnesium BLOSR6x89.01 BSOSR6x89.01
OSR6x32 Total Protein BLOSR6x32.01 BSOSR6x32.01
OSR6x118 Triglyceride BLOSR6x118.01 BSOSR6x118.01
OSR6x24 UIBC BLOSR6x24.01 BSOSR6x24.01
OSR6x205 UIBC BLOSR6x205.01 BSOSR6x205.01
OSR6x34 Urea BLOSR6x34.01 BSOSR6x34.02
OSR6x41 Urea - STAT BLOSR6x41.02 BSOSR6x41.02
OSR6x98 Uric Acid BLOSR6x98.01 BSOSR6x98.01
OSR6x70 Urinary/CSF Protein BLOSR6x70.01 BSOSR6x70.01
Specific Protein
OSR6x62 α-1 Acidglycoprotein BLOSR6x62.01 BSOSR6x62.01
OSR6x63 α-1 Antitrypsin BLOSR6x63.01 BSOSR6x63.01
OSR6x42 Apo A1 BLOSR6x42.01 BSOSR6x42.01
OSR6x43 Apo B BLOSR6x43.01 BSOSR6x43.01
OSR6x94 ASO BLOSR6x94.01 BSOSR6x94.02
OSR6x51 ß-2 Microglobulin BLOSR6x51.01 BSOSR6x51.01
OSR6x59 C3 BLOSR6x59.01 BSOSR6x59.01
OSR6x60 C4 BLOSR6x60.01 BSOSR6x60.01
OSR6x64 Ceruloplasmin BLOSR6x64.01 BSOSR6x64.01
OSR6x47 CRP BLOSR6x47.01 BSOSR6x47.01
OSR6x99 CRP Latex BLOSR6x99.01 BSOSR6x99.01
OSR6x135 D-Dimer BLOSR6x135.01 BSOSR6x135.01
OSR6x50 Ferritin BLOSR6x50.01 BSOSR6x50.01
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IFU Reference Setting Sheet Reference
Specific Protein
Code Code
OSR6x65 Haptoglobin BLOSR6x65.01 BSOSR6x65.01
OSR6x92 HbA1c (Hemoglobin A1c) BLOSR6x92.02 BSOSR6x92.02
HbA1c APT (Hemoglobin A1c,
OSR6x177 BLOSR6x177.01 BSOSR6x177.01
Whole Blood Application)
OSR6x171 IgA BLOSR6x171.01 BSOSR6x171.01
OSR6x172 IgG BLOSR6x172.02 BSOSR6x172.02
OSR6x173 IgM BLOSR6x173.01 BSOSR6x173.01
OSR6x67 Microalbumin BLOSR6x67.01 BSOSR6x67.01
OSR6x68 Myoglobin BLOSR6x68.01 BSOSR6x68.01
OSR6x75 Prealbumin BLOSR6x75.01 BSOSR6x75.01
OSR6x105 RF Latex BLOSR6x105.01 BSOSR6x105.01
OSR6x52 Transferrin BLOSR6x52.01 BSOSR6x52.01
Therapeutic Drug Monitoring (TDM)

OSR6414 Carbamazepine BLOSR6414.01 BSOSR6414.01
OSR6403 Digitoxin BLOSR6403.01 BSOSR6403.01
OSR6404 Digoxin BLOSR6404.01 BSOSR6404.01
OSR6420 Gentamycin BLOSR6420.01 BSOSR6420.01
OSR61202 Paracetamol BLOSR6x202.01 BSOSR6x202.01
OSR6413 Phenobarbital BLOSR6413.01 BSOSR6413.01
OSR6411 Phenytoin BLOSR6411.01 BSOSR6411.01
OSR6412 Theophylline BLOSR6412.01 BSOSR6412.01
OSR6415 Valproic Acid BLOSR6415.01 BSOSR6415.01
Drugs of Abuse in Urine (DAU)

OSR6323 Amphetamines / Ecstasy BLOSR6323.01 BSOSR6323.01
OSR6315 Barbiturates BLOSR6315.01 BSOSR6315.01
OSR6316 Benzodiazepines BLOSR6316.01 BSOSR6316.01
OSR6317 Cocaine BLOSR6317.01 BSOSR6317.01
OSR6318 EDDP BLOSR6318.01 BSOSR6318.01
OSR6319 Methadone BLOSR6319.01 BSOSR6319.01
OSR6320 Opiates BLOSR6320.01 BSOSR6320.01
OSR6322 THC BLOSR6322.01 BSOSR6322.01
ISE
66320 ISE Buffer BL66320.02
66316 ISE High Serum Standard BL66320.02
66314 ISE Internal Reference BL66320.02
66317 ISE Low Serum Standard BL66320.02
66315 ISE Low/High Urine Standard BL66320.02
66319 ISE Mid Standard BL66320.02
+ +
66313 ISE Na /K Selectivity Check BL66320.02
66318 ISE Reference BL66320.02
Calibrator
ODC6413 Antibiotic TDM Multi-Calibrator BLODC6413.01
ODR3005 Apo A1 & B Calibrator BLODR3005.01
ODR3022 Apo A1 & B Calibrator BLODR3022.01
ODR3013 ASO Calibrator BLODR3013.01
ODC0019 Bicarbonate Calibrator BLODC0019.01
ODR30034 CK-MB Calibrator BLODR30034.01
ODC6411 Core TDM Multi-Calibrator BLODC6411.01
ODC0027 CRP Latex Calibrator Highly Sensitive Set BLODC0027.01
ODC0026 CRP Latex Calibrator Normal Set BLODC0026.01
ODR3033 D-Dimer Calibrator BLODR3033.01
ODC6326 DAU Low Intermediate Multi-Drug Calibrator BLODC6326.01 BSODC6326.01
ODC6319 DAU Methadone Cut-off Calibrator BLODC6319.01
ODC6320 DAU Methadone Intermediate Calibrator BLODC6319.01
ODC6321 DAU Methadone High Calibrator BLODC6319.01
ODC6315 DAU Primary Cut-off Multi-Drug Calibrator BLODC6315.01
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IFU Reference
Calibrator Setting Sheet Reference Code
Code
ODC6316 DAU Secondary Cut-off Multi-Drug Calibrator BLODC6315.01
ODC6317 DAU Intermediate Multi-Drug Calibrator BLODC6315.01
ODC6318 DAU High Multi-Drug Calibrator BLODC6315.01
ODC6314 DAU Negative Calibrator BLODC6314.01
ODC6322 DAU THC 25 Calibrator BLODC6322.01
ODC6403 Digitoxin Calibrator BLODC6403.01
ODC6404 Digoxin Calibrator BLODC6404.01
ODR3032 HbA1c Calibrator BLODR3032.01
ODC0011 HDL-Cholesterol Calibrator BLODC0011.01
ODC0012 LDL-Cholesterol Calibrator BLODC0012.01
ODR30037 MC Cal A BLODR30037.01
ODR3024 Microalbumin Calibrator BLODR3024.01
ODR3025 Myoglobin Calibrator BLODR3025.01
ODR3029 Prealbumin Calibrator BLODR3029.01
ODC0028 RF Latex Calibrator BLODR0028.01
ODR3021 Serum Protein Multi-Calibrator BLODR3021.01
ODR3023 Serum Protein Multi-Calibrator 2 BLODR3023.01
66300 System Calibrator BL66300.01
ODC0025 Urine Calibrator BLODC0025.01
Control
ODR2041 CK-MB Control Serum BLODR2041.01
ODR30035 CK-MB Control Level 1 BLODR30035.01
ODR30036 CK-MB Control Level 2 BLODR30036.01
ODC0003 Control Serum 1 BLODC0003.01
ODC0004 Control Serum 2 BLODC0004.01
ODC0013 CRP (Latex) Control Serum BLODC0013.01
ODC0029 D-Dimer Control BLODC0029.01
ODC0006 DAU Multi-Drug Control BLODC0006.01
ODC0007 DAU Speciality Control BLODC0007.01
ODC0008 DAU THC 25 Control BLODC0008.01
ODC0009 DAU THC 50 Control BLODC0008.01
ODC0022 HbA1c Control BLODC0022.01
ODC0005 HDL/LDL-Cholesterol Control Serum BLODC0005.01
ODC0014 ITA Control Serum 1 BLODC0014.01
Miscellaneous
ODR20067 Cleaning Solution BLODR20067.01
OE66039 Cleaning Solution BLOE66039.01
OSR0004 Hemoglobin Denaturant BLOSR0004.01
OSR62166 LIH BLOSR6x166.01 BSOSR6x166.01
OSR0001 Wash Solution BLOSR0001.01
ODR2000 Wash Solution BLOSR0001.01
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-x–
APPENDIX 2 CALIBRATORS AND CONTROLS
Parameter name as
Code appears on kit Calibrator Control
Enzyme
OSR6x01 ACP 66300 ODC0003, ODC0004
OSR6x03 ALP 66300 ODC0003, ODC0004
OSR6x04 ALP 66300 ODC0003, ODC0004
OSR6x07 ALT 66300 ODC0003, ODC0004
Serum/Plasma: 66300 Serum/Plasma: ODC0003, ODC0004
OSR6x06 α-Amylase
Urine: ODC0025 Urine: Biorad Liquichek Cat. No: 397 & 398
OSR6x82 α-Amylase
OSR6x09 AST 66300 ODC0003, ODC0004
OSR6x14 Cholinesterase 66300 ODC0003, ODC0004
OSR6x79 CK (NAC) 66300 ODC0003, ODC0004
OSR6x155 CK-MB ODR30034 ODR30035, ODR30036
OSR6x20 GGT 66300 ODC0003, ODC0004
OSR6x29 HBDH 66300 ODC0003, ODC0004
OSR6x26 LDH 66300 ODC0003, ODC0004
OSR6x28 LDH 66300 ODC0003, ODC0004
OSR6x30 Lipase OSR6x30 or 66300 ODC0003, ODC0004
Metabolite
OSR6x02 Albumin 66300 ODC0003, ODC0004
OSR6x90 Bicarbonate ODC0019 *
OSR6x11 Direct Bilirubin 66300 ODC0003, ODC0004
OSR6x12 Total Bilirubin 66300 ODC0003, ODC0004
OSR6x13 Calcium oCPC
OSR6x117 Calcium Arsenazo III
OSR6x16 Cholesterol 66300 ODC0003, ODC0004
OSR6x78 Creatinine
OSR6x204 Creatinine (Enzymatic)
OSR6x21 Glucose
OSR6x40 Glucose-STAT 66300 ODC0003, ODC0004
OSR6x87 HDL-Cholesterol ODC0011 ODC0005
OSR6x22 Inorganic Phosphorous
OSR6x86 Iron 66300 ODC0003, ODC0004
OSR6x93 Lactate 66300 ODC0003, ODC0004
OSR6x83 LDL Cholesterol ODC0012 ODC0005
OSR6x89 Magnesium
OSR6x32 Total Protein 66300 ODC0003, ODC0004
OSR6x118 Triglyceride 66300 ODC0003, ODC0004
OSR6x24 UIBC 66300 ODC0003, ODC0004
OSR6x205 UIBC 66300 ODC0003, ODC0004
OSR6x34 Urea
OSR6x41 Urea-STAT 66300 ODC0003, ODC0004
OSR6x98 Uric Acid
OSR6x70 Urinary/CSF Protein OSR6x70 *
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Specific Protein
OSR6x62 α-1 Acidglycoprotein ODR3023 ODC0014, ODC0015, ODC0016
OSR6x63 α-1 Antitrypsin ODR3023 ODC0014, ODC0015, ODC0016
OSR6x42 Apo A1 ODR3005 (3 pt) / ODR3022 (5 pt) ODC0003, ODC0004
OSR6x43 Apo B ODR3005 (3 pt) / ODR3022 (5 pt) ODC0003, ODC0004

OSR6x94 ASO ODR3021 or ODR3013 / ODR30037 ODC0014, ODC0015, ODC0016
OSR6x51 β-2 Microglobulin ODR3023 ODC0014, ODC0015, ODC0016
OSR6x59 C3 ODR3021 / ODR30037 ODC0014, ODC0015, ODC0016
OSR6x60 C4 ODR3021 / ODR30037 ODC0014, ODC0015, ODC0016
OSR6x64 Ceruloplasmin ODR3023 ODC0014, ODC0015, ODC0016
OSR6x47 CRP ODR3021 ODC0014, ODC0015, ODC0016
ODC0014, ODC0015, ODC0016
ODC0026 (Normal), (Normal, Highly sensitive),ODC0003
OSR6x99 CRP Latex
ODC0027 (Highly sensitive) (Normal)
ODC0013 (Highly sensitive)
OSR6x135 D-Dimer ODR3033 ODC0029
OSR6x50 Ferritin ODR3021 ODC0014, ODC0015, ODC0016
OSR6x65 Haptoglobin ODR3023 ODC0014, ODC0015, ODC0016
OSR6x92 HbA1c (Hemoglobin A1c) ODR3032 ODC0022
HbA1c APT (Hemoglobin A1c,
OSR6x177 ODR3032 ODC0022
Whole Blood Application)
OSR6x171 IgA ODR3021 ODC0014, ODC0015, ODC0016
OSR6x172 IgG ODR3021 ODC0014, ODC0015, ODC0016
OSR6x173 IgM ODR3021 ODC0014, ODC0015, ODC0016
OSR6x67 Microalbumin ODR3024 *
OSR6x68 Myoglobin ODR3025 *
OSR6x75 Prealbumin ODR3029 ODC0014, ODC0015, ODC0016
OSR6x105 RF Latex ODC0028 ODC0014, ODC0015, ODC0016
OSR6x52 Transferrin ODR3021 / ODR30037 ODC0014, ODC0015, ODC0016
TDM
OSR6414 Carbamazepine ODC6411 *
OSR6403 Digitoxin ODC6403 *
OSR6404 Digoxin ODC6404 *
OSR6420 Gentamycin ODC6413 *
OSR6x202 Paracetamol OSR6x202 *
OSR6413 Phenobarbital ODC6411 *
OSR6411 Phenytoin ODC6411 *
OSR6412 Theophylline ODC6411 *
OSR6415 Valproic Acid ODC6411 *
DAU
Amphetamines/ Qualitative: ODC6315 (1000 µg/L) or ODC6316 (500 µg/L). 1000 µg/L Cut off: ODC0006
OSR6323
Ecstasy Semi Quantitative: ODC6314-ODC6318 500 µg/L Cut-off: ODC0007
Qualitative: ODC6315 (300 µg/L) or ODC6316 (200 µg/L). 300 µg/L Cut-off: ODC0006
OSR6315 Barbiturates Semi Quantitative: ODC6314-ODC6318 200 µg/L Cut-off: ODC0007
OSR6316 Benzodiazepines Semi Quantitative: ODC6314 ODC6318 200 µg/L Cut-off: ODC0007
Qualitative: ODC6315 (300 µg/L) or ODC6316 (150 µg/L). 300 µg/L Cut off: ODC0006
OSR6317 Cocaine Semi Quantitative: ODC6314-ODC6318 150 µg/L Cut-off: ODC0007
Qualitative: ODC6315 or ODC6316 (both 100 µg/L).
OSR6318 EDDP Semi Quantitative: ODC6314, ODC6315 or ODC6316, ODC0006 or ODC0007
ODC6317, ODC6318
Qualitative: ODC6319 (300 µg/L).
OSR6319 Methadone Semi Quantitative: ODC6314, ODC6319-ODC6321 ODC0006
Qualitative: ODC6316 (300 µg/L).

OSR6320 Opiates Semi Quantitative: ODC6314, ODC6316-ODC6318 ODC0007
OSR6322 THC Semi Quantitative: ODC6314, ODC6322-ODC6325 50 µg/L Cut-off: ODC0009
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ACP
OSR6101 2 x 15 mL R1-1 ACP-T
2 x 15 mL R1-2 ACP-NP
4x R1-S ACP-S
1 x 10 mL ACP-St
Intended Use
Kinetic colour test for the quantitative determination of acid phosphatase, EC 3.1.3.2 (ACP), in human serum on Beckman Coulter analysers. For in vitro
diagnostic use only.
Summary1,2
The term acid phosphatase refers to a group of similar enzymes which cleave phosphate esters optimally at a pH below 7.0. These enzymes are present
in the lysosomes of all body cells with the highest activities in the liver, spleen, bone marrow, prostate, erythrocytes and platelets. Because the activities
from the prostate are inhibited by tartrate, the tartrate inhibited proportion of total ACP is also referred to as prostatic ACP. Elevations of the enzymatic
activity of prostatic ACP (and thus, generally, of total ACP activity) may be found in the sera of men with metastatic prostate cancer. The frequency of such
findings in a population of men with prostate cancer increases as the cancer progresses, thus prostatic ACP is useful in the staging of prostate carcinoma.
Serum prostatic ACP measurement is useful in monitoring remission or relapse of a prostatic malignancy and in assessing the effectiveness of various
treatment regimens.
Other diseases which can be associated with elevations of tartrate inhibited ACP include leukemia, polycythemia vera, primary thrombocythemia and
megaloblastic anemia.The majority of the normally low ACP activity of serum is of a tartrate-resistant type and probably originates mainly in osteoclasts.
Activities of this fraction are physiologically increased in growing children and pathologically in conditions of increased osteolysis and bone remodelling
such as Paget's disease, hyperparathyroidism, multiple myeloma, osteosarcoma, osteogenesis imperfecta and renal osteodystrophy. Other diseases
associated with elevated tartrate resistant ACP are Gaucher's disease and Niemann-Pick disease.
Test Principle3
Hillmann reaction
ACP
1-Naphthyl phosphate + H2O Phosphate + 1-Naphthol
1-Naphthol + FRTR salt Azo dye
Contents, Reagent Composition in the Test

Final concentration of reactive ingredients:
Total ACP method
Citrate buffer (pH 4.8) 140 mmol/L
1-Naphthyl Phosphate 11 mmol/L
Fast red TR salt 0.6 mmol/L
Pentanediol 210 mmol/L
Non-Prostatic ACP method
Tartrate 125 mmol/L
Citrate buffer (pH 4.8) 140 mmol/L
1-Naphthyl Phosphate 11 mmol/L
Fast Red TR salt 0.6 mmol/L
Pentanediol 210 mmol/L
Precautions and Warnings

Hazard Warnings and Risk Phrases:
R1-S (ACP substrate). Harmful, contains Fast Red TR salt. R68: Possible risk of irreversible effects.
Safety Phrases:
S36/37, S60. Wear suitable protective clothing and gloves. This material and its container must be disposed of as hazardous waste.
Exercise the normal precautions required for handling all laboratory reagents.
Dispose of all waste material in accordance with local guidelines.
Refer to Safety Data Sheets for further information.
Reagent Preparation
R1-1 (Total ACP)
To perform a total ACP (ACP-T) assay dissolve the contents of one R1-S vial (ACP substrate) completely with the contents of one bottle of R1-1 (Total
ACP buffer).
R1-2 (Non-Prostatic-ACP)
To perform a non-prostatic ACP (ACP-NP) assay dissolve the contents of one R1-S vial (ACP substrate) completely with the contents of one bottle of
R1-2 (Non-prostatic ACP buffer).
Attach the adaptor to the buffer bottle then connect the substrate powder vial to the other end. Gently mix the linked bottles until the powder is completely
dissolved. The solution can then be collected in the buffer bottle and placed on board the instrument.
Storage and Stability

The reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C. Once prepared, reagents stored on board the instrument are
stable for 14 days.
Specimen
Serum. Do not use plasma.
Serum should be separated as soon as possible and 1 drop of stabiliser (ACP-St) added to 1mL of the sample.
Separated serum, without stabiliser, is stable for 15 min when stored at 15…25°C and 3-4 hours when stored at 2…8°C. Stabilised serum can be stored
4
for 8 days when stored at 2…25°C.
Lipemic, icteric and haemolysed samples should be avoided.
EN.01 BLOSR6x01.01 Enzyme

2009-08
Test Procedure
Refer to the appropriate User’s Guide and the accompanying Instrument Setting Sheet for analyser-specific assay instructions.
Calibration
The test is run in MB-mode, two analyser specific MB-factors (ACP-T & ACP-NP) are required. To provide a robust approach to generate the analyser
specific MB factor, it is recommended that 5 separate calibration events should be used. A fresh vial of calibrator, utilising System Calibrator Cat No. 66300
in the AB calibration mode, should be used for each of these runs. When calculating the mean factor from the separate runs the data should be examined
for obvious outliers which should be repeated and replaced. For the AU2700/AU5400 this procedure needs to be performed for each ring. Quality control
procedures should be undertaken immediately following calibration in accordance with good laboratory practice.
The calibrator value is traceable to a Beckman Coulter Master Calibrator.
Re-establishment of the analyser specific MB factor is recommended when a critical part of the analyser is replaced.
Reagent blank measurement is recommended when changing to a new lot of reagent.
Quality Control
controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter system may be used.
Each laboratory should establish its own control frequency however good laboratory practice suggests that controls be tested each day patient
samples are tested and each time calibration/blanking is performed.
The results obtained by any individual laboratory may vary from the given mean value. It is therefore recommended that each laboratory
generates analyte specific control target values and intervals based on multiple runs according to their requirements. These target values should
fall within the corresponding acceptable ranges given in the relevant product literature.
If any trends or sudden shifts in values are detected, review all operating parameters.
Each laboratory should establish guidelines for corrective action to be taken if controls do not recover within the specified limits.
Calculation
The Beckman Coulter analysers automatically compute the ACP-T and ACP-NP activity of each sample. The activity of prostatic ACP can be generated by
the analyser when set as a calculated test:
Prostatic-ACP = Total-ACP – Non-Prostatic-ACP.
Reference Intervals1
Adults 37°C-Total ACP
Male ≤ 6.6 Pentanediol activation U/L (0.11 µkat/L)
Female ≤ 6.5 Pentanediol activation U/L (0.11 µkat/L)
Adults Tartrate-inhibited ACP (Prostatic ACP)
≤ 3.0 Pentanediol activation U/L (0.05 µkat/L)
Values obtained with different prostatic acid phosphatase assays cannot be used interchangeably. Before changing assays, laboratories must confirm
baseline values for patients being serially monitored.
Expected values may vary with age, sex, sample type, diet and geographical location. Each laboratory should verify the transferability of the expected
values to its own population, and if necessary determine its own reference interval according to good laboratory practice. For diagnostic purposes, results
should always be assessed in conjunction with the patient's medical history, clinical examinations and other findings.
Specific performance characteristics

Data contained within this section is representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ from these
values.
Linearity
The test is linear within an enzyme activity range of 0 - 100 U/L (0 - 1.67 µkat/L) for Total ACP and 0 - 30 U/L (0 - 0.5 μkat/L) for Prostatic ACP.
Precision
The following data was obtained on an AU600 using 3 serum pools analysed over 10 days.
Total ACP
n = 60 Within Run Total
Mean U/L SD CV% SD CV%
7.10 0.08 1.07 0.13 1.82
17.55 0.15 0.87 0.23 1.31
34.28 0.28 0.83 0.48 1.39
Prostatic ACP
5.68 0.10 1.76 0.40 7.01
7.46 0.17 2.24 0.23 3.05
15.77 0.32 2.01 0.42 2.69
Sensitivity
The lowest detectable level on an AU600 analyser was estimated at 0.05 U/L for Total ACP and 0.10 U/L for Prostatic ACP.
The lowest detectable level represents the lowest measurable level of ACP that can be distinguished from zero. It is calculated as the absolute mean plus
three standard deviations of 20 replicates of an analyte free sample.
Method Comparison
Patient serum samples were used to compare this Total ACP OSR6101 assay on the AU600 against another commercially available Total ACP assay.
Results of linear regression analysis were as follows:
y = 0.944x – 0.271 r = 0.999 n = 116 Sample range = 0.58 – 58.51 U/L
Patient serum samples were used to compare this Prostatic OSR6101 assay on the AU600 against another commercially available Prostatic ACP assay.
y = 0.915x – 0.060 r = 0.966 n = 93 Sample range = 0.19 – 3.57 U/L
Interfering Substances
Results of studies conducted to evaluate the susceptibility of the Total ACP method to interference were as follows:
Icterus: Interference less than 10% up to 0.6 mg/dL or 10.3 µmol/L bilirubin
Ascorbate: Interference less than 10% up to 8 mg/dL ascorbate
Haemolysis: Interference less than 10% up to 4 g/L haemoglobin
®
Lipemia: Interference less than 10% up to 300 mg/dL Intralipid
Enzyme BLOSR6x01.01 EN.01

2009-08
Results of studies conducted to evaluate the susceptibility of the Prostatic ACP method to interference were as follows:
Icterus: Interference less than 10% up to 1 mg/dL or 17.1 µmol/L bilirubin
®
5
Refer to Young for further information on interfering substances.
Limitations
Prostatic acid phosphatase results may be influenced by a number of factors other than malignancy and should always be interpreted in conjunction with
the patients clinical history and other diagnostic procedures. Do not use plasma.
Setting Sheet Footnotes

‡ For determination of Prostatic ACP the above parameters must be entered twice using test names ACPT (Total ACP) and ACPNP (Non-Prostatic
ACP). Set the test as a CALCULATED TEST in the INTER TESTS menu. See leaflet.
# User defined ¤ Analyser default value
§ For use in AB mode only, refer to leaflet for further instruction. Ensure that the correct calibrator value is entered for ACPT and ACPNP as
appropriate. Set factor ranges for Total ACP 800-1200* and Non-Prostatic ACP 1000-1550
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60.
ж Set this test as COMMON TEST PARAMETER TEST NAME in CALCULATED TEST. Enter the formula (A-B), where A = ACPT and
B = ACPNP in Parameters Specific Test Parameters CALCULATED TEST.
BIBLIOGRAPHY
1. Thomas L. Saüre Phosphatase (SP). In:Thomas L, ed. Labor und Diagnose 6 Auflage:TH-Books Verlagsgesellschaft mbH Frankfürt/Main, 2005:118-120.
2. Moss DW, Henderson RA. Clinical enzymology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders Company,
1999;711-14.
3. Hillmann G. Fortlaufende photometrische messung der sauren prostataphosphatase-aktivität. Z Klin Chem u Klin Biochem 1971;9(3):273-274.
4. Young DS. Effects of preanalytical variables on clincal laboratory tests, 2nd ed.Washington: AACC press 1997:3-5pp.
5. Young DS. Effects of drugs on clinical laboratory tests, 5th ed. AACC Press, 2000.

2009-08
ACP, AU400/AU640 Serum Application ACP, AU600 Serum Application
System Reagent: OSR6101 System Reagent: OSR6101
Reagent ID: TOTAL (ACPT) 001, NON-PROSTATIC (ACPNP) 008 Reagent ID: TOTAL (ACPT) 001, NON-PROSTATIC (ACPNP) 008
Specific Test Parameters Specific test parameters
General LIH ISE Range
Test No # Test name ACP‡ ∇ Sample type Ser ∇ Page 1/2
Test Name: ACP‡ ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 10 Dil. vol 0 μL Min. OD Max. OD
Sample: Volume 10 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 150 Dil. vol 0 μL L 0 H 2.5
Reagents: R1 Volume 150 μL Dilution 0 μL Min OD Max OD Reagent 2 vol 0 Dil. vol 0 μL Reagent OD limit
R2 Volume 0 μL Dilution 0 μL L 0 H 2.5 Fst. L -0.1 Fst. H 0.3
Reagent OD limit: Wave Main 410 Sub 800 ∇ Lst. L -0.1 Lst. H 0.3
Wavelength: Pri. 410 ∇ Sec. 800 ∇ First L -0.1 First H 0.3 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L -0.1 Last H 0.3 Reaction + ∇ L 0* H 100*
Reaction slope: + ∇ Dynamic Range: Point 1 Fst 12 Lst 27 Correlation factor A 1
Measuring Point 1: First 12 Last 27 L 0* H 100* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: NO ∇ On-board stability period: 14 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 14
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name ACP‡ ∇ Sample type Ser ∇ Page 2/2
Test Name: ACP‡ ∇ < > Type: Serum ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse
SERUM APPLICATION
Calibration specific
Calibration Specific
General ISE Test No # Test name ACP‡ ∇
Test Name: ACP‡ ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Formula 1 Y=AX+B ∇ Process Conc ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § § §
Point 1: § § § §
Point 2 ∇
Point 2:
Point 3: Point 3 ∇
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor #
MB Type Factor: # Calibration Stability Period: Calibrator stability period
‡ For determination of Prostatic ACP the above parameters must be entered twice using test names ACPT (Total ‡ For determination of Prostatic ACP the above parameters must be entered twice using test names ACPT (Total
ACP) and ACPNP (Non-Prostatic ACP). Set the test as a CALCULATED TEST in the INTER TESTS menu, See leaflet. ACP) and ACPNP (Non-Prostatic ACP). Set the test as a CALCULATED TEST in the INTER TESTS menu. See leaflet.
# User defined # User defined ¤ Analyser default value
§ For use in AB mode only. Refer to IFU for further instruction. Ensure that the correct calibrator value is entered for § For use in AB mode only. Refer to IFU for further instruction. Ensure that the correct calibrator value is entered for
ACPT and ACPNP as appropriate. Set factor ranges for Total ACP 800-1200* and Non-Prostatic ACP 1000-1550* ACPT and ACPNP as appropriate. Set factor ranges for Total ACP 800-1200* and Non-Prostatic ACP 1000-1550*
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
Enzyme BSOSR6x01.01
2009-08
ACP, AU2700/AU5400 Serum Application
System Reagent: OSR6101
Reagent ID: TOTAL (ACPT) 001, NON-PROSTATIC (ACPNP) 008
Test Name: ACP‡ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 8 μL Dilution 0 μL Pre-Dilution Rate: 1

Reagents: R1 Volume 120 μL Dilution 10 μL Min OD Max OD
R2 Volume 0 μL Dilution 0 μL L 0 H 2.5
Reagent OD limit: 0.3
Wavelength: Pri. 410 ∇ Sec. 800 ∇ First L -0.1 First H 0.3
Method: RATE ∇ Last L -0.1 Last H 0.3
Reaction slope: + ∇ Dynamic Range:
Measuring Point 1: First 12 Last 27 L 0* H 100*
Measuring Point 2: First Last Correlation Factor:
No Lag Time: NO ∇ On-board stability period: 14

Test Name: ACP‡ ∇ < > Type: Serum ∇
Value/Flag: # ∇ Level L: # Level H: #

Normal Ranges: Age L Age H
Sex Year Month Year Month L H
ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
7. None Selected # #
8. Out of Range L H # #
Panic Value: # # Unit: U/L* Decimal places: #
SERUM APPLICATION
General ISE
Test Name: ACP‡ ∇ < > Type Serum ∇

Point 1: # § § §
Point 2:
Point 3:
Point 4:
Point 5:
Point 6:
Point 7:
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇
MB Type Factor: # Calibration Stability Period:
‡ For determination of Prostatic ACP the above parameters must be entered twice using test names ACPT (Total ACP)
and ACPNP (Non-Prostatic ACP). Set the test as a CALCULATED TEST in the INTER TESTS menu. See eaflet.
# User defined.
§ For use in AB mode only. Refer to IFU for further instruction Ensure that the correct calibrator value is entered for
ACPT and ACPNP as appropriate. Set factor ranges for Total ACP 800-1200* and Non-Prostatic ACP 1000-1550*
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
Enzyme BSOSR6x01.01
2009-08
ACP, AU680/AU480 Serum Application
System Reagent: OSR6101 Reagent ID: TOTAL (ACPT) 001, NON-PROSTATIC (ACPNP) 008
Parameters Specific Test Parameters
General LIH ISE HbA1c Calculated Test Range
Test Name: ACPж ∇ < > Type: Serum ∇ Operation Yes ∇
Sample Volume 8 μL Dilution 0 μL OD Limit

Pre-Dilution Rate 1 ∇ Min.OD 0 Max.OD 2.5
Rgt. Volume R1(R1-1) 120 μL Dilution 10 μL Reagent OD Limit
First Low -0.1 High 0.3
Last Low -0.1 High 0.3
R2(R2-1) 0 μL Dilution 0 μL
Dynamic Range Low 0* High 100*
Common Rgt. Type NoneΦ Name ф Correlation Factor A 1 B 0
Wavelength Pri 410 ∇nm Sec. 800 ∇nm Factor for Maker A 1 B 0
Method RATE ∇
Reaction Slope + ∇ Onboard Stability Period 14 Day 0 Hour
Measuring Point1 First 12 Last 27 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus + ∇
Lag Time Check NO ∇ Hemolysis ++++ ∇

Test Name: ACPж ∇ < > Type: Serum ∇
Value/Flag: # ∇ Low High

Level # # Panic Value
Specificl Ranges: From To Low High
Sex Year Month Year Month Low High # #
ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
7. No demographics # #
8. Not within expected values # #
Unit U/L* Decimal Places #
Parameters Calibration Parameters
Calibrators Calibration Specific STAT Table Calibration
SERUM APPLICATION
General ISE
Test Name: ACPж ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇

<Calibrator Parameters> Range
Calibrator OD Conc Low High Slope Check None ∇
Point 1: # ∇ § § §
Point 2: ∇ Allowance Range Check
Point 3: ∇
Point 4: ∇ ο Reagent Blank
Point 5: ∇ ο Calibration
Point 6: ∇
Point 7 ∇ Advanced Calibration
Point 8 ∇ Operation # ∇
Point 9 ∇
Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration
Master Curve> OD Range ж Set this test as Common Test Parameter test name in CALCULATED TEST. Enter the formula (A-B) , where A=ACPT
Calibrator OD Conc Low High Stability and B=ACPNP in Parameters Specific Test Parameters CALCULATED TEST
Point-1 ∇ Reagent Blank Day Hour # User defined.
Point-2 ∇ Calibration Day Hour § For use in AB mode only. Refer to IFU for further instruction Ensure that the correct calibrator value is entered for
MB Type Factor: # 1-Point Calibration Point ∇ ο with Conc-0 ACPT and ACPNP as appropriate. Set factor ranges for Total ACP 800-1200* and Non-Prostatic ACP 1000-1550*
Ф AU680
Enzyme BSOSR6x01.01
2009-08
ALP
OSR6103 4 x 30 mL R1
4 x 30 mL R2
Intended Use
Kinetic colour test for the quantitative determination of alkaline phosphatase, EC 3.1.3.1 (ALP), in human serum and plasma on Beckman Coulter
analysers. For in vitro diagnostic use only.
Summary1,2
Alkaline phosphatase (ALP) is present in almost all body tissues, located at or in cell membranes. It occurs at particularly high levels in interstitial
epithelium, kidney tubules, bone (osteoblasts), liver and placenta. The precise metabolic function of ALP has not yet been fully elucidated, however the
enzyme is associated with intestinal lipid transport and bone calcification. ALP originates in approximately equal proportions from the liver and the skeletal
system. Approximately 25% of healthy individuals also have intestinal ALP which accounts for approximately 10% of the total ALP in a fasting sample.
Increases in total ALP are either due to physiological causes, or are caused by diseases of the liver or bone. Physiological increases in ALP are found in
pregnancy from the 2nd trimester onwards due to placental ALP, in growing children due to bone ALP and postprandially in individuals with blood groups
B and O, who are secretors of blood group substance H (intestinal ALP). The most common cause of elevated ALP is hepatobiliary disease, with
pathological ALP levels found in approximately 60% of patients with disease of the liver or biliary tract. ALP levels may also be elevated in primary bone
diseases, such as osteomalacia, osteogenesis imperfecta, vitamin D intoxication and primary bone tumours. ALP levels may also be increased in
secondary bone diseases, such as skeletal metastases, and in diseases such as multiple myeloma, acromegaly, renal insufficiency, hyperthyroidism,
ectopic ossification, sarcoidosis, bone tuberculosis and healing fractures. In bone diseases such as Paget’s disease, vitamin D deficiency rickets and
metastatic bone disease, ALP activity is a good indicator of bone activity, in the absence of co-existing chronic liver disease. Total ALP is only occasionally
elevated in some metabolic bone diseases such as hyperparathyroidism, osteopenia or osteoporosis. Reduced levels of ALP are found in familial
hypophosphatasia, hypoparathyroidism, achondroplasia, adynamic bone disease in dialysis patients, pituitary dwarfism, chronic radiation sickness and
malnutrition.
Test Principle3
Method based on the recommendations of the “German Society for Clinical Chemistry” (GSCC).
Alkaline phosphatase activity is determined by measuring the rate of conversion of p-Nitrophenyl phospate (pNPP) to p-Nitrophenol (pNP) in the presence
of magnesium ions and diethanolamine as phosphate acceptor at pH 9.8.
The rate of increase in absorbance due to the formation of pNP is measured at 410/480 nm and is directly proportional to the ALP activity in the sample.
Reaction Principle
ALP
p-Nitrophenyl phosphate + H2O Phosphate + p-Nitrophenol
2+
Mg
Diethanolamine buffer, pH 9.8 1.0 mol/L
Magnesium chloride 0.5 mmol/L
p-Nitrophenyl phosphate 10 mmol/L
Preservative
R1 reagent: Harmful, contains 2,2’-iminodiethanol and a mixture of: 5-chloro-2-methyl-4-isothiazolin-3-one [EC No 247-500-7] and 2-methyl-4-isothiazolin-
3-one [EC No 220-239-6] (3:1). R22, R38, R41, R43, R48/22. Harmful if swallowed. Irritating to skin. Risk of serious damage to eyes. May cause
sensitisation by skin contact, Harmful: danger of serious damage to health by prolonged exposure if swallowed.
R2 reagent: Irritant, contains a mixture of: 5-chloro-2-methyl-4-isothiazolin-3-one [EC No 247-500-7] and 2-methyl-4-isothiazolin-3-one [EC No 220-239-6]
(3:1).R43. May cause sensitisation by skin contact.
Safety Phrases:
S24, S26, S37/39, S60.Avoid contact with skin, In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable
gloves and eye/face protection. This material and its container must be disposed of as hazardous waste.
During the reaction p-nitrophenol is produced. This is harmful when inhaled, swallowed or absorbed through skin.
Reagent Preparation
The reagents are ready for use and can be placed directly on board the instrument.
The reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C.
Absorption of atmospheric CO2 by the reagent on board the analyser can impair its stability. This effect will vary depending upon the rate of use. Bottle
replacement is recommended when one of the following conditions are encountered:
14 days have elapsed on board the analyser,
Significant shift in control values (>7%) following local QC procedures,
Major preventative maintenance was performed on the analyser or a critical part was replaced.
Specimen
4
Serum and heparinised plasma. Complexing anticoagulants such as citrate, oxalate and EDTA should be avoided.
Haemolysed samples should be avoided.
5
Stable in serum and plasma for 7 days when stored at 2…25°C.
Test Procedure
Refer to the appropriate User Guide and Setting Sheet for analyser-specific assay instructions for the sample type as listed in the Intended Use statement.
The paediatric application is suitable for use with small volume serum/plasma samples.

2009-08
Calibration
The test is run in MB-mode. To provide a robust approach to generate the analyser specific MB factor, it is recommended that 5 separate calibration events
should be used. A fresh vial of calibrator, utilising System Calibrator Cat No. 66300 in the AB calibration mode, should be used for each of these runs.
When calculating the mean factor from the separate runs the data should be examined for obvious outliers which should be repeated and replaced. For the
AU2700/AU5400 this procedure needs to be performed for each ring. Quality control procedures should be undertaken immediately following calibration in
accordance with good laboratory practice.
The calibrator value is traceable to a Beckman Coulter Master Calibrator
Quality Control
Controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter system may be used.
Each laboratory should establish its own control frequency however good laboratory practice suggests that controls be tested each day patient samples
are tested and each time calibration/blanking is performed.
The results obtained by any individual laboratory may vary from the given mean value. It is therefore recommended that each laboratory generates analyte
specific control target values and intervals based on multiple runs according to their requirements. These target values should fall within the corresponding
acceptable ranges given in the relevant product literature.
Calculation
The Beckman Coulter analysers automatically compute the ALP activity of each sample.
Reference Intervals6,7
Women 64 - 300 U/L (1.0 – 5.0 µkat/L)
Men 80 – 300 U/L (1.3 – 5.0 µkat/L)
Children up to 15 years Up to 640 U/L (10.6 µkat/L)
Children 15-17 years Up to 480 U/L (8.0 µkat/L)
Specific Performance Characteristics
values.
Linearity
The test is linear within an enzyme activity range of 5 - 1500 U/L (0.1 – 25.0 µkat/L).
Precision
38 0.60 1.55 1.68 4.36
362 2.26 0.62 9.99 2.76
1088 5.81 0.53 23.40 2.15
Sensitivity
The lowest detectable level on an AU600 analyser was estimated at 1 U/L.
The lowest detectable level represents the lowest measurable level of ALP that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison
Patient serum samples were used to compare this ALP OSR6103 assay on the AU600 against another commercially available ALP assay. Results of
linear regression analysis were as follows:
y = 0.947x - 12 r = 0.999 n = 120 Sample range = 67 – 1404 U/L
Results of studies conducted to evaluate the susceptibility of the method to interference were as follows:
Icterus: Interference less than 5% up to 40 mg/dL or 684 µmol/L bilirubin
®
8
§ For use in AB mode only, refer to leaflet for further instruction.
‡ Depends on usage pattern in the laboratory.
BIBLIOGRAPHY
1. Moss DW, Henderson RA. Clinical Enzymology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders Company,
1999; 676-684.
2. Thomas L. Alkaline phosphatase (ALP). In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
TH-Books Verlagsgesellschaft, 1998:36-46.
3. Empfehlungen der Deutschen Gesellschaft für Klinische Chemie. Z Klin Chem u Klin Biochem 10 Jg 1972,182-192.
4. Moss DW, Henderson RA, Kachmar JF. Enzymes In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:387pp.
5. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of Anticoagulants in Diagnostic Laboratory Investigations and Stability of Blood, Plasma and
Serum Samples. WHO/DIL/LAB/99.1 Rev.2:21pp.
6. Schlebusch H, Rick W, Lang H, Knedel M. Standards in the activities of clinically important enzymes. Dtsch Med Wochenschr 1974;Apr 12;99(15):765-6.
7. Rick W, ed. Klinische chemie und mikroskopie, 6th ed. Berlin: Springer-Verlag,1990: 294pp.

2009-08
ALP, AU400/AU640 Serum/Plasma Application ALP, AU600 Serum/Plasma Application
System Reagent: OSR6103 Reagent ID: 003 System Reagent: OSR6103 Reagent ID: 003

Test No # Test name ALP03 ∇ Sample type Ser ∇ Page ½
Test Name: ALP03 ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 3 Dil. Vol 0 μL Min. OD Max. OD
Sample: Volume 3 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 60 Dil. Vol 60 μL L -0.1 H 2.5
Reagents: R1 Volume 60 μL Dilution 60 μL Min OD Max OD Reagent 2 vol 60 Dil. Vol 60 μL Reagent OD limit
R2 Volume 60 μL Dilution 60 μL L -0.1 H 2.5 Fst. L -0.1 Fst. H 0.8
General LIH ISE Range Test No # Test name ALP03 ∇ Sample type Ser ∇ Page 2/2
Test Name: ALP03 ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name ALP03 ∇
Test Name: ALP03 ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
SERUM/PLASMA APPLICATION
Formula 1 Y=AX+B ∇ Process ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇
Point 1 § ∇ § §4300* §7000*
Point 1: § § §4300* §7000*
Point 2 ∇
Point 2:
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. Point
MB type factor #
MB Type Factor: # Calibration Stability Period: ‡ ‡
Calibrator stability period
# User defined
§ For use in AB mode only, refer to leaflet for further instruction
‡ Depends on usage pattern in the Laboratory
Enzyme BSOSR6x03.01
2009-08
ALP, AU2700/AU5400 Serum/Plasma Application ALP, AU680/AU480 Serum/Plasma Application
Specific Test Parameters General LIH ISE HbA1c Calculated Test Range
Test Name: ALP03 ∇ < > Type: Serum ∇ Operation Yes ∇
Sample: Volume 1.8 Dilution 0 Pre-Dilution Rate: 1 Sample Volume 1.8 μL Dilution 0 μL OD Limit
μL μL
Pre-Dilution Rate 1 ∇ Min.OD -0.1 Max.OD 2.5
R2 Volume 36 μL Dilution 36 μL L -0.1 H 2.5
Reagent OD limit:
Reaction slope: + ∇ Dynamic Range: Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Measuring Point 1: First 16 Last 27 L 5* H 1500* Wavelength Pri 410 ∇nm Sec. 480 ∇nm Factor for Maker A 1 B 0
Measuring Point 2: First Last Correlation Factor: Method RATE ∇
Linearity : 15 % A 1 B 0 Reaction Slope + ∇ Onboard Stability Period 14 Day 0 Hour
No Lag Time: NO ∇ On-board stability period: 14 Measuring Point1 First 16 Last 27 LIH Influence Check # ∇
Linearity Limit 15 % Icterus +++++ ∇
Specific Test Parameters Lag Time Check NO ∇ Hemolysis ++ ∇
Test Name: ALP03 ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: ALP03 ∇ < > Type: Serum ∇
Sex Year Month Year Month L H Value/Flag: # ∇ Low High
ο 1. # ∇ # # # # # # Level # # Panic Value
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: U/L* Decimal places: # 7. No demographics # #
General ISE
General ISE
Test Name: ALP03 ∇ < > Type Serum ∇ Test Name: ALP03 ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibrator OD Conc Low High Slope Check None ∇
Point 1: § § §4300* §7000* Point 1: § ∇ § §4300* §7000*
Point 2: Point 2: ∇ Allowance Range Check
Point 3: Point 3: ∇
Point 4: Point 4: ∇ ο Reagent Blank
Point 5: Point 5: ∇ ο Calibration
Point 7: Point 7 ∇ Advanced Calibration
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
MB Type Factor: # Calibration Stability Period: ‡
# User defined Master Curve> OD Range
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 Calibrator OD Conc Low High Stability
§ For use in AB mode only, refer to leaflet for further instruction Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
‡ Depends on usage pattern in the Laboratory Point-2 ∇ Calibration Day Hour
ф AU680 MB Type Factor: # 1-Point Calibration Point ∇ ο with Conc-0
Enzyme BSOSR6x03.01
2009-08
ALP, AU400/AU640 Paediatric Application ALP, AU600 Paediatric Application

Test No # Test name ALP3P ∇ Sample type Ser ∇ Page 1/2
Test Name: ALP3P ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 3 μL Dilution 10 μL Pre-Dilution Rate: 1 Reagent 1 vol 60 Dil. vol 60 μL L -0.1 H 2.5
General LIH ISE Range Test No # Test name ALP3P ∇ Sample type Ser ∇ Page 2/2
Test Name: ALP3P ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name ALP3P
PAEDIATRIC APPLICATION
∇
Test Name: ALP3P ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §4300* §7000*
Point 1: § § §4300* §7000*
Point 2 ∇
Point 2:
MB type factor #
# User defined
Enzyme BSOSR6x03.01
2009-08
ALP, AU2700/AU5400 Paediatric Application ALP, AU680/AU480 Paediatric Application
Test Name: ALP3P ∇ < > Type: Serum ∇ Operation Yes ∇
μL μL
Reagent OD limit:
Test Name: ALP3P ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: ALP3P ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: ALP3P ∇ < > Type Serum ∇ Test Name: ALP3P ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: § § §4300* §7000* Point 1: § ∇ § §4300* §7000*
Point 9 ∇
§ For use in AB mode only, refer to leaflet for further instruction Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
Enzyme BSOSR6x03.01
2009-08
ALP
OSR6004 4 x 12 mL R1
4 x 12 mL R2
OSR6104 4 x 30 mL R1
4 x 30 mL R2
OSR6204 4 x 53 mL R1
4 x 53 mL R2
Intended Use
Kinetic colour test for the quantitative determination of alkaline phosphatase, EC 3.1.3.1 (ALP), in human serum and plasma on Beckman Coulter
Summary1,2
Alkaline phosphatase (ALP) is present in almost all body tissues, located at or in cell membranes. It occurs at particularly high levels in interstitial
epithelium, kidney tubules, bone (osteoblasts), liver and placenta. The precise metabolic function of ALP has not yet been fully elucidated, however the
enzyme is associated with intestinal lipid transport and bone calcification. ALP originates in approximately equal proportions from the liver and the skeletal
system. Approximately 25% of healthy individuals also have intestinal ALP which accounts for approximately 10% of the total ALP in a fasting sample.
Increases in total ALP are either due to physiological causes, or are caused by diseases of the liver or bone. Physiological increases in ALP are found in
pregnancy from the 2nd trimester onwards due to placental ALP, in growing children due to bone ALP and postprandially in individuals with blood groups B
and O, who are secretors of blood group substance H (intestinal ALP). The most common cause of elevated ALP is hepatobiliary disease, with
pathological ALP levels found in approximately 60% of patients with disease of the liver or biliary tract. ALP levels may also be elevated in primary bone
diseases, such as osteomalacia, osteogenesis imperfecta, vitamin D intoxication and primary bone tumours. ALP levels may also be increased in
secondary bone diseases, such as skeletal metastases, and in diseases such as multiple myeloma, acromegaly, renal insufficiency, hyperthyroidism,
ectopic ossification, sarcoidosis, bone tuberculosis and healing fractures. In bone diseases such as Paget’s disease, vitamin D deficiency rickets and
metastatic bone disease, ALP activity is a good indicator of bone activity, in the absence of co-existing chronic liver disease. Total ALP is only occasionally
elevated in some metabolic bone diseases such as hyperparathyroidism, osteopenia or osteoporosis. Reduced levels of ALP are found in familial
hypophosphatasia, hypoparathyroidism, achondroplasia, adynamic bone disease in dialysis patients, pituitary dwarfism, chronic radiation sickness and
malnutrition.
Test Principle3
Method based on the recommendations of the “International Federation for Clinical Chemistry” (IFCC).
Alkaline phosphatase activity is determined by measuring the rate of conversion of p-nitro-phenylphosphate (pNPP) to p-nitrophenol (pNP) in the presence
of magnesium and zinc ions and of 2-amino-2-methyl-1-propanol (AMP) as phosphate acceptor at
pH 10.4. The rate of change in absorbance due to the formation of pNP is measured bichromatically at 410/480 nm and is directly proportional to the ALP
activity in the sample.
Reaction Principle
ALP
pNPP + AMP pNP + AMP-PO4
2+
Mg
2-Amino-2-Methyl-1-Propanol (AMP) pH 10.4 0.35 mol/L
p-Nitrophenyl phosphate 16 mmol/L
HEDTA 2 mmol/L
Zinc Sulphate 1 mmol/L
Magnesium Acetate 2 mmol/L
Preservative
R1 reagent: Irritant. R36/38. Irritating to eyes and skin.
R2 reagent: Irritant, contains a mixture of 5-chloro-2-methyl-4-isothiazolin-3-one [EC No 247-500-7] and 2-methyl-4-isothiazolin-3-one [EC No
220-239-6] (3:1). R43; May cause sensitisation by skin contact.
Safety Phrases:
S24,S26,S37,S60. Avoid contact with skin. In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable
gloves. This material and its container must be disposed of as hazardous waste.
To avoid the possible build-up of azide compounds, flush waste-pipes with water after the disposal of undiluted reagent.
During the reaction p-nitrophenol is produced. This is harmful when inhaled, swallowed or absorbed through skin.
Reagent Preparation
The reagents are stable, unopened, up to the stated expiry date when stored at 2…8 °C.
Absorption of atmospheric CO2 by the reagent on board the analyser can impair its stability. This effect will vary depending upon the rate of use. Bottle
replacement is recommended when one of the following conditions are encountered:
14 days have elapsed on board the analyser;
Significant shift in control values (>7 %) following local QC procedures;
Specimen
4
Serum and heparinised plasma. Complexing anticoagulants such as citrate, oxalate and EDTA should be avoided.
Strongly haemolysed samples should be avoided.
5
Stable in serum and plasma for 7 days when stored at 2…25 °C.
2009-08
Test Procedure
Refer to the appropriate User Guide and Setting Sheet for analyser-specific assay instructions for the sample type as listed in the Intended Use
statement. The paediatric application is suitable for use with small volume serum/plasma samples.
Calibration
Re-establishment of the analyser specific MB factor is recommended when a critical part of the analyser is replaced;
Quality Control
Each laboratory should establish its own control frequency.
Good laboratory practice suggests that controls be tested each day patient samples are tested and each time calibration is performed. Values obtained for
the controls should fall within specified limits as defined by the user. If any trends or sudden shifts in values are detected, review all operating parameters.
Calculation
The Beckman Coulter analysers automatically compute the alkaline phosphatase activity of each sample.
Adults > 17y : 30 – 120 U/L
Children Male (U/L) Female (U/L)

1 – 30d 75 – 316 48 – 406
30d – 1y 82 – 383 124 – 341
1 – 3y 104 – 345 108 – 317
4 – 6y 93 – 309 96 – 297
7 – 9y 86 – 315 69 – 325
10 – 12y 42 – 362 51 – 332
13 – 15y 74 – 390 50 – 162
16 – 18y 52 – 171 47 – 119
values.
Linearity
The test is linear within an enzyme activity range of 5 – 1500 U/L (0.1 – 25.0 µkat/L).
Precision
21 0.40 1.94 0.93 4.48
260 1.45 0.56 2.57 0.99
856 7.67 0.90 11.09 1.30
Sensitivity
The lowest detectable level represents the lowest measurable level of ALP that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison
Patient serum samples were used to compare this ALP OSR6104 assay on the AU600 against another commercially available ALP assay. Results of
y = 0.934x + 8 r = 0.999 n = 124 Sample range = 17 – 1500 U/L
Haemolysis: Interference less than 10% up to 4.5 g/L haemoglobin
®
6
BIBLIOGRAPHY
1. Moss DW, Henderson RA. Clinical Enzymology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders Company, 1999; 676-684.
2. Thomas L. Alkaline phosphatase (ALP). In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
3. Tietz NW, Rinker D, Shaw LM. IFCC methods for the measurement of catalytic concentration of enzymes Part 5. IFCC method for alkaline phosphatase.
J Clin Chem Clin Biochem 1983;21:731-48.
4. Moss DW, Henderson RA, Kachmar JF. Enzymes In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:387pp.
5. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of Anticoagulants in Diagnostic Laboratory Investigations and Stability of Blood, Plasma and Serum
Samples. WHO/DIL/LAB/99.1 Rev.2:21pp.

2009-08
ALP (IFCC), AU400/AU640 Serum/Plasma Application ALP (IFCC), AU600 Serum/Plasma Application
System Reagent: OSR6004, OSR6104, OSR6204 Reagent ID: 004 System Reagent: OSR6004, OSR6104, OSR6204 Reagent ID: 004
Test No # Test name ALP04 ∇ Sample type Ser ∇ Page ½
Sample: Volume 3 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 60 Dil. Vol 60 μL L 0.0 H 2.5
R2 Volume 60 μL Dilution 60 μL L 0.0 H 2.5 Fst. L 0.0 Fst. H 1.2
Reagent OD limit: Wave Main 410 Sub 480 ∇ Lst. L 0.0 Lst. H 1.2
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L 0.0 First H 1.2 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 0.0 Last H 1.2 Reaction + ∇ L 5* H 1500*
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil Rate ¤
General LIH ISE Range Test No # Test name ALP04 ∇ Sample type Ser ∇ Page 2/2
Test Name: ALP04 ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name ALP04 ∇
Test Name: ALP04 Type Serum
∇ < > ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § § 4000* § 6500*
Point 1: § § § 4000* § 6500*
Point 2 ∇
Point 2:
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration # ∇ 1-point cal. Point
MB type factor #
# User defined
§ For use in the AB mode only, refer to IFU for further instruction
Enzyme BSOSR6x04.01
2009-08
ALP (IFCC), AU2700/AU5400 Serum/Plasma Application ALP (IFCC), AU680/AU480 Serum/Plasma Application
Test Name: ALP04 ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: ALP04 < > Type: Serum ∇ Operation: Yes ∇
μL μL
Linearity Limit 15 % Icterus ++++ ∇
Specific Test Parameters Lag Time Check NO ∇ Hemolysis +++++ ∇
Test Name: ALP04 ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: ALP04 ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
SERUM/PLASMA
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: ALP04 ∇ < > Type Serum ∇
Test Name: ALP04 Type Serum Use Serum Cal.
APPLICATION
∇ < > ∇ ο
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
Point 1: § § § 4000* § 6500* Point 1: § ∇ § §4000* §6500*
Point 9 ∇
§ For use in the AB mode only, refer to IFU for further instruction Calibrator OD Conc Low High Stability
* Values set for working in U/L To work in SI units (µkat/L) divide by 60 Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
ф AU680
MB Type Factor: # 1-Point Calibration Point ∇ ο with Conc-0
Enzyme BSOSR6x04.01
2009-08
ALP (IFCC), AU400/AU640 Paediatric Application ALP (IFCC), AU600 Paediatric Application

Test No # Test name ALP4P ∇ Sample type Ser ∇ Page 1/2
Sample: Volume 3 μL Dilution 10 μL Pre-Dilution Rate: 1 Reagent 1 vol 60 Dil. vol 60 μL L 0.0 H 2.5
Method: RATE ∇ Last L 0.0 Last H 1.2 Reaction + ∇ L 5* H 1500*
General LIH ISE Range Test No # Test name ALP4P ∇ Sample type Ser ∇ Page 2/2
Test Name: ALP4P ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name ALP4P ∇
Test Name: ALP4P ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § § 4000* § 6500*
Point 1: § § § 4000* § 6500*
Point 2 ∇
Point 2:
MB type factor #
# User defined
Enzyme BSOSR6x04.01
2009-08
ALP (IFCC), AU2700/AU5400 Paediatric Application ALP (IFCC), AU680/AU480 Paediatric Application
Test Name: ALP4P ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: ALP4P < > Type: Serum ∇ Operation: Yes ∇

Sample Volume 1.8 μL Dilution 10 μL OD Limit
Sample: Volume 1.8 μL Dilution 10 μL Pre-Dilution Rate: 1 Pre-Dilution Rate 1 ∇ Min.OD -0.1 Max.OD 2.5
Reagents: R1 Volume 36 μL Dilution 36 μL Min OD Max OD Rgt. Volume R1(R1-1) 36 μL Dilution 36 μL Reagent OD Limit
R2 Volume 36 Dilution 26 L -0.1 H 2.5 First Low -0.1 High 1.2
μL μL
Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Method RATE ∇
No Lag Time: NO ∇ On-board stability period: 14
Lag Time Check NO ∇ Hemolysis +++++ ∇
General LIH ISE Range Parameters Specific Test Parameters
Test Name: ALP4P ∇ < > Type: Serum ∇
Test Name: ALP4P ∇ < > Type: Serum ∇
Normal Ranges: Age L Age H Value/Flag: # ∇ Low High
Sex Year Month Year Month L H Level # # Panic Value
ο 1. # ∇ # # # # # # Specificl Ranges: From To Low High
ο 2. # ∇ # # # # # #
ο 1. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 4. # ∇ # # # # # # # # # # # # #
ο 3. ∇
ο 5. # ∇ # # # # # # ο 4. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 5. # ∇ # # # # # #
7. None Selected # # ο 6. # ∇ # # # # # #
8. Out of Range L H # # 7. No demographics # #
Panic Value: # # Unit: U/L* Decimal places: # 8. Not within expected values # #
Calibration Specific Calibrators Calibration Specific STAT Table Calibration
General ISE
General ISE
Test Name: ALP4P ∇ < > Type Serum ∇ ο Use Serum Cal.
Test Name: ALP4P ∇ < > Type Serum ∇
Point 1: § ∇ § §4000* §6500*
Point 1: § § § 4000* § 6500*
Point 7: Point 8 ∇ Operation # ∇
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 9 ∇
MB Type Factor: # Calibration Stability Period: ‡ Point 10 ∇ Interval (RB/ACAL) # ∇
Master Curve> OD Range
# User defined Calibrator OD Conc Low High Stability
§ For use in the AB mode only, refer to IFU for further instruction Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 Point-2 ∇ Calibration Day Hour
‡ Depends on usage pattern in the Laboratory MB Type Factor: # 1-Point Calibration Point ∇ ο with Conc-0
ф AU680
Enzyme BSOSR6x04.01
2009-08
ALT
OSR6007 4 x 12 mL R1
4 x 6 mL R2
OSR6107 4 x 50 mL R1
4 x 25 mL R2
OSR6507 4 x 102 mL R1
4 x 52 mL R2
OSR60180 4 x 17 mL Liquid P-5-P R1-2
60106 6 x 4 mL Liquid Pyridoxal Phosphate
60100 4 x 25 Tablets Pyridoxal Phosphate
Intended Use
Kinetic UV test for the quantitative determination of alanine aminotransferase, EC 2.6.1.2 (ALT), in human serum and plasma on Beckman Coulter
OSR6507 for use on the AU2700 and AU5400 systems only. OSR60180 for use with 3-part-reagent enabled systems only.
Summary1,2,3
ALT is an aminotransferase, a group of enzymes which catalyse the reversible transformation of α-keto acids into amino acids by transfer of amino groups.
Since the specific activity of ALT in the liver is approximately 10 times that of heart and skeletal muscle, elevated serum ALT activity is mainly regarded as
an indicator of parenchymal liver disease. ALT is present in the cytosol of hepatocytes, and increased serum levels indicate deterioration in the integrity of
the hepatocyte plasma membrane. ALT has greater diagnostic sensitivity for hepatobiliary disease than AST. Activities >50 times the upper reference limit
are mainly associated with acute viral hepatitis, acute disorders of liver perfusion and acute liver necrosis due to ingestion of toxins including paracetamol
and carbon tetrachloride. Markedly elevated serum ALT levels may be found in a variety of diseases involving the liver, including hepatitis, mononucleosis
and cirrhosis. Elevated ALT levels may be detected in viral hepatitis and other forms of liver disease prior to development of overt clinical symptoms such
as jaundice. Levels greater than 15 times the upper reference limit are always indicative of acute hepatocellular necrosis of viral, toxic or circulatory origin.
Increased ALT levels may also be detected in cirrhosis and extrahepatic cholestasis. Slight or moderate increases in ALT levels may also be observed
after ingestion of alcohol, or administration of drugs including penicillin, salicylates or opiates.
Test Principle4
ALT transfers the amino group from alanine to 2-oxoglutarate to form pyruvate and glutamate. The addition of pyridoxal phosphate to the reaction mixture
ensures maximum catalytic activity of ALT. The pyruvate enters a lactate dehydrogenase (LDH) catalysed reaction with NADH to produce lactate and
+
NAD . The decrease in absorbance due to the consumption of NADH is measured at 340 nm and is proportional to the ALT activity in the sample.
Endogenous pyruvate is removed during the incubation period.
Reaction Principle
ALT
2-Oxoglutarate + L-Alanine L-Glutamate + Pyruvate
+ LDH +
Pyruvate + NADH + H L-lactate + NAD

Tris buffer, pH: 7.15 (37°C) 100 mmol/L
L- Alanine 500 mmol/L
2-Oxoglutarate 12 mmol/L
LDH ≥ 1.8 kU/L
NADH 0.20 mmol/L
Pyridoxal Phosphate (P-5-P) 0.1 mmol/L (when Cat. No. 60106, 60100 or OSR60180 is used)
Preservative

Safety data sheet available for professional user on request.
Reagent Preparation
With manual pyridoxal phosphate addition
Pyridoxal Phosphate Liquid (Cat. No. 60106) and Pyridoxal Phosphate tablets (Cat No. 60100) are supplied separately for pyridoxal phosphate activation.
Pipette Pyridoxal Phosphate Liquid 60106 into the R1 bottle according to the table below, and mix by gentle inversion. Alternatively, dissolve tablets
completely in the R1 bottle according to the table below, by mixing gently several times. R2 is ready for use and can be placed directly on board the
instrument. This method can also be used for 3-part-reagent enabled systems.
Cat. No. Volume Pyridoxal Phosphate Liquid No. of Pyridoxal Phosphate tablets
OSR6007 0.25 mL 1 tablet
OSR6107 1 mL 4 tablets
Without pyridoxal phosphate activation


2009-08
Reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C.
Once open, reagents stored on board the instrument are stable for 30 days.
With pyridoxal phosphate activation
After addition of pyridoxal phosphate, R1 stored on board the instrument is stable for 7 days.
R2 stored on board the instrument is stable for 30 days.
Pyridoxal Phosphate Liquid reagent (Cat. No 60106)
Once open, Pyridoxal Phosphate Liquid reagent is stable until the expiry date printed on the label, provided that contamination is avoided through
adherence to GLP, the cap is replaced immediately after use and the reagent is stored at 2…8°C.
3-part-reagent enabled systems with P-5-P activation
Reagent Preparation
P-5-P Liquid (Cat. No. OSR60180) is specifically for use on board 3-part-reagent enabled systems. The reagent is ready for use and can be placed directly
on board the instrument in the R1 carousel. R1 and R2 are also ready for use and can be placed directly on board the instrument.
Storage and stability
Once open, P-5-P (Cat no OSR60180) is stable on board the instrument for 60 days. R1 and R2 reagent stored on board the instrument are stable for
30 days.
Specimen
Serum, and EDTA or heparinised plasma.
5
Stable in serum and plasma for 7 days when stored at 2…8°C and 3 days when stored at 15…25°C.
Test Procedure
Calibration
The test is run in MB-mode. To provide a robust approach to generate the analyser specific MB factor it is recommended that 5 separate calibration events
With pyridoxal phosphate activation, the calibrator value is traceable to the IFCC reference method and IRMM/IFCC-454. Without pyridoxal phosphate
activation, the calibrator is traceable to a Beckman Coulter Master Calibrator.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the ALT activity of each sample.
Male (Adult) < 50 U/L (0.85 µkat/L)
Female (Adult) < 35 U/L (0.60 µkat/L)
Newborn/Infant 13 − 45 U/L (0.22 − 0.75 µkat/L)

values.
Linearity
Precision
18 0.53 2.95 0.57 3.16
56 0.80 1.43 1.00 1.78
459 2.88 0.63 3.67 0.80
Sensitivity
The lowest detectable level represents the lowest measurable level of ALT that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison with pyridoxal phosphate activation

Patient serum samples were used to compare ALT OSR6107 assay on the AU640 against the IFCC reference method. Results of linear regression
analysis were as follows:
y = 1.010x - 0.1 r = 0.999 n = 117 Sample range = 3 – 264 U/L
2009-08
®
Pyruvate: Interference less than 5% up to 1 mmol/L pyruvate
8
Limitations
Highly lipemic samples may exceed the reaction absorbance and will be flagged with a “@”. Such samples should be diluted and re-run.

BIBLIOGRAPHY
1. Thomas L. Alanine aminotransferase (ALT), Aspartate aminotransferase (AST). In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical
laboratory results. Frankfurt/Main: TH-Books Verlagsgesellschaft, 1998:55-65.
2. Moss DW, Henderson RA, Kachmar JF. Enzymes. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:369-373.
3. Schmidt E, Schmidt FW. Diagnosis of icteric diseases. Dtsch Med Wschr 1984; 109: 139-146.
4. Schumann G, Bonora R, Ceriotti F et al. IFCC Primary Reference Procedures for the Measurement of Catalytic Activity Concentrations of Enzymes at 37°C.
Part 4. Reference Procedure for the Measurement of Catalytic Concentration of Alanine Aminotransferase. Clin Chem Lab Med 2002;40:718–24.
5. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of anticoagulants in diagnostic laboratory investigations and stability of blood, plasma and
serum samples. WHO/DIL/LAB/99.1 Rev.2:21pp.
6. Thomas L, Müller M, Schumann G et al. Consensus of DGKL and VDGH for interim reference intervals on enzymes in serum. J Lab Med 2005;29:301-08.
7. Painter PC, Cope JY, Smith JL. Reference information for the clinical laboratory. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry.
Philadelphia:WB Saunders Company, 1999; 1800pp.

2009-08
ALT (IFCC, without Pyridoxal phosphate activation), AU400/AU640 ALT (IFCC, without Pyridoxal phosphate activation), AU600
Serum/Plasma Application Serum/Plasma Application
System Reagent: OSR6007, OSR6107 Reagent ID: 007 System Reagent: OSR6007, OSR6107 Reagent ID: 007
Test No # Test name ALT ∇ Sample type Ser ∇ Page ½
Test Name: ALT ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 10 Dil. Vol 0 µL Min. OD Max. OD
Sample: Volume 10 µL Dilution 0 µL Pre-Dilution Rate: 1 Reagent 1 vol 100 Dil. Vol 50 µL L 0.60 H 2.5
Reagents: R1 Volume 100 µL Dilution 50 µL Min OD Max OD Reagent 2 vol 50 Dil. Vol 50 µL Reagent OD limit
R2 Volume 50 µL Dilution 50 µL L 0.60 H 2.5 Fst. L 1.1 Fst. H 2.5
Method: RATE ∇ Last L 1.1 Last H 2.5 Reaction - ∇ L 3* H 500*
Reaction slope: - ∇ Dynamic Range: Point 1 Fst 14 Lst 27 Correlation factor A 1
No Lag Time: YES ∇ On-board stability period: 30 Linearity Fst 15 % Sec %
General LIH ISE Range Test No # Test name ALT ∇ Sample type Ser ∇ Page 2/2
Test Name: ALT ∇ < > Type: Serum ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name ALT ∇
Test Name: ALT ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470*
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. Point
MB type factor #
§ For use in AB mode only, refer to IFU for further instruction. § For use in AB mode only, refer to IFU for further instruction.
Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, without Pyridoxal phosphate activation), AU2700/AU5400 ALT (IFCC, without Pyridoxal phosphate activation), AU680/AU480
Test Name: ALT ∇ < > Type: Serum ∇ Operation Yes ∇
Sample: Volume 5 Dilution 0 Pre-Dilution Rate: 1 Sample Volume 5 µL Dilution 0 µL OD Limit

µL µL
Pre-Dilution Rate 1 ∇ Min.OD 0.60 Max.OD 2.5
Reagents: R1 Volume 50 µL Dilution 25 µL Min OD Max OD
Rgt. Volume R1(R1-1) 50 µL Dilution 25 µL Reagent OD Limit
R2 Volume 25 µL Dilution 25 µL L 0.60 H 2.5
First Low 1.1 High 2.5
Reagent OD limit:
Last Low 1.1 High 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.1 First H 2.5
R2(R2-1) 25 µL Dilution 25 µL
Method: RATE ∇ Last L 1.1 Last H 2.5
Reaction slope: - ∇ Dynamic Range: Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Linearity : 15 % A 1 B 0 Reaction Slope - ∇ Onboard Stability Period 30 Day 0 Hour
No Lag Time: YES ∇ On-board stability period: 30 Measuring Point1 First 14 Last 27 LIH Influence Check # ∇
Specific Test Parameters Lag Time Check YES ∇ Hemolysis +++++ ∇
Test Name: ALT ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: ALT ∇ < > Type: Serum ∇
R 1. # ∇ # # # # # # Level # # Panic Value
R 2. Specificl Ranges: From To Low High
# ∇ # # # # # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # # # # # # #
∇
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: ALT ∇ < > Type Serum ∇
Test Name: ALT ∇ < > Type Serum ∇ R Use Serum Cal.
Point 1: § § §3650* §5470* Point 1: § ∇ § §3650* §5470*
Point 4: Point 4: ∇ R Reagent Blank
Point 5: Point 5: ∇ R Calibration
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
§ For use in AB mode only, refer to IFU for further instruction. Point-1 ∇ Reagent Blank Day Hour
Ф AU680 Point-2 ∇ Calibration Day Hour
MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0
Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, with OE60106 activation), AU400/AU640 Serum/Plasma ALT (IFCC, with OE60106 activation), AU600
Application Serum/Plasma Application

Test No # Test name ALT ∇ Sample type Ser ∇ Page ½
General LIH ISE Range Test No # Test name ALT ∇ Sample type Ser ∇ Page 2/2
Level L Level H
Test Name: ALT ∇ < > Type: Serum ∇
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
Panic value # #
General ISE Test No # Test name ALT ∇
Test Name: ALT ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #

SERUM/PLASMA APPLICATION Formula 1
Y=AX+B ∇ Process ∇

Cal. No. OD CONC Factor/OD-L Factor/OD-H Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470* Point 2 ∇
Point 2:
Point 3 ∇
Point 3:
Point 4 ∇
Point 4:
Point 5 ∇
Point 5:
Point 6 ∇
Point 6:
1-point cal. Point
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇
MB type factor #
Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, with OE60106 activation), AU2700/AU5400
Serum/Plasma Application
System Reagent: OSR6007, OSR6107, OSR6507 Reagent ID: 007

Sample: Volume 5 µL Dilution 0 µL Pre-Dilution Rate: 1

Reagent OD limit:
Reaction slope: - ∇ Dynamic Range:
No Lag Time: YES ∇ On-board stability period: 7

Test Name: ALT ∇ < > Type: Serum ∇

R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # #
General ISE
Test Name: ALT ∇ < > Type Serum ∇

Point 1: § § §3650* §5470*
Point 2:
Point 3:
Point 4:
Point 5:
Point 6:
Point 7:
# User defined
§ For use in AB mode only, refer to IFU for further instruction.
Ф AU680
Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, with OE60106 activation), AU680/AU480 Serum/Plasma Application ALT (IFCC, with OSR60180 activation), AU680 Serum/Plasma Application
Parameters Specific Test Parameters Parameters Specific Test Parameters
General LIH ISE HbA1c Calculated Test Range General LIH ISE HbA1c Calculated Test Range
Test Name: ALT ∇ < > Type: Serum ∇ Operation Yes ∇ Test Name: ALT ∇ < > Type: Serum ∇ Operation Yes ∇
Sample Volume 5 µL Dilution 0 µL OD Limit Sample Volume 5 µL Dilution 0 µL OD Limit

Pre-Dilution Rate 1 ∇ Min.OD 1.0 Max.OD 2.5 Pre-Dilution Rate 1 ∇ Min.OD 1.0 Max.OD 2.5
Rgt. Volume R1(R1-1) 50 µL Dilution 25 µL Reagent OD Limit Rgt. Volume R1(R1-1) 50 µL Dilution 10 µL Reagent OD Limit
First Low 1.3 High 2.5 R1-2 5 µL Dilution 10 µL First Low 1.3 High 2.5
Last Low 1.3 High 2.5 Last Low 1.3 High 2.5
R2(R2-1) 25 µL Dilution 25 µL R2(R2-1) 25 µL Dilution 25 µL
Dynamic Range Low 3* High 500* Dynamic Range Low 3* High 500*
Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0 Common Rgt. Type R1-2 Name # Correlation Factor A 1 B 0
Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0 Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0
Method RATE ∇ Method RATE ∇
Reaction Slope - ∇ Onboard Stability Period 7 Day 0 Hour Reaction Slope - ∇ Onboard Stability Period 30 Day 0 Hour
Measuring Point1 First 14 Last 27 LIH Influence Check # ∇ Measuring Point1 First 14 Last 27 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇ Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus +++++ ∇ Linearity Limit 15 % Icterus +++++ ∇
Lag Time Check YES ∇ Hemolysis +++++ ∇ Lag Time Check YES Hemolysis +++++
∇ ∇

Test Name: ALT ∇ < > Type: Serum ∇ Test Name: ALT ∇ < > Type: Serum ∇
Value/Flag: # ∇ Low High Value/Flag: # Low High
∇
Level # # Panic Value Level # # Panic Value
Specificl Ranges: From To Low High Specificl Ranges: From To Low High
Sex Year Month Year Month Low High # # Sex Year Month Year Month Low High # #
R 1. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # # R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # # R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 6. # ∇ # # # # # #
7. No demographics # # 7. No demographics # #
8. Not within expected values # # 8. Not within expected values # #
Unit U/L* Decimal Places # Unit U/L* Decimal Places #
Parameters Calibration Parameters Parameters Calibration Parameters
Calibrators Calibration Specific STAT Table Calibration Calibrators Calibration Specific STAT Table Calibration
General ISE General ISE
Test Name: ALT ∇ < > Type Serum ∇ R Use Serum Cal. R
Test Name: ALT ∇ < > Type Serum ∇ Use Serum Cal.
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
<Calibrator Parameters> Range <Calibrator Parameters> Range
Calibrator OD Conc Low High Slope Check None ∇ Calibrator OD Conc Low High Slope Check None ∇
Point 1: § ∇ § §3650* §5470* Point 1: § § §3650* §5470*
∇
Point 2: ∇ Allowance Range Check Point 2: Allowance Range Check
∇
Point 3: ∇ Point 3: ∇
Point 4: ∇ R Reagent Blank Point 4: ∇ R Reagent Blank
Point 5: ∇ R Calibration Point 5: ∇ R Calibration
Point 7 ∇ Advanced Calibration Point 7 Advanced Calibration
∇
Point 8 ∇ Operation # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇ Point 9 ∇
Point 10 ∇ Interval (RB/ACAL) # ∇ Point 10 ∇ Interval (RB/ACAL) # ∇
Master Curve> OD Range Master Curve> OD Range
Calibrator OD Conc Low High Stability Calibrator OD Conc Low High Stability
Point-1 ∇ Reagent Blank Day Hour Point-1 ∇ Reagent Blank Day Hour
Point-2 ∇ Calibration Day Hour Point-2 Calibration Day Hour
∇
MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0 MB Type Factor: # 1-Point Calibration Point R with Conc-0
∇
Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, without Pyridoxal phosphate activation), AU400/AU640 ALT (IFCC, without Pyridoxal phosphate activation), AU600
Paediatric Application Paediatric Application
Test No # Test name ALTP ∇ Sample type Ser ∇ Page 1/2
Test Name: ALTP ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 10 Dil. vol 10 µL Min. OD Max. OD
Sample: Volume 10 µL Dilution 10 µL Pre-Dilution Rate: 1 Reagent 1 vol 100 Dil. vol 50 µL L 0.60 H 2.5
Reagents: R1 Volume 100 µL Dilution 50 µL Min OD Max OD Reagent 2 vol 50 Dil. vol 40 µL Reagent OD limit
General LIH ISE Range Test No # Test name ALTP ∇ Sample type Ser ∇ Page 2/2
Test Name: ALTP ∇ < > Type: Serum ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name ALTP ∇
Test Name: ALTP ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470*
1-Point Cal. Point: R With CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor #
# User defined
Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, without Pyridoxal phosphate activation), AU2700/AU5400 ALT (IFCC, without Pyrioxal phosphate activation), AU680/AU480
Test Name: ALTP ∇ < > Type: Serum ∇ Operation Yes ∇
Sample: Volume 5 µL Dilution 10 µL Pre-Dilution Rate: 1 Sample Volume 5 µL Dilution 10 µL OD Limit

Reagents: R1 Volume 50 µL Dilution 25 µL Min OD Max OD Pre-Dilution Rate 1 ∇ Min.OD 0.60 Max.OD 2.5
R2 Volume 25 µL Dilution 15 µL L 0.60 H 2.5 Rgt. Volume R1(R1-1) 50 µL Dilution 25 µL Reagent OD Limit
Reagent OD limit: First Low 1.1 High 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.1 First H 2.5 Last Low 1.1 High 2.5
Method: RATE ∇ Last L 1.1 Last H 2.5 R2(R2-1) 25 µL Dilution 15 µL
Reaction slope: - ∇ Dynamic Range: Dynamic Range Low 3* High 500*
Measuring Point 1: First 14 Last 27 L 3* H 500* Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Measuring Point 2: First Last Correlation Factor: Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0
Linearity : 15 % A 1 B 0 Method RATE ∇
No Lag Time: YES ∇ On-board stability period: 30 Reaction Slope - ∇ Onboard Stability Period 30 Day 0 Hour
Test Name: ALTP ∇ < > Type: Serum ∇ Parameters Specific Test Parameters
Value/Flag: # ∇ Level L: # Level H: # Test Name: ALTP Type: Serum
∇ < > ∇
R 2. # ∇ # # # # # # Specificl Ranges: From To Low High
R 3. # ∇ # # # # # # Sex Year Month Year Month Low High # #
R 4. # # # # # # # R 1. # ∇ # # # # # #
∇
R 5. R 2. # ∇ # # # # # #
# ∇ # # # # # #
R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # #
R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
Calibration Specific Parameters Calibration Parameters
General ISE Calibrators Calibration Specific STAT Table Calibration
General ISE
Test Name: ALTP ∇ < > Type Serum ∇
Test Name: ALTP ∇ < > Type Serum ∇ R Use Serum Cal.
Cal. No. OD CONC Factor/OD-L Factor/OD-H <Calibrator Parameters> Range
Point 1: § § §3650* §5470* Calibrator OD Conc Low High Slope Check None ∇
Point 2: Point 1: § ∇ § §3650* §5470*
MB Type Factor: # Calibration Stability Period: Point 9 ∇
Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, with OE60106 activation), AU400/AU640 ALT (IFCC, with OE60106 activation), AU600

Test No # Test name ALTP ∇ Sample type Ser ∇ Page 1/2
General LIH ISE Range Test No # Test name ALTP ∇ Sample type Ser ∇ Page 2/2
Test Name: ALTP ∇ < > Type: Serum ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name ALTP ∇
Test Name: ALTP ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470*
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor #
# User defined
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 # User defined ¤ Analyser default value
§ For use in AB mode only, refer to IFU for further instruction. * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, with OE60106 activation), AU2700/AU5400
Paediatric Application
System Reagent: OSR6007, OSR6107, OSR6507 Reagent ID: 007


Reagent OD limit:

Test Name: ALTP ∇ < > Type: Serum ∇

R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # #
General ISE
Test Name: ALTP ∇ < > Type Serum ∇

Point 1: § § §3650* §5470*
Point 2:
Point 3:
Point 4:
Point 5:
Point 6:
Point 7:
# User defined
Ф AU680
Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, with OE60106 activation), AU680/AU480 Paediatric Application ALT (IFCC, with OSR60180 activation), AU680 Paediatric Application
Test Name: ALTP ∇ < > Type: Serum ∇ Operation Yes ∇ Test Name: ALTP ∇ < > Type: Serum ∇ Operation Yes ∇

Lag Time Check YES ∇ Hemolysis +++++ ∇ Lag Time Check YES Hemolysis +++++
∇ ∇

Test Name: ALTP ∇ < > Type: Serum ∇ Test Name: ALTP ∇ < > Type: Serum ∇
Value/Flag: # ∇ Low High Value/Flag: # Low High
∇
R 1. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # # R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # # R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 6. # ∇ # # # # # #
Test Name: ALTP ∇ < > Type Serum ∇ R Use Serum Cal. R
Test Name: ALTP ∇ < > Type Serum ∇ Use Serum Cal.
Point 1: § ∇ § §3650* §5470* Point 1: § § §3650* §5470*
∇
Point 2: ∇ Allowance Range Check Point 2: Allowance Range Check
∇
Point 7 ∇ Advanced Calibration Point 7 Advanced Calibration
∇
Point-2 ∇ Calibration Day Hour Point-2 Calibration Day Hour
∇
MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0 MB Type Factor: # 1-Point Calibration Point R with Conc-0
∇
Enzyme BSOSR6x07.02
2010-05
α-AMYLASE
OSR6006 4 x 10 mL R1
OSR6106 4 x 40 mL R1
Intended Use
Kinetic colour test for the quantitative determination of α-amylase, [1,4-α-D-glucan 4-glucanohydrolase, EC 3.2.1.1], in human serum, plasma and urine on
Beckman Coulter analysers. For in vitro diagnostic use only.
Summary1,2,3
Amylases are a group of hydrolases that split complex carbohydrates composed of alpha-D-glucose units linked through carbon atoms 1 and 4 located on
adjacent glucose residues. In the body, amylase is present in a number of organs and tissues. The greatest concentration is present in the pancreas,
where the enzyme is synthesised by the acinar cells and then secreted into the intestinal tract by way of the pancreatic duct system. The salivary glands
also secrete a potent amylase to initiate hydrolysis of starches while the food is still in the mouth and oesophagus.
Diseases resulting in elevation of plasma alpha-amylase include: acute pancreatitis, parotitis, alcoholism, renal insufficiency and diseases such as viral
hepatitis, AIDS, abdominal typhoid, sarcoidosis and trauma to the upper abdomen. There is also a detectable increase in amylase after an ERCP
procedure.
In acute pancreatitis, amylase increases 5-6 hours after the onset of symptoms and remains elevated for 2-5 days. The increase in plasma activity does
not reflect disease severity and conversely, extensive destruction of the pancreas may not cause a significant increase in the plasma concentration of
pancreatic alpha-amylase.
Alpha amylase is excreted by glomerular filtration and then 50% of it is reabsorbed by the tubules. This reabsorption is significantly reduced in transient
tubular damage, after burns, in the presence of diabetic ketoacidosis and acute pancreatitis as well as proteinuria resulting in an increase of alpha-amylase
clearance. The measurement of alpha amylase in urine is indicated in the investigation of hyperamylasemia associated with macroamylasemia or renal
insufficiency. Hypoamylasemia has been observed in advanced cystic fibrosis, severe liver disease and pancreatectomy. Also due to the decreased
concentration of the salivary fraction hypoamylasemia has been found in obese subjects.
Test Principle4
The α-amylase colour test employs 2-chloro-4-nitrophenyl-α-D-maltotrioside (CNPG3) as substrate. This substrate reacts directly with α-amylase and does
not require the presence of ancillary enzymes. The release of 2-chloro-4-nitrophenol (CNP) from the substrate and the resulting absorbance increase at
410 nm is directly proportional to the α-amylase activity in the sample.
Reaction Principle
CNPG3 + H2O α-amylase CNP + Maltotriose

MES (pH 6.05) 36.1 mmol/L
Calcium acetate 3.60 mmol/L
NaCl 37.2 mmol/L
Potassium thiocyanate 253 mmol/L
CNPG3 1.63 mmol/L
Preservative
Reagent Preparation
The reagent is ready for use and can be placed directly on board the instrument.
Care should be taken when handling this reagent to avoid contamination with skin and body fluids.
The reagent is stable, unopened, up to the stated expiry date when stored at 2...8°C. Once open the reagent stored on board the instrument is stable for
30 days.
Discard reagents if any discolouration is observed.
Specimen
Serum, heparinised plasma. Haemolysed and strongly icteric samples should be avoided, separate from blood cells as soon as possible.
5
1,6
Plasma using EDTA, oxalate and citrate should be avoided.
1 6
Urine: Timed or random sample. Adjust pH to approximately 7.0 before storage.
5
Stable in urine for 10 days when stored at 2…8°C and 2 days when stored at 15…25°C.
Test Procedure
Calibration
should be used. A fresh vial of calibrator, utilising System Calibrator Cat No. 66300 for serum application and Urine calibrator Cat no. ODC0025 for urine
application, in the AB calibration mode, should be used for each of these runs. When calculating the mean factor from the separate runs the data should be
examined for obvious outliers which should be repeated and replaced. For the AU2700/AU5400 this procedure needs to be performed for each ring.
Quality control procedures should be undertaken immediately following calibration in accordance with good laboratory practice.
The calibrator values are traceable to a Beckman Coulter master calibrator.

2009-08
Quality Control
Controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter system may be used for the
serum/plasma application.
Biorad Liquichek Urine Chemistry Controls Cat. No. 397 and 398 or other control materials with values determined by this Beckman Coulter system may
be used for the urine application.
Calculation
The Beckman Coulter analysers automatically compute the α-amylase activity of each sample.
Reference Intervals
3
Serum/Plasma 22 – 80 U/L (0.36 – 1.33 µkat/L)
7
Urine 42 – 321 U/L (0.7 – 5.35 µkat/L)
values.
Linearity
The test is linear within an enzyme activity range of 10 – 2000 U/L (0.2 – 33.3 µkat/L) for serum and plasma. The test is linear within an enzyme activity
range of 5 – 4800 U/L (0.1 – 80 µkat/L) for urine.
Precision
45 0.88 1.97 1.17 2.63
140 1.12 0.80 4.39 3.14
1562 12.62 0.81 28.82 1.84
The following data was obtained on an AU640 using 3 urine pools analysed over 20 days.
45.97 0.69 1.51 2.13 4.64
494.94 3.99 0.81 14.1 2.85
3207.84 50.73 1.58 101.63 3.17
Sensitivity
The lowest detectable level using serum settings on an AU600 analyser was calculated as 2 U/L.
The lowest detectable level using urine settings on an AU2700 analyser was calculated as 2 U/L.
The lowest detectable level represents the lowest measurable level of α-amylase that can be distinguished from zero. It is calculated as the absolute mean
plus three standard deviations of 20 replicates of an analyte free sample.
Method Comparison
Patient serum samples were used to compare this α-Amylase OSR6106 assay on the AU600 against another commercially available α-amylase assay.
y = 0.879x + 6.313 r = 0.999 n = 112 Sample range = 11 – 1530 U/L
Patient urine samples were used to compare this α-Amylase OSR6106 assay on the AU2700 against another commercially available α-amylase assay.
y = 1.067x – 4.492 r = 0.995 n = 125 Sample range = 18 – 339 U/L
Results of serum studies conducted to evaluate the susceptibility of the method to interference were as follows:
®
Results of urine studies conducted to evaluate the susceptibility of the method to interference were as follows:
8
BIBLIOGRAPHY
1. Lorentz K. α-Amylase. In: Thomas L, hrsg. Labor und Diagnose. Indikation und Bewertung von Laborbefunden für die Medizinische Diagnostik. Frankfurt/Main:
2. Tietz NW, ed. Clinical guide to laboratory tests, 3rd ed. Philadelphia: WB Saunders Company,1995:47pp.
1999;689-698.
4. IFCC methods for measurement of catalytic concentration of enzymes. Part 9. IFCC method for alpha-amylase [1,4-α-D-glucan 4-glucanohydrolase, EC 3.2.1.1].
International Federation of Clinical Chemistry. Clin Chim Acta. 1999;281(1-2):S5-39.
serum samples. WHO/DIL/LAB/99.1 Rev.2:22,46pp.
7. Ballsells D, Gella FJ, Gubern G, Canalias F, Reference values for α-amylase in human serum and urine using 2-chloro-4-nitrophenyl-α-D-maltotrioside as
substrate. Clin Chim Acta 1998;274:213–217.

2009-08
AMYLASE, AU400/AU640 Serum/Plasma Application AMYLASE, AU600 Serum/Plasma Application

Test No # Test name AMY ∇ Sample type Ser ∇ Page ½
Test Name: AMY ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name AMY ∇ Sample type Ser ∇ Page 2/2
Test Name: AMY ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name AMY ∇
Test Name: AMY ∇ < > Type Serum ∇ Cal type 1 MB Count #
∇
Point 1 § ∇ § §7120* §10680*
Point 1: § § §7120* §10680* Point 2 ∇
Point 2:
Point 3 ∇
Point 3:
Point 4 ∇
Point 4:
Point 5 ∇
Point 5:
1-point cal. Point
MB type factor #
§ For use in AB mode only, refer to leaflet for further instruction. § For use in AB mode only, refer to leaflet for further instruction.
Enzyme BSOSR6x06.01
2009-08
AMYLASE, AU2700/AU5400 Serum/Plasma Application AMYLASE, AU680/AU480 Serum/Plasma Application
Test Name: AMY ∇ < > Type: Serum ∇ Operation Yes ∇
μL μL
Reagent OD limit:
Specific Test Parameters Lag Time Check NO ∇ Hemolysis +++ ∇
Test Name: AMY ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: AMY ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: AMY ∇ < > Type Serum ∇
Test Name: AMY ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: § § §7120* §10680* Point 1: § ∇ § §7120* §10680*
Point 9 ∇
Point 10 ∇ Interval (RB/ACAL) ∇
# User defined
Calibrator OD Conc Low High Stability
Point-1 ∇ Reagent Blank Day Hour
Point-2 ∇ Calibration Day Hour
ф AU680
Enzyme BSOSR6x06.01
2009-08
AMYLASE, AU400/AU640 Urine Application AMYLASE, AU600 Urine Application
System Reagent: OSR6006, OSR6106 Reagent ID: 006 Reagent: OSR6006, OSR6106 Reagent ID: 006

Test No # Test name AMY ∇ Sample type Uri ∇ Page 1/2
Test Name: AMY ∇ < > Type: Urine ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name AMY ∇ Sample type Uri ∇ Page 2/2
Test Name: AMY ∇ < > Type: Urine ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
URINE APPLICATION
Test Name: AMY ∇ < > Type Urine ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §9300* §17300*
Point 1: § § §9300* §17300*
Point 2 ∇
Point 2:
Point 3 ∇
Point 3:
1-Point Cal. Point: With CONC-0 Slope Check: None # 1-point cal. point
ο ∇ Advanced Calibration: ∇
MB type factor #
§ For use in AB mode only, refer to leaflet for further instruction. § For use in AB mode only, refer to leaflet for further instruction.
Enzyme BSOSR6x06.01
2009-08
AMYLASE, AU2700/AU5400 Urine Application AMYLASE, AU680/AU480 Urine Application
Test Name: AMY ∇ < > Type: Urine ∇ Operation Yes ∇
μL μL
Reagent OD limit:
No Lag Time: NO ∇ On-board stability period: 30 Measuring Point1 First 5 Last 10
Measuring Point2 First Last
Linearity Limit 15 %
Specific Test Parameters Lag Time Check NO ∇
Test Name: AMY ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: AMY ∇ < > Type: Urine ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
URINE APPLICATION
General ISE
General ISE
Test Name: AMY ∇ < > Type Urine ∇ Test Name: AMY ∇ < > Type Urine ∇ ο Use Serum Cal.
Point 1: § § §9300* §17300* Point 1: § ∇ § §9300* §17300*
Point 9 ∇
# User defined
Point-2 ∇ Calibration Day Hour
ф AU680
Enzyme BSOSR6x06.01
2009-08
α-AMYLASE
OSR6182 4 x 40 mL R1
4 x 10 mL R2
Intended Use
Kinetic colour test for the quantitative determination of α-amylase, [1,4-α-D-glucan 4-glucanohydrolase, EC 3.2.1.1], in human serum, plasma
and urine on Beckman Coulter analysers. For in vitro diagnostic use only.
Summary1,2,3
Amylases are a group of hydrolases that split complex carbohydrates composed of alpha-D-glucose units linked through carbon atoms 1 and 4
located on adjacent glucose residues. In the body, amylase is present in a number of organs and tissues. The greatest concentration is present
in the pancreas, where the enzyme is synthesised by the acinar cells and then secreted into the intestinal tract by way of the pancreatic duct
system. The salivary glands also secrete a potent amylase to initiate hydrolysis of starches while the food is still in the mouth and oesophagus.
Diseases resulting in elevation of plasma alpha-amylase include: acute pancreatitis, parotitis, alcoholism, renal insufficiency and diseases such
as viral hepatitis, AIDS, abdominal typhoid, sarcoidosis and trauma to the upper abdomen. There is also a detectable increase in amylase after
an ERCP procedure.
In acute pancreatitis, amylase increases 5-6h after the onset of symptoms and remains elevated for 2-5 days. The increase in plasma activity
does not reflect disease severity and conversely, extensive destruction of the pancreas may not cause a significant increase in the plasma
concentration of pancreatic alpha-amylase.
Alpha amylase is excreted by glomerular filtration and then 50% of it is reabsorbed by the tubules. This reabsorption is significantly reduced in
transient tubular damage, after burns, in the presence of diabetic ketoacidosis and acute pancreatitis as well as proteinuria resulting in an
increase of alpha-amylase clearance. The measurement of alpha amylase in urine is indicated in the investigation of hyperamylasemia
associated with macroamylasemia or renal insufficiency.
Hypoamylasemia has been observed in advanced cystic fibrosis, severe liver disease and pancreatectomy. Also due to the decreased
concentration of the salivary fraction hypoamylasemia has been found in obese subjects.
Test Principle4
Method based on the recommendations of the “International Federation of Clinical Chemistry” (IFCC).
The α-amylase colour test employs 4,6-ethylidene(G7)-p-nitrophenyl(G1)-α-D-maltoheptaoside (ethylidene-G7PNP) as substrate. This substrate
reacts with α-amylase and the fragments with α-glucosidase to give a 100% release of p-Nitrophenol (PNP). The increase of absorbance at
410 nm is directly proportional to the α-amylase activity in the sample.
Reaction Principle (Simplified)4
α-amylase
5 ethylidene-G7PNP + 5H20 2 ethylidene-G5 + 2 G2PNP + 2 ethylidene-G4
+ 2 G3PNP + ethylidene-G3 + G4PNP
α-glucosidase
2 G2 PNP + 2 G3PNP + G4PNP + 14 H20 5PNP + 14G
(PNP = p-Nitrophenol; G = Glucose)
HEPES buffer (pH 7.15) 50 mmol/L
Sodium chloride 70 mmol/L
Calcium chloride 1 mmol/L
4,6-ethylidene-G7PNP > 1.4 mmol/L
α-Glucosidase ≥ 4.8 kU/L
Preservatives
R43. May cause sensitisation by skin contact.
S24,S37,S60. Avoid contact with skin. Wear suitable gloves. This material and its container must be disposed of as hazardous waste.
Reagent Preparation
Care should be taken when handling this reagent to avoid contamination with skin and body fluids.
The reagents are stable, unopened, up to the stated expiry date when stored at 2...8°C. Once open reagents stored on board the instrument are
stable for 90 days.
Discard reagents if any discolouration is observed.
Specimen
Serum or heparinised plasma.
5
1,6
Plasma using EDTA, oxalate and citrate should be avoided.
1 6
Urine: Timed or random sample. Adjust pH to approximately 7.0 before storage.
5
Test Procedure

2009-08
Calibration
The test is run in MB-mode. To provide a robust approach to generate the analyser specific MB factor, it is recommended that 5 separate
calibration events should be used. A fresh vial of calibrator, utilising System Calibrator Cat No. 66300 for serum application and Urine calibrator
Cat no. ODC0025 for urine application, in the AB calibration mode, should be used for each of these runs. When calculating the mean factor
from the separate runs the data should be examined for obvious outliers which should be repeated and replaced. For the AU2700/AU5400 this
procedure needs to be performed for each ring. Quality control procedures should be undertaken immediately following calibration in
The calibrator values are traceable to a manual IFCC reference method and IRMM/IFCC-456.
Re-establishment of the analyser specific MB factor is recommended when a critical part of the analyser is replaced or as deemed necessary by the user.
Quality Control
Controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter system may be used for
the serum application.
Biorad Liquichek Urine Chemistry Controls Cat. No. 397 and 398 or other control materials with values determined by this Beckman Coulter
system may be used for the urine application.
Calculation
The Beckman Coulter analysers automatically compute the α-amylase activity of each sample.
Serum/Plasma 28 - 100 U/L (0.46 – 1.66 µkat/L)
Urine
Male ≤ 490 U/L (8.16 µkat/L) or 280 U/g creatinine
Female ≤ 450 U/L (7.50 µkat/L) or 380 U/g creatinine
Expected values may vary with age, sex, sample type, diet and geographical location. Each laboratory should verify the transferability of the
expected values to its own population, and if necessary determine its own reference interval according to good laboratory practice. For
diagnostic purposes, results should always be assessed in conjunction with the patient's medical history, clinical examinations and other
findings.
Data contained within this section is representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ
from these values.
Linearity
The test is linear within an enzyme activity range of 10 – 1500 U/L (0.2 – 25.0 µkat/L) for serum and plasma.
The test is linear within an enzyme activity range of 10 – 4800 U/L (0 – 80.0 µkat/L) for urine.
Precision
51 0.36 0.71 0.59 1.16
132 0.73 0.56 1.82 1.38
1267 6.98 0.55 12.94 0.99
The following data was obtained on an AU640 using 3 urine pools analysed over 20 days
46 0.62 1.35 1.53 3.34
585 5.04 0.86 7.41 1.27
3732 21.11 0.57 44.65 1.20
Sensitivity
The lowest detectable level using serum settings on an AU640 analyser was calculated as 1 U/L.
The lowest detectable level using urine settings on an AU640 analyser was calculated as 2 U/L.
The lowest detectable level represents the lowest measurable level of α-amylase that can be distinguished from zero. It is calculated as the
absolute mean plus three standard deviations of 20 replicates of an analyte free sample.
Method Comparison
Patient serum samples were used to compare this α-Amylase OSR6182 assay on the AU640 against another commercially available amylase
assay. Results of linear regression analysis were as follows:
Patient urine samples were used to compare this α-Amylase OSR6182 assay on the AU640 against another commercially available amylase
y = 1.003x – 5.213 r = 1.000 n = 98 Sample range = 18 – 4470 U/L
®
8

2009-08
BIBLIOGRAPHY
1. Lorentz K. α-Amylase. In: Thomas L, hrsg. Labor und Diagnose.. Indikation und Bewertung von Laborbefunden für die Medizinische Diagnostik. Frankfurt/Main:
TH-Books Verlagsgellschaft, 2005:51-56.
1999; 689-692.
4. Lorentz K. Approved recommendation on IFCC methods for the measurement of catalytic concentration of enzymes. Part 9. IFCC method for α-amylase (1,4-α-
D-glucan 4-glucanohydrolase, EC 3.2.1.1). International Federation of Clinical Chemistry and Laboratory Medicine (IFCC). Committee on enzymes. Clin Chem
Lab Med 1998;36(3):185-203.
serum samples. WHO/DIL/LAB/99.1 Rev.2:22, 46pp.
6. Moss DW, Henderson RA, Kachmar JF. Enzymes. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia: WB Saunders Company, 1987:393-396.
7. Junge W, Wortmann W, Wilke B, Waldenstrom J, Weittenhiller A, Finke J, Klein G, Development and evaluation of assays for the determination of total and
pancreatic amylase at 37°C according to the principle recommended by the IFCC. Clin Biochem 2001;34:607–615.

2009-08
AMYLASE (EPS Liquid), AU400/AU640 Serum/Plasma Application AMYLASE (EPS Liquid), AU600 Serum/Plasma Application

Test No # Test name AMY ∇ Sample type Ser ∇ Page ½
General LIH ISE Range Test No # Test name AMY ∇ Sample type Ser ∇ Page 2/2
Test Name: AMY ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Test Name: AMY ∇ < > Type Serum ∇ Cal type 1 MB Count #
∇
Point 1 § ∇ § §5570* §8360*
Point 1: § § §5570* §8360*
Point 2 ∇
Point 2:
1-Point Cal. Point: Slope Check: None # 1-point cal. Point
ο with CONC-0 ∇ Advanced Calibration: ∇
MB type factor #
MB Type Factor: # Calibration Stability Period: 999 Calibrator stability period 999
§ For use in AB mode only, refer to IFU for further instruction § For use in AB mode only, refer to IFU for further instruction
Enzyme BSOSR6x82.01
2009-08
AMYLASE (EPS liquid), AU2700/AU5400 Serum/Plasma Application AMYLASE (EPS liquid), AU680/AU480 Serum/Plasma Application
Test Name: AMY ∇ < > Type: Serum ∇ Operation Yes ∇
Sample: Volume 2 Dilution 0 Pre-Dilution Rate: 1 Sample Volume 2 μL Dilution 0 μL OD Limit

μL μL
Reagent OD limit:
Test Name: AMY ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: AMY ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: AMY ∇ < > Type Serum ∇ Test Name: AMY ∇ < > Type Serum ∇ ο Use Serum Cal.
<Calibrator Parameters> Factor Range
Point 1: § § §6450* §9680* Point 1: § ∇ § §6450* §9680*
Point 9 ∇
MB Type Factor: # Calibration Stability Period: 999
§ For use in AB mode only, refer to IFU for further instruction Point-1 ∇ Reagent Blank 999 Day 0 Hour
ф AU680 Point-2 ∇ Calibration Day Hour
Enzyme BSOSR6x82.01
2009-08
AMYLASE (EPS Liquid), AU400/AU640 Urine Application AMYLASE (EPS Liquid), AU600 Urine Application

Test No # Test name AMY ∇ Sample type Uri ∇ Page 1/2
Sample vol. 1.5 Dil. vol 0 μL Min. OD Max. OD
Sample: Volume 1.5 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 150 Dil. vol 0 μL L -0.1 H 2.0
General LIH ISE Range Test No # Test name AMY ∇ Sample type Uri ∇ Page 2/2
Test Name: AMY ∇ < > Type: Urine ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
URINE APPLICATION
Test Name: AMY ∇ < > Type Urine ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §10500* §19500*
Point 1: § § §10500* §19500*
Point 2 ∇
Point 2:
1-Point Cal. Point: with CONC-0 Slope Check: None # 1-point cal. point
MB type factor #
MB Type Factor: # Calibration Stability Period: 999 Calibrator stability period 999
§ For use in AB mode only, refer to IFU for further instruction § For use in AB mode only, refer to IFU for further instruction
Enzyme BSOSR6x82.01
2009-08
AMYLASE (EPS liquid), AU2700/AU5400 Urine Application AMYLASE (EPS liquid), AU680/AU480 Urine Application
Test Name: AMY ∇ < > Type: Urine ∇ Operation Yes ∇
μL μL
Reagent OD limit:
No Lag Time: NO ∇ On-board stability period: 90 Measuring Point1 First 20 Last 27
Specific Test Parameters Lag Time Check NO ∇
Test Name: AMY ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: AMY ∇ < > Type: Urine ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
URINE APPLICATION
General ISE
General ISE
Test Name: AMY ∇ < > Type Urine ∇ Test Name: AMY ∇ < > Type Urine ∇ ο Use Serum Cal.
Point 1: § § §10500* §19500* Point 1: § ∇ § §10500* §19500*
1-Point Cal. Point: ο with CONC-0 Slope Check None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
§ For use in AB mode only, refer to IFU for further instruction Point-1 ∇ Reagent Blank 999 Day 0 Hour
Enzyme BSOSR6x82.01
2009-08
AST
OSR6009 4x 6 mL R1
4x 6 mL R2
OSR6109 4x 25 mL R1
4x 25 mL R2
OSR6209 4x 50 mL R1
4x 50 mL R2
OSR6509 4x 104 mL R1
4x 104 mL R2
OSR60180 4x 17 mL Liquid P-5-P R1-2
60106 6x 4 mL Liquid Pyridoxal Phosphate
60100 4x 25 Tablets Pyridoxal Phosphate
Intended Use
Kinetic UV test for the quantitative determination of aspartate aminotransferase, EC 2.6.1.1 (AST), in human serum and plasma on Beckman Coulter
OSR6509 for the use on the AU2700 and AU5400 systems only. OSR60180 for use with 3-part-reagent enabled systems only.
Summary1,2,3
AST occurs in a wide variety of tissues including liver, cardiac muscle, skeletal muscle, brain, kidneys, lungs, pancreas, erythrocytes and leucocytes, with
highest activities found in liver and skeletal muscle. Measurement of AST is indicated in the diagnosis, differentiation and monitoring of hepatobiliary
disease, myocardial infarction and skeletal muscle damage. AST measurement may also be performed as part of medical screening examinations. In
some cases, AST may be useful in monitoring the course of myocardial infarction. Where recent myocardial infarction is suspected, AST has a diagnostic
sensitivity of 96%, with a diagnostic sensitivity of 86% at 12 hours after onset of chest pain. AST levels may be increased in viral hepatitis and liver disease
associated with hepatic necrosis, with 20 to 50 fold elevations frequently encountered. The evaluation of AST activity in relation to ALT (De Ritis ratio;
AST/ALT) is a useful indicator of liver damage. Ratios <1.0 are indicative of mild liver damage, and are particularly associated with diseases of an
inflammatory nature. Ratios >1.0 are indicative of severe liver disease, usually involving necrosis. Increased AST levels may be detected in cirrhosis,
extrahepatic cholestasis, progressive muscular dystrophy, dermatomyositis, acute pancreatitis, haemolytic disease, gangrene, crushed muscle injuries and
pulmonary emboli. Slight or moderate increases in AST levels may also be observed after ingestion of alcohol, or administration of drugs including
penicillin, salicylates or opiates.
Test Principle4
In this method, aspartate aminotransferase (AST) catalyses the transamination of aspartate and 2-oxoglutarate, forming L-glutamate and oxalacetate. The
addition of pyridoxal phosphate to the reaction mixture ensures maximum catalytic activity of AST. The oxalacetate is reduced to L-malate by malate
dehydrogenase (MDH), while NADH is simultaneously converted to NAD+. The decrease in absorbance due to the consumption of NADH is measured at
340 nm and is proportional to the AST activity in the sample. Endogenous pyruvate is removed by the LDH-reaction during the incubation period.
Reaction Principle
AST
2-Oxoglutarate + L-Aspartate L-Glutamate + Oxalacetate
+ MDH +
Oxalacetate + NADH + H L-Malate + NAD

Tris buffer, pH 7.65 (37°C) 80 mmol/L
L-aspartate 240 mmol/L
2-Oxoglutarate 12 mmol/L
LDH ≥ 0.9 kU/L
MDH ≥ 0.6 kU/L
NADH 0.20 mmol/L
Pyridoxal phosphate (P-5-P) 0.1 mmol/L (when Cat. No. 60106, 60100 or OSR60180 is used)
Preservative
Reagent Preparation
With manual pyridoxal phosphate addition
Pyridoxal Phosphate Liquid (Cat. No. 60106), and Pyridoxal Phosphate tablets (Cat. No. 60100) are supplied separately for pyridoxal phosphate activation.
Pipette Pyridoxal Phosphate Liquid 60106 into the R1 bottle according to the table below, and mix by gentle inversion. Alternatively, dissolve tablets
completely in the R1 bottle according to the table below, by mixing gently several times. R2 is ready for use and can be placed directly on board the
instrument. This method can also be used for 3-part-reagent enabled systems.
Cat No. Volume Pyridoxal Phosphate Liquid No. of tablets
OSR6009 0.25 mL 1 tablet
2009-08

Reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C.
Once open, reagents stored on board the instrument are stable for 30 days.
With pyridoxal phosphate activation
After addition of pyridoxal phosphate, R1 stored on board the instrument is stable for 7 days.
R2 stored on board the instrument is stable for 30 days.
Pyridoxal Phosphate Liquid reagent (Cat no 60106)
Once open, Pyridoxal Phosphate Liquid reagent is stable until the expiry date printed on the label, provided that contamination is avoided through
adherence to GLP, the cap is replaced immediately after use and the reagent is stored at 2…8°C.
3-part-reagent enabled systems with P-5-P activation
Reagent Preparation
P-5-P Liquid (Cat. No. OSR60180) is specifically for use on board 3-part-reagent enabled systems. The reagent is ready for use and can be placed directly
on board the instrument in the R1 carousel. R1 and R2 are also ready for use and can be placed directly on board the instrument.
Storage and stability
Once open, P-5-P (Cat no OSR60180) is stable on board the instrument for 60 days. R1 and R2 reagent stored on board the instrument are stable for
30 days.
Specimen
5
Serum and heparinised plasma: Stable in serum for 7 days when stored at 2…8°C and 4 days when stored at 15...25°C.
Haemolysed samples should be avoided as the concentration of AST in erythrocytes is approximately 15 times higher than that of normal serum.
Test Procedure
Calibration
The test is run in MB-mode. To provide a robust approach to generate the analyser specific MB factor it is recommended that 5 separate calibration events
With pyridoxal phosphate activation the calibrator value is traceable to the IFCC reference method. Without pyridoxal phosphate activation,the calibrator
value is traceable to a Beckman Coulter Master Calibrator.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the AST activity of each sample.
Newborn 25 − 75 U/L (0.42 − 1.25 µkat/L)
Infant 15 − 60 U/L (0.25 − 1 µkat/L)
values.
Linearity
Precision
27 0.53 1.93 1.16 4.23
71 0.59 0.84 0.95 1.35
415 2.42 0.58 5.05 1.22
Sensitivity
The lowest detectable level represents the lowest measurable level of AST that can be distinguished from zero. It is calculated as the absolute mean plus

2009-08
Method Comparison with pyridoxal phosphate activation
Patient serum samples were used to compare this AST OSR6109 assay on the AU640 against the IFCC reference method. Results of linear regression
y = 1.032x - 0.2 r = 0.997 n = 112 Sample range = 9 – 250 U/L
®
Pyruvate: Interference less than 10% up to 1 mmol/L pyruvate
8
Limitations
BIBLIOGRAPHY
1. Thomas L. Alanine aminotransferase (ALT), Aspartate aminotransferase (AST). In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical
laboratory results. Frankfurt/Main: TH-Books Verlagsgesellschaft, 1998:55-65.
3. Schmidt E, Schmidt FW. Diagnosis of icteric diseases. Dtsch Med Wschr 1984;109:139-146.
4. Schumann G, Bonora R, Ceriotti F et al. IFCC Primary Reference Procedures for the Measurement of Catalytic Activity Concentrations of Enzymes at 37°C.
Part 5. Reference Procedure for the Measurement of Catalytic Concentration of Aspartate Aminotransferase. Clin Chem Lab Med 2002;40:725–733.
5. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of Anticoagulants in Diagnostic Laboratory Investigations and Stability of Blood, Plasma
and Serum Samples. WHO/DIL/LAB/99.1 Rev.2:23pp.
6. Thomas L, Müller M, Schumann G et al. Consensus of DGKL and VDGH for interim reference intervals on enzymes in serum. J Lab Med 2005;29:301-08.
Philadelphia:WB Saunders Company, 1999;1802pp.

2009-08
AST (IFCC, without Pyridoxal phosphate activation), AU400/AU640 AST (IFCC, without Pyridoxal phosphate activation), AU600

Test No # Test name AST ∇ Sample type Ser ∇ Page ½
Test Name: AST ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name AST ∇ Sample type Ser ∇ Page 2/2
Test Name: AST ∇ < > Type: Serum ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name AST ∇
Test Name: AST ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470*
Point 2 ∇
Point 2:
Point 3 ∇
Point 3:
1-Point Cal. Point: R With CONC-0 Slope Check: None # 1-point cal. Point
∇ Advanced Calibration: ∇
MB type factor #
§ For use in AB mode only, refer to leaflet for further instruction. § For use in AB mode only, refer to IFU for further instruction.
Enzyme BSOSR6x09.02
2010-05
AST (IFCC, without Pyridoxal phosphate activation), AU2700/AU5400 AST (IFCC, without Pyridoxal phosphate activation), AU680/AU480
Serum/Plasma Application
Serum/Plasma Application System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009
System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009 Parameters Specific Test Parameters
General LIH ISE Range Test Name: AST ∇ < > Type: Serum ∇ Operation Yes ∇

Sample Volume 5 µL Dilution 0 µL OD Limit
Sample: Volume 5 µL Dilution 0 µL Pre-Dilution Rate: 1 Pre-Dilution Rate 1 ∇ Min.OD 0.60 Max.OD 2.5
Reagents: R1 Volume 25 µL Dilution 25 µL Min OD Max OD Rgt. Volume R1(R1-1) 25 µL Dilution 25 µL Reagent OD Limit
R2 Volume 25 µL Dilution 50 µL L 0.60 H 2.5 First Low 1.1 High 2.5
Reagent OD limit: Last Low 1.1 High 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.1 First H 2.5 R2(R2-1) 25 µL Dilution 50 µL
Method: RATE ∇ Last L 1.1 Last H 2.5 Dynamic Range Low 3* High 1000*
Lag Time Check YES ∇ Hemolysis + ∇
General LIH ISE Range Parameters Specific Test Parameters
Test Name: AST ∇ < > Type: Serum ∇
Normal Ranges: Age L Age H Value/Flag: # ∇ Low High
Sex Year Month Year Month L H Level # # Panic Value
R 3. # # # # # # # R 1. # ∇ # # # # # #
∇
R 4. R 2. # ∇ # # # # # #
# ∇ # # # # # #
R 3. # ∇ # # # # # #
R 5. # ∇ # # # # # #
R 4. # ∇ # # # # # #
R 6. # ∇ # # # # # #
R 5. # ∇ # # # # # #
7. None Selected # # R 6. # ∇ # # # # # #
Panic Value: # # Unit: U/L* Decimal places: # 8. Not within expected values # #
Test Name: AST ∇ < > Type Serum ∇ Test Name: AST ∇ < > Type Serum ∇ R Use Serum Cal.
Point 1: § § §3650* §5470* Point 1: § ∇ § §3650* §5470*
Point 6: ∇
Point 6:
Point 7:
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 9 ∇
MB Type Factor: # Calibration Stability Period: Point 10 ∇ Interval (RB/ACAL) # ∇
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 Point-1 ∇ Reagent Blank Day Hour
§ For use in AB mode only. Refer to IFU for further instruction Point-2 ∇ Calibration Day Hour
ф AU680
Enzyme BSOSR6x09.02
2010-05
AST (IFCC, with OE60106 activation), AU400/AU640 Serum/Plasma AST (IFCC, with OE60106 activation), AU600
Application Serum/Plasma Application

Test No # Test name AST ∇ Sample type Ser ∇ Page 1/2
General LIH ISE Range Test No # Test name AST ∇ Sample type Ser ∇ Page 2/2
Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
Panic value # #
General ISE Test No # Test name AST ∇
Test Name: AST ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #

SERUM/PLASMA APPLICATION Formula 1
Y=AX+B ∇ Process ∇

Point 1: § § §3650* §5470* Point 2 ∇
Point 4:
Point 5 ∇
Point 5:
Point 6 ∇
Point 6:
Point 7 ∇
Point 7:
1-point cal. point
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ MB type factor #
Enzyme BSOSR6x09.02
2010-05
AST (IFCC, with OE60106 activation), AU2700/AU5400 Serum/Plasma
Application
System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009


Reagent OD limit:


R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # #
General ISE
Test Name: AST ∇ < > Type Serum ∇

Point 1: § § §3650* §5470*
Point 2:
Point 3:
Point 4:
Point 5:
Point 6:
Point 7:
# User defined
§ For use in AB mode only. Refer to IFU for further instruction
ф AU680
Enzyme BSOSR6x09.02
2010-05
AST (IFCC, with OE60106 activation ), AU680/AU480 AST (IFCC, with OSR60180 activation ), AU680
System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009 System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009
Test Name: AST ∇ < > Type: Serum ∇ Operation Yes ∇ Test Name: AST ∇ < > Type: Serum ∇ Operation Yes ∇

Lag Time Check YES ∇ Hemolysis + ∇ Lag Time Check YES ∇ Hemolysis + ∇

Test Name: AST ∇ < > Type: Serum ∇ Test Name: AST ∇ < > Type: Serum ∇
Value/Flag: # ∇ Low High Value/Flag: # ∇ Low High

R 1. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # # R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # # R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 6. # ∇ # # # # # #
Test Name: AST ∇ < > Type Serum ∇ R Use Serum Cal. Test Name: AST ∇ < > Type Serum ∇ R Use Serum Cal.
Point 1: § ∇ § §3650* §5470* Point 1: § ∇ § §3650* §5470*
Point 2: ∇ Allowance Range Check Point 2: ∇ Allowance Range Check
Point 7 ∇ Advanced Calibration Point 7 ∇ Advanced Calibration
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration <Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
Point-2 ∇ Calibration Day Hour Point-2 ∇ Calibration Day Hour
MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0 MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0
Enzyme BSOSR6x09.02
2010-05
AST (IFCC, without Pyridoxal phosphate activation), AU400/AU640 AST (IFCC, without Pyridoxal phosphate activation), AU600
Test No # Test name ASTP ∇ Sample type Ser ∇ Page ½
Test Name: ASTP ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name ASTP ∇ Sample type Ser ∇ Page 2/2
Test Name: ASTP ∇ < > Type: Serum ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name ASTP ∇
Test Name: ASTP ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470*
Point 2 ∇
Point 2:
Point 3 ∇
Point 3:
1-Point Cal. Point: R with CONC-0 Slope Check: None # 1-point cal. Point
MB type factor #
Enzyme BSOSR6x09.02
2010-05
AST (IFCC, without Pyridoxal phosphate activation), AU2700/AU5400 AST (IFCC, without Pyridoxal phosphate activation), AU680/AU480
Paediatric Application System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009
System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009 Parameters Specific Test Parameters
General LIH ISE Range Test Name: ASTP Type: Serum
∇ < > ∇ Operation Yes ∇
Reagent OD limit: Last Low 1.1 High 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.1 First H 2.5 R2(R2-1) 25 Dilution 40
µL µL
Method: RATE ∇ Last L 1.1 Last H 2.5 Dynamic Range Low 3* High 1000*
Specific Test Parameters Lag Time Check YES ∇ Hemolysis + ∇
Test Name: ASTP ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: ASTP ∇ < > Type: Serum ∇
R 1. Level # # Panic Value
# ∇ # # # # # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # ∇ # # # # # #
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
Test Name: ASTP ∇ < > Type Serum ∇ Test Name: ASTP ∇ < > Type Serum ∇ R Use Serum Cal.
Point 1: § § §3650* §5470* Point 1: § ∇ § §3650* §5470*
Point 9 ∇
# User defined
Enzyme BSOSR6x09.02
2010-05
AST (IFCC, with OE60106 activation), AU400/AU640 AST (IFCC, with OE60106 activation), AU600

Test No # Test name ASTP ∇ Sample type Ser ∇ Page 1/2
General LIH ISE Range Test No # Test name ASTP ∇ Sample type Ser ∇ Page 2/2
Test Name: ASTP ∇ < > Type: Serum ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name ASTP ∇
Test Name: ASTP ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470*
Point 2 ∇
Point 2:
Point 3 ∇
Point 3:
1-Point Cal. Point: R with CONC-0 Slope Check: None # 1-point cal. point
MB type factor #
Enzyme BSOSR6x09.02
2010-05
AST (IFCC, with OE60106 activation), AU2700/AU5400
System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009


Reagent OD limit:

Test Name: ASTP ∇ < > Type: Serum ∇

R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # #
General ISE
Test Name: ASTP ∇ < > Type Serum ∇

Point 1: § § §3650* §5470*
Point 2:
Point 3:
Point 4:
Point 5:
Point 6:
Point 7:
1-Point Cal. Point: R with CONC-0 Slope Check: None Advanced Calibration: # ∇
# User defined
ф AU680
Enzyme BSOSR6x09.02
2010-05
AST (IFCC, with OE60106 activation), AU680/AU480 Paediatric Application AST (IFCC, with OSR60180 activation), AU680 Paediatric Application
System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009 System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009
Test Name: ASTP ∇ < > Type: Serum ∇ Operation Yes ∇ Test Name: ASTP ∇ < > Type: Serum ∇ Operation Yes ∇

Lag Time Check YES ∇ Hemolysis + ∇ Lag Time Check YES ∇ Hemolysis + ∇

Test Name: ASTP ∇ < > Type: Serum ∇ Test Name: ASTP ∇ < > Type: Serum ∇
Value/Flag: # ∇ Low High Value/Flag: # ∇ Low High

R 1. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # # R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # # R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 6. # ∇ # # # # # #
Test Name: ASTP ∇ < > Type Serum ∇ R Use Serum Cal. Test Name: ASTP ∇ < > Type Serum ∇ R Use Serum Cal.
Point 1: § ∇ § §3650* §5470* Point 1: § ∇ § §3650* §5470*
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration <Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
Point-2 ∇ Calibration Day Hour Point-2 ∇ Calibration Day Hour
MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0 MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0
Enzyme BSOSR6x09.02
2010-05
CHOLINESTERASE
OSR6114 4 x 30 mL R1
4 x 6 mL R2
Intended Use
Kinetic colour test for the quantitative determination of cholinesterase, EC 3.1.1.8 (CHE) in human serum and plasma on Beckman Coulter
Summary1,2
Cholinesterase is responsible for the rapid hydrolysis of acetylcholine released at nerve endings to mediate the transmission of the neural
impulse across the synapse. Degradation of acetylcholine is necessary for the depolarisation of the nerve so that it can undergo repolarisation in
the next conduction event. Serum cholinesterase levels are useful as an indicator of insecticide poisoning, for detection of patients with atypical
forms of the enzyme, or as a test of liver function. Cholinesterase is competitively inhibited by the alkaloids prostigmine and physostigmine.
Cholinesterase activity is also inhibited by organic phosphorous compounds including Parathion, Sarin and tetraethyl pyrophosphate, chemicals
which may be inhaled or absorbed by workers in the agriculture or organic chemical industries. A wide variety of other compounds inhibit
cholinesterase, including morphine, quinine, tertiary amines, phenothiazines, pyrophosphate, bile salts, citrate, fluoride and borate. If sufficient
material is absorbed to inactivate all the acetylcholinesterase of nervous tissue, death will result. A 40% drop in serum cholinesterase activity
occurs before the first symptoms are noted, and a drop of 80% is required before neuromuscular effects are apparent. Near zero levels of
enzyme activity require emergency treatment of the patient with enzyme regulators such as pyridine-2-aldoxime.
Patients with low levels or weakly active acetylcholine may undergo a period of prolonged apnoea on administration of succinyldicholine
(suxamethonium), a muscle relaxant used during surgical procedures. Sensitivity to succinyldicholine is dependent on the phenotype of the
patient, and preoperative screening has been advocated to identify patients in whom complications may arise. In these cases, patients may be
characterised by measurement of serum cholinesterase activity, in addition to the calculation of inhibition of enzyme activity toward specified
substrates in the presence of dibucaine or fluoride. Measurement of serum cholinesterase activity may also be used as a measure of the
synthetic capacity of the liver. A 30-50% decrease in level is observed in acute hepatitis, with decreases of 50-70% observed in advanced
cirrhosis and carcinoma with metastases to the liver. Increased cholinesterase levels may be found in diabetes mellitus, coronary heart disease,
hyperlipoproteinaemia Type IV, Gilbert’s syndrome, and those with Cynthiana variant, a familial variant which results in cholinesterase levels of
2-3 times normal . Increased cholinesterase activity is not considered to be of clinical significance. Normal levels are noted in chronic hepatitis,
mild cirrhosis and obstructive jaundice.
Test Principle3
Method based on the recommendations of the “German Society for Clinical Chemistry” (GSCC, 1994).
Cholinesterase catalyses the hydrolysis of butyrylthiocholine to butyrate and thiocholine. Thiocholine reduces yellow hexacyanoferrate (III) to
colourless hexacyanoferrate (II). The decrease in absorbance at 410 nm is directly proportional to the cholinesterase activity in the sample.
Reaction principle
CHE
Butyrylthiocholine + H2O Butyric acid + Thiocholine
- 3- 4-
2 Thiocholine + 2 OH + 2 [Fe(CN)6] (Yellow) Dithiobis(choline) + H2O + 2 [Fe(CN)6] (Colourless)

Tetra sodium diphosphate (pH 6.2) 75 mmol/L
Ferricyanide (III) 2.0 mmol/L
Butyrylthiocholine 15 mmol/L
Preservative

Reagent Preparation

The reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C. Once open, reagents stored on board the instrument
are stable for 30 days. Protect R1 from light.
Specimen
4
Stable in serum and plasma for 1 year when stored at 2…25°C.
Test Procedure
Calibration
The test is run in MB-mode. To provide a robust approach to generate the analyser specific MB factor, it is recommended that 5 separate calibration
events should be used. A fresh vial of calibrator, utilising System Calibrator Cat No. 66300 in the AB calibration mode, should be used for each of these
runs. When calculating the mean factor from the separate runs the data should be examined for obvious outliers which should be repeated and replaced.
For the AU2700/AU5400 this procedure needs to be performed for each ring. Quality control procedures should be undertaken immediately
following calibration in accordance with good laboratory practice.
3
The calibrator value is traceable to a standard method.

2009-08
Quality Control
Good laboratory practice suggests that controls be tested each day patient samples are tested and each time calibration is performed. Values
obtained for the controls should fall within specified limits as defined by the user. If any trends or sudden shifts in values are detected, review all
operating parameters.
Calculation
The Beckman Coulter analysers automatically compute the CHE activity of each sample.
Male 4.62 – 11.5 kU/L ( 77 – 192 µkat/L)
Female 3.93 – 10.8 kU/L ( 65 – 180 µkat/L)

from these values.
Linearity
The test is linear within an enzyme activity range of 1 – 15 kU/L (17 – 250 µkat/L).
Precision
Mean kU/L SD CV% SD CV%
5.2 0.03 0.58 0.05 0.90
11.7 0.06 0.48 0.43 3.65
14.8 0.07 0.46 0.26 1.77
Sensitivity
The lowest detectable level on an AU600 analyser was estimated at 0.1 kU/L.
The lowest detectable level represents the lowest measurable level of CHE that can be distinguished from zero. It is calculated as the absolute
mean plus three standard deviations of 20 replicates of an analyte free sample.
Method Comparison
Patient serum samples were used to compare this CHE OSR6114 assay on the AU600 against another commercially available CHE assay.
y = 1.009x + 0.127 r = 1.000 n = 64 Sample range = 1.5 – 14.5 kU/L
®
5
Limitations
This product is not suitable for the determination of red cell cholinesterase.

* Values set for working in kU/L. To work in SI units (µkat/L) multiply by 16.66.
BIBLIOGRAPHY
1. Moss DW, Henderson RA. Clinical Enzymology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders
Company, 1999;708-711.
2. Thomas L. Cholinesterase. In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
3. Schmidt E. et al. Proposal of standard methods for the determination of enzyme catalytic concentrations in serum and plasma at 37°C. Eur J Clin
Chem Biochem 1992;30:163-170.
4. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of Anticoagulants in Diagnostic Laboratory Investigations and Stability of Blood,
Plasma and Serum Samples. WHO/DIL/LAB/99.1 Rev.2:26pp.

2009-08
CHOLINESTERASE, AU400/AU640 Serum/Plasma Application CHOLINESTERASE, AU600 Serum/Plasma Application

Test No # Test name CHE ∇ Sample type Ser ∇ Page 1/2
Test Name: CHE ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name CHE ∇ Sample type Ser ∇ Page 2/2
Test Name: CHE ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic Value: # # Unit: kU/L* Decimal places: # Panic value # #
General ISE Test No # Test name CHE ∇
Test Name: CHE ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §52* §78*
Point 1: § § §52* §78*
Point 2 ∇
Point 2:
MB type factor #
# User defined
* Values set for working in kU/L. To work in SI units (µkat/L) multiply by 16.66.
* Values set for working in kU/L. To work in SI units (µkat/L)multiply by 16.66.
Enzyme BSOSR6x14.01
2009-08
CHOLINESTERASE, AU2700/AU5400 Serum/Plasma Application CHOLINESTERASE, AU680/AU480 Serum/Plasma Application
Test Name: CHE ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CHE ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
R2 Volume 30 μL Dilution 0 μL L 0.4 H 2.5
Reagent OD limit:
Test Name: CHE ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CHE ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: kU/L* Decimal places: # 7. No demographics # #
Unit kU/L* Decimal Places #
General ISE
General ISE
Test Name: CHE Type Serum
∇ < > ∇ Test Name: CHE Type Serum Use Serum Cal.
∇ < > ∇ ο
Point 1: § § §52* §78* Point 1: § ∇ § §52* §78*
Point 9 ∇
# User defined. Calibrator OD Conc Low High Stability
* Values set for working in kU/L. To work in SI units (µkat/L) multiply by 16.66 Point-1 ∇ Reagent Blank Day Hour
§ For use in AB mode only, refer to IFU for further instruction. Point-2 Calibration Day Hour
∇
Ф AU680
Enzyme BSOSR6x14.01
2009-08
CK (NAC)
OSR6179 4 x 22 mL R1-1
4 x 4 mL R1-2
4 x 6 mL R2
OSR6279 4 x 44 mL R1-1
4 x 8 mL R1-2
4 x 13 mL R2
Intended Use
Kinetic UV test for the quantitative determination of creatine kinase, EC 2.7.3.2 (CK), in human serum and plasma on Beckman Coulter
Summary1,2,3,4
Creatine kinase (CK), a dimer composed of M-muscle and /or B-brain subunits which associate to form the isoenzymes CK-MM, CK-MB and
CK-BB, catalyses the reversible phosphorylation of creatine by ATP. Measurements of CK are primarily used in the diagnosis and treatment of
myocardial infarction as well as being the most sensitive indicator of muscle damage. CK is increased whenever there is necrosis or
regeneration of muscle and is therefore elevated in most myopathies such as Duchenne-muscular dystrophy and in conditions associated with
muscle necrosis such as rhabdomyolysis. Total CK can also be increased in diseases of the CNS such as Reyes Syndrome where a 70 fold
increase in CK activity indicates the severity of the encephalopathy.
CK-BB predominates in the brain, prostate, gut, lung, kidney, bladder, uterus, liver, thyroid and the placenta. CK-MM predominates in skeletal
and cardiac muscle. In healthy individuals the total activity consists mainly of CK-MM while the other CK isoenymes and variants are only
present in trace amounts or are undetectable. CK-MB is present to varying degrees in heart muscle and also to a minor degree in skeletal
muscle.
CK activity rises following myocardial damage, with a significant increase in both the CK-MM and CK-MB fractions. The proportional rise in the
CK-MB fraction to some extent depends on the size of the myocardial damage and on a history of previous myocardial damage. Changes in the
ratio of CK-MB to CK-MM may be used to diagnose a myocardial infarct (MI), the ratio reaching a peak within 1.5 hours post MI. The diagnostic
sensitivity and specificity of total CK estimation for the diagnosis of an MI can be improved by determining the rate of increase ("slope") of CK on
serial samples obtained on admission and at 4, 8 and 12 hours thereafter. A 50% incremental increase per hour over the time period
differentiates between an acute MI and non-infarction with an overall efficiency of 94%.
For patients in need of an early diagnosis of a myocardial infarction a rapidly appearing biomarker such as CK-MB plus a biomarker that rises
later e.g. cardiac troponin is recommended for confirmation of the diagnosis.
Test Principle5
CK reversibly catalyses the transfer of a phosphate group from creatine phosphate to adenosine diphosphate (ADP) to give creatine and
adenosine triphosphate (ATP) as products. The ATP formed is used to produce glucose-6-phosphate and ADP from glucose. This reaction is
catalysed by hexokinase (HK) which requires magnesium ions for maximum activity. The glucose-6-phosphate is oxidised by the action of the
enzyme glucose-
6-phosphate dehydrogenase (G6P-DH) with simultaneous reduction of the coenzyme nicotinamide adenine dinucleotide (NADP) to give NADPH
and 6-phosphogluconate. The rate of increase of absorbance at 340/660 nm due to the formation of NADPH is directly proportional to the
activity of CK in the sample.
Reaction Principle
CK
Creatine phosphate + ADP Creatine + ATP
HK
ATP + Glucose ADP + Glucose-6-phosphate
+ G6P-DH +
Glucose-6-phosphate + NADP 6-Phosphogluconate + NADPH + H

Final concentration of reactive ingredients
Immidazole (pH 6.5 @ 37°C) 100 mmol/L
NADP 2.0 mmol/L
ADP 2.0 mmol/L
AMP 5.0 mmol/L
EDTA 2.0 mmol/L
Glucose 20 mmol/L
Creatine phosphate 30 mmol/L
N-acetylcysteine 0.2 mmol/L
Activator 26 mmol/L
2+
Mg 10 mmol/L
Diadenosine pentaphosphate 0.01 mmol/L
HK ≥ 4.0 kU/L
G6P-DH ≥ 2.8 kU/L
Stabilisers
Preservative

R2: Irritant, contains a mixture of 5-chloro-2-methyl-4-isothiazolin-3-one [EC No 247-500-7] and 2-methyl-4-isothiazolin-3-one
[EC No 220-239-6] (3:1). R43: May cause sensitisation by skin contact.

2009-08
Safety Phrases:
S24, S37, S60. Avoid contact with skin. Wear suitable gloves. This material and its container must be disposed of as hazardous waste.
Reagent Preparation
R1: The entire contents of bottle R1-2 must be transferred into the entire volume of R1-1. Mix by gentle inversion before placing on board the
instrument.
R2: The reagent is ready for use and can be placed directly on board the instrument.

are stable for 30 days.
Specimen
Serum is the recommended specimen. Haemolysed samples should be avoided. Allow specimen to clot and remove serum from cells promptly
to minimise haemolysis and contamination by adenylate kinase from the red cells.
5,6
Stable in serum, protected from light, for 8-12 hours when stored at 2…8°C and 4 hours when stored at 15…25°C.
Heparinised plasma, free from haemolysis, can also be used. Plasma samples may occasionally produce unpredictable rate reactions resulting
5
in false low results. Plasma with EDTA, oxalate or citrate is not recommended.
Test Procedure
Calibration
calibration events should be used. A fresh vial of calibrator, utilising System Calibrator Cat No. 66300 in the AB calibration mode, should be
used for each of these runs. When calculating the mean factor from the separate runs the data should be examined for obvious outliers which
should be repeated and replaced. For the AU2700/AU5400 this procedure needs to be performed for each ring. Quality control procedures
should be undertaken immediately following calibration in accordance with good laboratory practice.
The calibrator value is traceable to the IFCC reference method.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the creatine kinase activity of each sample.
Male ≤ 171 U/L (2.85 µkat/L)
Female ≤ 145 U/L (2.42 µkat/L)
findings.

from these values.
Linearity
Precision
99 2.35 2.37 4.51 4.55
270 2.70 1.00 8.64 3.20
810 5.22 0.64 26.59 3.28
Sensitivity
The lowest detectable level on an AU640 analyser was estimated at 3 U/L
The lowest detectable level represents the lowest measurable level of CK that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient serum samples were used to compare this CK-Nac OSR6179 assay on the AU640 against another commercially available
CK-Nac assay. Results of linear regression analysis were as follows:
y = 0.992 x + 0.026 r = 1.000 n = 109 Sample range = 22 - 1903 U/L
®
8
2009-08
BIBLIOGRAPHY
1. Mayne PD, ed. Clinical chemistry in diagnosis and treatment, 6th ed. London: Arnold,1994:304-310.
2. Thygesen K, Alpert JS et al. Myocardial infarction redefined-A consensus document of the Joint European Society of Cardiology/American College of
Cardiology Committee for the redefinition of myocardial infarction. JACC 2000;36:959-969.
3. Stein W. Creatine kinase (total activity). Creatine kinase isoenzymes and variants. In:Thomas L, ed. Clinical laboratory diagnostics. Use and
assessment of clinical laboratory results. Frankfurt/Main: TH-Books Verlagsgesellschaft, 1998:71-79.
4. Moss DW, Henderson RA. Clinical enzymology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders
Company, 1999; 657-662.
5. Horder M, Elser R, Gerhardt W, Mathieu M, Sampson E.J. International Federation of Clinical Chemistry, Scientific division committee on enzymes:
Approved recommendation on IFCC methods for the measurement of catalytic concentration of enzymes. Part 7. IFCC method for creatine kinase.
Eur J Clin Chem Biochem 1991;29(7):435-56.
6. Moss DW, Henderson RA, Kachmar JF. Enzymes. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company,
1987:376pp.
7. Schumann G, Klauke R. New IFCC reference procedures for the determination of catalytic activity concentrations of five enzymes in serum:
preliminary upper reference limits obtained in hospitalised subjects. Clin Chim Acta 2003;327:69-79.

2009-08
CK (NAC), AU400/AU640 Serum/Plasma Application CK (NAC), AU600 Serum/Plasma Application

Test No # Test name CK ∇ Sample type Ser ∇ Page 1/2
Test Name: CK ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 3 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 120 Dil. vol 0 μL L 0 H 2
R2 Volume 30 μL Dilution 0 μL L 0 H 2 Fst. L 0 Fst. H 0.3
Reagent OD limit: Wave Main 340 Sub 660 ∇ Lst. L 0 Lst. H 0.3
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 0 First H 0.3 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 0 Last H 0.3 Reaction + ∇ L 10* H 2000*
General LIH ISE Range Test No # Test name CK ∇ Sample type Ser ∇ Page 2/2
Test Name: CK ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name CK ∇
Test Name: CK ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §6475* §9715*
Point 1: § § §6475* §9715*
Point 2 ∇
Point 2:
MB type factor #
# User defined
Enzyme BSOSR6x79.01
2009-08
CK (NAC), AU2700/AU5400 Serum/Plasma Application CK (NAC), AU680/AU480 Serum/Plasma Application
Test Name: CK ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CK ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: CK ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CK ∇ < > Type: Serum ∇

ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: CK ∇ < > Type Serum ∇ Test Name: CK ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: § § §6475* §9715* Point 1: § ∇ § §6475* §9715*
Point 9 ∇
# User defined. Master Curve> OD Range
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60. Calibrator OD Conc Low High Stability
Enzyme BSOSR6x79.01
2009-08
CK-MB
OSR61155 2 x 22 mL R1-1
2 x 4 mL R1-2
2 x 6 mL R2
Intended Use
Enzymatic immuno-inhibition test for the quantitative determination of the creatine kinase-MB isoenzyme (CK-MB) in human serum and plasma on
Beckman Coulter analysers. For in vitro diagnostic use only.
Summary1,2,3
Creatine kinase (CK) EC 2.7.3.2, a dimer composed of M-muscle and /or B-brain subunits which associate to form the isoenzymes CK-MM, CK-MB and
CK-BB, catalyses the reversible phosphorylation of creatine by ATP. Measurements of CK are primarily used in the diagnosis and treatment of myocardial
infarction as well as being the most sensitive indicator of muscle damage. CK is increased whenever there is necrosis or regeneration of muscle and is
therefore elevated in most myopathies such as Duchenne-muscular dystrophy and in conditions associated with muscle necrosis such as rhabdomyolysis.
Total CK can also be increased in diseases of the CNS such as Reye’s Syndrome where a 70 fold increase in CK activity indicates the severity of the
encephalopathy.
CK-BB predominates in the brain, prostate, gut, lung, kidney, bladder, uterus, liver, thyroid and the placenta. CK-MM predominates in skeletal and cardiac
muscle. In healthy individuals the total serum activity consists mainly of CK-MM while the other CK isoenymes and variants are only present in trace
amounts or are undetectable. CK-MB is present to varying degrees in heart muscle and also to a minor degree in skeletal muscle.
CK activity rises following myocardial damage, with a significant increase in both the CK-MM and CK-MB fractions. The proportional rise in the CK-MB
fraction to some extent depends on the size of the myocardial damage and on a history of previous myocardial damage. Changes in the ratio of CK-MB to
CK-MM may be used to diagnose a myocardial infarction (MI), the ratio reaching a peak within 1.5 hours post MI. The diagnostic sensitivity and specificity
of total CK estimation for the diagnosis of an MI can be improved by determining the rate of increase ("slope") of CK on serial samples obtained on
admission and at 4, 8 and 12 hours thereafter. A 50% incremental increase per hour over the time period differentiates between an acute MI and non-
infarction with an overall efficiency of 94%.
For patients in need of an early diagnosis of a myocardial infarction a rapidly appearing biomarker such as CK-MB plus a biomarker that rises later e.g.
cardiac troponin is recommended for confirmation of the diagnosis.
Test Principle4,5
R1 contains an antibody which binds to the M subunit of CK in the serum sample thereby inhibiting the activity of the M subunit. The B subunit of the
enzyme remains free to act on the substrate present in R2. CK reversibly catalyses the transfer of a phosphate group from creatine phosphate to
adenosine diphosphate (ADP) to give creatine and adenosine triphosphate (ATP) as products. The ATP formed is used to produce glucose-6-phosphate
and ADP from glucose. This reaction is catalysed by hexokinase (HK) which requires magnesium ions for maximum activity. The glucose-6-phosphate is
oxidised by the action of the enzyme glucose-6-phosphate dehydrogenase (G6P-DH) with simultaneous reduction of the coenzyme nicotinamide adenine
dinucleotide phosphate (NADP) to give NADPH and 6-phosphogluconate. The rate of increase of absorbance at 340 nm due to the formation of NADPH is
directly proportional to the activity of CK-MB in the sample.
Reaction Principle5
CK
Creatine Phosphate + ADP Creatine + ATP
2+
HK, Mg
ATP + Glucose ADP + Glucose-6-phosphate (G-6-P)
G6P-DH
+ +
G-6-P + NADP 6-phosphogluconate + NADPH + H

Imidazole buffer (pH 6.7) 100 mmol/L Diadenosine-pentaphosphate 0.01 mmol/L
Hexokinase (HK) ≥ 4.0 kU/L EDTA 2.0 mmol/L
NADP 2.0 mmol/L Glucose 20 mmol/L
G6P-DH ≥ 2.8 kU/L Creatine phosphate 30 mmol/L
ADP 2.0 mmol/L N-Acetylcysteine 0.2 mmol/L
Mg-Acetate 10 mmol/L Activator 26 mmol/L
AMP 5.0 mmol/L Antibody to CK-M-subunit Variable
Preservative

Safety Phrases:
Reagent Preparation
R1: The entire contents of bottle R1-2 must be transferred into the entire volume of R1-1. Mix by gentle inversion before placing on board the
instrument.


2009-08
Specimen
Serum is the recommended specimen. Lipaemic, haemolysed and strongly icteric samples should be avoided. Allow specimen to clot and remove serum
from cells promptly to minimise haemolysis and contamination by adenylate kinase from the red cells.
CK/CK-MB is stable in serum, protected from light, for 7 days when stored at 2…8°C, for 2 days when stored at 20…25°C and up to 1 year when stored at
5,6,7
-20°C.
Heparinised plasma, free from haemolysis, can also be used. Plasma samples may occasionally produce unpredictable rate reactions resulting in false low
6
results. Plasma with EDTA, oxalate or citrate is not recommended.
Test Procedure
Refer to the appropriate User’s Guide and Setting Sheet for analyser-specific assay instructions for the sample type as listed in the Intended
Use statement.
Calibration
The test is run in MB-mode. To provide a robust approach to generate the analyser specific MB factor, it is recommended that 5 separate calibration
events should be used. A fresh vial of calibrator, utilising CK-MB Calibrator Cat no ODR30034 in the AB calibration mode, should be used for each of these
runs. When calculating the mean factor from the separate runs the data should be examined for obvious outliers which should be repeated and replaced.
For the AU2700/AU5400 this procedure needs to be performed for each ring. Quality control procedures should be undertaken immediately
following calibration in accordance with good laboratory practice.
Traceability: This method has been standardised against the CKtotal IFCC Reference Method with addition of antibody, performed manually and
calculated via molar absorption coefficient ε.
Quality Control
CK-MB Control Level 1 ODR30035 and Level 2 ODR30036 or other control materials with values determined by this Beckman Coulter system may be
used.
Please note that the recovery of non-Beckman Coulter controls may vary with reagent lots of immunoassay products, due to the use of non-human
materials in the controls.
Calculation
The Beckman Coulter analysers automatically compute the CK-MB activity of each sample.
Reference Intervals
2,8
Adults (37°C) < 24 U/L (0.4 µkat/L)
2
Myocardial infarction: the probability of myocardial damage is high when the following conditions are fulfilled .
U/L µkat/L
1. CK total > 250 > 4.17
2. CK-MB > 24 > 0.4
3. CK-MB activity is between 6 and 25 % of total CK
If myocardial infarction is suspected and the values found are below the stated limits, the infarction may be fresh. In this case the determinations
should be repeated after 4 hours with a fresh sample.
A fraction of less than 6% indicates skeletal muscle damage. A fraction above 25 % may indicate the presence of Macro-CK and requires further
2
clarification.

from these values.
Linearity
Precision
17 0.69 4.03 0.86 5.05
86 0.65 0.75 0.99 1.15
194 1.04 0.54 1.76 0.90
Sensitivity
The lowest detectable level on an AU640 analyser was calculated as 5 U/L.
The lowest detectable level represents the lowest measurable level of CK-MB that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient serum samples were used to compare this CK-MB OSR61155 assay on the AU640 against another commercially available
CK-MB assay. Results of linear regression analysis were as follows:
®
9

2009-08
Limitations
Macro-CK is an atypical form of CK that is composed of immunoglobulin complexes of normal isoenzymes. It migrates electrophoretically between MM
and MB and is found mainly in elderly women. It is of no clinical significance, but its presence may cause falsely elevated results. If Macro-CK contribution
is suspected, its presence should be confirmed by electrophoresis.
In very rare cases, gammopathy, especially monoclonal IgM (Waldenström’s macroglobulinemia) can cause unreliable results.
For inhibition of adenylate kinase the recommended inhibitors AMP/Ap5A are included, but as the inhibition can never be completely 100 % a residual
activity could affect low CK-MB activity results.
The inhibition capacity of the anti-CK-MM antibody is >99.75% at a CK-MM concentration of 2000 U/L and >99 % at a CK-MM concentration of 8000 U/L.
In samples where the total CK activity exceeds 8000 U/L, CK-MB should be measured using a pre-diluted sample to ensure adequate inhibition of CK-M.

§ For use in AB mode only. Refer to leaflet for further instruction.
BIBLIOGRAPHY
1. Thygesen K, Alpert JS et al. Myocardial infarction redefined-A consensus document of the Joint European Society of Cardiology/American College of
Cardiology Committee for the redefinition of myocardial infarction. JACC 2000;36:959-969.
2. Stein W. Creatine kinase (total activity). Creatine kinase isoenzymes and variants. In:Thomas L, ed. Clinical laboratory diagnostics. Use and
assessment of clinical laboratory results. Frankfurt/Main: TH-Books Verlagsgesellschaft, 5th ed. 1998:71-79./ 6th ed. 2005 in German.
3. Moss DW, Henderson RA. Clinical enzymology. In: Burtis CA, Ashwood ER, eds. Tietz Textbook of Clinical Chemistry. Philadelphia:WB Saunders
Company, 1999;657-662.
4. Würzburg U, Hennrich N, Lang H. Bestimmung der Aktivität von Kreatinkinase MB im Serum unter Verwendung inhibierender Antikörper. Klin Wschr
1976; 54:357-360.
5. Ferard F, Franck PFH, Gella F-J et al. IFCC Primary Reference Procedures for the Measurement of Catalytic Activity Concentrations of Enzymes at
37°C. Clin Chem Lab Med 2002; 40:635-42.
6. Horder M, Elser R, Gerhardt W, Mathieu M, Sampson E.J. International Federation of Clinical Chemistry, Scientific division committee on enzymes:
Approved recommendation on IFCC methods for the measurement of catalytic concentration of enzymes. Part 7. IFCC method for creatine kinase.
Appendix A. Eur J Clin Chem Biochem 1991;29(7):435-56.
7. Guder WG,Narayanan S, Wisser H, Zawta B. The Quality of Diagnostic Samples in: Samples: From the Patient to the Laboratory. Weinheim:
Wiley-VCH Verlag GmbH & Co, KgaA, 3rd ed. 2003.
8. Kairisto V, Hänninen KP, Leino A, Pulkki K, Peltola O, Näntö V et al. Generation of reference values for cardiac enzymes from hospital admission
laboratory data. Eur J Clin Chem Clin Biochem 1994;32:789-96.

2009-08
CK-MB, AU400/AU640 Serum/Plasma Application CK-MB, AU600 Serum/Plasma Application

General LIH ISE Range Test No # Test name CK-MB ∇ Sample type Ser ∇ Page 1/2
Test Name: CK-MB ∇ < > Type: Serum ∇ Operation: Yes ∇ Sample vol. 6 Dil. vol 0 μL Min. OD Max. OD
Sample: Volume 6 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 120 Dil. vol 0 μL L 0 H 2
R2 Volume 30 μL Dilution 0 μL L 0 H 2 Fst. L 0 Fst. H 0.3
Reagent OD limit: Wave Main 340 Sub 660 ∇ Lst. L 0 Lst. H 0.3
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 0 First H 0.3 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 0 Last H 0.3 Reaction + ∇ L 10* H 2000*
General LIH ISE Range Test No # Test name CK-MB ∇ Sample type Ser ∇ Page 2/2
Level L Level H
Test Name: CK-MB ∇ < > Type: Serum ∇
Sex Age L Age H L H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic value # #
Calibration Specific Test No # Test name CK-MB ∇
General ISE
Cal type 1 MB ∇ Count #
Test Name: CK-MB ∇ < > Type Serum ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇ Selection calibrator
Point 1: § § §5900* §10900* Point 2 ∇
Point 7: 1-point cal. point
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ MB type factor #

Enzyme BSOSR6x155.01
2009-08
CK-MB, AU2700/AU5400 Serum/Plasma Application CK-MB, AU680/AU480 Serum/Plasma Application
Test Name: CK-MB ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CK-MB ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Specific Test Parameters Lag Time Check NO ∇ Hemolysis + ∇
Test Name: CK-MB ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CK-MB ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: CK-MB ∇ < > Type Serum ∇ Test Name: CK-MB ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: § § §5900* §10900* Point 1: § ∇ § §5900* §10900*
Point 9 ∇
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60. Calibrator OD Conc Low High Stability
Enzyme BSOSR6x155.01
2009-08
GGT
OSR6020 4 x 18 mL R1
4 x 18 mL R2
OSR6120 4 x 40 mL R1
4 x 40 mL R2
OSR6520 4 x 104 mL R1
4 x 104 mL R2
Intended Use
Kinetic colour test for the quantitative determination of gamma-glutamyltransferase, EC 2.3.2.2 (GGT) in human serum and plasma on Beckman
Coulter analysers. For in vitro diagnostic use only.
Summary1,2,3
Gamma glutamyltransferase (GGT) belongs to a group of peptidases which catalyse the transfer of amino acids from one peptide to another
and thus act as amino acid transferases. The enzyme only reacts with peptides or peptide-like compounds containing a terminal glutamate
residue joined to the remainder of the compound through the terminal carboxyl. GGT is present in all cells of the body except those in muscle
however the enzyme present in serum appears to originate primarily from the hepatobiliary system. An increase in GGT is always a sign of liver
damage if liver specific enzymes such as ALT, GLDH or CHE are also abnormal. GGT is however of little value in attempting to discriminate
between different kinds of liver disease.
GGT is dramatically increased in cases of intrahepatic or posthepatic biliary obstruction. It is more sensitive than alkaline phosphatase in
detecting obstructive jaundice, cholangitis and cholecystitis and its rise occurs earlier and persists longer. GGT is also increased in patients with
infectious hepatitis, fatty livers, in acute and chronic pancreatitis and in patients receiving anticonvulsant drugs such as phenytoin and
phenobarbital.
As elevated levels of GGT are observed in patients with alcoholic cirrhosis, and in the majority of sera from people who are heavy drinkers, GGT
plays a role in the detection of alcoholism, alcoholic liver damage and in monitoring alcohol abstinence. The enzyme is also useful in ratio with
HDL-cholesterol in cases of alcohol abuse, alkaline phosphatase in cases of alcoholic liver disease and aspartate aminotransferase for
distinguishing neonatal hepatitis from biliary atresia.
Test Principle4
GGT catalyses the transfer of the gamma-glutamyl group from the substrate, gamma-glutamyl-3-carboxy-4-nitroanlide, to glycylglycine, yielding
5-amino-2-nitrobenzoate. The change in absorbance at 410/480 nm is due to the formation of 5-amino-2-benzoate and is directly proportional to
the GGT activity in the sample.
Reaction Principle
γ-GT
L-γ-Glutamyl-3-carboxy-4-nitroanilide + Glycylglycine L-γ-Glutamylglycylglycine + 5-Amino-2-nitrobenzoate

Glycylglycine, pH 7.7 (37°C) 150 mmol/L
L-γ-glutamyl-3-carboxy-4-nitroanilide 6 mmol/L
Preservative
R1 and R2 reagents: Irritant, contains a mixture of 5-chloro-2-methyl-4-isothiazolin-3-one [EC No 247-500-7] and 2-methyl-4-isothiazolin-3-one
[EC No 220-239-6] (3:1). R43; May cause sensitisation by skin contact.
Safety Phrases:
Reagent Preparation
Specimen
Serum and EDTA or heparinised plasma.
5
Test Procedure
Calibration
calibration events should be used. A fresh vial of calibrator,utilising System Calibrator Cat No. 66300 in the AB calibration mode, should be used
for each of these runs. When calculating the mean factor from the separate runs the data should be examined for obvious outliers which should
be repeated and replaced. For the AU2700/AU5400 this procedure needs to be performed for each ring. Quality control procedures should be
undertaken immediately following calibration in accordance with good laboratory practice.

2009-08
The calibrator value is traceable to the IFCC reference method and IRMM/IFCC-452.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the GGT activity of each sample.
Reference Intervals
6
1
Children Male Female
1-182 days 12-122 U/L (0.2-2.03 µkat/L) 15-132 U/L (0.25-2.2 µkat/L)
183-365 days 1-39 U/L (0.02-0.65 µkat/L) 1-39 U/L (0.02-0.65 µkat/L)
1-12 years 3-22 U/L (0.05-0.37 µkat/L) 4-22 U/L (0.07-0.37 µkat/L)
13-18 years 2-42 U/L (0.03-0.7 µkat/L) 4-24 U/L (0.07-0.4 µkat/L)
findings.
from these values.
Linearity
Precision
35 0.27 0.79 0.33 0.95
70 0.71 1.01 0.80 1.13
378 3.57 0.95 4.73 1.25
Sensitivity
The lowest detectable level represents the lowest measurable level of GGT that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient serum samples were used to compare this GGT OSR6120 assay on the AU640 against the IFCC Reference method. Results of linear
regression analysis were as follows:
y = 1.026x - 3 r = 1.000 n = 94 Sample range = 11 - 269 U/L
In very rare cases gammopathy, especially monoclonal IgM (Waldenström’s macroglobulinemia), may cause unreliable results.
7
BIBLIOGRAPHY
1. Thomas L. Gamma glutamyltransferase (GGT). In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results.
Frankfurt/Main: TH-Books Verlagsgesellschaft, 1998:80-86.
2. Tietz NW, ed. Clinical guide to laboratory tests, 3rd ed. Philadelphia: WB Saunders Company,1995:286-87.
3. Moss DW, Henderson RA. Clinical enzymology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders
Company, 1999; 686-89.
4. Shaw M, Stromme H. London L, Theodorsen L. Part 4 IFCC method for γ-glutamyltransferase. Clin Chem Clin Biochem 1983;21(10):633-646.
6. Schumann G, Klauke R. New IFCC reference procedures for the determination of catalytic activity concentrations of five enzymes in serum:
preliminary upper reference limits obtained in hospitalised subjects. Clin Chim Acta 2003;327:69-79.

2009-08
GGT (IFCC), AU400/AU640 Serum/Plasma Application GGT (IFCC), AU600 Serum/Plasma Application

Test No # Test name GGT20 ∇ Sample type Ser ∇ Page ½
Test Name: GGT20 ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name GGT20 ∇ Sample type Ser ∇ Page 2/2
Test Name: GGT20 ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name GGT20 ∇
Test Name: GGT20 ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §3540* §5755*
Point 1: § § §3540* §5755*
Point 2 ∇
Point 2:
1-Point Cal. Point: ο With CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. Point
MB type factor #
# User defined
Enzyme BSOSR6x20.01
2009-08
GGT (IFCC), AU2700/AU5400 Serum/Plasma Application GGT (IFCC), AU680/AU480 Serum/Plasma Application
Test Name: GGT20 ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: GGT20 ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Test Name: GGT20 ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: GGT20 ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: GGT20 ∇ < > Type Serum ∇
Test Name: GGT20 ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: § § §4080* §6630* Point 1: § ∇ § §4080* §6630*
Point 9 ∇
Enzyme BSOSR6x20.01
2009-08
GGT (IFCC), AU400/AU640 Paediatric Application GGT (IFCC), AU600 Paediatric Application

Test No # Test name GT20P ∇ Sample type Ser ∇ Page 1/2
Test Name: GT20P ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name GT20P ∇ Sample type Ser ∇ Page 2/2
Test Name: GT20P ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name GT20P ∇
Test Name: GT20P ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §4216* §6325*
Point 1: § § §4216* §6325*
Point 2 ∇
Point 2:
MB type factor #
# User defined
Enzyme BSOSR6x20.01
2009-08
GGT (IFCC), AU2700/AU5400 Paediatric Application GGT (IFCC), AU680/AU480 Paediatric Application
Test Name: GT20P ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: GT20P ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Test Name: GT20P ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: GT20P ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: GT20P ∇ < > Type Serum ∇ Test Name: GT20P ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: § § §4080* §6630* Point 1: § ∇ § §4080* §6630*
Point 9 ∇
Enzyme BSOSR6x20.01
2009-08
HBDH
OSR6129 4 x 15 mL R1
4 x 15 mL R2
Intended Use
Kinetic UV test for the quantitative determination of 2-hydroxybutyrate dehydrogenase EC 2.2.4.3 (HBDH) in human serum and plasma on Beckman
Summary1
2-Hydroxybutyrate dehydrogenase (HBDH) activity is defined as the catalytic oxidation of 2-hydroxybutyrate to 2-oxobutyrate. HBDH measurement, and
calculation of the LDH/HBDH ratio is advocated as an alternative to the measurement of LDH isoenzymes. LDH-1 and LDH-2, which constitute the largest
proportions of monomer H, are relatively more active with 2–oxobutyrate than with pyruvate. The LDH/HBDH ratio in healthy individuals varies from 1.2 to
1.6. Patients with parenchymal disease have greater LDH-5 activity resulting in an increase in the LDH/HBDH ratio ranging from 1.6 to 2.5. In myocardial
infarction, LDH-1 and LDH-2 activity is increased, with a resultant decrease in the LDH/HBDH ratio to levels ranging from 0.8 to 1.2. Exact ratios are
dependent on assay conditions.
Test Principle2
Photometric UV determination based on the recommendations of the “German Society for Clinical Chemistry” (GSCC).
The anodal fraction of LDH isoenzymes (LDH-1 and LDH-2) shows a high activity with 2-oxobutyrate as substrate under suboptimal reaction conditions.
The LDH activity obtained with 2-oxobutyrate as substrate is called 2-hydroxybutyrate dehydrogenase (2-HBDH).
2-HBDH catalyses the reversible reduction of 2-oxobutyrate by NADH to 2-hydroxybutyrate. The decrease in NADH concentration is measured at 340 nm
and is directly proportional to the 2-HBDH activity in the sample.
Reaction Principle
+ HBDH +
2-Oxobutyrate + NADH + H 2-Hydroxybutyrate + NAD

Phosphate buffer (pH 7.5) 50 mmol/L
2-Oxobutyrate 3.0 mmol/L
NADH 0.18 mmol/L
Preservative

Reagent Preparation

The reagents are stable, unopened, up to the stated expiry date when stored at 2...8°C. Once open, reagents stored on board the instrument are stable for
30 days.
Specimen
3
Serum and heparinised plasma: Stable in serum for 4 days when stored at 2…8°C and 7 days when stored at 15…25°C.
Test Procedure
Calibration
The calibrator value is traceable to a Beckman Coulter master calibrator.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the HBDH activity of each sample.
Adults 90 – 180 U/L (1.5 – 3 µkat/L)

2009-08
values.
Linearity
Precision
70 1.03 1.48 1.38 1.98
167 1.76 1.06 3.55 2.13
970 8.58 0.88 12.98 1.34
Sensitivity
The lowest detectable level represents the lowest measurable level of HBDH that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison
Patient serum samples were used to compare this HBDH OSR6129 assay on the AU600 against another commercially available HBDH assay. Results of
y = 1.081x -11 r = 0.997 n = 118 Sample range = 88 – 574 U/L
Bilirubin: Interference less than 3% up to 40 mg/dL or 684 µmol/L bilirubin
®
5
Limitations

BIBLIOGRAPHY
1. Moss DW, Henderson RA, Kachmar JF. Enzymes. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:381pp.
2. Bergmeyer HU. Empfehlungen der Deutschen Gesellschaft für Klinische Chemie. Clin Chem Biochem 1970;8:658-660 & 1972;10:82-192.
4. Thomas L. Enzyme. Labor und Diagnose, 4 Auflage. Marburg: Die Medizinische Verlagsgesellschaft, 1992:136pp.

2009-08
HBDH, AU400/AU640 Serum/Plasma Application HBDH, AU600 Serum/Plasma Application

General LIH ISE Range Test No # Test name HBDH Sample type Ser Page 1/2
∇ ∇
Test Name: HBDH ∇ < > Type: Serum ∇ Operation: Yes ∇ Sample vol. 3 Dil. vol 0 μL Min. OD Max. OD
Specific Test Parameters Test No # Test name HBDH ∇ Sample type Ser ∇ Page 2/2
Level L Level H
Test Name: HBDH ∇ < > Type: Serum ∇ Value/flag # ∇ # #
Normal range
Value/Flag: # ∇ Level L: # Level H: # Sex Age L Age H L H
Normal Ranges: Age L Age H 1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # # 3 # # Y # M # Y # # #
∇ M→
ο 2. # ∇ # # # # # # 4 # # Y # M # Y # # #
∇ M→
ο 3. # ∇ # # # # # # 5 # # Y # M # Y # # #
∇ M→
ο 4. # ∇ # # # # # # 6 # # Y # M # Y # # #
∇ M→
ο 5. # ∇ # # # # # # 7 Non select # #
ο 6. # ∇ # # # # # # 8 Out of range # #
7. None Selected # # L H
8. Out of Range L H # # Panic value # #
Panic Value: # # Unit: U/L* Decimal places: # Select the function using the Function key or the Mouse
Calibration Specific Test No # Test name HBDH ∇
General ISE
Cal type 1 MB ∇ Count #
Test Name: HBDH Type Serum
∇ < > ∇ Formula 1 Y=AX+B Process
∇ ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇ Selection calibrator
Point 1: § § §8025* §13375* Point 2 ∇
1-Point Cal. Point: ο With CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ MB type factor #

# User defined
§ For use in AB mode only. Refer to IFU for further instruction.
§ For use in AB mode only. Refer to IFU for further instruction.
Enzyme BSOSR6x29.01
2009-08
HBDH, AU2700/AU5400 Serum/Plasma Application HBDH, AU680/AU480 Serum/Plasma Application
Test Name: HBDH ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: HBDH ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: HBDH ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: HBDH ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: HBDH Type Serum
∇ < > ∇ Test Name: HBDH Type Serum Use Serum Cal.
∇ < > ∇ ο
Calibration Type: MB ∇ Formula: Y=AX=B ∇ Counts: # Process: ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
Point 1: § § §5850* §9750* Point 1: § ∇ § §5850 §9750
Point 9 ∇
§ For use in AB mode only. Refer to IFU for further instruction. Point-1 ∇ Reagent Blank Day Hour
Enzyme BSOSR6x29.01
2009-08
LDH
OSR6126 4 x 50 mL R1
4 x 25 mL R2
Intended Use
Kinetic UV test for the quantitative determination of lactate dehydrogenase, EC 1.1.1.27 (LDH) in human serum and plasma on Beckman Coulter
Summary1,2,3
+ +
Lactate dehydrogenase, an NAD oxidoreductase, catalyses the reversible oxidation of L-lactate to pyruvate using NAD as a hydrogen acceptor. It is
present in all cells of the body and is invariably found only in the cytoplasm of the cell. The enzyme has a molecular weight of 134,000 and is composed of
4 peptide chains of 2 types: M and H. The total LDH measurable in serum consists of the activities of the 5 isoenzymes LDH-1 to LDH-5 which are
differentiated on the basis of their subunit composition. Because the concentration of LDH in the tissues is 500 times higher than that in plasma, damage to
even a small amount of tissue can lead to a significant increase in the activity of LDH in serum. The main role of total LDH is therefore in the detection of
minor tissue damage. High specific activities of the enzyme are found in the liver, cardiac muscle, skeletal muscle, kidneys and erythrocytes.
Myocardial infarction is usually associated with a 3-4 fold elevation of total LDH; similar increases in LDH can occur in myocarditis, cardiac dysrhythmias,
electrical cardioversion and prosthetic valve replacement. After prosthetic valve replacement there is a close correlation between the LDH level and
shortened erythrocyte survival time. Determination of LDH is therefore a reliable method for quantification of the extent of haemolysis. Elevations of LDH
activity are observed in liver damage, but these elevations are not as great as the increases in aminotransferase activity. Elevations are especially high
(10 times upper limit of normal) in toxic hepatitis with jaundice; slightly lower levels are observed in viral hepatitis and infectious mononucleosis. The
LDH/AST ratio can be used to differentiate between prehepatic jaundice caused by haemolysis or dyserythropoiesis from hepatic jaundice. In Duchenne
muscular dystrophy elevations of LDH activity are found years before the appearance of clinical symptoms, in the course of the disease the activity can
increase approximately 5 fold. Elevated LDH may also be seen in Aran-Duchenne and Kugelberg-Welander spinal muscular atrophy, dermatomyositis,
polymyositis and as a result of strenuous physical exercise. Other diseases that can cause an increased activation of LDH include renal infarction, Korean
haemorrhagic fever, chronic glomerular disease, shock, pulmonary embolism/infarction and haemolytic and megaloblastic anemias.
Test Principle4
Method based on the recommendations of the Scandinavian Committee on Enzymes.
+
LDH catalyses the reduction of pyruvate to lactate at a neutral pH. This reaction is coupled with the oxidation of NADH to NAD . The decrease of NADH is
measured at 340 nm and is directly proportional to the enzyme activity in the sample.
Reaction Principle
+ +
LDH
Pyruvate + NADH + H Lactate + NAD

Bis-Tris-Propane, pH 7.4 (37°C) 25 mmol/L
NADH 0.18 mmol/L
Pyruvate 1.2 mmol/L
Preservative

Reagent Preparation

The reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C. Once open, reagents stored on board the instrument are stable for
30 days.
Specimen
Serum and heparinised plasma. Separate serum from the clot as soon as possible. Haemolysed serum must not be used because erythrocytes contain
150 times more LDH activity than serum.
5
Stable in serum for 4 days when stored at 2…8°C and 7 days when stored at 15…25°C.
Where plasma is used, care should be taken to avoid contamination with platelets which contain high concentrations of LDH.
Test Procedure
Calibration

2009-08
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the LDH activity of each sample.
Adult 208 - 378 U/L (3.47 – 6.30 µkat/L)
Children
1 day <1327 U/L (22.1 µkat/L)
2 − 5 day <1732 U/L (28.9 µkat/L)
6 day − 6 month <975 U/L (16.3 µkat/L)
4 − 6 year <615 U/L (10.3 µkat/L)

values.
Linearity
The test is linear within an enzymatic activity range of 50 - 3000 U/L (0.8 – 50.0 µkat/L).
Precision
116 2.46 2.13 5.10 4.41
402 6.64 1.65 10.02 2.49
1992 29.22 1.47 39.11 1.96
Sensitivity
The lowest detectable level represents the lowest measurable level of LDH that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison
Patient serum samples were used to compare this LDH OSR6126 assay on the AU600 against another commercially available LDH assay. Results of
y = 1.009x + 1 r = 0.998 n = 82 Sample range = 211 – 700 U/L
Results of studies conducted to evaluate the susceptibility of the LDH method to interference were as follows:
®
7

‡ Set min OD at 0.6 and reagent OD limit low at 0.8 on AU640
BIBLIOGRAPHY
1. Thomas L. Lactate dehydrogenase (LD). In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
1999;668-673.
4. The Committee on Enzymes of the Scandinavian Society for Clinical Chemistry and Clinical Physiology. Recommended methods for the determination of four
enzymes in blood. Scand J Clin Lab Invest 1974;33:291-306.
6. Fischbach F, Zawta B. Age-Dependent Reference Limits in Plasma. Klin. Lab 1992;38,555.

2009-08
LDHP (SCE), AU400/AU640 Serum/Plasma Application LDHP (SCE), AU600 Serum/Plasma Application

Test No # Test name LDH26 ∇ Sample type Ser ∇ Page ½
Test Name: LDH26 ∇ < > Type: Serum ∇ Operation: Yes ∇
R2 Volume 50 μL Dilution 75 μL L 0.7‡ H 2.5 Fst. L 0.8 Fst. H 2.5
Wavelength: Pri. 340 ∇ Sec. 380 ∇ First L 1.0‡ First H 2.5 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 1.0‡ Last H 2.5 Reaction - ∇ L 50* H 3000*
No Lag Time: YES ∇ On-board stability period: 30 Linearity Fst 15 % Sec 0 %
General LIH ISE Range Test No # Test name LDH26 ∇ Sample type Ser ∇ Page 2/2
Test Name: LDH26 ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name LDH26 ∇
Test Name: LDH26 ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §13230* §19840*
Point 1: § § §13230* §19840*
Point 2 ∇
Point 2:
1-Point Cal. Point: ο With CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. Point
MB type factor #
‡ Set min OD at 0.6 and reagent OD limit low at 0.8 on AU640 * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 § For use in AB mode only, refer to IFU for further instruction
§ For use in AB mode only, refer to IFU for further instruction
Enzyme BSOSR6x26.01
2009-08
LDHP (SCE), AU2700/AU5400 Serum/Plasma Application LDHP (SCE), AU680/AU480 Serum/Plasma Application
Test Name: LDH26 ∇ < > Type: Serum ∇ Operation Yes ∇
μL μL
Reagent OD limit:
Test Name: LDH26 ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: LDH26 ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: LDH26 ∇ < > Type Serum ∇ Test Name: LDH26 ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: § § §10700* §16060* Point 1: § ∇ § §10700* §16060*
Point 9 ∇
§ For use in AB mode only, refer to IFU for further instruction Point-2 Calibration Day Hour
∇
ф AU680
Enzyme BSOSR6x26.01
2009-08
LDHP (SCE), AU400/AU640 Paediatric Application LDHP (SCE), AU600 Paediatric Application

Test No # Test name LD26P ∇ Sample type Ser ∇ Page 1/2
Test Name: LD26P ∇ < > Type: Serum ∇ Operation: Yes ∇
R2 Volume 50 μL Dilution 65 μL L 0.7‡ H 2.5 Fst. L 0.8 Fst. H 2.5
Wavelength: Pri. 340 ∇ Sec. 380 ∇ First L 1.0‡ First H 2.5 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 1.0‡ Last H 2.5 Reaction - ∇ L 50* H 3000*
No Lag Time: YES ∇ On-board stability period: 30 Linearity Fst 15 % Sec 0 %
General LIH ISE Range Test No # Test name LD26P ∇ Sample type Ser ∇ Page 2/2
Test Name: LD26P ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name LD26P ∇
Test Name: LD26P ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §13230* §19840*
Point 1: § § §13230* §19840*
Point 2 ∇
Point 2:
MB type factor #
‡ Set min OD at 0.6 and reagent OD limit low at 0.8 on AU640 * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 § For use in AB mode only, refer to IFU for further instruction
§ For use in AB mode only, refer to IFU for further instruction
Enzyme BSOSR6x26.01
2009-08
LDHP (SCE), AU2700/AU5400 Paediatric Application LDHP (SCE), AU680/AU480 Paediatric Application
Test Name: LD26P ∇ < > Type: Serum ∇ Operation Yes ∇
μL μL
Reagent OD limit:
Test Name: LD26P ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: LD26P ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: LD26P ∇ < > Type Serum ∇ Test Name: LD26P ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: § § §10700* §16060* Point 1: § ∇ § §10700* §16060*
Point 9 ∇
§ For use in AB mode only, refer to IFU for further instruction Point-2 Calibration Day Hour
∇
ф AU680
Enzyme BSOSR6x26.01
2009-08
LDH
OSR6128 4 x 40 mL R1
4 x 20 mL R2
Intended Use
Kinetic UV test for the quantitative determination of lactate dehydrogenase, EC 1.1.1.27 (LDH) in human serum and plasma on Beckman Coulter
Summary1,2,3
+ +
Lactate dehydrogenase, an NAD oxidoreductase, catalyses the reversible oxidation of L-lactate to pyruvate using NAD as a hydrogen acceptor. It is
present in all cells of the body and is invariably found only in the cytoplasm of the cell. The enzyme has a molecular weight of 134,000 and is composed of
4 peptide chains of 2 types: M and H. The total LDH measurable in serum consists of the activities of the 5 isoenzymes LDH-1 to LDH-5 which are
differentiated on the basis of their subunit composition. Because the concentration of LDH in the tissues is 500 times higher than that in plasma, damage to
even a small amount of tissue can lead to a significant increase in the activity of LDH in serum. The main role of total LDH is therefore in the detection of
minor tissue damage. High specific activities of the enzyme are found in the liver, cardiac muscle, skeletal muscle, kidneys and erythrocytes.
Myocardial infarction is usually associated with a 3-4 fold elevation of total LDH; similar increases in LDH can occur in myocarditis, cardiac dysrhythmias,
electrical cardioversion and prosthetic valve replacement. After prosthetic valve replacement there is a close correlation between the LDH level and
shortened erythrocyte survival time. Determination of LDH is therefore a reliable method for quantification of the extent of haemolysis. Elevations of LDH
activity are observed in liver damage, but these elevations are not as great as the increases in aminotransferase activity. Elevations are especially high
(10 times upper limit of normal) in toxic hepatitis with jaundice; slightly lower levels are observed in viral hepatitis and infectious mononucleosis. The
LDH/AST ratio can be used to differentiate between prehepatic jaundice caused by haemolysis or dyserythropoiesis from hepatic jaundice. In Duchenne
muscular dystrophy elevations of LDH activity are found years before the appearance of clinical symptoms, in the course of the disease the activity can
increase approximately 5 fold. Elevated LDH may also be seen in Aran-Duchenne and Kugelberg-Welander spinal muscular atrophy, dermatomyositis,
polymyositis and as a result of strenuous physical exercise. Other diseases that can cause an increased activation of LDH include renal infarction, Korean
haemorrhagic fever, chronic glomerular disease, shock, pulmonary embolism/infarction and haemolytic and megaloblastic anemias.
Test Principle4
+
LDH catalyses the oxidation of lactate to pyruvate coupled with the reduction of NAD to NADH. The increase of NADH is measured at 340nm and is
directly proportional to the enzyme activity in the sample.
Reaction Principle
+ LDH +
Lactate + NAD Pyruvate + NADH + H

D(-)N-Methylglucamin buffer, pH 9.4 (37°C) 325 mmol/L
Lactate 50 mmol/L
+
NAD 10 mmol/L
Preservative
R1 and R2 reagents: Irritant, contains a mixture of 5-chloro-2-methyl-4-isothiazolin-3-one [EC No 247-500-7] and 2-methyl-4-isothiazolin-3-one [EC No
220-239-6] (3:1). R43; May cause sensitisation by skin contact.
Safety Phrases:
Reagent Preparation
30 days.
Specimen
Serum and heparinsied plasma. Separate serum from the clot as soon as possible. Haemolysed samples must not be used because erythrocytes contain
150 times more LDH activity than serum/plasma.
5
Stable in serum for 4 days when stored at 2…8°C and 7 days when stored at 15…25°C.
Where plasma is used, care should be taken to avoid contamination with platelets which contain high concentrations of LDH.
Test Procedure
Calibration
should be used. A fresh vial of calibrator,utilising System Calibrator Cat No. 66300 in the AB calibration mode, should be used for each of these runs.

2009-08
The calibrator value is traceable to the IFCC reference method and IRMM/IFCC-453.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the lactate dehydrogenase activity of each sample.
Female < 247 U/L (4.12 μkat/L)
Male < 248 U/L (4.13 μkat/L)
Children
0 − 4 day 290 − 775 U/L (4.83 – 12.92 μkat/L)
4 − 10 day 545 − 2000 U/L (9.1 – 33.3 μkat/L)
10 day − 24 month 180 − 430 U/L (3.0 – 7.2 μkat/L)
24 month − 12 year 110 − 295 U/L (1.83 – 4.92 μkat/L)
values.
Linearity
The test is linear within an enzyme activity range of 25 - 1200 U/L (0.4 - 20.0 µkat/L).
Precision
157 1.78 1.13 2.42 1.54
237 2.12 0.89 3.23 1.36
430 3.28 0.76 6.16 1.43
Sensitivity
The lowest detectable level represents the lowest measurable level of LDH that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison
Patient serum samples were used to compare this LDH OSR6128 assay on the AU640 against the IFCC reference method. Results of linear regression
y = 1.011x + 5.192 r = 0.989 n = 111 Sample range = 73 - 549 U/L
Icterus: Interference less than 3% up to 40 mg/dL or 684 μmol/L bilirubin
®
8
Limitations
* Values set for working in U/L. To work in SI units (μkat/L) divide by 60.
BIBLIOGRAPHY
1. Thomas L. Lactate dehydrogenase (LD). In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-
Books Verlagsgesellschaft, 1998:89-94.
1999;668-673.
4. Bais R, Philcox M. International Federation of Clinical Chemistry (IFCC) Approved recommendation on IFCC methods for the measurement of catalytic
concentration of enzymes. Part 8. IFCC method for lactate dehydrogenase. Eur J Clin Chem Clin Biochem 1994;32:639-55.
6. Schumann G, Klauke R. New IFCC reference procedures for the determination of catalytic activity concentrations of five enzymes in serum: preliminary upper
reference limits obtained in hospitalised subjects. Clin Chim Acta 2003;327:69-79.

2009-08
LDH (IFCC), AU400/AU640 Serum/Plasma Application LDH (IFCC), AU600 Serum/Plasma Application

Test No # Test name LDH28 ∇ Sample type Ser ∇ Page ½
General LIH ISE Range Test No # Test name LDH28 ∇ Sample type Ser ∇ Page 2/2
Test Name: LDH28 ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name LDH28 ∇
Test Name: LDH28 Type Serum
∇ < > ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §8500* §12800*
Point 1: § § §8500* §12800*
Point 2 ∇
Point 2:
MB type factor #
# User defined
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 # User defined ¤ Analyser default value
§ For use in AB mode only, refer to IFU for further instruction. * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
‡ Depends on usage pattern in the Laboratory § For use in AB mode only, refer to IFU for further instruction.
Enzyme BSOSR6x28.01
2009-08
LDH (IFCC), AU2700/AU5400 Serum/Plasma Application LDH (IFCC), AU680/AU480 Serum/Plasma Application
Test Name: LDH28 ∇ < > Type: Serum ∇ Operation Yes ∇
μL μL
Reagent OD limit:
Test Name: LDH28 ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: LDH28 ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: LDH28 ∇ < > Type Serum ∇
Test Name: LDH28 ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: § § §6900* §11640* Point 1: § ∇ § §6900* §11640*
Point 9 ∇
§ For use in AB mode only, refer to IFU for further instruction. Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
‡ Depends on usage pattern in the Laboratory Point-2 ∇ Calibration ‡ Day ‡ Hour
Enzyme BSOSR6x28.01
2009-08
LDH (IFCC), AU400/AU640 Paediatric Application LDH (IFCC), AU600 Paediatric Application

Test No # Test name LD28P ∇ Sample type Ser ∇ Page 1/2
General LIH ISE Range Test No # Test name LD28P ∇ Sample type Ser ∇ Page 2/2
Test Name: LD28P ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE
Test No # Test name LD28P ∇
Test Name: LD28P ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Point 1 § ∇ § §8500* §12800*
Point 1: § § §8500* §12800*
Point 2 ∇
Point 2:
MB type factor #
# User defined
Enzyme BSOSR6x28.01
2009-08
LDH (IFCC), AU2700/AU5400 Paediatric Application LDH (IFCC), AU680/AU480 Paediatric Application
Test Name: LD28P ∇ < > Type: Serum ∇ Operation Yes ∇
μL μL
Reagent OD limit:
Test Name: LD28P ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: LD28P ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: LD28P ∇ < > Type Serum ∇ Test Name: LD28P ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: § § §6900* §11640* Point 1: § ∇ § §6900* §11640*
Point 9 ∇
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
§ For use in AB mode only, refer to IFU for further instruction. Point-2 Calibration ‡ Day ‡ Hour
∇
Enzyme BSOSR6x28.01
2009-08
LIPASE
OSR6130 4x 10 mL R1 Buffer
4x R1 Lyo
4x 3.3 mL R2
2x Calibrator
OSR6230 4x 30 mL R1 Buffer
4x R1 Lyo
4x 10 mL R2
2x Calibrator
Intended Use
Kinetic colour test for the quantitative determination of lipase in human serum and plasma on Beckman Coulter analysers. For in vitro diagnostic
use only.
Summary1,2
Lipase is produced in the acinar cells of the pancreas and is responsible for the hydrolysis of water-insoluble long chain fatty acid esters of
glycerol. Lipase measurement in serum and plasma is used exclusively for the investigation of pancreatic disorders, usually pancreatitis. Serum
lipase may be elevated in acute pancreatitis, acute episodes of chronic pancreatitis and obstructive pancreatitis, with levels up to 80 times the
upper reference limit found in acute serious inflammation. However it should be noted that the severe destruction of the acinar cells in the later
stages of chronic pancreatitis results in a reduction of the amount of enzyme entering the circulation. Marginal or no increase of lipase is
therefore not unusual in this disease. In acute upper quadrant abdominal syndrome, hyperlipasemia of up to 5 times the upper reference limit
may be found in penetrating duodenal ulcer, duodenal diverticulum, cholecystitis, and ileus, where there is pancreatic involvement. Lipase levels
are also elevated in renal insufficiency, particularly where dialysis is required. Investigation of the biliary tract by endoscopic retrograde
pancreatography, or treatment with opiates, may also result in serum lipase elevation. Slight elevations are also frequently present in diabetic
ketoacidosis, viral hepatitis, epidemic parotiditis, abdominal typhoid and sarcoidosis, due to involvement of the pancreas.
Test Principle3
Pancreatic lipase hydrolyses esters of long chain fatty acids from their triglycerides. The enzyme activity requires the presence of co-lipase.
Pancreatic specific 1,2-Diglyceride is hydrolysed to 2-Monoglyceride and fatty acid. The 2-Monoglyceride is then measured by coupled enzyme
reactions catalysed by monoglyceride lipase (MGLP), glycerol kinase (GK), glycerol phosphate oxidase (GPO) and peroxidase (POD).
Reaction Principle
Lipase
1,2-Diglyceride + H2O 2-Monoglyceride + fatty acid
MGLP
2-Monoglyceride + H2O Glycerol + fatty acid
GK
Glycerol + ATP Glycerol-3-phosphate + ADP
GPO
Glycerol-3-phosphate + O2 Dihydroxyacetone-P + H2O2
POD
2 H2O2 + 4-aminophenazone + TOOS Quinonediimine-dye + 4 H2O

Buffer MES/BES (pH 6.8) 27 mmol/L
1,2-Diglyceride substrate 0.04 mmol/L
Monoglyceride lipase > 400 U/L
Glycerol kinase > 100 U/L
POD > 500 U/L
4-Aminophenazone 0.25 mmol/L
TAPS (pH 8.7) 50 mmol/L
TOOS 1.0 mol/L
Co-lipase > 15 kU/L
GPO > 15 kU/L
ATP > 0.85 mol/L
Preservative
Calibrator: Human serum containing porcine lipase.

Biological materials of human origin contained in this product were tested for Anti-HCV, HbsAg and Anti-HIV 1/2 on a single donor basis using
FDA approved methods and were found to be non-reactive. As there is no known test method that can offer complete assurance that products
derived from human blood will not transmit infectious agents, this product should be handled as a potentially infectious material.
Reagent Preparation
R1: Dissolve the contents of one vial of R1 Lyo completely with the contents of one vial of R1 buffer. Mix gently by inversion and place on board the
instrument.
R2 is ready for use and can be placed directly on board the instrument.

2009-08
Calibrator Preparation
1. Carefully remove the cap and rubber stopper from the bottle, avoiding any loss of lyophilised material.
2. Add 3.0 mL of sterile deionised water at 15…25°C to the lyophilised material using a volumetric pipette calibrated to deliver exactly 3.0 mL.
3. With the rubber stopper back in place, dissolve the contents completely by gently mixing for 30 minutes. Avoid foaming.
4. Continue mixing until the solution is homogeneous and all lyophilized material is reconstituted.
5. Record the date the calibrator was reconstituted on the bottle label.

The reagents are stable, unopened, up to the stated expiry date when stored at 2...8°C. Once open or prepared, reagents stored on board the
instrument are stable for 21 days.
The lipase calibrator is stable, unopened, up to the stated expiry date when stored at 2...8°C. Reconstituted lipase calibrator is stable for 60
days when stored at 2...8°C.
Specimen
4
Stable in serum and plasma for 3 weeks when stored at 2…8°C and 7 days when stored at 15…25°C.
Lipemic and icteric samples should be avoided.
Test Procedure
Calibration
Calibrator provided in the kit: For value assigned to the calibrator provided in the kit, please refer to bottle label.
The calibrator value is traceable to a Beckman Coulter master calibrator. Recalibrate the assay every 7 days, or when the following occur:
Change in reagent bottle number or significant shift in control values;
Note: System Calibrator Cat. No.: 66300 can also be used. Refer to System Calibrator Cat. No.: 66300 instructions for use for further
information.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the lipase activity of each sample.
Reference Intervals
5
Adult < 67 U/L (< 1.12 µkat/L)
6
Children < 1y 0 – 8 U/L (0 – 0.13 µkat/L)
1 – 9y 5 – 31 U/L (0.08 – 0.52 µkat/L)
10 – 18y 7 – 39 U/L (0.12 – 0.65 µkat/L)

values.
Linearity
The test is linear within an enzyme activity range of 3 – 600 U/L (0.05 – 10 µkat/L).
Precision
25 0.45 1.83 0.91 3.68
51 0.72 1.42 1.30 2.56
242 1.79 0.74 5.51 2.28
Sensitivity
The lowest detectable level represents the lowest measurable level of lipase that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient serum samples were used to compare this Lipase OSR6130 assay on the AU600 against another commercially available lipase assay. Results of
y = 0.984x – 3 r = 0.999 n = 84 Sample range = 5 – 715 U/L
®
7

2009-08
Limitations
Carry over from Triglyceride, HDL-Cholesterol and LDL-Cholesterol reagents to Lipase reagent results in elevated lipase values. Please refer to
contamination parameters. For AU400/AU480/AU640/AU680 customers who use Lipase OSR6x30, HDL-Cholesterol OSR6x87 and
LDL-Cholesterol OSR6x83, it is recommended that the programming of these tests is set up such that Lipase is the first test to be run followed
by LDL and then HDL. For example: Lipase – Channel 1, LDL - Channel 2 and HDL – Channel 3. All other tests can be programmed in the
remaining test channels.

† For use in AB mode only, refer to leaflet for further instruction.
BIBLIOGRAPHY
1. Lorentz K. Lipase. In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
Verlagsgesellschaft, 1998:95-97.
1999; 698-704.
3. Imamura S, Hirayama T, Arai T, Takao K, Misaki H. An enzymatic method using 1,2-Diglyceride for pancreatic lipase test in serum. Clin Chem 1989; 35:1126.
5. Data on File.
6. Lorentz K. Lipase. In:Thomas L, hrsg. Labor und Diagnose. Indikation und bewertung von laborbefunden für die medizinische diagnostik. Frankfurt/Main:

2009-08
LIPASE, AU400/AU640 Serum/Plasma Application LIPASE, AU600 Serum/Plasma Application

Test No # Test name LIP ∇ Sample type Ser ∇ Page 1/2
Test Name: LIP ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name LIP ∇ Sample type Ser ∇ Page 2/2
Test Name: LIP ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name LIP ∇
Test Name: LIP Type Serum
∇ < > ∇ Cal type 8 AB ∇ Count #
Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇
Point 1 # ∇ † 5000* 10000*
Point 1: # † 5000* 10,000*
Point 2 ∇
Point 2:
MB type factor
MB Type Factor: Calibration Stability Period: 7 Calibrator stability period 7
# User defined
† For use in AB mode only, refer to IFU for further instructions
† For use in AB mode only, refer to IFU for further instrucation
Enzyme BSOSR6x30.01
2009-08
LIPASE, AU2700/AU5400 Serum/Plasma Application LIPASE, AU680/AU480 Serum/Plasma Application
Test Name: LIP ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: LIP ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Measuring Point2 First Last Lipemia + ∇
Linearity Limit 15 % Icterus +++ ∇
Test Name: LIP ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: LIP ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: LIP ∇ < > Type Serum ∇
Test Name: LIP ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # ∇
Point 1: # † 5000* 10000* Point 1: # ∇ † 5000* 10000*
Point 9 ∇
MB Type Factor: ICF: Calibration Stability Period: 7
† For use in AB mode only, refer to IFU for further instructions Calibrator OD Conc Low High Stability
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 Point-1 ∇ Reagent Blank 7 Day 0 Hour
ф AU680 Point-2 ∇ Calibration 7 Day 0 Hour
MB Type Factor: 1-Point Calibration Point ∇ ο with Conc-0
Enzyme BSOSR6x30.01
2009-08
ALBUMIN
OSR6102 4 x 29 mL R1
OSR6202 4 x 54 mL R1
Intended Use
Photometric colour test for the quantitative determination of albumin in human serum and plasma on Beckman Coulter analysers. For in vitro diagnostic
use only.
Summary1,2
Albumin is the most abundant protein in human plasma, representing 55-65% of the total protein. Its primary biological functions are to transport and store
a wide variety of ligands, to maintain the plasma oncotic pressure and to serve as a source of endogenous amino acids. Albumin binds and solubilises
non-polar compounds such as plasma bilirubin and long-chain fatty acids as well as binding numerous pharmaceuticals.
Hyperalbuminemia is infrequent and is caused by severe dehydration and excessive venous stasis. Hypoalbuminemia may be caused by impaired
synthesis e.g. in liver disease or in protein deficient diets; increased catabolism as a result of tissue damage and inflammation; reduced absorption of
amino acids caused by malabsorption syndromes or malnutrition; protein loss to the exterior as observed in nephrotic syndrome, enteropathy or burns; and
altered distribution e.g. in ascites. Severe hypoalbuminemia results in a serious imbalance of intravascular oncotic pressure causing the development of
edema.
Measurements of albumin concentrations are vital to the understanding and interpretation of calcium and magnesium levels because these ions are bound
to albumin, and so decreases of albumin are also directly responsible for depression of their concentrations.
Test Principle3
A coloured complex is formed when bromocresol green reacts with albumin. The absorbance of the albumin-BCG complex is measured bichromatically
(600/800nm) and is proportional to the albumin concentration in the sample.
Reaction Principle
pH 4.2
Albumin + Bromocresol Green Green Complex

Succinate buffer (pH 4.2) 100 mmol/L
Bromocresol green 0.2 mmol/L
Preservative

Safety Phrases:
Reagent Preparation

The reagent is stable, unopened, up to the stated expiry date when stored at 2…25°C. Once open, reagent stored on board the instrument is stable for
90 days.
Specimen
4
Separate from cells immediately. Separated plasma and serum is stable at 2…8°C for 30 days and at 15…25°C for 7 days.
Test Procedure
Calibration
System Calibrator Cat. No. 66300.
The calibrator albumin value is traceable to IFCC (International Federation of Clinical Chemistry) standard CRM 470. Recalibrate the assay when the
following occur:
Change in reagent lot number or significant shift in control values;
Quality Control
EN.01 BLOSR6x02.01 Metabolite

2009-08
Calculation
The Beckman Coulter analysers automatically compute the albumin concentration of each sample.
Serum (Adults) 35 – 52 g/L (3.5 – 5.2 g/dL)
Serum (Newborn 0 – 4 day) 28 – 44 g/L (2.8 – 4.4 g/dL)

values.
Linearity
The test is linear within a concentration range of 15 – 60 g/L (1.5 – 6.0 g/dL)
Precision
Mean g/L SD CV% SD CV%
24.04 0.43 1.80 0.63 2.62
42.93 0.50 1.17 0.67 1.55
57.77 0.53 0.92 1.03 1.79
Sensitivity
The lowest detectable level in serum on an AU600 analyser was estimated at 0.07 g/L.
The lowest detectable level represents the lowest measurable level of albumin that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient serum samples were used to compare this Albumin OSR6102 assay on the AU600 against another commercially available albumin assay. Results
of linear regression analysis were as follows:
y = 0.982x – 3.860 r = 0.994 n = 121 Sample range = 11.70 – 50.85 g/L
®
7

† System Calibrator Cat. No.: 66300
* Values set for working in SI units (g/L). To work in g/dL divide by 10.
BIBLIOGRAPHY
1. Grant GH, Silverman LM, Christenson RH. Amino acids and proteins. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company,
1987:328-329.
2. McPherson RA. Specific proteins. In: Henry JB, ed. Clinical diagnosis and management by laboratory methods. Philadelphia:WB Saunders Company, 1996:244-
245.
3. Doumas BT, Watson WA, Biggs HG. Albumin standards and the measurement of serum albumin with bromcresol green. Clin Chim Acta 1971;31:87-96.
4. Young DS, ed. Effects of preanalytical variables on clinical laboratory tests, 2nd ed. Washington:AACC Press, 1997:3-15-3-16.
5. Baudner S, Dati F. Standardization of the measurement of 14 proteins in human serum based on the new IFCC/BCR/CAP international reference material CRM
470. J Lab Med 1996;20:145-152.
Metabolite BLOSR6x02.01 EN.01

2009-08
ALBUMIN, AU400/AU640 Serum/Plasma Application ALBUMIN, AU600 Serum/Plasma Application

Test No # Test name ALB ∇ Sample type Ser ∇ Page ½
Test Name: ALB ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 2 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 58 Dil. Vol 242 μL L H
R2 Volume 0 μL Dilution 0 μL L H Fst. L -0.1 Fst. H 0.5
Wavelength: Pri. 600 ∇ Sec. 800 ∇ First L -0.1 First H 0.5 Method END ∇ Dynamic range
Method: END ∇ Last L -0.1 Last H 0.5 Reaction + ∇ L 15* H 60*
Linearity : % A 1 B 0
No Lag Time: ∇ On-board stability period: 90 Linearity Fst % Sec %
No lag time ∇ On-board stability period 90
General LIH ISE Range Test No # Test name ALB ∇ Sample type Ser ∇ Page 2/2
Test Name: ALB ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic Value: # # Unit: g/L* Decimal places: # Panic value # #
General ISE Test No # Test name ALB ∇
Test Name: ALB Type Serum
∇ < > ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 54* 88*
Point 1: # † 54* 88*
Point 2 ∇
Point 2:
1-Point Cal. Point: Slope Check: None Advanced Calibration: # 1-point cal. Point
ο with CONC-0 ∇ ∇
MB type factor
† System Calibrator Cat. No.: 66300 † System Calibrator Cat. No.: 66300
* Values set for working in SI units (g/L). To work in g/dL divide by 10 * Values set for working in SI units (g/L). To work in g/dL divide by 10
Metabolite BSOSR6x02.01
2009-08
ALBUMIN, AU2700/AU5400 Serum/Plasma Application ALBUMIN, AU680/AU480 Serum/Plasma Application
Test Name: ALB ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: ALB ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Pre-Dilution Rate 1 ∇ Min.OD Max.OD
R2 Volume 0 μL Dilution 0 μL L H
Reagent OD limit:
Method: END ∇ Last L -0.1 Last H 0.5
Measuring Point 2: First Last Correlation Factor: Method END ∇
Linearity : % A 1 B 0 Reaction Slope + ∇ Onboard Stability Period 90 Day 0 Hour
No Lag Time: ∇ On-board stability period: 90 Measuring Point1 First 0 Last 1 LIH Influence Check # ∇
Linearity Limit % Icterus +++++ ∇
Specific Test Parameters Lag Time Check ∇ Hemolysis +++++ ∇
Test Name: ALB ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: ALB ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: g/L* Decimal places: # 7. No demographics # #
Unit g/L* Decimal Places #
General ISE
General ISE
Test Name: ALB Type Serum
∇ < > ∇ Test Name: ALB Type Serum Use Serum Cal.
∇ < > ∇ ο
Point 1: # † 54* 96* Point 1: # ∇ † 54* 96*
1-Point Cal. Point: ο with Conc-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
MB Type Factor: Calibration Stability Period: 999
† System Calibrator Cat. No.: 66300 Point-1 ∇ Reagent Blank 999 Day 0 Hour
* Values set for working in SI units (g/L). To work in g/dL divide by 10 Point-2 Calibration 999 Day 0 Hour
∇
ф AU680
2009-08
ALBUMIN, AU400/AU640 Paediatric Application ALBUMIN, AU600 Paediatric Application

Test No # Test name ALBP ∇ Sample type Ser ∇ Page 1/2
Test Name: ALBP ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 2 μL Dilution 10 μL Pre-Dilution Rate: 1 Reagent 1 vol 58 Dil. vol 232 μL L H
General LIH ISE Range Test No # Test name ALBP ∇ Sample type Ser ∇ Page 2/2
Test Name: ALBP ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE
Test No # Test name ALBP ∇
Test Name: ALBP ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 54* 88*
Point 1: # † 54* 88*
Point 2 ∇
Point 2:
MB type factor
# User defined
* Values set for working in SI units (g/L). To work in g/dL divide by 10
* Values set for working in SI units (g/L). To work in g/dL divide by 10
2009-08
ALBUMIN, AU2700/AU5400 Paediatric Application ALBUMIN, AU680/AU480 Paediatric Application
Test Name: ALBP ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: ALBP ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: ALBP ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: ALBP ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: ALBP ∇ < > Type Serum ∇ Test Name: ALBP ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † 54* 96* Point 1: # ∇ † 54* 96*
Point 9 ∇
† System Calibrator Cat. No.: 66300 Calibrator OD Conc Low High Stability
* Values set for working in SI units (g/L). To work in g/dL divide by 10 Point-1 ∇ Reagent Blank 999 Day 0 Hour
2009-08
BICARBONATE
OSR6190 4 x 25 mL R1
OSR6290 4 x 50 mL R1
Intended Use
System reagent for the quantitative determination of bicarbonate in human serum and plasma on Beckman Coulter analysers. For in vitro diagnostic use
only.
Summary
Bicarbonate measurements are used in the diagnosis and treatment of numerous potentially serious disorders associated with changes in body acid-base
-
balance. The determination of bicarbonate (HCO3 ) is used in conjunction with other clinical and laboratory information for the evaluation of acid-base
- -
status. An elevation of the HCO3 level may be observed in compensated respiratory acidosis and metabolic alkalosis. Low HCO3 levels may be observed
1
in compensated respiratory alkalosis and metabolic acidosis. Additional laboratory determinations will permit differentiation between metabolic and
respiratory conditions.
Test Principle
-
The Bicarbonate reagent utilizes an enzymatic method to measure bicarbonate in human serum and plasma. In this procedure bicarbonate (HCO3 ) and
phosphoenolpyruvate (PEP) are converted to oxaloacetate and phosphate in the reaction catalyzed by phosphoenolpyruvate carboxylase (PEPC). Malate
dehydrogenase (MD) catalyzes the reduction of oxaloacetate to malate with the concomitant oxidation of a reduced nicotinamide adenine dinucleotide
(NADH). This oxidation of the NADH results in a decrease in absorbance of the reaction mixture measured bichromatically at 380/410 nm proportional to
-
the HCO3 content of the sample.
Reaction Principle
PEPC
- -
PEP + HCO3 Oxaloacetate + H2PO4
Magnesium
MD
+ +
Oxaloacetate + NADH + H Malate + NAD
Contents, reagent composition in the test

MD (microbial) > 2000 U/L
PEPC (microbial) > 572 U/L
NADH 1.6 mmol/L
PEP 8.2 mmol/L
Magnesium 2.8 mmol/L
Also contains preservatives.

Reagent Preparation

The reagent is stable unopened, up to the stated expiry date when stored at 2…8°C. Once open, reagent stored on board the instrument is stable for
7 days.
Specimen
Serum and heparinised plasma samples free from haemolysis are the recommended specimens. Separate serum or plasma from cells promptly to
minimise haemolysis.
- 1
Once separated from cells, HCO3 in serum is stable for several hours when stored at 2…8°C and protected from exposure to air.
Test Procedure
Calibration
Bicarbonate Calibrator Cat No. ODC0019.
The calibrator bicarbonate value is traceable to the National Institute of Standards and Technology (NIST) Standard Reference Material (SRM) 351.
Recalibrate the assay every day, or when the following occur:
Absorption of atmospheric CO2 by the reagent on board the analyser can impair calibration stability. This effect will vary depending upon the rate of use.
Consequently each laboratory should set a calibration frequency in the instrument parameters appropriate to their usage pattern.
Quality Control
Control materials with values determined by this Beckman Coulter system may be used.
2009-08
Calculation
The Beckman Coulter analysers automatically compute the bicarbonate concentration of each sample.
Adult 21 − 31 mmol/L
Newborn 17 − 24 mmol/L
Infant 19 − 24 mmol/L
2 months − 2 years 16 − 24 mmol/L

values.
Linearity
The test is linear within a concentration range of 2.0 to 45.0 mmol/L.
Precision
4
n = 80 Within run Total
Mean, mmol/L SD CV% SD CV%
9.0 0.37 4.15 0.63 7.02
25.1 0.29 1.15 0.58 2.31
34.9 0.37 1.05 0.71 2.02
Sensitivity
The lowest detectable level on an AU640 analyser was estimated at 0.71 mmol/L.
The lowest detectable level represents the lowest measurable level of bicarbonate that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient samples were used to compare this Bicarbonate Reagent OSR6190 on an AU640 to a blood gas analyser method.
y = 0.999x – 1.207 r = 0.98 n = 102 Sample range = 9.4 – 32.2 mmol/L
5
AU600/AU640
Lipemia: Interference less than 5% up to 1000 mg/dL Intralipid.®
Haemolysis: Interference less than 10% up to 500 mg/dL haemoglobin
Icterus: Interference less than 5% up to 40 mg/dL billirubin
6

† Bicarbonate Calibrator Catalogue Number ODC0019
* Values set for working in mmol/L
BIBLIOGRAPHY
1. Tietz, N.W., Hg., Fundamentals of Clinical Chemistry, 3rd edition, W.B. Saunders Company, 1999.
2. Kaplan, L.A. and Pesce, A.J. Clinical Chemistry Theory, Analysis, Corelation 3rd Edition, CV Mosbey, 1996.
3. Painter PC, Cope JY, Smith JL. Reference information for the clinical laboratory. In:Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry.
4. NCCLS, Evaluation Protocol EP5-A, 1999.
5. NCCLS, Interference Testing in Clinical Chemistry EP7-P, 1986.
.

2009-08
BICARBONATE (CO2), AU400/AU640 Serum/Plasma Application BICARBONATE (CO2), AU600 Serum/Plasma Application

Test No # Test name CO2 ∇ Sample type Ser ∇ Page 1/2
Test Name: CO2 ∇ < > Type: Serum ∇ Operation: Yes ∇
R2 Volume 0 μL Dilution 0 μL L 0.14 H 2.5 Fst. L -2.0 Fst. H 2.5
Wavelength: Pri. 380 ∇ Sec. 410 ∇ First L -2.0 First H 2.5 Method FIXED ∇ Dynamic range
Method: FIXED ∇ Last L -2.0 Last H 2.5 Reaction - ∇ L 2* H 45*
General LIH ISE Range Test No # Test name CO2 ∇ Sample type Ser ∇ Page 2/2
Test Name: CO2 ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic Value: # # Unit: mmol/L* Decimal places: # Panic value # #
General ISE Test No # Test name CO2 ∇
Test Name: CO2 ∇ < > Type Serum ∇ Cal type 10 2AB ∇ Count #
Formula 3 POLYGONAL ∇ Process Conc ∇
Calibration Type: 2AB ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇
Point 1 # ∇ 20 0.050* 0.200*
Point 1: # 20 0.050* 0.200*
Point 2 # ∇ 40 0.100* 0.300*
Point 2: # 40 0.100* 0.300*
1-Point Cal. Point: With CONC-0 Slope Check: + # 1-point cal. point
MB type factor
MB Type Factor: Calibration Stability Period: ‡ Calibrator stability period ‡
† Bicarbonate Calibrator Cat. No.: ODC0019 † Bicarbonate Calibrator Cat. No.: ODC0019
* Values set for working in mmol/L * Values set for working in mmol/L
‡ Depends on usage pattern in the Laboratory ‡ Depends on usage pattern in the Laboratory
2009-08
BICARBONATE (CO2), AU2700/AU5400 Serum/Plasma Application BICARBONATE (CO2), AU680/AU480 Serum/Plasma Application
Test Name: CO2 ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CO2 ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Method: FIXED ∇ Last L -2.0 Last H 2.5
Measuring Point 2: First Last Correlation Factor: Method FIXED ∇
Linearity : % A 1 B 0 Reaction Slope - ∇ Onboard Stability Period 7 Day 0 Hour
Test Name: CO2 ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CO2 ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: mmol/L* Decimal places: # 7. No demographics # #
Unit mmol/L* Decimal Places #
General ISE
General ISE
Test Name: CO2 ∇ < > Type Serum ∇ Test Name: CO2 Type Serum Use Serum Cal.
∇ < > ∇ ο
Calibration Type: 2AB ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇ Calibration Type: 2AB ∇ Formula: POLYGONAL ∇ Counts: # ∇
<Calibrator Parameters> OD Range
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibrator OD Conc Low High Slope Check + ∇
Point 1: # 20 0.050* 0.200* Point 1: # ∇ 20 0.050* 0.200*
Point 2: # 40 0.100* 0.300* Point 2: # ∇ 40 0.100* 0.300* Allowance Range Check
1-Point Cal. Point: ο with CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
MB Type Factor: Calibration Stability Period: ‡
† Bicarbonate Calibrator Cat. No.: ODC0019
* Values set for working in mmol/L Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
ф AU680 MB Type Factor: 1-Point Calibration Point None ∇ ο with Conc-0
2009-08
BICARBONATE (CO2), AU400/AU640 Paediatric Application BICARBONATE (CO2), AU600 Paediatric Application

Test No # Test name CO2P ∇ Sample type Ser ∇ Page 1/2
Test Name: CO2P ∇ < > Type: Serum ∇ Operation: Yes ∇
R2 Volume 0 μL Dilution 0 μL L 0.14 H 2.5 Fst. L -2.0 Fst. H 2.5
Method: FIXED ∇ Last L -2.0 Last H 2.5 Reaction - ∇ L 2* H 45*
General LIH ISE Range Test No # Test name CO2P ∇ Sample type Ser ∇ Page 2/2
Test Name: CO2P ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name CO2P ∇
Test Name: CO2P ∇ < > Type Serum ∇ Cal type 10 2AB ∇ Count #
Point 1 # ∇ 20 0.050* 0.200*
Point 1: # 20 0.050* 0.200*
Point 2 # ∇ 40 0.100* 0.300*
Point 2: # 40 0.100* 0.300*
1-Point Cal. Point: with CONC-0 Slope Check: + Advanced Calibration: # 1-point cal. point
ο ∇ ∇
MB type factor
† Bicarbonate Calibrator Cat. No.: ODC0019 † Bicarbonate Calibrator Cat. No.: ODC0019
* Values set for working in mmol/L * Values set for working in mmol/L
2009-08
BICARBONATE (CO2), AU2700/AU5400 Paediatric Application BICARBONATE (CO2), AU680/AU480 Paediatric Application
Test Name: CO2P ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CO2P ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: CO2P ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CO2P ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: CO2P ∇ < > Type Serum ∇ Test Name: CO2P ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # 20 0.050* 0.200* Point 1: # ∇ 20 0.050* 0.200*
Point 2: # 40 0.100* 0.300* Point 2: # ∇ 40 0.100* 0.300* Allowance Range Check
Point 9 ∇
† Bicarbonate Calibrator Cat. No.: ODC0019
* Values set for working in mmol/L Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
2009-08
DIRECT BILIRUBIN
OSR6111 4 x 6 mL R1 DBILC
4 x 6 mL R1 DBILB
OSR6211 4 x 20 mL R1 DBILC
4 x 20 mL R1 DBILB
Intended Use
Photometric colour test for the quantitative determination of direct bilirubin in human serum and plasma on Beckman Coulter analysers. For
in vitro diagnostic use only.
Summary1,2
80 – 85% of bilirubin produced daily originates from haemoglobin released by the breakdown of senescent erythrocytes, the remaining
15 – 20% results from the breakdown of haem-containing proteins such as myoglobin, cytochromes, catalases and from bone marrow as a
result of ineffective erythropoiesis.
Because of its poor solubility in water unconjugated bilirubin (indirect bilirubin) is transferred to the liver bound to albumin. Inside the
hepatocytes it is rapidly conjugated with glucuronic acid to produce bilirubin mono- and diglucuronide (direct bilirubin) which are then excreted in
bile together with all other normal biliary constituents.
Whereas prehepatic jaundice (e.g. haemolytic anemia and neonatal jaundice) is primarily associated with an increase in unconjugated bilirubin,
the assessment of direct bilirubin is helpful in the determination of hepatic and post-hepatic jaundice. Diseases of hepatic origin with
predominantly conjugated hyperbilirubinemia include acute and chronic viral hepatitis, liver cirrhosis and hepatocellular carcinoma. Diseases of
post hepatic origin with predominantly conjugated hyperbilirubinemia include extrahepatic cholestasis and liver transplant rejection. Chronic
congenital conjugated hyperbilirubinemias include Dubin-Johnson and Rotor syndrome. The differentiation between chronic congenital
hyperbilirubinemias and acquired types of bilirubininemia is accomplished via the measurement of bilirubin fractions and the detection of normal
liver enzyme activities.
Test Principle3
A stabilised diazonium salt, 3,5 Dichlorophenyl diazonium tetrafluoroborate (DPD) couples directly with direct (conjugated) bilirubin in an acid
medium to form azobilirubin. The absorbance at 570 nm is proportional to the direct bilirubin concentration in the sample.
Reaction Principle
Bilirubin + DPD Azobilirubin

3,5 Dichlorophenyl diazonium tetrafluoroborate 0.07 mmol/L

DBILB and DBILC: Corrosive, Contains sulphuric acid. R35; Causes severe burns.
Safety Phrases:
S20, S26, S30, S36/37/39, S45, S60. When using do not eat or drink. In case of contact with eyes, rinse immediately with plenty of water and
seek medical advice. Never add water to this product. Wear suitable protective clothing, gloves and eye/face protection. In case of accident or if
you feel unwell, seek medical advice immediately (show the label where possible). This material and its container must be disposed of as
hazardous waste.
Reagent Preparation

The reagents are stable, protected from light, unopened, up to the stated expiry date when stored at 2…8°C. Once open, reagents stored on
board the instrument are stable for 21 days.
Specimen
1
Serum and heparinised plasma: stable for 3 days when protected from light and stored 15…25°C.
Even slight haemolysis can cause a reduction in value and such samples should be avoided.
Lipemic samples should also be avoided.
Test Procedure
Calibration
Recalibrate the assay every 21 days, or when the following occur:
Change in reagent lot or significant shift in control values;

2009-08
Quality Control
samples are tested and each time calibration is performed.
Calculation
The Beckman Coulter analysers automatically compute the direct bilirubin concentration of each sample.
Adults and Children < 3.4 µmol/L (< 0.2 mg/dL)
expected values to its own population, and if necessary determine its own reference interval according to good laboratory practice.
For diagnostic purposes, results should always be assessed in conjunction with the patient's medical history, clinical examinations and other
findings.

from these values.
Linearity
The test is linear within a concentration range of 0 – 171 µmol/L (0 – 10 mg/dL).
Precision
Mean µmol/L SD CV% SD CV%
10 0.18 1.78 0.33 3.24
17 0.26 1.51 0.50 2.92
116 1.34 1.16 2.55 2.20
Sensitivity
The lowest detectable level on an AU600 analyser was calculated as 0.24 µmol/L.
The lowest detectable level represents the lowest measurable level of direct bilirubin that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this Direct Bilirubin (OSR6111) assay on the AU640 against another commercially available direct
bilirubin assay. Results of linear regression analysis were as follows:
y = 0.789x – 0.7 r = 0.998 n = 99 Sample range = 0.2 – 151.5 µmol/L
®
4

‡ The above parameters must be entered twice using test names DBILC (colour) and DBILB (blank). Set the test as SAMPLE BLANK in the
INTER RELATED TEST menu
* Values set for working in SI units (µmol/L). To work in mg/dL divide by 17.1
§ Set the factor range for the blank reagent at –99999 to 99999
ж Set the test as SAMPLE BLANK in the COMMON TEST PARAMETERS TEST NAME SAMPLE BLANK menu.
N
BIBLIOGRAPHY
1. Thomas L. Bilirubin. In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
2. Balistreri WF, Shaw LM. Liver Function. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:733-737.
3. Hymans van den Bergh AA, Mueller P. Ueber eine direkte and indirekte diazoreaktion auf bilirubin. Biochem Z 1916, 77:90.

2009-08
DIRECT BILIRUBIN, AU400/AU640 Serum/Plasma Application DIRECT BILIRUBIN, AU600 Serum/Plasma Application
System Reagent: OSR6111, OSR6211 System Reagent: OSR6111, OSR6211
Reagent ID: R1 COLOUR (DBILC) 011, R1 BLANK (DBILB) 010 Reagent ID: R1 COLOUR (DBILC) 011, R1 BLANK (DBILB) 010
Test No # Test name DBIL‡ ∇ Sample type Ser ∇ Page ½
Test Name: DBIL‡ ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name DBIL‡ ∇ Sample type Ser ∇ Page 2/2
Test Name: DBIL‡ ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic Value: # # Unit: µmol/L* Decimal places: # Panic value # #
General ISE Test No # Test name DBIL‡ ∇
Test Name: DBIL‡ ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Selection calibrator
Cal. No. OD CONC Factor/OD-L Factor/OD-H Point 1 # ∇ † 580*§ 960*§
Point 1: # † 580*§ 960*§ Point 2 ∇
Point 3:
Point 4 ∇
Point 4:
Point 5 ∇
Point 5:
Point 6 ∇
Point 6:
Point 7 ∇
Point 7:
1-point cal. Point
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibrator: # ∇ MB type factor
‡ The above parameters must be entered twice using test names DBILC (Colour Rg) and DBILB (Blank Rg). Set ‡ The above parameters must be entered twice using test names DBILC (Colour Rg) and DBILB (Blank Rg). Set
the test as SAMPLE BLANK in the INTER RELATED TEST menu. the test as SAMPLE BLANK in the INTER RELATED TEST menu
* Values set for working in SI units (μmol/L). To work in mg/dL divide by 17.1 * Values set for working in SI units (μmol/L). To work in mg/dL divide by 17.1
§ Set the factor range for the blank reagent at –99999 to 99999 § Set the factor range for the blank reagent at –99999 to 99999
2009-08
DIRECT BILIRUBIN, AU2700/AU5400 Serum/Plasma Application
System Reagent: OSR6111, OSR6211
Reagent ID: R1 COLOUR (DBILC) 011, R1 BLANK (DBILB) 010
Test Name: DBIL‡ ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 2.5 μL Dilution 0 μL Pre-Dilution Rate: 1

Reagent OD limit:
No Lag Time: ∇ On-board stability period: 21

Test Name: DBIL‡ ∇ < > Type: Serum ∇

ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
Panic Value: # # Unit: µmol/L* Decimal places: #
General ISE
Test Name: DBIL‡ ∇ < > Type Serum ∇

Point 1: # † 580*§ 960*§
Point 2:
Point 3:
Point 4:
Point 5:
Point 6:
Point 7:
‡ The above parameters must be entered twice using test names DBILC (Colour Rg) and DBILB (Blank Rg). Set
the test as SAMPLE BLANK in the INTER RELATED TEST menu.
# User defined
* Values set for working in SI units (μmol/L). To work in mg/dL divide by 17.1
2009-08
DIRECT BILIRUBIN, AU680/AU480 Serum/Plasma Application DIRECT BILIRUBIN, AU680/AU480 Serum/Plasma Application
System Reagent: OSR6111, OSR6211 Reagent ID: R1 COLOR (DBILC) 011 System Reagent: OSR6111, OSR6211 Reagent ID: R1 BLANK (DBILB) 010
Test Name: DBILCж ∇ < > Type: Serum ∇ Operation Yes ∇ Test Name: DBILBж ∇ < > Type: Serum ∇ Operation Yes ∇
Sample Volume 2.5 μL Dilution 0 μL OD Limit Sample Volume 2.5 μL Dilution 0 μL OD Limit
Pre-Dilution Rate 1 ∇ Min.OD Max.OD Pre-Dilution Rate 1 ∇ Min.OD Max.OD
Rgt. Volume R1(R1-1) 25 μL Dilution 100 μL Reagent OD Limit Rgt. Volume R1(R1-1) 25 μL Dilution 100 μL Reagent OD Limit
First Low -0.1 High 0.1 First Low -0.1 High 0.1
Last Low -0.1 High 0.1 Last Low -0.1 High 0.1
R2(R2-1) 0 μL Dilution 0 μL R2(R2-1) 0 μL Dilution 0 μL
Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0 Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Method END ∇ Method END ∇
Reaction Slope + ∇ Onboard Stability Period 21 Day 0 Hour Reaction Slope + ∇ Onboard Stability Period 21 Day 0 Hour
Linearity Limit % Icterus +++++ ∇ Linearity Limit % Icterus +++++ ∇
Lag Time Check ∇ Hemolysis + ∇ Lag Time Check ∇ Hemolysis + ∇

Test Name: DBILCж ∇ < > Type Serum ∇ ο Use Serum Cal. Test Name: DBILBж ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # ∇ Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # ∇
<Calibrator Parameters> Factor Range <Calibrator Parameters> Factor Range
Point 1: # ∇ † 580* 960* Point 1: # ∇
Point 4: ∇ ο Reagent Blank Point 4: ∇ ο Reagent Blank
Point 5: ∇ ο Calibration Point 5: ∇ ο Calibration
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration <Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration
Point-1 ∇ Reagent Blank 21 Day 0 Hour Point-1 ∇ Reagent Blank 21 Day 0 Hour
Point-2 ∇ Calibration 21 Day 0 Hour Point-2 ∇ Calibration 21 Day 0 Hour
MB Type Factor: 1-Point Calibration Point ∇ ο with Conc-0 MB Type Factor: 1-Point Calibration Point ∇ ο with Conc-0
# User defined ж Set the test as SAMPLE BLANK in the COMMON TEST PARAMETERS TEST NAME SAMPLE BLANK menu.
† System Calibrator Cat. No.: 66300 # User defined
* Values set for working in SI units (μmol/L). To work in mg/dL divide by 17.1 † System Calibrator Cat. No.: 66300
ф AU680 * Values set for working in SI units (μmol/L). To work in mg/dL divide by 17.1
ф AU680
2009-08
DIRECT BILIRUBIN, AU400/AU640 Paediatric Application DIRECT BILIRUBIN, AU600 Paediatric Application
Reagent ID: R1 COLOUR (DBILC) 011, R1 BLANK (DBILB) 010 Reagent ID: R1 COLOUR (DBILC) 011, R1 BLANK (DBILB) 010
Test No # Test name DBILP‡ ∇ Sample type Ser ∇ Page 1/2
Test Name: DBILP‡ ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 3.5 μL Dilution 10 μL Pre-Dilution Rate: 1 Reagent 1 vol 35 Dil. vol 130 μL L H
General LIH ISE Range Test No # Test name DBILP‡ ∇ Sample type Ser ∇ Page 2/2
Level L Level H
Test Name: DBILP‡ ∇ < > Type: Serum ∇
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic value # #
General ISE Test No # Test name DBILP‡ ∇
Test Name: DBILP‡ ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Cal. No. OD CONC Factor/OD-L Factor/OD-H Point 1 # ∇ † §530* §804*
Point 1: # † §530* §804* Point 2 ∇
Point 3:
Point 4 ∇
Point 4:
Point 5 ∇
Point 5:
Point 6 ∇
Point 6:
Point 7 ∇
Point 7:
1-point cal. point
1-Point Cal. Point: ο with CONC-0 Slope Check: None Advanced Calibration: # ∇ MB type factor
‡ The above parameters must be entered twice using test names DBILC (Colour Rg) and DBILB (Blank Rg). Set ‡ The above parameters must be entered twice using test names DBILC (Colour Rg) and DBILB (Blank Rg). Set
2009-08
DIRECT BILIRUBIN, AU2700/AU5400 Paediatric Application
Reagent ID: R1 COLOUR (DBILC) 011, R1 BLANK (DBILB) 010
Test Name: DBILP‡ ∇ < > Type: Serum ∇ Operation: Yes ∇

Reagent OD limit:

Test Name: DBILP‡ ∇ < > Type: Serum ∇

ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
General ISE
Test Name: DBILP‡ ∇ < > Type Serum ∇

Point 1: # † §530* §804*
Point 2:
Point 3:
Point 4:
Point 5:
Point 6:
Point 7:
‡ The above parameters must be entered twice using test names DBILC (Colour Rg) and DBILB (Blank Rg). Set
# User defined
* Values set for working in SI units (μmol/L). To work in mg/dL divide by 17.1
2009-08
DIRECT BILIRUBIN, AU680/AU480 Paediatric Application DIRECT BILIRUBIN, AU680/AU480 Paediatric Application
System Reagent: OSR6111, OSR6211 Reagent ID: R1 COLOR (DBILC) 011 System Reagent: OSR6111, OSR6211 Reagent ID: R1 BLANK (DBILB) 010
Test Name: DBILPCж ∇ < > Type: Serum ∇ Operation Yes ∇ Test Name: DBILPBж ∇ < > Type: Serum ∇ Operation Yes ∇
Sample Volume 2.7 μL Dilution 10 μL OD Limit Sample Volume 2.7 μL Dilution 10 μL OD Limit

Test Name: DBILPCж ∇ < > Type Serum ∇ ο Use Serum Cal. Test Name: DBILPBж ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # ∇ † 530* 804* Point 1: # ∇
ж Set the test as SAMPLE BLANK in the COMMON TEST PARAMETERS TEST NAME SAMPLE BLANK menu. ж Set the test as SAMPLE BLANK in the COMMON TEST PARAMETERS TEST NAME SAMPLE BLANK menu.
# User defined # User defined
ф AU680 ф AU680
2009-08
TOTAL BILIRUBIN
OSR6112 4 x 15 mL R1 TBILC
4 x 15 mL R1 TBILB
OSR6212 4 x 40 mL R1 TBILC
4 x 40 mL R1 TBILB
Intended Use
Photometric colour test for the quantitative determination of total bilirubin in human serum and plasma on Beckman Coulter analysers. For
Summary1,2
80 – 85% of bilirubin produced daily originates from haemoglobin released by the breakdown of senescent erythrocytes, the remaining
15 – 20% results from the breakdown of haem-containing proteins such as myoglobin, cytochromes, catalases and from bone marrow as a
result of ineffective erythropoiesis. A number of diseases affect one or more of the steps involved in the production, uptake, storage, metabolism
and excretion of bilirubin. Depending on the disorder unconjugated or conjugated bilirubin or both are major contributors to the resulting
hyperbilirubinemia. Hyperbilirubinemia can be classified as follows:
Prehepatic Jaundice: Diseases of prehepatic origin with predominantly unconjugated hyperbilirubinemia include corpuscular haemolytic
anemias e.g. thalassemia and sickle cell anemia; extracorpuscular haemolytic anemia e.g. blood transfusion reaction due to ABO and Rh
incompatibility; neonatal jaundice and haemolytic disease of the newborn.
Hepatic Jaundice: Diseases of hepatic origin with predominantly conjugated hyperbilirubinemia include acute and chronic viral hepatitis, liver
cirrhosis and hepatocellular carcinoma.
Post hepatic Jaundice: Diseases of post-hepatic origin with predominantly conjugated hyperbilirubinemia include extrahepatic cholestasis and
liver transplant rejection.
Chronic congenital hyperbilirubinemias include the unconjugated hyperbilirubinemias Crigler-Najjar syndrome and Gilbert's syndrome as well as
the conjugated hyperbilirubinemias Dubin-Johnson syndrome and Rotor syndrome. The differentiation between chronic congenital
hyperbilirubinemias and acquired types of bilirubinemia is accomplished via the measurement of bilirubin fractions and the detection of normal
liver enzyme activities.
Test Principle3
A stabilised diazonium salt, 3,5-dichlorophenyldiazonium tetrafluoroborate (DPD), reacts with conjugated bilirubin directly and with unconjugated
bilirubin in the presence of an accelerator to form azobilirubin. The absorbance at 540 nm is proportional to the total bilirubin concentration.
A separate sample blank is performed to reduce endogenous serum interference.
Reaction Principle
Caffeine
Bilirubin + DPD Azobilirubin
Surfactant

Caffeine 2.1 mmol/L
3,5-dichlorophenyldiazonium tetrafluoroborate 0.31 mmol/L
Surfactant
Preservative

Reagent Preparation

are stable for 90 days. Protect TBILC from light.
Specimen
Serum and EDTA or heparinised plasma. Protect samples from light.
4
Stable in serum and plasma for 7 days when stored at 2…8°C and 1 day when stored at 15…25°C.
Test Procedure
Calibration
The calibrator total bilirubin value is traceable to the National Institute of Standards and Technology (NIST) Standard Reference Material (SRM)
916a.
Recalibrate the assay when the following occur:

2009-08
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the total bilirubin concentration of each sample.
Serum (Adults) 5 – 21 µmol/L (0.3 – 1.2 mg/dL)
Serum (Children)
0 − 1 day 24 – 149 µmol/L (1.4 – 8.7 mg/dL)
1 − 2 days 58 – 197 µmol/L (3.4 – 11.5 mg/dL)
3 − 5 days 26 – 205 µmol/L (1.5 – 12.0 mg/dL)
expected values to its own population, and if necessary determine its own reference interval according to good laboratory practice.
For diagnostic purposes, results should always be assessed in conjunction with the patient's medical history, clinical examinations and other
findings.

from these values.
Linearity
The test is linear within a concentration range of 0 – 513 µmol/L (0 – 30 mg/dL).
Precision
7.58 0.10 1.35 0.14 1.79
17.76 0.17 0.94 0.37 2.06
222.75 1.12 0.50 3.15 1.41
Sensitivity
The lowest detectable level on an AU600 analyser was estimated at 0.39 µmol/L.
The lowest detectable level represents the lowest measurable level of total bilirubin that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this Total Bilirubin OSR6112 assay on the AU600 against another commercially available total
bilirubin assay. Results of linear regression analysis were as follows:
y = 0.942x + 0.392 r = 0.998 n = 111 Sample range = 0.86 – 447.34 µmol/L
®
6

‡ The above parameters must be entered twice using test names TBILC (colour) and TBILB (blank). Set the test as SAMPLE BLANK in the
INTER RELATED TEST menu
* Values set for working in SI units (µmol/L ). To work in mg/dL divide by 17.1
BIBLIOGRAPHY
1. Thomas L. Bilirubin. In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
2 Balistreri WF, Shaw LM. Liver Function. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:733-737.
3. Tolman KG, Rej R. Liver Function. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia: WB Saunders Company,
1999;1136-1137.
4. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of anticoagulants in diagnostic laboratory investigations and stability of blood,
plasma and serum samples. WHO/DIL/LAB/99.1 Rev.2: 24pp.

2009-08
TOTAL BILIRUBIN, AU400/AU640 Serum/Plasma Application TOTAL BILIRUBIN, AU600 Serum/Plasma Application
Reagent ID: RI COLOUR (TBILC) 012, R1 BLANK (TBILB) 025 Reagent ID: RI COLOUR (TBILC) 012, R1 BLANK (TBILB) 025
Test No # Test name TBIL‡ ∇ Sample type Ser ∇ Page ½
Test Name: TBIL‡ ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name TBIL‡ ∇ Sample type Ser ∇ Page 2/2
Test Name: TBIL‡ ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name TBIL‡ ∇
Test Name: TBIL‡ ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Cal. No. OD CONC Factor/OD-L Factor/OD-H Point 1 # ∇ † §428* §752*
Point 1: # † §428* §752* Point 2 ∇
Point 3:
Point 4 ∇
Point 4:
Point 5 ∇
Point 5:
Point 6 ∇
Point 6:
Point 7 ∇
Point 7:
1-point cal. Point
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibrator: # ∇ MB type factor
‡ The above parameters must be entered twice using test names TBILC (Colour) and TBILB (Blank). Set ‡ The above parameters must be entered twice using test names TBILC (Colour) and TBILB (Blank). Set
* Values set for working in SI units (µmol/L). To work in mg/dL divide by 17.1 * Values set for working in SI units (µmol/L). To work in mg/dL divide by 17.1
2009-08
TOTAL BILIRUBIN, AU2700/AU5400 Serum/Plasma Application
Reagent ID: RI COLOUR (TBILC) 012, R1 BLANK (TBILB) 025
Test Name: TBIL‡ ∇ < > Type: Serum ∇ Operation: Yes ∇

Reagent OD limit:

Test Name: TBIL‡ ∇ < > Type: Serum ∇

ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
General ISE
Test Name: TBIL‡ ∇ < > Type Serum ∇

Point 1: # † §428* §752*
Point 2:
Point 3:
Point 4:
Point 5:
Point 6:
Point 7:
‡ The above parameters must be entered twice using test names TBILC (Colour) and TBILB (Blank). Set
# User defined
2009-08
TOTAL BILIRUBIN, AU680/AU480 Serum/Plasma Application TOTAL BILIRUBIN, AU680/AU480 Serum/Plasma Application
System Reagent: OSR6112, OSR6212 Reagent ID: R1 COLOUR (TBILC) 012 System Reagent: OSR6112, OSR6212 Reagent ID: R1 BLANK (TBILB) 025
Test Name: TBILCж ∇ < > Type: Serum ∇ Operation Yes ∇ Test Name: TBILBж ∇ < > Type: Serum ∇ Operation Yes ∇
Sample Volume 5 μL Dilution 0 μL OD Limit Sample Volume 5 μL Dilution 0 μL OD Limit


Test Name: TBILCж ∇ < > Type Serum ∇ ο Use Serum Cal. Test Name: TBILBж ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # ∇ † 428* 752* Point 1: # ∇
# User defined ж Set the test as SAMPLE BLANK in the COMMON TEST PARAMETERS TEST NAME SAMPLE BLANK menu.
† System Calibrator Cat. No.: 66300 # User defined
* Values set for working in SI units (µmol/L). To work in mg/dL divide by 17.1 † System Calibrator Cat. No.: 66300
ф AU680 * Values set for working in SI units (µmol/L). To work in mg/dL divide by 17.1
ф AU680
2009-08
TOTAL BILIRUBIN, AU400/AU640 Paediatric Application TOTAL BILIRUBIN, AU600 Paediatric Application
Reagent ID: RI COLOUR (TBILC) 012, R1 BLANK (TBILB) 025 Reagent ID: RI COLOUR (TBILC) 012, R1 BLANK (TBILB) 025
Test No # Test name TBILP‡ ∇ Sample type Ser ∇ Page 1/2
Test Name: TBILP‡ ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name TBILP‡ ∇ Sample type Ser ∇ Page 2/2
Test Name: TBILP‡ ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Calibration Specific Calibration specific
General ISE
Test No # Test name TBILP‡ ∇
Test Name: TBILP‡ ∇ < > Type Serum ∇
Cal type 8 AB ∇ Count #
Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Formula 1 Y=AX+B ∇ Process Conc ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H Cal No OD Conc Factor/OD-L Factor/OD-H
Point 1: # † §1120* §1680* Point 1 # ∇ † §1120* §1680*
MB Type Factor: Calibration Stability Period: 999 MB type factor
Calibrator stability period 999
‡ The above parameters must be entered twice using test names TBILC (Colour) and TBILB (Blank). Set Select the function using the Function key or the Mouse
the test as SAMPLE BLANK in the INTER RELATED TEST menu. ‡ The above parameters must be entered twice using test names TBILC (Colour) and TBILB (Blank). Set
# User defined the test as SAMPLE BLANK in the INTER RELATED TEST menu
† System Calibrator Cat. No.: 66300 # User defined ¤ Analyser default value
* Values set for working in SI units (µmol/L). To work in mg/dL divide by 17.1 † System Calibrator Cat. No.: 66300
2009-08
TOTAL BILIRUBIN, AU2700/AU5400 Paediatric Application
Reagent ID: RI COLOUR (TBILC) 012, R1 BLANK (TBILB) 025
Test Name: TBILP‡ ∇ < > Type: Serum ∇ Operation: Yes ∇

Reagent OD limit:

Test Name: TBILP‡ ∇ < > Type: Serum ∇

ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
General ISE
Test Name: TBILP‡ ∇ < > Type Serum ∇

Point 1: # † §1120* §1680*
Point 2:
Point 3:
Point 4:
Point 5:
Point 6:
Point 7:
‡ The above parameters must be entered twice using test names TBILC (Colour) and TBILB (Blank). Set
# User defined
2009-08
TOTAL BILIRUBIN, AU680/AU480 Paediatric Application TOTAL BILIRUBIN, AU680/AU480 Paediatric Application
System Reagent: OSR6112, OSR6212 Reagent ID: R1 COLOUR (TBILC) 012 System Reagent: OSR6112, OSR6212 Reagent ID: R1 BLANK (TBILB) 025
Test Name: TBILPCж ∇ < > Type: Serum ∇ Operation Yes ∇ Test Name: TBILPBж ∇ < > Type: Serum ∇ Operation Yes ∇
Sample Volume 5 μL Dilution 10 μL OD Limit Sample Volume 5 μL Dilution 10 μL OD Limit


Test Name: TBILPCж ∇ < > Type Serum ∇ ο Use Serum Cal. Test Name: TBILPBж ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # ∇ † 1120* 1680* Point 1: # ∇
Point 6: ∇ Point 6:
∇
ж Set the test as SAMPLE BLANK in the COMMON TEST PARAMETERS TEST NAME SAMPLE BLANK menu. ж Set the test as SAMPLE BLANK in the COMMON TEST PARAMETERS TEST NAME SAMPLE BLANK menu.
* Values set for working in SI units (µmol/L). To work in mg/dL divide by 17.1 * Values set for working in SI units (µmol/L). To work in mg/dL divide by 17.1
ф AU680 ф AU680
2009-08
CALCIUM oCPC
OSR6113 4 x 27 mL R1
4 x 27 mL R2
OSR6213 4 x 50 mL R1
4 x 50 mL R2
Intended Use
Photometric colour test for the quantitative determination of total calcium in human serum, plasma and urine on Beckman Coulter analysers. For
Summary1
Measurement of calcium is used in the diagnosis and treatment of parathyroid disease, a variety of bone diseases, chronic renal disease,
urolithiasis and tetany (intermittent muscular contractions or spasms).
Total serum calcium is composed of three fractions: free or ionised calcium, 50%; protein bound calcium most of which is bound to albumin with
only a small portion bound to globulins, 45%; and complex-bound calcium, mainly to phosphate, citrate, and bicarbonate, 5%. The ionised
calcium is physiologically most significant, but has proven difficult to assay directly. It may be estimated from total calcium given knowledge of
the protein content and pH of the blood, which strongly affect the level of ionised calcium.
Calcium ions are important in the transmission of nerve impulses, as a cofactor in several enzyme reactions, in the maintenance of normal
muscle contractility, and in the process of coagulation. A significant reduction in calcium ion concentration results in muscle tetany. A higher
than normal concentration of calcium ions produces lowered neuromuscular excitability and muscle weakness along with other more complex
symptoms.
Test Principle2,3
Calcium ions react with o-Cresolphthalein-complexone in an alkaline medium to form a purple coloured complex. In this method the absorbance
of the Ca-oCPC complex is measured bichromatically at 570/660 nm. The resulting increase in absorbance of the reaction mixture is directly
proportional to the calcium concentration in the sample.
Reaction Principle
8-Hydroxyquinoline
2+ 2+
Ca + oCPC Ca-oCPC Complex (purple)
pH 10.6

Ethanolamine (pH 10.6) 0.375 mol/L
8-Hydroxyquinoline 7.16 mmol/L
o-Cresolphthalein complexone 82.0 µmol/L
R1 reagent: Irritant, contains 2-aminoethanol. R36/37/38. Irritating to eyes, respiratory system and skin.
Safety Phrases:
S26 – S36/37/39, S45, S60. In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable
protective clothing, gloves and eye/face protection. In case of accident or if you feel unwell, seek medical advice immediately (show the label
where possible). This material and its container must be disposed of as hazardous waste.
Reagent Preparation
Specimen
Serum or heparinised plasma should be promptly separated to avoid the uptake of calcium by erythrocytes. Do not use the following
4
anticoagulants in collecting blood for use in this test: EDTA, Sodium Citrate, Sodium Fluoride or Oxalate.
5
Stable in serum when stored at 2...8°C for up to 1 week.
6
Urine : Acidified with 6M HCl. Collect timed 24 hour specimen using standard laboratory procedures.
Store at 2...8°C.
Test Procedure
Calibration
Use System Calibrator Cat. No. 66300 for serum/plasma application and Urine Calibrator Cat. No. ODC0025 for urine application.
The calcium values of both calibrators are traceable to the National Institute of Standards and Technology (NIST) Standard Reference Material
(SRM) 909b Level 1.
Recalibrate the assay every day or, when the following occur:
Change in reagent bottle or significant shift in control values;
Absorption of atmospheric CO2 by the reagent on board the analyser can impair calibration stability. This effect will vary depending upon the
rate of use. Consequently each laboratory should set a calibration frequency in the instrument parameters appropriate to their usage pattern.

2009-08
Quality Control
the serum/plasma application.
Calculation
The Beckman Coulter analysers automatically compute the calcium concentration of each sample.
Reference Intervals
1
Serum, plasma – Adults 2.20 – 2.65 mmol/L or 8.8 – 10.6 mg/dL
7
Serum, Children 0 – 10 day 1.90 – 2.60 mmol/L or 7.6 – 10.4 mg/dL
Serum, Children 2 – 12 year 2.20 – 2.70 mmol/L or 8.8 – 10.8 mg/dL
1
Urine :
24h urine 2h urine
Female < 6.2 mmol (250 mg) Male and Female ≤ 0.57 mmol/mmol (0.2g/g) of creatinine
Male < 7.5 mmol (300mg)
Male and Female ≤ 0.1 mmol (4 mg)/kg of body weight
Small Children: ≤ 0.8 g/g creatinine
findings.
from these values.
Linearity
The test is linear within a concentration range of 0 – 4.5 mmol/L (0 – 18 mg/dL) for serum and plasma. The test is linear within a concentration
range of 0 – 10 mmol/L (0 – 40 mg/dL) for urine.
Precision
The following data was obtained on an AU640 using 3 serum control materials analysed over 20 days.
Mean mmol/L SD CV% SD CV%
2.27 0.01 0.60 0.04 1.72
2.62 0.02 0.63 0.03 1.14
3.49 0.02 0.51 0.05 1.40
1.13 0.02 2.02 0.04 3.93
4.42 0.03 0.75 0.06 1.33
9.65 0.05 0.53 0.11 1.17
Sensitivity
The lowest detectable level in serum on an AU640 analyser was estimated at 0.03 mmol/L.
The lowest detectable level in urine on the AU2700 was estimated at 0.10 mmol/L.
The lowest detectable level represents the lowest measurable level of calcium that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this Calcium oCPC OSR6113 assay on the AU640 against a flame photometry method. Results
y = 0.986x + .002 r = 0.985 n = 106 Sample range = 1.37 – 3.57mmol/L
Patient urine samples were used to compare this Calcium oCPC OSR6113 assay on the AU2700 against another commercially available
calcium assay. Results of linear regression analysis were as follows:
y = 0.971x + .004 r = 0.997 n = 125 Sample range = 0.07 – 7.73mmol/L
®
Magnesium: Interference less than 3% up to 40mg/dL magnesium
8
Limitations
Care should be taken when interpreting calcium results from patients who have received gadolinium containing contrast medium within the
9, 10, 11
previous 24 hours, especially if the patient has impaired renal function. Such samples should be assayed using non-colorimetric
techniques e.g. ion selective electrodes or emission spectroscopy. If non-colorimetric assays are unavailable, samples should be drawn prior to
administration of such contrast media.

2009-08
† System Calibrator Cat. No.: 66300/ Urine Calibrator Cat. No.:ODC0025
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 4.
BIBLIOGRAPHY
1. Thomas L, ed. Clinical Laboratory Diagnostics Use and Assessment of Clinical Laboratory Results, 1st ed. Frankfurt/Main: TH-Books
2. Gitelman HJ. An improved automated procedure for the determination of calcium in biochemical specimen. Anal. Biochem 1967;18: 521-531.
3. Pollard FH, Martin JV. The Spectrophotometric Determination of the Alkaline-earth Metals with Murexide, Eriochrome Black T and with o-
Cresolphthalein-Complexone. Analyst 1956; 81:348-353.
4. Kaplan LA, Pesce AJ, eds. Clinical Chemistry Theory, analysis, and correlation, 3rd ed. St Louis: Mosby, 1996:550pp.
5. Heins M, Heil W, Withold W. Storage of serum or whole blood samples. Effects of time and temperature on 22 serum analytes. Eur J Clin Chem Clin
Biochem 1995;33:231-238.
6. NCCLS. Urinalysis and Collection, Transportation, and Preservation of Urine Specimens; Approved Guideline. NCCLS Document GP16-A2, 2nd ed.
Pennsylvania: NCCLS, 2001.
7. Painter PC, Cope JY, Smith JL. Reference information for the clinical laboratory. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical
chemistry. Philadelphia:WB Saunders Company, 1999;1804pp.
8. Young DS. Effects of Drugs on Clinical Laboratory Tests, AACC, 5th ed. AACC Press, 2000.
9. Lin J, Idee JM, Port M, Diai A, Berthommier C, Robert M, et al. Interference of magnetic resonance imaging contrast agents with the serum calcium
measurement technique using colorimetric reagents. J Pharm Biomed Analysis 1999;21:931-943.
10. Normann PT, Frøysa A, Svaland M. Interference of gadodiamide injection (OMNISCAN®) on the colorimetric determination of serum calcium. Scand
J Clin Lab Invest 1995;55:421-426.
11. As communicated to Beckman Coulter Biomedical Ltd by Amersham Health, Summary of Product Characteristics OMNISCAN, June 1999.

2009-08
CALCIUM oCPC, AU400/AU640 Serum/Plasma Application CALCIUM oCPC, AU600 Serum/Plasma Application

Test No # Test name CA ∇ Sample type Ser ∇ Page ½
Test Name: CA ∇ < > Type: Serum ∇ Operation: Yes ∇
Method: END ∇ Last L -0.1 Last H 1.0 Reaction + ∇ L 0* H 4.5*
Measuring Point 1: First 0 Last 27 L 0* H 4.5* Point 2 Fst 0 Lst 10 B 0
Measuring Point 2: First 0 Last 10 Correlation Factor: Sample Pre-dil. Rate ¤
General LIH ISE Range Test No # Test name CA ∇ Sample type Ser ∇ Page 2/2
Test Name: CA ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name CA ∇
Test Name: CA ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 2* 8*
Point 1: # † 2* 8*
Point 2 ∇
Point 2:
1-Point Cal. Point: Slope Check: None # 1-point cal. Point
MB type factor
# User defined
† System Calibrator Cat. No.: 66300 # User defined ¤ Analyser default value
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 4 † System Calibrator Cat. No.: 66300
‡ Depends on usage pattern in the Laboratory * Values set for working in SI units (mmol/L). To work in mg/dL multiply by 4
2009-08
CALCIUM oCPC, AU2700/AU5400 Serum/Plasma Application CALCIUM oCPC, AU680/AU480 Serum/Plasma Application
Specific Test Parameters Parameters Specific Test Parameters
General LIH ISE Range General LIH ISE HbA1c Calculated Test Range
Test Name: CA ∇ < > Type: Serum ∇ Operation: Yes ∇ Test Name: CA ∇ < > Type: Serum ∇ Operation Yes ∇

Reagent OD limit:
Method: END ∇ Last L -0.1 Last H 1.0 R2(R2-1) 45 Dilution 10
μL μL
Reaction slope: + ∇ Dynamic Range: Dynamic Range Low 0* High 4.5*
Measuring Point 1: First 0 Last 27 L 0* H 4.5* Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Measuring Point 2: First 0 Last 10 Correlation Factor: Wavelength Pri 570 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0
Linearity : % A 1 B 0 Method END ∇
No Lag Time: ∇ On-board stability period: 30 Reaction Slope + ∇ Onboard Stability Period 30 Day 0 Hour
Measuring Point2 First 0 Last 10 Lipemia +++++ ∇
Specific Test Parameters Linearity Limit % Icterus +++++ ∇
General LIH ISE Range Lag Time Check ∇ Hemolysis +++++ ∇
Test Name: CA ∇ < > Type: Serum ∇ Parameters Specific Test Parameters
Normal Ranges: Age L Age H Test Name: CA ∇ < > Type: Serum ∇
ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # Sex Year Month Year Month Low High # #
ο 4. # ∇ # # # # # # ο 1. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 3. # ∇ # # # # # #
7. None Selected # # ο 4. # ∇ # # # # # #
8. Out of Range L H # # ο 5. # ∇ # # # # # #
Panic Value: # # Unit: mmol/L* Decimal places: # ο 6. # ∇ # # # # # #
General ISE
Test Name: CA ∇ < > Type Serum ∇ General ISE
Test Name: CA ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H <Calibrator Parameters> Factor Range
Point 1: # † 2* 8* Calibrator OD Conc Low High Slope Check None ∇
Point 2: Point 1: # ∇ † 2* 8*
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 7 ∇ Advanced Calibration
Point 9 ∇
# User defined. <Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration
† System Calibrator Cat. No.: 66300 Master Curve> OD Range
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 4
‡ Depends on usage pattern in the Laboratory Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
ф AU680 Point-2 ∇ Calibration ‡ Day ‡ Hour
2009-08
CALCIUM oCPC, AU400/AU640 Urine Application CALCIUM oCPC, AU600 Urine Application

Test No # Test name CA ∇ Sample type Uri ∇ Page 1/2
Test Name: CA ∇ < > Type: Urine ∇ Operation: Yes ∇
Measuring Point 1: First 0 Last 27 L 0* H 10* Point 2 Fst 0 Lst 10 B 0
General LIH ISE Range Test No # Test name CA ∇ Sample type Uri ∇ Page 2/2
Test Name: CA ∇ < > Type: Urine ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
URINE APPLICATION
General ISE Test No # Test name CA ∇
Test Name: CA ∇ < > Type Urine ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 5* 20*
Point 1: # † 5* 20*
Point 2 ∇
Point 2:
1-Point Cal. Point: Slope Check: None # 1-point cal. point
MB type factor
† Urine Calibrator Cat. No.: ODC0025 † Urine Calibrator Cat. No.: ODC0025
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 4 * Values set for working in SI units (mmol/L). To work in mg/dL multiply by 4
‡ Depends on the usage pattern in the laboratory ‡ Depends on the usage pattern in the laboratory
2009-08
CALCIUM oCPC, AU2700/AU5400 Urine Application CALCIUM oCPC, AU680/AU480 Urine Application
Test Name: CA ∇ < > Type: Urine ∇ Operation: Yes ∇ Test Name: CA ∇ < > Type: Urine ∇ Operation Yes ∇

Reagent OD limit:
μL μL
Reaction slope: + ∇ Dynamic Range: Dynamic Range Low 0* High 10*
Measuring Point1 First 0 Last 27
Specific Test Parameters Linearity Limit %
General LIH ISE Range Lag Time Check ∇
Test Name: CA ∇ < > Type: Urine ∇ Parameters Specific Test Parameters
Normal Ranges: Age L Age H Test Name: CA ∇ < > Type: Urine ∇
ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 4. # ∇ # # # # # # ο 1. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 3. # ∇ # # # # # #
7. None Selected # # ο 4. # ∇ # # # # # #
8. Out of Range L H # # ο 5. # ∇ # # # # # #
URINE APPLICATION
General ISE
Test Name: CA ∇ < > Type Urine ∇ General ISE
Test Name: CA ∇ < > Type Urine ∇ ο Use Serum Cal.

Point 2: Point 1: # ∇ † 5* 20*
Point 9 ∇
† Urine Calibrator Cat. No.: ODC0025 Master Curve> OD Range
‡ Depends on the usage pattern in the laboratory Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
2009-08
CALCIUM oCPC, AU400/AU640 Paediatric Application CALCIUM oCPC, AU600 Paediatric Application

Test No # Test name CAP ∇ Sample type Ser ∇ Page 1/2
Test Name: CAP ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name CAP ∇ Sample type Ser ∇ Page 2/2
Test Name: CAP ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE
Test No # Test name CAP ∇
Test Name: CAP ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 2* 8*
Point 1: # † 2* 8*
Point 2 ∇
Point 2:
1-Point Cal. Point: Slope Check: None # 1-point cal. point
MB type factor
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 4 * Values set for working in SI units (mmol/L). To work in mg/dL multiply by 4
2009-08
CALCIUM oCPC, AU2700/AU5400 Paediatric Application CALCIUM oCPC, AU680/AU480 Paediatric Application
Test Name: CAP ∇ < > Type: Serum ∇ Operation: Yes ∇ Test Name: CAP ∇ < > Type: Serum ∇ Operation Yes ∇

Reagent OD limit:
μL μL
Reaction slope: + ∇ Dynamic Range: Dynamic Range Low 0* High 4.5*
Measuring Point 1: First 0 Last 27 L 0* H 4.5* Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Test Name: CAP ∇ < > Type: Serum ∇ Parameters Specific Test Parameters
Normal Ranges: Age L Age H Test Name: CAP ∇ < > Type: Serum ∇
ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 4. # ∇ # # # # # # ο 1. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 3. # ∇ # # # # # #
7. None Selected # # ο 4. # ∇ # # # # # #
8. Out of Range L H # # ο 5. # ∇ # # # # # #
General ISE
Test Name: CAP ∇ < > Type Serum ∇ General ISE
Test Name: CAP ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 2: Point 1: # ∇ † 2* 8*
Point 9 ∇
† System Calibrator Cat. No.: 66300 Master Curve> OD Range
‡ Depends on usage pattern in the Laboratory Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
2009-08
CALCIUM ARSENAZO III
OSR60117 4 x 15 mL R1
OSR61117 4 x 29 mL R1
Intended Use
Photometric colour test for the quantitative determination of total calcium in human serum, plasma and urine on AU Beckman Coulter analysers. For in vitro
Summary1
Measurement of calcium is used in the diagnosis and treatment of parathyroid disease, a variety of bone diseases, chronic renal disease, urolithiasis and
tetany (intermittent muscular contractions or spasms).
Total serum calcium is composed of three fractions: free or ionised calcium, 50%; protein bound calcium most of which is bound to albumin with only a
small portion bound to globulins, 45%; and complex-bound calcium, mainly to phosphate, citrate, and bicarbonate, 5%. The ionised calcium is
physiologically most significant, but has proven difficult to assay directly. It may be estimated from total calcium given knowledge of the protein content and
pH of the blood, which strongly affect the level of ionised calcium.
Calcium ions are important in the transmission of nerve impulses, as a cofactor in several enzyme reactions, in the maintenance of normal muscle
contractility, and in the process of coagulation. A significant reduction in calcium ion concentration results in muscle tetany. A higher than normal
concentration of calcium ions produces lowered neuromuscular excitability and muscle weakness along with other more complex symptoms.
Test Principle2,3
2+
This Calcium procedure is based on calcium ions (Ca ) reacting with Arsenazo III (2,2’-[1,8-Dihydroxy-3,6-disulphonaphthylene-2,7-bisazo]-
bisbenzenearsonic acid) to form an intense purple coloured complex. In this method the absorbance of the Ca-Arsenazo III complex is measured
bichromatically at 660/700 nm. The resulting increase in absorbance of the reaction mixture is directly proportional to the calcium concentration in the
sample.
Magnesium does not significantly interfere in calcium determination using Arsenazo III.
Reaction Principle
2+
Ca + Arsenazo III Ca-Arsenazo III complex (purple)
pH 6.9

Imidazole (pH 6.9)
Arsenazo III 0.1 – 0.2%
Triton X-100
Preservative
To avoid the possible build-up of azide compounds, flush waste pipes with water after the disposal of undiluted reagent.
Reagent Preparation
The reagent is stable, unopened, up to the stated expiry date when stored at 2…8°C. Once open, reagent stored on board the instrument is stable for
90 days.
Specimen
Serum or heparinised plasma should be promptly separated to avoid the uptake of calcium by erythrocytes. Do not use the following anticoagulants in
4
collecting blood for use in this test: EDTA, Sodium Citrate, Sodium Fluoride or Oxalate.
5
6
Urine: Acidified with 6M HCl. Collect timed 24 hr specimen using standard laboratory procedures. Samples with urine pH below 1.5 may result in a
negative bias.
Store at 2...8°C.
5
Test Procedure
Calibration
The calcium values of both calibrators are traceable to the National Institute of Standards and Technology (NIST) Standard Reference Material (SRM)
909b Level 1. Recalibrate the assay when the following occur:
Change in reagent lot number or significant shift in control values,
Quality Control

2010-04
Calculation
The Beckman Coulter analysers automatically compute the calcium concentration of each sample.
Reference Intervals
1
Serum, plasma – Adults 2.20 – 2.65 mmol/L (8.8 – 10.6 mg/dL)
7
Serum, Children 0 – 10 day 1.90 – 2.60 mmol/L (7.6 – 10.4 mg/dL)
7
Serum, Children 10 day – 24 months 2.25 – 2.75 mmol/L (9.0 – 11.0 mg/dL)
7
Serum, Children 2 – 12 year 2.20 – 2.70 mmol/L (8.8 – 10.8 mg/dL)
1
Urine:
24h urine 2h urine
Female < 6.2 mmol (250 mg) Male and Female ≤ 0.57 mmol/mmol (0.2 g/g) of creatinine
Male < 7.5 mmol (300 mg)
Male and Female ≤ 0.1 mmol (4 mg)/kg of body weight
Small Children: 2.28 mmol/mmol (≤ 0.8 g/g) creatinine
values.
Linearity
The test is linear within a concentration range of 1 – 5 mmol/L (4 – 20 mg/dL) for serum and plasma. The test is linear within a concentration range of
0 – 10 mmol/L (0 – 40 mg/dL) for urine.
Precision
1.61 0.01 0.57 0.02 0.95
2.52 0.02 0.65 0.03 0.96
4.19 0.02 0.48 0.04 0.94
0.11 0.00 2.05 0.00 2.60
5.52 0.06 1.00 0.07 1.21
9.33 0.10 1.09 0.13 1.40
Sensitivity
The lowest detectable level using serum settings on an AU640 analyser was estimated as 0.01 mmol/L.
The lowest detectable level using urine settings on the AU2700 was estimated as 0.03 mmol/L.
The lowest detectable level represents the lowest measurable level of calcium that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient serum samples were used to compare this Calcium Arsenazo III reagent OSR61117 on the AU640 against Calcium Arsenazo III OSR6176.
y = 1.003x + 0.015 r = 0.999 n = 105 Sample range = 1.03 – 3.83 mmol/L
Patient urine samples were used to compare this Calcium Arsenazo III reagent OSR61117 on the AU640 against Calcium Arsenazo III OSR6176. Results
y = 1.000x + -0.038 r = 0.999 n = 94 Sample range = 0.50 – 9.62 mmol/L
®
Magnesium: Interference less than 10% up to 4 mmol/L magnesium
Ascorbate : Interference less than 3% up to 50 mg/dL ascorbate
Magnesium: Interference less than 3% up to 4 mmol/L magnesium
8
Limitations
Care should be taken when interpreting calcium results from patients who have received gadolinium containing contrast medium within the previous
9, 10, 11
24 hours, especially if the patient has impaired renal function. Such samples should be assayed using non-colourimetric techniques e.g. ion selective
electrodes or emission spectroscopy. If non-colourimetric assays are unavailable, samples should be drawn prior to administration of such contrast media.

2010-04
BIBLIOGRAPHY
1. Thomas L. Calcium (Ca). In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-
Books Verlagsgesellschaft, 1998:231-241.
2. Bauer PJ. Affinity and stoichiometry of calcium binding by arsenazo III. Anal Biochem 1981;110:61-72.
3. Michaylova V, Ilkova P. Photometric determination of micro amounts of calcium with arsenazo III. Anal Chim Acta 1971;53:194-198.
4. Kazmierczak SC. Methods of analysis-calcium. In: Kaplan LA, Pesce AJ, eds. Clinical chemistry theory, analysis, and correlation. St Louis: Mosby,
1996:550pp.
Plasma and Serum Samples. WHO/DIL/LAB/99.1 Rev.2:26,46pp.
6. NCCLS. Urinalysis and collection, transportation, and preservation of urine specimens; approved guideline. NCCLS Document GP16-A2, 2nd ed.
Philadelphia:WB Saunders Company,1999;1804pp.
8. Young DS. Effects of drugs on clinical laboratory tests, AACC, 5th ed. AACC Press, 2000.
9. Lin J, Idee JM, Port M, Diai A, Berthommier C, Robert M, et al. Interference of magnetic resonance imaging contrast agents with the serum calcium
measurement technique using colorimetric reagents. J Pharm Biomed Analysis 1999;21:931-943.
10. Normann PT, Frøysa A, Svaland M. Interference of gadodiamide injection (OMNISCAN®) on the colorimetric determination of serum calcium. Scand
J Clin Lab Invest 1995;55:421-426.
11. As communicated to Beckman Coulter Biomedical Ltd by Amersham Health, Summary of product characteristics OMNISCAN, June 1999.

2010-04
CALCIUM ARSENAZO, AU400/AU640 Serum/Plasma Application CALCIUM ARSENAZO, AU600 Serum/Plasma Application

Test No # Test name CALA ∇ Sample type Ser ∇ Page 1/2
Test Name: CALA ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name CALA ∇ Sample type Ser ∇ Page 2/2
Test Name: CALA ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name CALA ∇
Test Name: CALA ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 2.5* 4.6*
Point 1: # † 2.5* 4.6*
Point 2 ∇
Point 2:
MB type factor
# User defined
2009-08
CALCIUM ARSENAZO, AU2700/AU5400 Serum/Plasma Application CALCIUM ARSENAZO, AU680/AU480 Serum/Plasma Application
Test Name: CALA ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CALA ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: CALA ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CALA ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: CALA ∇ < > Type Serum ∇
Test Name: CALA ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † 2.5* 4.6* Point 1: # ∇ † 2.5* 4.6*
Point 9 ∇
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 4 Point-1 ∇ Reagent Blank 999 Day 0 Hour
2009-08
CALCIUM ARSENAZO, AU400/AU640 Urine Application CALCIUM ARSENAZO, AU600 Urine Application
System Reagent: OSR60117, 61117 Reagent ID: 117 System Reagent: OSR60117, 61117 Reagent ID: 117

Test No # Test name CALA ∇ Sample type Urine ∇ Page 1/2
Test Name: CALA ∇ < > Type: Urine ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name CALA ∇ Sample type Uri ∇ Page 2/2
Test Name: CALA ∇ < > Type: Urine ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
URINE APPLICATION
General ISE Test No # Test name CALA ∇
Test Name: CALA ∇ < > Type Urine ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 5.0* 9.3*
Point 1: # † 5.0* 9.3*
Point 2 ∇
Point 2:
1-Point Cal. Point: ο With CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor
# User defined
† Urine Calibrator Cat. No.: ODC0025
2009-08
CALCIUM ARSENAZO, AU2700/AU5400 Urine Application CALCIUM ARSENAZO, AU680/AU480 Urine Application
System Reagent: OSR60117, 61117 Reagent ID: 117 System Reagent: OSR60117, OSR61117 Reagent ID: 117
Test Name: CALA ∇ < > Type: Urine ∇ Operation Yes ∇
Test Name: CALA ∇ < > Type: Urine ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
No Lag Time: ∇ On-board stability period: 90 Measuring Point1 First 0 Last 10
Linearity Limit %
Specific Test Parameters Lag Time Check ∇
Test Name: CALA ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CALA ∇ < > Type: Urine ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
URINE APPLICATION
General ISE
General ISE
Test Name: CALA ∇ < > Type Urine ∇ Test Name: CALA ∇ < > Type Urine ∇ ο Use Serum Cal.
Point 1: # † 5.0* 9.3* Point 1: # ∇ † 5.0* 9.3*
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
† Urine Calibrator Cat. No.: ODC0025 Calibrator OD Conc Low High Stability
2009-08
CALCIUM ARSENAZO, AU400/AU640 Paediatric Application CALCIUM ARSENAZO, AU600 Paediatric Application

Test No # Test name CALAP ∇ Sample type Ser ∇ Page 1/2
Test Name: CALAP ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name CALAP ∇ Sample type Ser ∇ Page 2/2
Test Name: CALAP ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name CALAP
∇
Test Name: CALAP ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 2.5* 4.6*
Point 1: # † 2.5* 4.6*
Point 2 ∇
Point 2:
MB type factor
# User defined
2009-08
CALCIUM ARSENAZO, AU2700/AU5400 Paediatric Application CALCIUM ARSENAZO, AU680/AU480 Paediatric Application
Test Name: CALAP ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CALAP ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: CALAP ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CALAP ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: CALAP ∇ < > Type Serum ∇ Test Name: CALAP ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † 2.5* 4.6* Point 1: # ∇ † 2.5* 4.6*
Point 9 ∇
2009-08
CHOLESTEROL
OSR6116 4 x 22.5 mL R1
OSR6216 4x 45 mL R1
*OSR6516 4 x 107 mL R1
Intended Use
Enzymatic colour test for the quantitative determination of cholesterol in human serum and plasma on Beckman Coulter AU analysers. For in vitro
*Cholesterol reagent OSR6516 for use on the AU2700 and AU5400 systems only.
Summary1
Cholesterol is synthesised ubiquitously throughout the body and is an essential component of cell membranes and lipoproteins as well as being a
precursor for the synthesis of steroid hormones and bile acids.
The individual predictive value of total cholesterol concentration with regard to coronary risk is low. Cholesterol is mainly transported in two lipoprotein
classes (LDL and HDL), both of which play a contradictory role in the pathogenesis of lipid disorders. The total cholesterol concentration therefore provides
only a baseline value that indicates whether further laboratory investigations of lipoprotein metabolism should be carried out (HDL, LDL, and triglycerides).
Test Principle2
The Cholesterol reagent utilises an enzymatic method to measure cholesterol in human serum and plasma. In this procedure cholesterol esters in a
sample are hydrolysed by cholesterol esterase (CHE). The free cholesterol produced is oxidised by cholesterol oxidase (CHO) to cholestene-3-one with
the simultaneous production of hydrogen peroxide (H2O2), which oxidatively couples with 4-aminoantipyrine and phenol in the presence of peroxidase
(POD) to yield a chromophore.
The red quinoneimine dye formed can be measured spectrophotometrically at 540/600 nm as an increase in absorbance.
Reaction Principle
CHE
2 Cholesterol esters + 2 H2O 2 Cholesterol + 2 Fatty acids
CHO
2 Cholesterol + 2 O2 2 Cholestene-3-one + 2 H2O2
POD
2 H2O2 + 4-Aminoantipyrine + Phenol Quinoneimine + 4 H2O

4-Aminoantipyrine 0.31 mmol/L
Phenol 5.2 mmol/L
Cholesterol esterase ≥ 0.2 kU/L (3.3 µkat/L)
Cholesterol oxidase ≥ 0.2 kU/L (3.3 µkat/L)
Peroxidase ≥ 10.0 kU/L (166.7 µkat/L)
Preservative

Reagent Preparation

The reagent is stable, unopened, up to the stated expiry date when stored at 2…8 °C. Once open, reagent stored on board the instrument is stable for
90 days.
Specimen3
Serum and EDTA or heparinised plasma. Icteric samples should be avoided.
Plasma is not recommended using anticoagulants such as oxalate, citrate or fluoride.
Stable in serum and plasma for 7 days when stored at 2...8 °C.
Test Procedure
Calibration
The calibrator cholesterol value is traceable to the National Institute of Standards and Technology (NIST) Standard Reference Material (SRM) 909b
4
Level 1 (Isotope Dilution Mass Spectrometry). This method is also certified against the Centers of Disease Control Reference Method (Abell-Kendall).

2010-04
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the cholesterol concentration of each sample.
5
Total cholesterol levels in plasma should be corrected by multiplying the result obtained by 1.03 to be equivalent to serum levels of total cholesterol.
Reference Intervals
6
National Cholesterol Education Program Adult Treatment Panel III recommendations:
< 5.2 mmol/L (200 mg/dL) Desirable
5.2 – 6.2 mmol/L (200 – 239 mg/dL) Borderline High
≥ 6.2 mmol/L (240 mg/dL) High
7
European Atherosclerosis Society recommendations:
Cholesterol < 5.2 mmol/L (< 200 mg/dL) No Lipid metabolism disorder
Triglyceride < 2.3 mmol/L (< 200 mg/dL)
Cholesterol 5.2 – 7.8 mmol/L (200 – 300 mg/dL) Lipid metabolism disorder if HDL-Cholesterol is < 0.9 mmol/L (< 35 mg/dL)
Cholesterol > 7.8 mmol/L (> 300 mg/dL) Lipid metabolism disorder.
Triglyceride > 2.3 mmol/L (> 200mg/dL)
National and regional guidelines for interpretation and treatment may differ from the above recommendations. Please follow the guidelines which are
applicable to your population.

values.
Linearity
The test is linear within a concentration range of 0.5 – 18.0 mmol/L (20 – 700 mg/dL).
Precision
3.07 0.03 0.91 0.03 1.06
5.92 0.04 0.72 0.09 1.45
11.18 0.08 0.72 0.13 1.13
Sensitivity
The lowest detectable level represents the lowest measurable level of cholesterol that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient serum samples were used to compare this Cholesterol OSR6116 assay on the AU2700 against another commercially available cholesterol assay.
®
8

* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 38.7.
BIBLIOGRAPHY
1. Riesen WF. Lipid Metabolism. In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
Verlagsgesellschaft, 1998:167–169.
2. Allain CC, Poon LS, Chan CSG, Richmond W, Fu PC. Enzymatic determination of total serum cholesterol. Clin Chem 1974;20:470-475.
3. World Health Organization (2002). Use of Anticoagulants in Diagnostic Laboratory Investigations (Stability of Blood, Plasma and serum samples);
WHO/DIL/LAB/99.1 Rev.2: 1-64
4. Data on file.
5. Current status of blood cholesterol measurement in clinical laboratories in the United States: a report from the Laboratory Standardization Panel of the National
Cholesterol Education Program. Clin Chem 1988;34:193-201.
6. National cholesterol education program expert panel. Executive summary of the third report of the National Cholesterol Education Program (NCEP) expert panel
on detection, evaluation, and treatment of high blood cholesterol in adults (Adult Treatment Panel III). JAMA 2001;285:2486-2497.
7. Study Group, European Atherosclerosis Society, Strategies for the prevention of coronary heart disease: A policy statement of the European Atherosclerosis
Society. European Heart Journal 1987; 8, 77-88.

2010-04
CHOLESTEROL, AU400/AU640 Serum/Plasma Application CHOLESTEROL, AU600 Serum/Plasma Application

Test No # Test name CHOL ∇ Sample type Ser ∇ Page 1/2
Test Name: CHOL ∇ < > Type: Serum ∇ Operation: Yes ∇
Method: END ∇ Last L -0.1 Last H 0.3 Reaction + ∇ L 0.5* H 18.0*
Measuring Point 1: First 0 Last 27 L 0.5* H 18.0* Point 2 Fst Lst B 0
General LIH ISE Range Test No # Test name CHOL ∇ Sample type Ser ∇ Page 2/2
Test Name: CHOL ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name CHOL ∇
Test Name: CHOL ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 16* 30*
Point 1: # † 16* 30*
Point 2 ∇
Point 2:
MB type factor
# User defined
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 38.7
2009-08
CHOLESTEROL, AU2700/AU5400 Serum/Plasma Application CHOLESTEROL, AU680/AU480 Serum/Plasma Application
Test Name: CHOL ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CHOL ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Dynamic Range Low 0.5* High 18.0*
Measuring Point 1: First 0 Last 27 L 0.5* H 18.0* Wavelength Pri 540 ∇nm Sec. 600 ∇nm Factor for Maker A 1 B 0
Linearity Limit % Icterus ++ ∇
Specific Test Parameters Lag Time Check ∇ Hemolysis ++++ ∇
Test Name: CHOL ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CHOL ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: CHOL ∇ < > Type Serum ∇ Test Name: CHOL ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † 16* 30* Point 1: # ∇ † 16* 30*
Point 9 ∇
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 38.7 Point-1 ∇ Reagent Blank 999 Day 0 Hour
2009-08
CREATININE
OSR6178 4 x 51 mL R1
4 x 51 mL R2
OSR6578 4 x 103 mL R1
4 x 103 mL R2
Intended Use
Kinetic colour test (Jaffé method) for the quantitative determination of creatinine in human serum, plasma and urine on Beckman Coulter AU analysers. For
in vitro diagnostic use only. The OSR6x78 reagents may be used in two ways for the serum application:
Method A : Kinetic Jaffe (compensated method) traceable to the IDMS reference method.
Method B : Kinetic Jaffe uncompensated.
Creatinine reagent OSR6578 for use on the AU2700 and AU5400 systems only.
Summary1,2,3
Creatinine is a metabolic product of creatine and phosphocreatine, which are both found almost exclusively in muscle. Thus, creatinine production is
proportional to muscle mass and varies little from day to day.
Measurements of creatinine are used in the diagnosis and treatment of renal disease and prove useful in the evaluation of kidney glomerular function and
in monitoring renal dialysis. However, the serum level is not sensitive to early renal damage and responds more slowly than blood urea nitrogen (BUN) to
haemodialysis during treatment of renal failure. Both serum creatinine and BUN are used to differentiate prerenal and postrenal (obstructive) azotemia. An
increase in serum BUN without concomitant increase of serum creatinine is key to identifying prerenal azotemia. In post renal conditions where obstruction
to the flow of urine is present e.g. malignancy, nephrolithiasis and prostatism, both the plasma creatinine and urea levels will be increased; in these
situations the rise is disproportionately greater for BUN due to the increased back diffusion of urea.
Serum creatinine varies with the subject’s age, body weight, and sex. It is sometimes low in subjects with relatively small muscle mass, cachectic patients,
amputees, and in older persons. A serum creatinine level that would usually be considered normal does not rule out the presence of impaired renal
function.
Test Principle4,5,6
Creatinine forms a yellow-orange coloured compound with picric acid in an alkaline medium. The rate of change in absorbance at 520/800 nm is
proportional to the creatinine concentration in the sample.
Reaction Principle
Creatinine + picric acid Creatinine picrate complex

Sodium hydroxide 120 mmol/L
Picric acid 2.9 mmol/L
Preservative

R1 reagent: Irritant. R36/38 Irritating to eyes and skin.
Safety Phrases:
S26, S37, S60: In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable gloves. This material and its
container must be disposed of as hazardous waste.
R2 contains picric acid. Risk of explosion by shock, friction, fire or other sources of ignition. Dilute any spills with water and wipe up immediately.
Reagent Preparation

The reagents are stable, protected from light, unopened, up to the stated expiry date when stored at 2…8°C. Once open, reagents stored on board the
Specimen
7
Strongly lipemic samples should be avoided.
8
Urine: Collect urine without using preservatives. Store at 2...8°C.
Test Procedure
Two distinct test procedures are available for the serum application of this reagent:
A: Interference from protein is mathematically corrected by subtracting 18 µmol/L from each test result (Specific Test Parameters: Correlation
factor B = -18).
B: Uncompensated Jaffe (no protein compensation, B = 0)
Both methods require different calibrator set points and deliver slightly different results for patients and control sera at low creatinine concentrations which
results in different reference ranges.

2009-11
Calibration
Use System Calibrator Cat. No. 66300 for serum and plasma application and Urine Calibrator Cat. No. ODC0025 for urine application.
The serum calibrator creatinine value for method A is traceable to the Isotope Dilution Mass Spectroscopy (IDMS) method via National Institute of
Standards and Technology (NIST) Standard Reference Material (SRM) 967.
The serum calibrator creatinine value for method B is traceable to the National Institute of Standards and Technology (NIST) Standard Reference Material
(SRM) 909b Level 2.
The urine calibrator creatinine value is traceable to the IDMS method.
Recalibrate the assay every day, or when the following occur:
Absorption of atmospheric CO2 by the reagent on board the analyser can impair calibration stability. This effect will vary depending upon the rate of use.
Consequently each laboratory should set a calibration frequency in the instrument parameters appropriate to their usage pattern.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the creatinine concentration of each sample.
Reference Intervals
Method A (IDMS traceable) Method B (Uncompensated Jaffe)
9,10 3,12
Serum/Plasma Serum/Plasma
Male 59 – 104 µmol/L (0.67 – 1.17 mg/dL) Male < 50 years 74 – 110 µmol/L (0.84 – 1.25 mg/dL)
Female 45 – 84 µmol/L (0.51 – 0.95 mg/dL) Male >50 years 72 – 127 µmol/L (0.81 – 1.44 mg/dL)
Neonate 27 – 87 µmol/L (0.31 – 0.98 mg/dL) Female 58 – 96 µmol/L (0.66 – 1.09 mg/dL)
Infant 14 – 34 µmol/L (0.16 – 0.39 mg/dL) Neonate 45 – 105 µmol/L (0.5 – 1.2 mg/dL)
Child 23 – 68 µmol/L (0.26 – 0.77 mg/dL) Infant 35 – 62 µmol/L (0.4 – 0.7 mg/dL)
Child 45 – 105 µmol/L (0.5 – 1.2 mg/dL)
11 11
Urine Urine
Male 124 – 230 µmol/kg/d (14 – 26 mg/kg/d) Male 124 – 230 µmol/kg/d (14 – 26 mg/kg/d)
Female 97 – 177 µmol/kg/d (11 – 20 mg/kg/d) Female 97 – 177 µmol/kg/d (11 – 20 mg/kg/d)

values.
Linearity
Method A is linear within a concentration range of 5 – 2200 µmol/L (0.06 – 25.0 mg/dL) for serum and plasma.
Method B is linear within a concentration range of 18 – 2200 µmol/L (0.2 – 25.0 mg/dL) for serum and plasma.
The test is linear within a concentration range of 88 – 35360 µmol/L (1 – 400 mg/dL) for urine.
Precision
105.06 1.26 1.19 2.10 2.00
152.70 1.45 0.95 2.47 1.62
1049.87 9.00 0.86 16.93 1.61
2,600 24.82 0.96 64.53 2.48
13,100 97.30 0.74 202.22 1.54
34,560 215.88 0.62 454.32 1.31
Sensitivity
The lowest detectable level of method A using serum settings on an AU400 analyser was established at 2.4 µmol/L (0.027 mg/dL).
The lowest detectable level of method B using serum settings on an AU640 analyser was established at 3.15 µmol/L (0.036 mg/dL).
The lowest detectable level using urine settings on an AU2700 analyser was established at 0.1 µmol/L (0.001 mg/dL).
The lowest detectable level represents the lowest measurable level of creatinine that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient serum samples were used to compare this Creatinine OSR6178 assay (method A) on the AU2700 against the Creatinine OSR6178 assay
(method B). Results of linear regression analysis were as follows:
y = 1.04x – 17 r = 0.999 n = 701 Sample range = 26.5 – 1024 µmol/L
Patient serum samples were used to compare this Creatinine OSR6178 assay (method A) on the AU2700 against a commercially available enzymatic
creatinine assay which has demonstrated equivalence to the IDMS reference method. Results of linear regression analysis were as follows:
y = 1.01x + 2.8 r = 0.997 n = 701 Sample range = 13.3 – 1007 µmol/L
2009-11
Patient urine samples were used to compare this Creatinine OSR6178 assay on the AU2700 against another commercially available creatinine assay.
y = 0.916x + 266.607 r = 0.998 n = 124 Sample range = 1246 – 32562 µmol/L
®
Lipemia: Interference less than 10% up to 600 mg/dL Intralipid.
Protein: Interference less than 6% between 3 and 10 g/dL protein for method A
Interference less than 20% between 3 and 12 g/dL protein for method B
Glucose: Interference less than 3% up to 3000mg/dL glucose
13

* Values set for working in SI units (µmol/L). To work in mg/dL divide by 88.4.
^ Equivalent to 5 µmol/L due to B = -18.
BIBLIOGRAPHY
1. Newman DJ, Price CP. Renal function and nitrogen metabolites. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia: WB
Saunders Company, 1999;1239-1242.
2. Mayne PD, ed. Clinical chemistry in diagnosis and treatment, 6th ed. London: Arnold,1994:18pp.
3. Thomas L. Creatinine. In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
4. Folin O. Beitrag zur chemie des kreatinins und kreatins im harne. Physiol Chem 1904;41:223-42.
5. Cook JGH. Creatinine assay in the presence of protein. Clin Chim Acta 1971;32:485-6.
6. Larsen K. Creatinine assay in the presence of protein with LKB 8600 reaction rate analyser. Clin Chim Acta 1972;38:475-6.
nd
8. NCCLS. Urinalysis and collection, transportation, and preservation of urine specimens; approved guideline. NCCLS Document GP16-A2, 2 ed.
9. Mazzachi BC, Peake MJ, Ehrhard V.; Reference range and method comparison studies for enzymatic and Jaffe creatinine assays in plasma and
serum and early morning urine. Clin. Lab. 2000; 46: 53-55.
10. Schlebusch H, Liappis N, Klein G. Creatinine and ultrasensitive CRP: Reference intervals from infancy to childhood. Clin. Chem. Lab. Med. 2001; 39
Special supplement pp S1-S448, May 2001. PO-T042.
12. Soldin SJ, Brugnara C, Wong EC, eds. Pediatric Reference Ranges. 4th ed. AACC Press, 2003:71-72.
th
13. Young DS. Effects of drugs on clinical laboratory tests, 5 ed. AACC Press, 2000.

2009-11
CREATININE, AU400/AU640 Serum/Plasma Application CREATININE, AU600 Serum/Plasma Application
Method A Method A
Test No # Test name CRE ∇ Sample type Ser ∇ Page 1/2
Test Name: CRE ∇ < > Type: Serum ∇ Operation: Yes ∇
Method: FIXED ∇ Last L -0.2 Last H 0.2 Reaction + ∇ L 23*^ H 2200*
Measuring Point 1: First 13 Last 24 L 23*^ H 2200* Point 2 Fst Lst B -18*
Linearity : % A 1 B -18*
Method B Select using Space key, or select from list displayed by Guide key
Specific Test Parameters Method B
General LIH ISE Range Specific test parameters
Test Name: CRE ∇ < > Type: Serum ∇ Operation: Yes ∇ Test No # Test name CRE ∇ Sample type Ser ∇ Page 1/2
Sample: Volume 20 μL Dilution 0 μL Pre-Dilution Rate: 1 Sample vol. 20 Dil. vol 0 μL Min. OD Max. OD
Reagents: R1 Volume 120 μL Dilution 90 μL Min OD Max OD Reagent 1 vol 120 Dil. vol 90 μL L -0.2 H 2.5
R2 Volume 120 μL Dilution 0 μL L -0.2 H 2.5 Reagent 2 vol 120 Dil. vol 0 μL Reagent OD limit
Reagent OD limit: Fst. L -0.2 Fst. H 0.2
Wavelength: Pri. 520 ∇ Sec. 800 ∇ First L -0.2 First H 0.2 Wave Main 520 Sub 800 ∇ Lst. L -0.2 Lst. H 0.2
Method: FIXED ∇ Last L -0.2 Last H 0.2 Method FIXED ∇ Dynamic range
Reaction slope: + ∇ Dynamic Range: Reaction + ∇ L 18* H 2200*
Measuring Point 1: First 13 Last 24 L 18* H 2200* Point 1 Fst 13 Lst 24 Correlation factor A 1
Measuring Point 2: First Last Correlation Factor: Point 2 Fst Lst B 0
Linearity : % A 1 B 0 Sample Pre-dil Rate ¤
Linearity Fst % Sec %
Method A and/or Method B
Calibration Specific Select using Space key, or select from list displayed by Guide key
General ISE Method A and/or Method B
Test Name: CRE ∇ < > Type Serum ∇
Test No # Test name CRE ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H Formula 1 Y=AX+B Process Conc
∇ ∇
Point 1: # † 4900* 14000*
Point 2: Cal No OD Conc Factor/OD-L Factor/OD-H
Point 3: Point 1 # ∇ † 4900* 14000*
Point 5:
Point 3 ∇
Point 6:
Point 4 ∇
Point 7:
Point 5 ∇
1-Point Cal. Point: ο With CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 6 ∇
MB Type Factor: Calibration Stability Period: ‡ Point 7 ∇
1-point cal. point
# User defined MB type factor
† System Calibrator Cat. No.: OE66300 Calibrator stability period ‡
‡ Depends on usage pattern in the Laboratory Select the function using the Function key or the Mouse
^ Equivalent to 5µmol/L due to B = -18 # User defined ¤ Analyser default value
† System Calibrator Cat. No.: OE66300
‡ Depends on usage pattern in the Laboratory ^Equivalent to 5 µmol/L due to B = -18
2009-08
CREATININE, AU2700/AU5400 Serum/Plasma Application CREATININE, AU680/AU480 Serum/Plasma Application Method A
Method A Parameters Specific Test Parameters
General LIH ISE Range Test Name: CRE Type: Serum
Test Name: CRE ∇ < > Type: Serum ∇ Operation: Yes ∇
Reagent OD limit: Last Low -0.2 High 0.2
Wavelength: Pri. 520 ∇ Sec. 800 ∇ First L -0.2 First H 0.2 R2(R2-1) 48 Dilution 0
μL μL
Method: FIXED ∇ Last L -0.2 Last H 0.2 Dynamic Range Low 23*^ High 2200*
Measuring Point 1: First 13 Last 24 L 23*^ H 2200* Wavelength Pri 520 ∇nm Sec. 800 ∇nm Factor for Maker A 1 B -18*
Linearity : % A 1 B -18* Reaction Slope + ∇ Onboard Stability Period 14 Day 0 Hour
Method B
Lag Time Check ∇ Hemolysis +++++ ∇
Test Name: CRE ∇ < > Type: Serum ∇ Operation: Yes ∇ General LIH ISE HbA1c Calculated Test Range
Sample: Volume 8 μL Dilution 0 μL Pre-Dilution Rate: 1 Test Name: CRE ∇ < > Type: Serum ∇
R2 Volume 48 μL Dilution 0 μL L -0.2 H 2.5 Level # # Panic Value
Reagent OD limit: Specificl Ranges: From To Low High
Wavelength: Pri. 520 ∇ Sec. 800 ∇ First L -0.2 First H 0.2 Sex Year Month Year Month Low High # #
Method: FIXED ∇ Last L -0.2 Last H 0.2 ο 1. # ∇ # # # # # #
Reaction slope: + ∇ Dynamic Range: ο 2. # ∇ # # # # # #
Measuring Point 1: First 13 Last 24 L 18* H 2200* ο 3. # ∇ # # # # # #
Measuring Point 2: First Last Correlation Factor: ο 4. # ∇ # # # # # #
Linearity : % A 1 B 0 ο 5. # ∇ # # # # # #
No Lag Time: ∇ On-board stability period: 14 ο 6. # ∇ # # # # # #
Method A and/or Method B Unit µmol/L* Decimal Places #
General ISE
Test Name: CRE Type Serum General ISE
∇ < > ∇
Test Name: CRE ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 2: Point 1: # ∇ † 4500* 13000*
Point 9 ∇
# User defined <Point Cal. For No. of Correction Points Use Master Curve
∇ ∇ ο Lot Calibration
* Values set for working in SI units (µmol/L). To work in mg/dL divide by 88.4. Master Curve> OD Range
‡ Depends on usage pattern in the Laboratory Calibrator OD Conc Low High Stability
^ Equivalent to 5µmol/L due to B = -18 Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
2009-08
CREATININE, AU680/AU480 Serum/Plasma Application Method B
System Reagent: OSR6178, OSR6578 Reagent ID: 078
Test Name: CRE ∇ < > Type: Serum ∇ Operation Yes ∇

Method FIXED ∇

Test Name: CRE ∇ < > Type: Serum ∇

ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
Unit µmol/L* Decimal Places #
General ISE
Test Name: CRE ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # ∇ † 4500* 13000*
Point 3: ∇
Point 6: ∇
Point 9 ∇
Calibrator OD Conc Low High Stability # User defined
Point-1 ∇ Reagent Blank ‡ Day ‡ Hour † System Calibrator Cat. No.: OE66300
Point-2 Calibration ‡ Day ‡ Hour * Values set for working in SI units (µmol/L). To work in mg/dL divide by 88.4.
∇
MB Type Factor: 1-Point Calibration Point ∇ ο with Conc-0 ^ Equivalent to 5µmol/L due to B = -18
ф AU680
2009-08
CREATININE, AU400/AU640 Urine Application CREATININE, AU600 Urine Application

Test No # Test name CRE ∇ Sample type Uri ∇ Page 1/2
Test Name: CRE ∇ < > Type: Urine ∇ Operation: Yes ∇
Method: FIXED ∇ Last L -2.0 Last H 2.5 Reaction + ∇ L 88.4* H 35360*
Measuring Point 1: First 13 Last 24 L 88.4* H 35360* Point 2 Fst Lst B 0
General LIH ISE Range Test No # Test name CRE ∇ Sample type Uri ∇ Page 2/2
Test Name: CRE ∇ < > Type: Urine ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
URINE APPLICATION
General ISE Test No # Test name CRE ∇
Test Name: CRE ∇ < > Type Urine ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 21000* 70000*
Point 1: # † 21000* 70000*
Point 2 ∇
Point 2:
MB type factor
* Values set for working in SI units (µmol/L). To work in mg/dL divide by 88.4. * Values set for working in SI units (µmol/L). To work in mg/dL divide by 88.4.
‡ Depends on usage pattern in the laboratory. ‡ Depends on usage pattern in the laboratory.
2009-08
CREATININE, AU2700/AU5400 Urine Application CREATININE, AU680/AU480 Urine Application
Test Name: CRE ∇ < > Type: Urine ∇ Operation Yes ∇
Test Name: CRE ∇ < > Type: Urine ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Dynamic Range Low 88.4* High 35360*
Measuring Point 1: First 13 Last 24 L 88.4* H 35360* Wavelength Pri 520 ∇nm Sec. 800 ∇nm Factor for Maker A 1 B 0
Linearity Limit %
Test Name: CRE ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CRE ∇ < > Type: Urine ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: µmol/L* Decimal places: # 7. No demographics # #
URINE APPLICATION
General ISE
General ISE
Test Name: CRE ∇ < > Type Urine ∇ Test Name: CRE ∇ < > Type Urine ∇ ο Use Serum Cal.
Point 1: # † 21000* 70000* Point 1: # ∇ † 21000* 70000*
Point 9 ∇
* Values set for working in SI units (µmol/L). To work in mg/dL divide by 88.4. Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
‡ Depends on usage pattern in the laboratory. Point-2 ∇ Calibration ‡ Day ‡ Hour
Ф AU680 MB Type Factor: 1-Point Calibration Point ∇ ο with Conc-0
2009-08
CREATININE, AU400/AU640 Paediatric Application CREATININE, AU600 Paediatric Application
Method A Method A
Test No # Test name CREP ∇ Sample type Ser ∇ Page 1/2
Test Name: CREP ∇ < > Type: Serum ∇ Operation: Yes ∇
Method: FIXED ∇ Last L -0.2 Last H 0.2 Reaction + ∇ L 23*^ H 2200*
Measuring Point 1: First 13 Last 24 L 23*^ H 2200* Point 2 Fst Lst B -18*
Linearity : % A 1 B -18*
Method B Select using Space key, or select from list displayed by Guide key
Specific Test Parameters Method B
General LIH ISE Range Specific test parameters
Test Name: CREP ∇ < > Type: Serum ∇ Operation: Yes ∇ Test No # Test name CREP ∇ Sample type Ser ∇ Page 1/2
Sample: Volume 20 μL Dilution 10 μL Pre-Dilution Rate: 1 Sample vol. 20 Dil. vol 10 μL Min. OD Max. OD
Reagents: R1 Volume 120 μL Dilution 80 μL Min OD Max OD Reagent 1 vol 120 Dil. vol 80 μL L -0.2 H 2.5
R2 Volume 120 μL Dilution 0 μL L -0.2 H 2.5 Reagent 2 vol 120 Dil. vol 0 μL Reagent OD limit
Reagent OD limit: Fst. L -0.2 Fst. H 0.2
Wavelength: Pri. 520 ∇ Sec. 800 ∇ First L -0.2 First H 0.2 Wave Main 520 Sub 800 ∇ Lst. L -0.2 Lst. H 0.2
Method: FIXED ∇ Last L -0.2 Last H 0.2 Method FIXED ∇ Dynamic range
Reaction slope: + ∇ Dynamic Range: Reaction + ∇ L 18* H 2200*
Measuring Point 1: First 13 Last 24 L 18* H 2200* Point 1 Fst 13 Lst 24 Correlation factor A 1
Linearity : % A 1 B 0 Sample Pre-dil Rate ¤
Method A and/or Method B No lag time ∇ On-board stability period 14
Calibration Specific Select using Space key, or select from list displayed by Guide key
General ISE Method A and/or Method B
Test Name: CREP Type Serum Calibration specific
∇ < > ∇
Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Test No # Test name CREP ∇

Point 1: # † 4900* 14000*
Point 2:
Point 3:
Point 1 # ∇ † 4900* 14000*
Point 4:
Point 2 ∇
Point 5:
Point 3 ∇
Point 6:
Point 5 ∇
Point 6 ∇
MB Type Factor: Calibration Stability Period: ‡ Point 7 ∇
1-point cal. point
# User defined MB type factor
† System Calibrator Cat. No.: OE66300 Calibrator stability period ‡
‡ Depends on usage pattern in the Laboratory Select the function using the Function key or the Mouse
^ Equivalent to 5µmol/L due to B = -18 # User defined ¤ Analyser default value
‡ Depends on usage pattern in the Laboratory ^Equivalent to 5µmol/L due to B = -18
2009-08
CREATININE, AU2700/AU5400 Paediatric Application CREATININE, AU680/AU480 Paediatric Application Method A
Method A Parameters Specific Test Parameters
General LIH ISE Range Test Name: CREP Type: Serum
Test Name: CREP ∇ < > Type: Serum ∇ Operation: Yes ∇
Reagent OD limit: Last Low -0.2 High 0.2
Wavelength: Pri. 520 ∇ Sec. 800 ∇ First L -0.2 First H 0.2 R2(R2-1) 48 Dilution 0
μL μL
Method: FIXED ∇ Last L -0.2 Last H 0.2 Dynamic Range Low 23*^ High 2200*
Measuring Point 1: First 13 Last 24 L 23*^ H 2200* Wavelength Pri 520 ∇nm Sec. 800 ∇nm Factor for Maker A 1 B -18*
Linearity : % A 1 B -18* Reaction Slope + ∇ Onboard Stability Period 14 Day 0 Hour
Method B
Test Name: CREP ∇ < > Type: Serum ∇ Operation: Yes ∇ General LIH ISE HbA1c Calculated Test Range
Sample: Volume 8 μL Dilution 10 μL Pre-Dilution Rate: 1 Test Name: CREP ∇ < > Type: Serum ∇
R2 Volume 48 μL Dilution 0 μL L -0.2 H 2.5 Level # # Panic Value
Reagent OD limit: Specificl Ranges: From To Low High
Wavelength: Pri. 520 ∇ Sec. 800 ∇ First L -0.2 First H 0.2 Sex Year Month Year Month Low High # #
Method: FIXED ∇ Last L -0.2 Last H 0.2 ο 1. # ∇ # # # # # #
Reaction slope: + ∇ Dynamic Range: ο 2. # ∇ # # # # # #
Measuring Point 1: First 13 Last 24 L 18* H 2200* ο 3. # ∇ # # # # # #
Measuring Point 2: First Last Correlation Factor: ο 4. # ∇ # # # # # #
Linearity : % A 1 B 0 ο 5. # ∇ # # # # # #
No Lag Time: ∇ On-board stability period: 14 ο 6. # ∇ # # # # # #
Method A and/ or Method B Unit µmol/L* Decimal Places #
General ISE
Test Name: CREP Type Serum General ISE
∇ < > ∇
Test Name: CREP ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 2: Point 1: # ∇ † 4500* 13000*
Point 9 ∇
# User defined <Point Cal. For No. of Correction Points Use Master Curve
* Values set for working in SI units (µmol/L). To work in mg/dL divide by 88.4. Master Curve> OD Range
‡ Depends on usage pattern in the Laboratory Calibrator OD Conc Low High Stability
^ Equivalent to 5µmol/L due to B = -18 Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
2009-08
CREATININE, AU680/AU480 Paediatric Application Method B
Test Name: CREP ∇ < > Type: Serum ∇ Operation Yes ∇

Method FIXED ∇

Test Name: CREP ∇ < > Type: Serum ∇

ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
General ISE
Test Name: CREP ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # ∇ † 4500* 13000*
Point 3: ∇
Point 6: ∇
Point 9 ∇
# User defined
Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
Point-2 ∇ Calibration ‡ Day ‡ Hour * Values set for working in SI units (µmol/L). To work in mg/dL divide by 88.4.
MB Type Factor: 1-Point Calibration Point ∇ ο with Conc-0 ^ Equivalent to 5µmol/L due to B = -18
ф AU680
2009-08
CREATININE (ENZYMATIC)
OSR61204 4 x 45 mL R1
4 x 15 mL R2
Intended Use
Enzymatic assay for the quantitative determination of creatinine in human serum, plasma and urine on Beckman Coulter analysers. For in vitro diagnostic
use only.
Summary1,2,3
Creatinine is a metabolic product of creatine and phosphocreatine, which are both found almost exclusively in muscle. Thus, creatinine production is
proportional to muscle mass and varies little from day to day.
Measurements of creatinine are used in the diagnosis and treatment of renal disease and prove useful in the evaluation of kidney glomerular function and
in monitoring renal dialysis. However, the serum level is not sensitive to early renal damage and responds more slowly than blood urea nitrogen (BUN) to
haemodialysis during treatment of renal failure. Both serum creatinine and BUN are used to differentiate prerenal and postrenal (obstructive) azotemia. An
increase in serum BUN without concomitant increase of serum creatinine is key to identifying prerenal azotemia. In post renal conditions where obstruction
to the flow of urine is present e.g. malignancy, nephrolithiasis and prostatism, both the plasma creatinine and urea levels will be increased; in these
situations the rise is disproportionately greater for BUN due to the increased back diffusion of urea.
Serum creatinine varies with the subject’s age, body weight, race and sex. It is sometimes low in subjects with relatively small muscle mass, cachectic
patients, amputees, and in older persons. A serum creatinine level that would usually be considered normal does not rule out the presence of impaired
renal function.
Test Principle
Creatinine is hydrolysed by creatininase to creatine. The creatine formed is hydrolysed by creatinase to sarcosine and urea. Sarcosine oxidase catalyzes
the oxidative demethylation of the sarcosine to yield glycine, formaldehyde and hydrogen peroxide. In the presence of peroxidase (POD), the hydrogen
peroxide formed reacts by quantitative oxidation condensation with N-(3-sulfopropyl)-3-methoxy-5-methylaniline (HMMPS) and 4-aminoantipyrine to yield a
blue pigment. The creatinine concentration is proportional to the change in absorbance at 600/700 nm.
Reaction Principle
Creatininase
Creatinine + H2O Creatine
Creatinase
Creatine + H2O Sarcosine + Urea
Sarcosine Oxidase
Sarcosine + H2O + O2 Glycine + Formaldehyde + H2O2
POD
H2O2 + 4-aminoantipyrine + HMMPS Blue Pigment

Good's Buffer 50 mmol/L
Creatinase 56.3 IU/mL
Sarcosine oxidase 15 IU/mL
HMMPS 0.68 mmol/L
Creatininase 100 IU/mL
Peroxidase 12.5 U/mL
Preservative

Reagent Preparation

60 days.
Specimen
Serum and heparinised plasma.
4
5
Urine: Collect urine without using preservatives. Store at 2…8°C.
Test Procedure
Refer to the appropriate User Guide and Setting Sheet for analyser-specific assay instructions for the sample type as listed in the Intended Use Statement.

2009-08
Calibration
Use System Calibrator Cat. No. 66300 for the serum and plasma application and Urine Calibrator Cat. No. ODC0025 for the urine application.
The serum calibrator creatinine value is traceable to the Isotope Dilution Mass Spectroscopy (IDMS) method via National Institute of Standards and
Technology (NIST) Standard Reference Material (SRM) 967.
The urine calibrator creatinine value is traceable to the IDMS method.
Recalibrate the serum and plasma application every 14 days and the urine application every 30 days, or when the following occur:
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the creatinine concentration of each sample.
Reference Intervals
6
Serum/Plasma
Male 64 – 104 µmol/L (0.72 – 1.18 mg/dL)
Female 49 –90 µmol/L (0.55 – 1.02 mg/dL)
Neonate 22 – 90 µmol/L (0.26 – 1.01 mg/dL)
Infant (2 months – < 3 years) 11 – 34 µmol/L (0.15 – 0.37 mg/dL)
Child (3 – < 15 years) 21 – 65 µmol/L (0.24 – 0.73 mg/dL)
7
Urine
Male 124 – 230 µmol/kg/day (14 – 26 mg/kg/day)
Female 97 – 177 µmol/kg/day (11 – 20 mg/kg/day)

values.
Linearity
The test is linear within a concentration range of 4.4 – 4420 µmol/L (0.05 – 50.0 mg/dL) for serum and plasma.
The test is linear within a concentration range of 88 – 44200 µmol/L (1 – 500 mg/dL) for urine.
Precision
62.1 0.7 1.2 1.4 2.3
180.2 1.2 0.6 2.9 1.6
908.3 6.7 0.7 14.0 1.5
2335 37 1.6 51 2.2
8601 111 1.3 189 2.2
15146 159 1.1 331 2.2
Sensitivity
The limit of detection for creatinine using serum settings on an AU640 analyser was established at 0.88 µmol/L; limit of blank = 0.11 µmol/L.
The limit of detection for creatinine using urine settings on an AU640 analyser was established at 13.9 µmol/L; limit of blank = 3.1 µmol/L.
8
The limit of detection was determined consistent with the guidelines in the NCCLS protocol EP17-A with proportions of false positives less than 5% and
false negatives less than 5%; based on 150 determinations, with 60 blank and 90 low-level samples.
Method Comparison
Patient serum samples were used to compare this Creatinine (Enzymatic) OSR61204 assay on the AU2700 against another commercially available
enzymatic creatinine assay. Results of linear regression analysis were as follows:
y = 1.014x + 1.768 r = 1.000 n = 237 Sample range = 12.4 – 1966 µmol/L
Patient urine samples were used to compare this Creatinine (Enzymatic) OSR61204 assay on the AU2700 against another commercially available IDMS
traceable creatinine assay. Results of linear regression analysis were as follows:
y = 0.986x - 104.577 r = 0.997 n = 151 Sample range = 707 – 19793 µmol/L
®
Creatine: Interference less than 5% up to 30 mg/dL creatine
2009-08
Icterus: Interference less than 5% up to 50 mg/dL conjugated bilirubin
Glucose: Interference less than 5% up to 3000 mg/dL glucose
9

† System Calibrator Cat. No.: 66300 / Urine Calibrator Cat. No.:ODC0025
BIBLIOGRAPHY
1. Newman DJ, Price CP. Renal function and nitrogen metabolites. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia: WB
th
2. Mayne PD, ed. Clinical chemistry in diagnosis and treatment, 6 ed. London: Arnold,1994:18pp.
3. Thomas L. Creatinine. In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
nd
5. NCCLS. Urinalysis and collection, transportation, and preservation of urine specimens; approved guideline. NCCLS Document GP16-A2, 2 ed.
6. Ceriotti F, Boyd JC, Klein G, Henny J, Queraltó J, Kairisto V, Panteghini M. Reference intervals for serum creatinine concentrations: Assessment of
available data for global application. Clin Chem. 2008; 54(3): 559-566.
8. NCCLS. Protocols for determination of limits of detection and limits of quantitation; approved guideline. NCCLS Document EP17-A. Pennsylvania:
NCCLS, 2004.
th

2009-08
CREATININE (ENZYMATIC), AU400/AU640 Serum/Plasma Application CREATININE (ENZYMATIC), AU600 Serum/Plasma Application

Test No # Test name CREZ ∇ Sample type Ser ∇ Page 1/2
Test Name: CREZ ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 7 µL Dilution 10 µL Pre-Dilution Rate: 1 Reagent 1 vol 150 Dil. vol 0 µL L H
R2 Volume 50 µL Dilution 0 µL L H Fst. L -0.1 Fst. H 0.2
Method: END ∇ Last L -0.1 Last H 0.2 Reaction + ∇ L 4.4* H 4420*
Measuring Point 1: First 0 Last 27 L 4.4* H 4420* Point 2 Fst 0 Lst 10 B 0
General LIH ISE Range Test No # Test name CREZ ∇ Sample type Ser ∇ Page 2/2
Test Name: CREZ ∇ < > Type: Serum ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name CREZ ∇
Test Name: CREZ ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 3900* 5900*
Point 1: # † 3900* 5900*
Point 2 ∇
Point 2:
1-Point Cal. Point: R with CONC-0 Slope Check: # 1-point cal. point
MB type factor
† System Calibrator Cat. No.: OE66300 † System Calibrator Cat. No.: OE66300
2009-11
CREATININE (ENZYMATIC), AU2700/AU5400 Serum/Plasma Application CREATININE (ENZYMATIC), AU680/AU480 Serum/Plasma Application
Test Name: CREZ ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CREZ ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 5.6 µL Dilution 10 µL Pre-Dilution Rate: 1 Sample Volume 5.6 µL Dilution 10 µL OD Limit
Reagents: R1 Volume 120 µL Dilution 0 µL Min OD Max OD Pre-Dilution Rate 1 ∇ Min.OD Max.OD
R2 Volume 40 µL Dilution 0 µL L H Rgt. Volume R1(R1-1) 120 µL Dilution 0 µL Reagent OD Limit
Reagent OD limit: First Low -0.1 High 0.2
Wavelength: Pri. 600 Sec. 700 First L -0.1 First H 0.2 Last Low -0.1 High 0.2
∇ ∇
Common Rgt. Type ф Name ф Correlation Factor A 1 B 0
Measuring Point 1: First 0 Last 27 L 4.4* H 4420*
Measuring Point 2: First 0 Last 10 Correlation Factor:
Method END ∇
Test Name: CREZ ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CREZ ∇ < > Type: Serum ∇
R 3. Sex Year Month Year Month Low High # #
# ∇ # # # # # #
R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # ∇ # # # # # #
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # # # # # # #
∇
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
Panic Value: # # Unit: µmol/L * Decimal places: # 7. No demographics # #
Unit µmol/L * Decimal Places #
General ISE
Test Name: CREZ ∇ < > Type Serum ∇
Test Name: CREZ ∇ < > Type Serum ∇ R Use Serum Cal.
Point 1: # † 3900* 5900*
Point 1: # ∇ † 3900* 5900*
Point 2:
Point 3:
Point 3: ∇
Point 4:
Point 4: ∇ R Reagent Blank
Point 5: Point 5: R Calibration
∇
Point 7: Point 7 Advanced Calibration
∇
1-Point Cal. Point: R with CONC-0 Slope Check: ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
MB Type Factor: ICF: Calibration Stability Period: 14 Point 9 ∇
* Values set for working in SI units (µmol/L). To work in mg/dL divide by 88.4. Calibrator OD Conc Low High Stability
† System Calibrator Cat. No.: OE66300 Point-1 ∇ Reagent Blank 14 Day 0 Hour
MB Type Factor: 1-Point Calibration Point ∇ R with Conc-0
2009-11
CREATININE (ENZYMATIC), AU400/AU640 Urine Application CREATININE (ENZYMATIC), AU600 Urine Application

Test No # Test name CREZ ∇ Sample type Uri ∇ Page 1/2
Test Name: CREZ ∇ < > Type: Urine ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name CREZ ∇ Sample type Uri ∇ Page 2/2
Test Name: CREZ ∇ < > Type: Urine ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
URINE APPLICATION
General ISE Test No # Test name CREZ ∇
Test Name: CREZ ∇ < > Type Urine ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 24000* 41000*
Point 1: # † 24000* 41000*
Point 2 ∇
Point 2:
1-Point Cal. Point: R with CONC-0 Slope Check: # 1-point cal. point
MB type factor
2009-11
CREATININE (ENZYMATIC), AU2700/AU5400 Urine Application CREATININE (ENZYMATIC), AU680/AU480 Urine Application
Test Name: CREZ ∇ < > Type: Urine ∇ Operation Yes ∇
Test Name: CREZ ∇ < > Type: Urine ∇ Operation: Yes ∇

µL µL
R2 Volume 50 µL Dilution 0 µL L H
Reagent OD limit:
Reaction slope: + ∇ Dynamic Range: Common Rgt. Type ф Name ф Correlation Factor A 1 B 0
Measuring Point 2: First 0 Last 10 Correlation Factor: Method END ∇
Linearity Limit %
Test Name: CREZ ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CREZ ∇ < > Type: Urine ∇
# ∇ # # # # # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # # # # # # #
∇
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
Panic Value: # # Unit: µmol/L * Decimal places: # 7. No demographics # #
URINE APPLICATION
Unit µmol/L * Decimal Places #
General ISE
General ISE
Test Name: CREZ ∇ < > Type Urine ∇ R
Test Name: CREZ ∇ < > Type Urine ∇ Use Serum Cal.
Point 1: # † 24000* 41000* Point 1: # ∇ † 24000* 41000*
1-Point Cal. Point: R with CONC-0 Slope Check: ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
MB Type Factor: ICF: Calibration Stability Period: 30
* Values set for working in SI units (µmol/L). To work in mg/dL divide by 88.4. Calibrator OD Conc Low High Stability
† Urine Calibrator Cat. No.: ODC0025 Point-1 ∇ Reagent Blank 30 Day 0 Hour
2009-11
GLUCOSE
OSR6121 4 x 25 mL R1
4 x 12.5 mL R2
OSR6221 4 x 53 mL R1
4 x 27 mL R2
OSR6521 4 x 102 mL R1
4 x 52 mL R2
Intended Use
Enzymatic UV test (hexokinase method) for the quantitative determination of glucose in human serum, plasma, urine, haemolysate and
cerebrospinal fluid on Beckman Coulter AU analysers. For in vitro diagnostic use only.
Glucose reagent OSR6521 for use on the AU2700 and AU5400 systems only.
Summary1,2,3
In the fasting state, blood sugar levels are regulated by the liver, which ensures that levels are maintained within precise limits. The rapid and
precise manner in which fasting blood sugar levels are regulated is in marked contrast to the rapid increase in blood sugar, which occurs during
ingestion of carbohydrates. A fall in blood glucose to a critical level (approximately 2.5 mM) leads to dysfunction of the central nervous system.
This manifests as hypoglycaemia, and is characterised by muscle weakness, lack of coordination and mental confusion. Further decrease in
blood glucose levels leads to hypoglycaemic coma. Blood glucose concentrations show intra-individual fluctuations, which are dependent on
muscular activity and the time interval since food intake. These fluctuations are increased further where there is dysregulation, such as occurs in
a number of pathological conditions in which blood glucose may be elevated (hyperglycaemia) or depressed (hypoglycaemia). Hyperglycaemia
most commonly occurs as a result of a deficiency in either the amount or efficiency of insulin, a condition known as diabetes mellitus. This
disease is characterised by the elevation of blood glucose to such an extent that the renal threshold is exceeded and sugar appears in the urine
(glycosuria). Blood glucose measurement is used as a screening test for diabetes mellitus, where there is suspected hyperglycaemia,
monitoring of therapy in diabetes mellitus, evaluation of carbohydrate metabolism, for example in gestational diabetes acute hepatitis, acute
pancreatitis and Addison’s disease. Hypoglycaemia is associated with a range of pathological conditions including neonatal respiratory distress
syndrome, toxaemia of pregnancy, congenital enzyme defects, Reye’s syndrome, alcohol ingestion, hepatic dysfunction, insulin-producing
pancreatic tumours (insulinomas), insulin antibodies, nonpancreatic neoplasms, septicaemia and chronic renal failure.
CSF glucose may be low or undetectable in patients with acute bacterial, cryptococcal, tubular or carcinomatous meningitis, or in cerebral
abscess, probably due to consumption of glucose by leucocytes or other rapidly metabolising cells. In meningitis or encephalitis due to viral
infections, it is usually normal.
Test Principle4
Glucose is phosphorylated by hexokinase (HK) in the presence of adenosine triphosphate (ATP) and magnesium ions to produce glucose-6-
phosphate and adenosine diphosphate (ADP). Glucose-6-phosphate dehydrogenase (G6P-DH) specifically oxidises glucose-6-phosphate to
+
gluconate-6-phosphate with the concurrent reduction of NAD to NADH. The increase in absorbance at 340nm is proportional to the glucose
concentration in the sample.
Reaction principle
HK, Mg2+
Glucose + ATP Glucose-6-phosphate + ADP
G6P-DH
+ +
Glucose-6-Phosphate + NAD Gluconate-6-P + NADH + H
PIPES buffer (pH 7.6) 24.0 mmol/L
ATP ≥ 2.0 mmol/L
+
NAD ≥ 1.32 mmol/L
2+
Mg 2.37 mmol/L
Hexokinase ≥ 0.59 kU/L
G6P-DH ≥ 1.58 kU/L
Preservative
Reagent Preparation
Specimen
5,6
Serum, EDTA or heparinised plasma. To minimise loss of glucose through glycolysis serum should be removed from red cells as soon as
possible. Specimens that cannot be rapidly separated should be collected into tubes containing fluoride, monoiodoacetate or mannose. Glucose
in stabilised haemolysate and plasma is stable for up to 7 days when stored at 2…8°C and 2 days when stored at 15...25°C. Icteric and strongly
lipemic samples should be avoided.
7
Urine: Fresh, random collections are recommended for urine specimens. Stable in urine for 2 hours when stored at 2…25°C. Analyse as soon
5
as possible.
7
Cerebrospinal fluid: Process immediately to avoid falsely low results.

2010-04
Test Procedure
For Haemolysate application use either of the following reagents; Rolf Greiner Cat No. H10582 or Hatado Cat No. 60690201.
Calibration
Serum/plasma/haemolysate/CSF: Use System Calibrator Cat. No. 66300.
Urine: Use Urine Calibrator Cat. No. ODC0025.
The glucose values of both calibrators are traceable to the National Institute of Standards and Technology (NIST) Standard Reference Material
(SRM) 965.
Quality Control
Serum/plasma/haemolysate: controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman
Coulter system may be used.
Urine: Biorad Liquichek Urine Chemistry Controls Cat. No. 397 and 398 or other control materials with values determined by this Beckman
Coulter system may be used.
CSF: Control materials with values determined by this Beckman Coulter system may be used.
Calculation
The Beckman Coulter analysers automatically compute the glucose concentration of each sample.
Serum/Plasma (fasting) Adults 4.1 – 5.9 mmol/L (74 – 106 mg/dL)
Children 3.3 – 5.6 mmol/L (60 – 100 mg/dL)
Haemolysate Adults 3.3 – 5.5 mmol/L (60 – 100 mg/dL)
Urine 0.1 – 0.8 mmol/L (1 – 15 mg/dL)
CSF Adult 2.2 – 3.9 mmol/L (40 – 70 mg/dL) ≈ 60% of plasma value
9
The generally accepted cut-off levels for the diagnosis of diabetes are:
(a) random plasma glucose of ≥ 11.1 mmol/L
(b) fasting plasma glucose (FPG) ≥ 7.0 mmol/L or
(c) 2-h postload glucose ≥ 11.1 mmol/L during an oral glucose tolerance test (OGTT).
If any one of these criteria is met, results must be confirmed by repeat testing on a subsequent day, unless there is unequivocal hyperglycaemia with acute
metabolic decompensation.
findings.
from these values.
Linearity
The test is linear within a concentration range of 0.6 – 45.0 mmol/L (10 – 800 mg/dL) for serum, plasma, haemolysate and CSF. The test is
linear within a concentration range of 0 – 45 mmol/L (1 – 800 mg/dL) for urine.
Precision
3.27 0.02 0.70 0.04 1.25
6.27 0.03 0.54 0.06 0.97
16.36 0.08 0.51 0.18 1.11
0.46 0.01 1.39 0.01 2.53
11.40 0.09 0.82 0.17 1.46
42.45 0.13 0.31 0.52 1.22
The following data was obtained on an AU2700 using 3 haemolysate analysed over 20 days.
2.25 0.05 2.30 0.09 4.15
5.94 0.09 1.54 0.20 3.41
18.6 0.12 0.67 0.35 1.90
Sensitivity
The lowest detectable level, using serum settings, on an AU600 analyser was estimated at 0.04 mmol/L.
The lowest detectable level, using urine settings, on an AU2700 analyser was estimated at 0.04 mmol/L.
The lowest detectable level represents the lowest measurable level of glucose that can be distinguished from zero. It is calculated as the

2010-04
Method Comparison
Patient serum samples were used to compare this Glucose assay OSR6121 on the AU600 against another commercially available glucose
Patient urine samples were used to compare this Glucose assay OSR6121 on the AU2700 against another commercially available glucose
Patient CSF samples were used to compare this Glucose assay OSR6121 on the AU600 against another commercially available glucose assay.
Results of linear regression analysis were as follows.
y = 0.97x – 0.02 r = 0.991 n = 101 Sample range 1.8 – 7.7 mmol/L
®
Results of haemolysate studies conducted to evaluate the susceptibility of the method to interference were as follows:
®
10
Limitations
Please note that there is a requirement to dedicate a separate test channel specifically for Haemolysate sample settings when assigning this Glucose
application on AU680/480/2700/5400 analyers.

‡ Pretreat all samples: 20 µL sample and 1000 µL haemolysing reagent.
∂ Separate the Haemolysate application from other setting types.
BIBLIOGRAPHY
1. Thomas L. Blood glucose. In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
2. Sacks DB. Carbohydrates. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders Company, 1999; 766-85.
3. Smith AF, Beckett GJ, Walker SW, Rae PWH, eds. Lecture notes on clinical biochemisty, 6th ed.Oxford: Blackwell Science,1998:283pp.
4. Czok R, Barthelmai W. Enzymatische Bestimmungen der Glucose in Blut, Liquor und Harn. Klin Wschr 1962;40:585-589.
serum samples. WHO/DIL/LAB/99.1 Rev.2:32pp, 47pp.
6. Dods RF. Diabetes mellitis. In: Kaplan LA, Pesce AJ, eds. Clinical chemistry theory, analysis, and correlation. St Louis: Mosby, 1996:635pp.
9. Sacks DB, Bruns DE, Goldstein DE, MacLaren NK, McDonald JM, Parrott M. Guidelines and recommendations for laboratory analysis in the diagnosis and
management of diabetes mellitus. Clin Chem 2002;48:436-72.

2010-04
GLUCOSE, AU400/AU640 Serum/Plasma Application GLUCOSE, AU600 Serum/Plasma Application

Test No # Test name GLUC ∇ Sample type Ser ∇ Page ½
Test Name: GLUC ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 2 µL Dilution 0 µL Pre-Dilution Rate: 1 Reagent 1 vol 50 Dil. Vol 150 µL L H
Measuring Point 1: First 0 Last 27 L 0.6* H 45.0* Point 2 Fst 0 Lst 10 B 0
General LIH ISE Range Test No # Test name GLUC ∇ Sample type Ser ∇ Page 2/2
Test Name: GLUC ∇ < > Type: Serum ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name GLUC ∇
Test Name: GLUC ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 21* 35*
Point 1: # † 21* 35*
Point 2 ∇
Point 2:
MB type factor
# User defined
2010-06
GLUCOSE, AU2700/AU5400 Serum/Plasma Application GLUCOSE, AU680/AU480 Serum/Plasma Application
Test Name: GLUC ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: GLUC ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 1.6 Dilution 0 Pre-Dilution Rate: 1 Sample Volume 1.6 µL Dilution 0 µL OD Limit
µL µL
Reagent OD limit:
Measuring Point2 First 0 Last 10 Lipemia ++++ ∇
Linearity Limit % Icterus ++++ ∇
Test Name: GLUC ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: GLUC ∇ < > Type: Serum ∇
# ∇ # # # # # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # # # # # # #
∇
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: GLUC ∇ < > Type Serum ∇ R
Test Name: GLUC ∇ < > Type Serum ∇ Use Serum Cal.
Point 1: # † 18* 27* Point 1: # ∇ † 18* 27*
Point 9 ∇
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 18. Point-1 ∇ Reagent Blank 30 Day 0 Hour
Ф AU680 Point-2 ∇ Calibration 30 Day 0 Hour
2010-06
GLUCOSE, AU400/AU640 Urine Application GLUCOSE, AU600 Urine Application

Test No # Test name GLUC ∇ Sample type Uri ∇ Page 1/2
Test Name: GLUC ∇ < > Type: Urine ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name GLUC ∇ Sample type Uri ∇ Page 2/2
Test Name: GLUC ∇ < > Type: Urine ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
URINE APPLICATION
Test Name: GLUC ∇ < > Type Urine ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 20* 36*
Point 1: # † 20* 36*
Point 2 ∇
Point 2:
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibrations: # ∇ 1-point cal. point
MB type factor
# User defined
2010-06
GLUCOSE, AU2700/AU5400 Urine Application GLUCOSE, AU680/AU480 Urine Application
Test Name: GLUC ∇ < > Type: Urine ∇ Operation Yes ∇
Test Name: GLUC ∇ < > Type: Urine ∇ Operation: Yes ∇
µL µL
Reagent OD limit:
Dynamic Range Low 0* High 44.44*
Measuring Point 1: First 0 Last 27 L 0* H 44.44* Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0
Linearity Limit %
Test Name: GLUC ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: GLUC ∇ < > Type: Urine ∇
# ∇ # # # # # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # # # # # # #
∇
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
URINE APPLICATION
General ISE
General ISE
Test Name: GLUC ∇ < > Type Urine ∇ R
Test Name: GLUC ∇ < > Type Urine ∇ Use Serum Cal.
Point 1: # † 15* 27* Point 1: # ∇ † 15* 27*
Point 9 ∇
2010-06
GLUCOSE, AU400/AU640 Paediatric Application GLUCOSE, AU600 Paediatric Application

Test No # Test name GLUCP ∇ Sample type Ser ∇ Page ½
Test Name: GLUCP ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name GLUCP ∇ Sample type Ser ∇ Page 2/2
Test Name: GLUCP ∇ < > Type: Serum ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
General ISE
Test No # Test name GLUCP ∇
Test Name: GLUCP ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 21* 35*
Point 1: # † 21* 35*
Point 2 ∇
Point 2:
MB type factor
# User defined
2010-06
GLUCOSE, AU2700/AU5400 Paediatric Application GLUCOSE, AU680/AU480 Paediatric Application
Test Name: GLUCP ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: GLUCP ∇ < > Type: Serum ∇ Operation: Yes ∇
µL µL
Reagent OD limit:
Test Name: GLUCP ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: GLUCP ∇ < > Type: Serum ∇
# ∇ # # # # # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # # # # # # #
∇
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: GLUCP ∇ < > Type Serum ∇ R
Test Name: GLUCP ∇ < > Type Serum ∇ Use Serum Cal.
Point 1: # † 18* 27* Point 1: # ∇ † 18* 27*
Point 9 ∇
2010-06
GLUCOSE, AU400/AU640 CSF Application GLUCOSE, AU600 CSF Application

Test No # Test name GLUC ∇ Sample type Others ∇ Page 1/2
Test Name: GLUC ∇ < > Type: Others ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name GLUC ∇ Sample type Others ∇ Page 2/2
Test Name: GLUC ∇ < > Type: Others ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
CSF APPLICATION
Test Name: GLUC ∇ < > Type Others ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 21* 35*
Point 1: # † 21* 35*
Point 2 ∇
Point 2:
MB type factor
# User defined
2010-06
GLUCOSE, AU2700/AU5400 CSF Application GLUCOSE, AU680/AU480 CSF Application
Test Name: GLUC ∇ < > Type: Others ∇ Operation Yes ∇
µL µL
Reagent OD limit:
Linearity Limit %
Test Name: GLUC ∇ < > Type: Others ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: GLUC ∇ < > Type: Others ∇
# ∇ # # # # # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # # # # # # #
∇
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
CSF APPLICATION
General ISE
General ISE
Test Name: GLUC ∇ < > Type Others ∇ R
Test Name: GLUC ∇ < > Type Others ∇ Use Serum Cal.
Point 1: # † 18* 27* Point 1: # ∇ † 18* 27*
1-Point Cal. Point: R With CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
2010-06
GLUCOSE, AU400/AU640 Haemolysate Application GLUCOSE, AU600 Haemolysate Application
Test No # Test name GLUC ∇ Sample type Others ∇ Page 1/2
General LIH ISE Range Test No # Test name GLUC ∇ Sample type Others ∇ Page 2/2
Test Name: GLUC ∇ < > Type: Others ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
Test Name: GLUC ∇ < > Type Others ∇ Cal type 8 AB ∇ Count #
HAEMOLYSATE APPLICATION
Point 1 # ∇ † 140* 184*
Point 1: # † 140* 184*
Point 2 ∇
Point 2:
1-Point Cal. Point: R With CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor
† System Calibrator Cat. No.: 66300. † System Calibrator Cat. No.: 66300
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 18. * Values set for working in SI units (mmol/L). To work in mg/dL multiply by 18.
‡ Pretreat all samples: 20µL + 1000µL Haemolysing Reagent ‡ Pretreat all samples: 20µL + 1000µL Haemolysing Reagent
2010-06
GLUCOSE, AU2700/AU5400 Haemolysate Application∂ GLUCOSE, AU680/AU480 Haemolysate Application∂
Test Name: GLUC ∇ < > Type: Others ∇ Operation: Yes ∇ Test Name: GLUC ∇ < > Type: Others ∇ Operation Yes ∇

Reagent OD limit:
µL µL
Reaction slope: + ∇ Dynamic Range: Dynamic Range Low 0.6* High 45.0*
Measuring Point 1: First 0 Last 27 L 0.6* H 45.0* Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Specific Test Parameters Linearity Limit %
General LIH ISE Range Lag Time Check ∇
Test Name: GLUC ∇ < > Type: Others ∇ Parameters Specific Test Parameters
Normal Ranges: Age L Age H Test Name: GLUC ∇ < > Type: Others ∇
R 1. Value/Flag: # ∇ Low High
# ∇ # # # # # #
# ∇ # # # # # #
R 4. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 5. # ∇ # # # # # # R 2. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 3. # ∇ # # # # # #
7. None Selected # # R 4. # ∇ # # # # # #
8. Out of Range L H # # R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # #
Panic Value: # # Unit: mmol/L* Decimal places: #
General ISE
HAEMOLYSATE APPLICATION
Test Name: GLUC ∇ < > Type Others ∇ General ISE
Test Name: GLUC ∇ < > Type Others ∇ R Use Serum Cal.
Point 2: Point 1: # ∇ † 140* 184*
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 7 ∇ Advanced Calibration
Point 9 ∇
# User defined. <Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
† System Calibrator Cat. No.: 66300. Master Curve> OD Range
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 18. Calibrator OD Conc Low High Stability
‡ Pretreat all samples: 20µL + 1000µL Haemolysing Reagent Point-1 ∇ Reagent Blank 30 Day 0 Hour
∂ Separate the Haemolysate application from other setting types(see IFU for further instruction) MB Type Factor: 1-Point Calibration Point ∇ R with Conc-0
2010-06
GLUCOSE - STAT
OSR6140 4 x 25 mL R1
4 x 12.5 mL R1-2
Intended Use
Enzymatic UV test (hexokinase method) for the quantitative determination of glucose in human serum and plasma on Beckman Coulter
Summary1,2
In the fasting state, blood sugar levels are regulated by the liver, which ensures that levels are maintained within precise limits. The rapid and
precise manner in which fasting blood sugar levels are regulated is in marked contrast to the rapid increase in blood sugar, which occurs during
ingestion of carbohydrates. A fall in blood glucose to a critical level (approximately 2.5 mM) leads to dysfunction of the central nervous system.
This manifests as hypoglycaemia, and is characterised by muscle weakness, lack of coordination and mental confusion. Further decrease in
blood glucose levels leads to hypoglycaemic coma. Blood glucose concentrations show intra-individual fluctuations, which are dependent on
muscular activity and the time interval since food intake. These fluctuations are increased further where there is dysregulation, such as occurs in
a number of pathological conditions in which blood glucose may be elevated (hyperglycaemia) or depressed (hypoglycaemia). Hyperglycaemia
most commonly occurs as a result of a deficiency in either the amount or efficiency of insulin, a condition known as diabetes mellitus. This
disease is characterised by the elevation of blood glucose to such an extent that the renal threshold is exceeded and sugar appears in the urine
(glycosuria). Blood glucose measurement is used as a screening test for diabetes mellitus, where there is suspected hyperglycaemia,
monitoring of therapy in diabetes mellitus, evaluation of carbohydrate metabolism, for example in gestational diabetes acute hepatitis, acute
pancreatitis and Addison’s disease. Hypoglycaemia is associated with a range of pathological conditions including neonatal respiratory distress
syndrome, toxaemia of pregnancy, congenital enzyme defects, Reye’s syndrome, alcohol ingestion, hepatic dysfunction, insulin-producing
pancreatic tumours (insulinomas), insulin antibodies, nonpancreatic neoplasms, septicaemia and chronic renal failure.
Test Principle3
Glucose is phosphorylated by hexokinase (HK) in the presence of adenosine triphosphate (ATP) and magnesium ions to produce glucose-6-
phosphate and adenosine diphosphate (ADP). Glucose-6-phosphate dehydrogenase (G6P-DH) specifically oxidises glucose-6-phosphate to
+
gluconate-6-phosphate with the concurrent reduction of NAD to NADH. The increase in absorbance at 340nm is proportional to the glucose
Reaction principle
2+
Glucose + ATP HK+ Mg Glucose-6-phosphate + ADP
+ G6P-DH +
Glucose-6-Phosphate + NAD Gluconate-6-P + NADH + H

PIPES buffer (pH 7.6) 24.0 mmol/L
ATP ≥2.0 mmol/L
+
NAD ≥1.32 mmol/L
2+
Mg 2.37 mmol/L
Hexokinase ≥0.59 kU/L
G6P-DH ≥1.58 kU/L
Preservative
Reagent Preparation
Slowly add the contents of the bottle labelled R1-2 to the bottle labelled R1. Mix gently by inversion and place on-board the instrument.
The reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C. Once prepared, reagents stored on board the
Specimen
4,5
Serum, EDTA or heparinised plasma : To minimise loss of glucose through glycolysis serum should be removed from red cells as soon as
possible. Specimens that cannot be rapidly separated should be collected into tubes containing fluoride, monoiodoacetate or mannose. Glucose
in stabilised haemolysate and plasma is stable for up to 7 days when stored at 2…8°C and 2 days when stored at 15…25°C. Lipemic and
strongly icteric samples should be avoided.
Test Procedure
Calibration
The calibrator glucose value provided in the calibrator package insert is traceable to the National Institute of Standards and Technology (NIST)
Standard Reference Material (SRM) 965.
Quality Control

2009-08
Calculation
The Beckman Coulter analysers automatically compute the glucose concentration of each sample.
Serum/Plasma (fasting) Adults 4.1 – 5.9 mmol/L (74 – 106 mg/dL)
Children 3.3 – 5.6 mmol/L (60 – 100 mg/dL)
7
The generally accepted cut-off levels for the diagnosis of diabetes are : a) random plasma glucose of ≥ 11.1 mmol/L, (b) fasting plasma glucose
(FPG) ≥ 7.0 mmol/L or (c) 2-h postload glucose ≥ 11.1 mmol/L during an oral glucose tolerance test (OGTT). If any one of these criteria is met,
results must be confirmed by repeat testing on a subsequent day, unless there is unequivocal hyperglycaemia with acute metabolic
decompensation.
findings.
from these values.
Linearity
Precision
3.40 0.03 0.74 0.04 1.06
6.30 0.04 0.55 0.06 1.02
16.13 0.07 0.46 0.14 0.90
Sensitivity
The lowest detectable level represents the lowest measurable level of glucose that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this Glucose OSR6140 assay on the AU600 against another commercially available glucose
y = 0.999x + 0.019 r = 0.997 n = 119 Sample range = 0 – 43.3 mmol/L
®
8
BIBLIOGRAPHY
1. Thomas L. Blood glucose. In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
2. Sacks DB. Carbohydrates. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders Company, 1999;766-85.
3. Czok R, Barthelmai W. Enzymatische Bestimmungen der glucose in blut, liquor und harn. Klin Wschr 1962;40:585-589.
5. Dods RF. Diabetes mellitus. In: Kaplan LA, Pesce AJ, eds. Clinical chemistry theory, analysis, and correlation. St Louis: Mosby, 1996:635pp.
7. Sacks DB, Bruns DE, Goldstein DE, MacLaren NK, McDonald JM, Parrott M. Guidelines and recommendations for laboratory analysis in the diagnosis and
management of diabetes mellitus. Clin Chem 2002;48:436-72.

2009-08
GLUCOSE STAT, AU400/AU640 Serum/Plasma Application GLUCOSE STAT, AU600 Serum/Plasma Application

Test No # Test name GLU-S ∇ Sample type Ser ∇ Page 1/2
Test Name: GLU-S ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name GLU-S ∇ Sample type Ser ∇ Page 2/2
Test Name: GLU-S ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name GLU-S ∇
Test Name: GLU-S ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 21* 35*
Point 1: # † 21* 35*
Point 2 ∇
Point 2:
MB type factor
# User defined
2009-08
GLUCOSE STAT, AU2700/AU5400 Serum/Plasma Application GLUCOSE STAT, AU680/AU480 Serum/Plasma Application
Test Name: GLU-S ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: GLU-S ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Measuring Point2 First Last Lipemia + ∇
Test Name: GLU-S ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: GLU-S ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: GLU-S ∇ < > Type Serum ∇ Test Name: GLU-S Type Serum Use Serum Cal.
∇ < > ∇ ο
Point 1: # † 24* 37* Point 1: # ∇ † 24* 37*
Point 9 ∇
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 18. Point-2 Calibration 30 Day 0 Hour
∇
Ф AU680
2009-08
HDL-CHOLESTEROL
OSR6187 4 x 27 mL R1
4 x 9 mL R2
OSR6287 4 x 51.3 mL R1
4 x 17.1 mL R2
OSR6587 4 x 107 mL R1
4 x 40 mL R2
Intended Use
Enzymatic colour test for the quantitative determination of HDL-cholesterol in human serum and plasma on Beckman Coulter analysers. For in vitro
HDL-Cholesterol reagent OSR6587 for use on the AU2700 and AU5400 systems only.
Summary1
Approximately 25% of total serum cholesterol is transported in the HDL fraction. Numerous clinical and epidemiological studies have demonstrated a
strong inverse association between HDL-cholesterol and the incidence of coronary heart disease. It has been proposed that the uptake and transport of
cholesterol from peripheral tissue to the liver acts as a protective factor against the development of atherosclerotic plaques. Determination of
HDL-cholesterol is therefore essential for the interpretation of individual cholesterol determinations. Low HDL-cholesterol is a risk factor independent of total
cholesterol concentration and is highly predictive of the risk of coronary heart disease. Measurement of HDL-cholesterol is used in the early recognition of
atherosclerosis risk, and may also be used in the monitoring of individuals during treatment with lipid lowering drugs.
Test Principle
Anti human-β-lipoprotein antibody in R1 binds to lipoproteins other than HDL (LDL, VLDL and chylomicrons). The antigen-antibody complexes formed
block enzyme reactions when R2 is added. HDL-cholesterol is quantified by the presence of an enzyme chromogen system.
Reaction Principle
Anti human - β - lipoprotein antibody
LDL,VLDL and chylomicrons Antigen–Antibody complexes
CHE and CHO
HDL-cholesterol + H2O + O2 Cholest-4-en-3-one + Fatty acids + H2O2
POD + -
H2O2 + 4 – AA + F–DAOS Blue dye + F + 2H2O
Anti human-β-lipoprotein antibody Variable
Cholesterol esterase (CHE) 0.8 lU/mL
Cholesterol oxidase (CHO) 4.4 lU/mL
Peroxidase (POD) 1.7 lU/mL
Ascorbate oxidase 2.0 lU/mL
Good’s buffer (pH 7.0) 30 mmol/L
N-Ethyl – N - (2-hydroxy-3-sulfopropyl) - 3.5– dimethoxy – 4 fluoroaniline (F–DAOS) 0.20 mmol/L
Preservative
Detergent
Reagent Preparation
The colour of R2 may turn to light green when stored on board the analyser. This does not affect the performance of the reagent.
Specimen
2
Serum and heparinised plasma (fasting and non-fasting): Stable for 7 days when stored at 2…8°C and 2 days when stored at 15…25°C.
Test Procedure
Calibration
HDL-Cholesterol Calibrator ODC0011.
The calibrator is traceable to the US CDC (Centre for Disease Control) HDL-cholesterol reference method.
Recalibrate the assay every 30 days and perform reagent blank every 7 days, or when the following occur:
Quality Control
HDL/LDL-Cholesterol Control Serum ODC0005 or other control materials with values determined by this method may be used.
2009-08
Calculation
The Beckman Coulter analysers automatically compute the HDL-cholesterol concentration of each sample.
National Cholesterol Education Program (NCEP) guidelines3
< 1.03 mmol/L (< 40 mg/dL) Low HDL-cholesterol (major risk factor for coronary heart disease)
≥ 1.55 mmol/L (≥ 60 mg/dL) High HDL-cholesterol (“negative” risk factor for coronary heart disease)
values.
Linearity
The test is linear within a concentration range of 0.05 - 4.65 mmol/L (2 -180 mg/dL).
Precision
0.69 0.006 0.85 0.013 1.92
1.09 0.007 0.62 0.018 1.69
2.08 0.013 0.61 0.027 1.32
Sensitivity
The lowest detectable level on an AU640 analyser was calculated as 0.002 mmol/L.
The lowest detectable level represents the lowest measurable level of HDL-cholesterol that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient serum samples were used to compare this HDL-Cholesterol OSR6187 assay on the AU640 against another commercially available
HDL-cholesterol assay. Results of linear regression analysis were as follows:
Bilirubin: Interference less than 3% up to 40 mg/dL or 684 µmol/L bilirubin
®
Lipemia: Interference less than 10% up to 900 mg/dL *Intralipid
* No significant interference was observed from samples containing native triglycerides up to 11.3 mmol/L (1000 mg/dL), however there is poor correlation
between lipemia and triglyceride concentration – see limitations section.
4
Limitations
When triglyceride in a sample exceeds 11.3 mmol/L (1000 mg/dL), dilute the sample with a saline solution, repeat assay and multiply result by dilution
factor.
† HDL Cholesterol Calibrator Cat. No.: ODC0011
‡ Perform reagent blank every 7 days
BIBLIOGRAPHY
1. Riesen WF. Lipid metabolism. In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
plasma and serum samples. WHO/DIL/LAB/99.1 Rev.2:26pp.
3. National cholesterol education program expert panel. Executive summary of the third report of the National Cholesterol Education Program (NCEP)
expert panel on detection, evaluation, and treatment of high blood cholesterol in adults (Adult Treatment Panel III). JAMA 2001;285:2486-2497.
th

2009-08
HDL CHOLESTEROL, AU400/AU640 Serum/Plasma Application HDL CHOLESTEROL, AU600 Serum/Plasma Application

Test No # Test name HDL-c ∇ Sample type Ser ∇ Page 1/2
Test Name: HDL-c ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name HDL-c ∇ Sample type Ser ∇ Page 2/2
Test Name: HDL-c ∇ < > Type: Serum ∇ Level L Level H

ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name HDL-c ∇
Test Name: HDL-c ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 5* 8*
Point 1: # † 5* 8*
Point 2 ∇
Point 2:
Point 3 ∇
Point 3:
Point 4 ∇
Point 4:
1-point cal. point
MB type factor
MB Type Factor: Calibration Stability Period: 30‡ Calibrator stability period 30‡
† HDL Cholesterol Calibrator Cat. No.: ODC0011 † HDL Cholesterol Calibrator Cat. No.: ODC0011
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 38.7 * Values set for working in SI units (mmol/L). To work in mg/dL multiply by 38.7
‡ Perform reagent blank every 7 days ‡ Perform reagent blank every 7 days
2009-08
HDL CHOLESTEROL, AU2700/AU5400 Serum/Plasma Application HDL CHOLESTEROL, AU680/AU480 Serum/Plasma Application
Test Name: HDL-C ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: HDL-C ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 1.6 μL Dilution 0 μL Pre-Dilution Rate: 1 Sample Volume 1.6 μL Dilution 0 μL OD Limit
Reagents: R1 Volume 144 Dilution 0 Min OD Max OD Pre-Dilution Rate 1 ∇ Min.OD Max.OD
μL μL
Reagent OD limit:
Measuring Point 1: First 0 Last 27 L 0.05* H 4.65*
Method END ∇
Measuring Point1 First 0 Last 27 LIH Influence Check: #
Measuring Point2 First 0 Last 10 Lipemia +++++
Linearity Limit % Icterus +++++
Specific Test Parameters Lag Time Check ∇ Hemolysis +++++
Test Name: HDL-C ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # Level L: # Level H: # Test Name: HDL-C ∇ < > Type: Serum ∇
∇
Normal Ranges: Age L Age H Value/Flag: # Low High
∇
ο 1. # ∇ # # # # # #
ο 3. # ∇ # # # # # # ο 1. # ∇ # # # # # #
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Test Name: HDL-C ∇ < > Type Serum ∇ Test Name: HDL-C Type Serum Use Serum Cal.
∇ < > ∇ ο
Point 1: # † 5* 8* Point 1: # ∇ † 5* 8*
Point 9 ∇
MB Type Factor: Calibration Stability Period: 30‡
† HDL Cholesterol Calibrator Cat. No.: ODC0011 Point-1 Reagent Blank 7 Day 0 Hour
∇
Point-2 ∇ Calibration 30 Day 0 Hour
‡ Perform reagent blank every 7 days
ф AU680 MB Type Factor: 1-Point Calibration Point ∇ ο with Conc-0
2009-08
INORGANIC PHOSPHOROUS
OSR6122 4 x 15 mL R1
4 x 15 mL R2
OSR6222 4 x 40 mL R1
4 x 40 mL R2
Intended Use
Photometric UV test for the quantitative determination of inorganic phosphorous in human serum, plasma and urine on Beckman Coulter
Summary1,2,3,4
In plasma and serum the majority of phosphate exists in the inorganic form (Pi), approximately 15% bound to protein and the remainder in
complexed and free forms. Serum phosphate concentrations are dependent on diet and variation in the secretion of hormones such as PTH.
Intracellularly phosphate occurs primarily as organic phosphate however a small but extremely important fraction exists as inorganic phosphate
which, because it is a substrate for oxidative phosphorylation, participates in reactions concerned with generation of metabolic energy. About
85% of extracellular phosphate occurs in the Pi form as hydroxyapatite thereby playing an important role in bone structure.
Hypophosphataemia (phosphate depletion) is relatively common in hospitalised patients and is found in up to 30% of surgical patients.
Hypophosphataemia is caused by a decreased intake or absorption of phosphate such as occurs in Vit D deficiency, malabsorption, use of oral
phosphate binders and primary PTH excess; increased excretion such as occurs in secondary PTH excess, post renal transplant and re-feeding
starved patients; and from redistribution of phosphate e.g. hyperalimentation, recovery from diabetic ketoacidosis and respitatory alkalosis.
Hyperphosphataemia is caused by increased intake such as occurs in intravenous therapy and phosphate enemas; reduced excretion such as
occurs in acute and chronic renal failure, low PTH or resistance to PTH and vitamin D toxicity; and redistribution of phosphate that occurs in
tumour lysis, rhabdomyolysis and heat stroke.
Test Principle5,6
Inorganic phosphorous reacts with molybdate to form a heteropolyacid complex. The use of a surfactant eliminates the need to prepare a
protein free filtrate. The absorbance at 340/380 nm is directly proportional to the inorganic phosphorous concentration in the sample.
Reaction principle
6- +
7 H3PO4 + 12 (Mo7O24) + 72 H 7 H3PO4(MoO3)12 + 36 H2O

Sulphuric acid 200 mmol/L
Ammoniumheptamolybdate 0.35 mmol/L
Glycine 50 mmol/L
Preservative

Irritant. R36/38 Irritating to eyes and skin.
Safety Phrases:
S26, S37, S45, S60: In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable gloves. In case
of accident or if you feel unwell, seek medical advice immediately (show the label where possible). This material and its container must be
disposed of as hazardous waste.
Reagent Preparation

The reagents are stable, unopened, up to the stated expiry date when stored at 2...8°C. Once open, reagents stored on board the instrument
Specimen
7
Serum and heparinised plasma: Stable in serum for 4 days when stored at 2…8°C and 1 day when stored at 15…25°C.
Strongly haemolysed samples should be avoided.
8
Urine: Acidified with 6M HCl. Collect timed 24-hour specimen using standard laboratory procedures. Store at 2...8°C.
Test Procedure
statement.
Calibration
Use System Calibrator Cat. No. 66300 for serum application and Urine Calibrator Cat. No. ODC0025 for urine application.
The inorganic phosphorous values of both calibrators are traceable to a Beckman Coulter Master Calibrator.

2009-08
Quality Control
the serum application.
Calculation
The Beckman Coulter analysers automatically compute the inorganic phosphorous concentration of each sample.
Serum Adults 0.81 – 1.45 mmol/L (2.5 – 4.5 mg/dL)
Children 1.29 – 2.26 mmol/L (4.0 – 7.0 mg/dL)
Urine On non-restricted diet 12.9 – 42.0 mmol/d (0.4 – 1.3 g/day)
findings.

from these values.
Linearity
The test is linear within a concentration range of 0.32 – 6.40 mmol/L (1 – 20 mg/dL) for serum.
The test is linear within a concentration range of 0 – 113 mmol/L (0 – 350 mg/dL) for urine.
Precision
0.96 0.01 1.03 0.01 1.55
1.64 0.01 0.63 0.02 1.33
3.36 0.02 0.61 0.04 1.23
9.54 0.13 1.41 0.28 2.99
32.96 0.23 0.71 0.51 1.55
87.35 0.62 0.71 1.14 1.30
Sensitivity
The lowest detectable level using serum settings on an AU600 analyser was estimated at 0.10 mmol/L.
The lowest detectable level using urine settings on an AU2700 analyser was estimated at 0.48 mmol/L.
The lowest detectable level represents the lowest measurable level of Inorganic phosphorous that can be distinguished from zero. It is
calculated as the absolute mean plus three standard deviations of 20 replicates of an analyte free sample.
Method Comparison
Patient serum samples were used to compare this Inorganic phosphorous assay on the AU600 against another commercially available inorganic
phosphorous assay. Results of linear regression analysis were as follows:
Patient urine samples were used to compare this Inorganic phosphorous assay on the AU2700 against another commercially available inorganic
phosphorous assay. Results of linear regression analysis were as follows:
®
9

‡ Dilute samples and Urine Calibrator Cat. No. ODC0025 1:10 with purified H2O.
† System Calibrator Cat. No.: 66300/ Urine Calibrator Cat. No.: ODC0025

2009-08
BIBLIOGRAPHY
1. Smith AF, Beckett GJ, Walker SW, Rae PWH, eds. Lecture notes on clinical biochemisty, 6th ed.Oxford: Blackwell Science,1998:75pp.
2. Endres DB, Rude RK. Mineral and bone metabolism. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia: WB Saunders Company,
1999;1406-1441.
3. Fraser D, Jones G, Kooh SW, Radde IC. Calcium and phosphate metabolism. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders
Company, 1987:706pp.
4. Thomas L. Phosphate. In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
5. Daly JA, Ertingshausen G. Direct method for determining inorganic phosphate in serum with the “CentrifiChem”. Clin Chem 1972;18(3):263-5.
6. Gamst O, Try K. Determination of serum-phosphate without deproteinization by ultraviolet spectrophotometry of the phosphomolybdic acid complex. Scand J Clin
Lab Invest 1980;40(5):483-6.
8. NCCLS. Urinalysis and collection, transportation, and preservation of urine specimens; approved guideline. NCCLS Document GP16-A2, 2nd ed. Pennsylvania:
NCCLS, 2001.

2009-08
INORGANIC PHOSPHOROUS, AU400/AU640 Serum/Plasma Application INORGANIC PHOSPHOROUS, AU600 Serum/Plasma Application

Test No # Test name PHOS ∇ Sample type Ser ∇ Page ½
Test Name: PHOS ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name PHOS ∇ Sample type Ser ∇ Page 2/2
Test Name: PHOS ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name PHOS ∇
Test Name: PHOS ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 8* 13*
Point 1: # † 8* 13*
Point 2 ∇
Point 2:
1-point cal. Point
1-Point Cal. Point: ο With CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ MB type factor
2009-08
INORGANIC PHOSPHOROUS, AU2700/AU5400 Serum/Plasma Application INORGANIC PHOSPHOROUS, AU680/AU480 Serum/Plasma Application
Test Name: PHOS ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: PHOS ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: PHOS ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: PHOS ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: PHOS Type Serum
∇ < > ∇ Test Name: PHOS Type Serum Use Serum Cal.
∇ < > ∇ ο
Point 1: # † 8* 13* Point 1: # ∇ † 8* 13*
Point 9 ∇
2009-08
INORGANIC PHOSPHOROUS, AU400/AU640 Urine Application INORGANIC PHOSPHOROUS, AU600
System Reagent: OSR6122, OSR6222 Reagent ID: 022 Manual Dilution Standard Mode Urine Application
Specific test parameters
Test No # Test name PHOS ∇ Sample type Uri ∇ Page 1/2
Test Name: PHOS ∇ < > Type: Urine ∇ Operation: Yes ∇
Sample vol. 2‡ Dil. vol 0 μL Min. OD Max. OD
Reagent 1 vol 30 Dil. vol 120 μL L H
Reagent 2 vol 30 Dil. vol 120 μL Reagent OD limit
Fst. L -0.1 Fst. H 0.1
Reagent OD limit:
Wave Main 340 Sub 380 ∇ Lst. L -0.1 Lst. H 0.5
Method END ∇ Dynamic range
Reaction + ∇ L 0* H 113*
Point 1 Fst 0 Lst 27 Correlation factor A 1
Point 2 Fst 0 Lst 10 B 0
Sample Pre-dil. Rate ¤
Specific Test Parameters Select using Space key, or select from list displayed by Guide key
Test No # Test name PHOS ∇ Sample type Uri ∇ Page 2/2
Test Name: PHOS ∇ < > Type: Urine ∇ Level L Level H
Normal range
Sex Age L Age H L H
1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # #
2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # #
3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # #
4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # #
5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # #
6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # #
7 Non select # #
8 Out of range # #
L H
URINE APPLICATION

General ISE
Test No # Test name PHOS ∇
Test Name: PHOS ∇ < > Type Urine ∇
Point 1: # † 120* 230* Point 1 # ∇ † 120* 230*
1-Point Cal. Point: ο With CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. point
# User defined
† Urine Calibrator Cat. No.: ODC0025 ‡ Dilute samples and Urine Calibrator Cat. No. ODC0025 1:10 with purified H2 0.
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 3.1 # User defined ¤ Analyser default value
2009-08
INORGANIC PHOSPHOROUS, AU2700/AU5400 Urine Application INORGANIC PHOSPHOROUS, AU680/AU480 Urine Application
Test Name: PHOS ∇ < > Type: Urine ∇ Operation Yes ∇
Test Name: PHOS ∇ < > Type: Urine ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Linearity Limit %
Test Name: PHOS ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: PHOS ∇ < > Type: Urine ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
URINE APPLICATION
General ISE
General ISE
Test Name: PHOS ∇ < > Type Urine ∇ Test Name: PHOS ∇ < > Type Urine ∇ ο Use Serum Cal.
Point 1: # † 120* 230* Point 1: # ∇ † 120* 230*
Point 9 ∇
2009-08
IRON
OSR6186 4 x 15 mL R1
4 x 15 mL R2
OSR6286 4 x 30 mL R1
4 x 30 mL R2
Intended Use
Photometric colour test for the quantitative determination of iron in human serum and plasma on Beckman Coulter analysers. For in vitro diagnostic use
only.
Summary1,2,3
Iron participates in a variety of vital processes in the body varying from cellular oxidative mechanisms to the transport and delivery of oxygen to body cells.
It is a constituent of the oxygen-carrying chromoproteins, haemoglobin and myoglobin, as well as various enzymes, such as cytochrome oxidase and
peroxidases. The remaining body iron is present in the flavoproteins, the iron-sulphur proteins, as well as storage iron-ferritin and transport iron-transferrin.
Measured serum iron concentration is principally the Fe (III) bound to serum transferrin and does not include the iron contained in serum as free
haemoglobin.
Serum iron concentration is decreased in many but not all patients with iron deficiency anemia; in acute or chronic inflammatory disorders such as acute
infection, immunisation, and myocardial infarction; acute or recent haemorrhage; malignancy; kwashiorkor; late pregnancy; menstruation and nephrosis.
Serum iron concentration diminishes markedly in patients who are beginning to respond to specific therapy for anemias of other causes e.g. treatment of
pernicious anemia with Vit B12. Greater than normal concentrations of serum iron occur in iron-overload disorders such as haemochromatosis and in acute
iron poisoning following oral or parenteral iron administration. Iron levels may also be increased in acute hepatitis, lead poisoning, acute leukemia,
thalassemia or oral contraception.
Test Principle4,5,6
The method utilises TPTZ [2,4,6-Tri-(2-pyridyl)-5-triazine] as the chromogen. In an acidic medium, transferrin-bound iron dissociates into free ferric ions and
apo-transferrin. Hydrochloric acid and sodium ascorbate reduce the ferric ions to the ferrous state. The ferrous ions then react with TPTZ to form a blue
coloured complex which can be measured bichromatically at 600/800 nm. The increase in absorbance is directly proportional to the amount of iron present.
Reaction Principle
3+ Buffer 3+
Transferrin 2(Fe ) 2(Fe ) + Apo-transferrin
3+ 2+ +
2 Fe + Ascorbic Acid + 2 H2O 2 Fe + Dehydroascorbic Acid + 2 H3O
2+ 2+
Fe + TPTZ Iron-complex (blue coloured complex)

Glycine buffer (pH 1.7) 215 mmol/L
L-ascorbic acid 4.7 mmol/L
2,4,6-Tri(2-pyridyl)-5-triazine 0.5 mmol/L
Preservative
Reagent Preparation
60 days.
Some discoloration may be observed in R1 as the reagent ages. This does not affect the performance of the reagent.
Specimen
Serum and heparinised plasma. Do not use EDTA, oxalate or citrate plasma.
7
Lipemic samples should be avoided. Haemolysed samples may react with the reagent to produce spuriously low results and such specimens should not
be used. Remove serum from red cells immediately to avoid haemolysis.
8
Samples should be taken in the morning from patients in a fasting state, since iron values can decrease by 30% during the course of the day.
Test Procedure
The paediatric application is suitable for use with small volume serum/plasma samples
Calibration
The calibrator iron value provided in the calibrator package insert is traceable to a Beckman Coulter Master Calibrator.
Quality Control
Control Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter system may be used.

2009-08
are tested and each time calibration is performed.
Calculation
The Beckman Coulter analysers automatically compute the iron concentration of each sample.
Serum (Adults) Male 12.5 – 32.2 µmol/L ( 70 – 180 µg/dL)
Female 10.7 – 32.2 µmol/L ( 60 – 180 µg/dL)
Serum (Children) Newborn 17.90 – 44.8 µmol/L (100 – 250 µg/dL)
Infant 7.2 – 17.9 µmol/L (40 – 100 µg/dL)
Child 9.0 – 21.5 µmol/L (50 – 120 µg/dL)
values.
Linearity
The test is linear within a concentration range of 2 – 179 µmol/L (10 – 1000 µg/dL).
Precision
9.59 0.10 1.02 0.20 2.09
28.34 0.19 0.66 0.50 1.77
105.54 0.68 0.65 1.30 1.23
Sensitivity
The lowest detectable level represents the lowest measurable level of iron that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison
Patient serum samples were used to compare this Iron OSR6186 assay on the AU600 against another commercially available iron assay. Results of linear
y = 1.002x – 0.304 r = 0.999 n = 96 Sample range = 1.9 – 41.6 µmol/L
®
Copper: Interference less than 10% up to 1 mg/dL or 0.157 mmol/L copper
Globulin: Interference less than 10% up to 2 g/dL or 20 g/L
Triglyceride: Interference less than 10% up to 300 mg/dL or 3.4 mmol/L triglyceride
11
Limitations
In rare instances, extremely high concentrations of monoclonal immunoglobulins, due to monoclonal gammopathies, may cause turbidity in the reaction
12
cuvette and elevate direct colorimetric iron assays.
* Values set for working in SI units (µmol/L). To work in µg/dL multiply by 5.585.
BIBLIOGRAPHY
rd
1. Tietz NW, ed. Clinical guide to laboratory tests, 3 ed. Philadelphia: WB Saunders Company,1995:374-375.
2. Fairbanks VF, Klee GG. Biochemical aspects of hematology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia: WB
3. Woo J, Henry JB. Metabolic intermediates and inorganic ions. In: Henry JB, ed. Clinical diagnosis and management by laboratory methods.
Philadelphia:WB Saunders Company, 1996:188-190.
4. Schade A, Ogama J, Reinhart R, MillerJ, ed. Proc Soc Exp Biol Med 1954; 87: 442.
5. Goodwin JF, Murphy B, Guillemette M. Direct measurement of serum iron and binding capacity. Clin Chem 1966;12:47-57.
6. Diehl H, SmithG.F. The Iron Reagents: Bathophenanthroline Bathophenanthroline-Disulfonic acid 2,4,6-Tripyridyl-s-triazine Phenyl-2-pyridyl Ketoxime.
nd
In:GF Smith 2 ed. Ohio: Chem Co 1960.
7. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of anticoagulants in Diagnostic laboratory investigations and stability of blood,
8. Perrotta G. Iron and total iron binding capacity. In: Kaplan LA, Pesce AJ, eds. Clinical chemistry theory, analysis, and correlation. St Louis: Mosby,
1996:714pp.
9. NCCLS. Determination of serum iron, total iron-binding capacity and percent transferrin saturation; approved standard. NCCLS Document HI7-A.
Pennsylvania: NCCLS, Dec 1998.
th
12. Bakker AJ. Influence of monoclonal immunoglobulins in direct determinations of iron in serum. Clin Chem 1991;37:690-4.

2009-08
IRON, AU400/AU640 Serum/Plasma Application IRON, AU600 Serum/Plasma Application

Test No # Test name IRON ∇ Sample type Ser ∇ Page 1/2
Test Name: IRON ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name IRON ∇ Sample type Ser ∇ Page 2/2
Test Name: IRON ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name IRON ∇
Test Name: IRON ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 680* 1038*
Point 1: # † 680* 1038*
Point 2 ∇
Point 2:
1-Point Cal. Point: with CONC-0 Slope Check: None Advanced Calibration: # 1-point cal. point
ο ∇ ∇
MB type factor
* Values set for working in SI units (µmol/L). To work in µg/dL multiply by 5.585 * Values set for working in SI units (µmol/L). To work in µg/dL multiply by 5.585
2009-08
IRON, AU2700/AU5400 Serum/Plasma Application IRON, AU680/AU480 Serum/Plasma Application
Test Name: IRON ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: IRON ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Measuring Point2 First 0 Last 10 Lipemia + ∇
Specific Test Parameters Lag Time Check ∇ Hemolysis + ∇
Test Name: IRON ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: IRON ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: IRON ∇ < > Type Serum ∇ ο Use Serum Cal.
Test Name: IRON ∇ < > Type Serum ∇
Point 1: # ∇ † 716* 1074*
Point 1: # † 716* 1074*
MB Type Factor: Calibration Stability Period: 999 Point 10 ∇ Interval (RB/ACAL) # ∇
# User defined.
* Values set for working in SI units (µmol/L). To work in µg/dL multiply by 5.585 Point-2 ∇ Calibration 999 Day 0 Hour
2009-08
IRON, AU400/AU640 Paediatric Application IRON, AU600 Paediatric Application

Test No # Test name IRONP ∇ Sample type Ser ∇ Page 1/2
Test Name: IRONP ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name IRONP ∇ Sample type Ser ∇ Page 2/2
Test Name: IRONP ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name IRONP ∇
Test Name: IRONP ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 680* 1038*
Point 1: # † 680* 1038*
Point 2 ∇
Point 2:
1-Point Cal. Point: With CONC-0 Slope Check: None # 1-point cal. point
MB type factor
* Values set for working in SI units (µmol/L). To work in µg/dL multiply by 5.585 * Values set for working in SI units (µmol/L). To work in µg/dL multiply by 5.585
2009-08
IRON, AU2700/AU5400 Paediatric Application IRON, AU680/AU480 Paediatric Application
Test Name: IRONP ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: IRONP ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Measuring Point2 First 0 Last 10 Lipemia + ∇
Specific Test Parameters Lag Time Check ∇ Hemolysis + ∇
Test Name: IRONP ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: IRONP ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: IRONP ∇ < > Type Serum ∇ ο Use Serum Cal.
Test Name: IRONP ∇ < > Type Serum ∇
Point 1: # ∇ † 716* 1074*
Point 1: # † 716* 1074*
MB Type Factor: Calibration Stability Period: 999 Point 10 ∇ Interval (RB/ACAL) # ∇
# User defined. Point-1 ∇ Reagent Blank 999 Day 0 Hour
† System Calibrator Cat. No.: 66300 Point-2 Calibration 999 Day 0 Hour
∇
* Values set for working in SI units (µmol/L). To work in µg/dL multiply by 5.585
2009-08
LACTATE
OSR6193 4 x 10 mL R1 Buffer
4x R1 Lyo
Intended Use
Enzymatic colour test for the quantitative determination of L-Lactate in human plasma and cerebrospinal fluid (CSF) on Beckman Coulter analysers. For
Summary1,2,3
L-lactate is the end product of anaerobic glycolysis. It is derived predominantly from white skeletal muscle, brain, skin, renal medulla and erythrocytes.
Lactate dehydrogenase catalyses the reduction of pyruvate to lactate. There are two major clinical settings in which lactic acidosis occurs (1) conditions
associated with hypoxia eg. shock, congestive heart failure, myocardial infarction, blood loss and pulmonary oedema (2) metabolic or drug/toxin related
disorders. Examples of metabolic disorders include diabetes mellitus, hepatic disease and neoplasia. Congenital metabolic disorders include type I
glycogen storage disease. Examples of drugs/toxins which give rise to elevated lactate are methanol, ethanol, epinephrine and acetaminophen.
Lactate levels in CSF will generally mirror those in blood/plasma. However, increased lactate levels in CSF in the absence of increased blood/plasma
lactate concentration have been reported in cases of bacterial meningitis, cerebral hypoxia, ischemia and in certain inborn errors of metabolism
eg. pyruvate dehydrogenase deficiency, mitochondrial myopathies and biotinidase deficiency.
Test Principle4,5
L-lactate is oxidised to pyruvate and hydrogen peroxide by lactate oxidase (LOD). A coloured product is produced by the reaction of peroxidase (POD),
hydrogen peroxide, 4 -aminoantipyrine and a hydrogen donor (TOOS). The coloured product is measured photometrically. The colour intensity is
proportional to the concentration of lactate in the sample.
Reaction Principle
LOD
L-Lactate + O2 Pyruvate + H2O2
POD
H2O2 + 4–AA + H donor Chromogen + 2H2O

Lactate oxidase ≥ 0.2 kU/L
Peroxidase ≥ 1 kU/L
Good’s Buffer (pH 7.0) 50 mmol/L
4-aminoantipyrine 0.1 mmol/L
TOOS* ≥ 0.3 mmol/L
* N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methylaniline
Preservative

Reagent Preparation
R1: Dissolve the contents of one vial of R1 Lyo completely with the contents of one vial of R1 buffer. Mix gently by inversion and place on board the
instrument.
A slight pink colouration of the working reagent will not influence performance. The reagent can still be used providing the reagent blank is within
specification.

30 days.
Specimen1,6,7,8,9
Plasma or cerebrospinal fluid. Do not use serum.
Plasma: Use plasma from blood collected into sodium fluoride-potassium oxalate tubes.
Sample Stability: Stable for 14 days when stored at 2…8°C and 8 hours when stored at 15…25°C.
Glycolysis resulting from physical exercise gives rise to an increased lactate concentration in the bloodstream. Therefore, the patient should be at rest
before taking the sample. In particular, movement of the hand or arm should be avoided. Keep the sample on ice and separate plasma from cells within
15 minutes of collection. Analyse the sample immediately. Note whether the sample is venous or arterial. Icteric and haemolysed samples should be
avoided.
Cerebrospinal Fluid: CSF collected into plain collection devices or sodium fluoride/potassium oxalate tubes.
Sample Stability: Analyse fresh otherwise stable stored at 2…8°C for up to 24 hours.
Test Procedure
Calibration
The calibrator is traceable to a primary standard which is gravimetrically prepared using reagent grade L-lactate. Recalibrate the assay every 30 days or
when the following occur:
Major preventative maintenance was performed on the analyser or a critical part was replaced

2009-08
Quality Control
Each laboratory should establish its own control frequency, however, good laboratory practice suggests that controls be tested each day patient samples
acceptable ranges given in the relevant product literature. If any trends or sudden shifts in values are detected, review all operating parameters.
Calculation
The Beckman Coulter analysers automatically compute the lactate concentration of each sample.
0.5 – 2.2 mmol/L (4.5 – 19.8 mg/dL) Plasma
1.1 – 6.7 mmol/L (10 – 60 mg/dL) CSF, neonate
1.1 – 4.4 mmol/L (10 – 40 mg/dL) CSF, 3 – 10 days old
1.1 – 2.8 mmol/L (10 – 25 mg/dL) CSF, >10 days old
1.1 – 2.4 mmol/L (10 – 22 mg/dL) CSF, adult

values.
Linearity
Precision
The following data was obtained on an AU640 using 3 pools analysed over 20 days.
1.15 0.007 0.6 0.021 1.8
4.35 0.034 0.8 0.090 2.1
13.50 0.113 0.8 0.231 1.7
Sensitivity
The lowest detectable level on an AU640 analyser was calculated at 0.001 mmol/L.
The lowest detectable level represents the lowest measurable level of lactate that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient plasma samples were used to compare this Lactate OSR6193 assay on the AU640 against another commercially available lactate assay. Results
Patient CSF samples were used to compare this Lactate OSR6193 assay on the AU400 against another commercially available lactate assay. Results of
Results of studies conducted to evaluate the susceptibility of the method to interference using an artificially manufactured matrix were as follows:
®
10
Limitations11
Ascorbate interference may be observed at concentrations greater than 10 mg/dL, although ascorbate is generally cleared via urinary excretion within
4 hours of ingestion. Plasma ascorbate concentrations at tissue saturation are reported to be 1 – 1.5 mg/dL. Studies conducted at Beckman Coulter show
that there is no observed interference from ascorbate at this concentration.

BIBLIOGRAPHY
1. Burtis CA, Ashwood ER, ed. Tietz textbook of clinical chemistry. 3rd ed. Philadelphia: WB Saunders Company, 1999:382pp.
2. Henry JB, ed. Clinical diagnosis and management by laboratory methods. 19th ed. Philadelphia: WB Saunders Company, 1999:787pp.
3. Hutchesson A, Preece MA, Gray G, Green A. Measurement of lactate in cerebrospinal fluid in investigation of inherited metabolic disease. Clin Chem
1997;43:158-161.
4. Trinder P. Determination of glucose in blood using glucose oxidase with an alternative oxygen acceptor. Ann Clin Biochem 1969;6:24-27.
5. Barham D, Trinder P. An improved colour reagent for the determination of blood glucose by the oxidase system. Analyst 1972;97:142-145.
6. Tietz NW. Clinical guide to laboratory tests, 3rd ed. Philadelphia, WB Saunders Co.;1995.
7. Astles R, Williams CP, Sedor F. Stability of plasma lactate in vitro in the presence of antiglycolytic agents. Clin Chem 1994;40:1327-1330.
8. Westgard JO, Lahmeyer BL, Birnbaum ML. Use of the Du Pont “automatic clinical analyzer” in direct determination of lactic acid in plasma stabilized with sodium
fluoride. Clin Chem 1972;18:1334-1338.
9. Thomas L. Lactate. In: Thomas L, 1st ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
Verlagsgesellschaft, 1998,160pp.
10. Young DS. Effects of drugs on clinical laboratory tests, 5th ed. AACC Press, Washington D.C, 2000.
11. Omaye ST, Turnbull JD, Sauberbich HE: Selected Methods for Determination of Ascorbic Acid in Animal Cells, Tissues and Fluids. In: Methods of
Enzymeology. 1979; 62: 3-11.

2009-08
LACTATE, AU400/AU640 Plasma/CSF Application LACTATE, AU600 Plasma/CSF Application

Test No # Test name LAC ∇ Sample type Ser ∇ Page 1/2
Test Name: LAC ∇ < > Type: Serum ∇ Operation: Yes ∇
Method: FIXED ∇ Last L -0.1 Last H 0.2 Reaction + ∇ L 0.22* H 13.32*
General LIH ISE Range Test No # Test name LAC ∇ Sample type Ser ∇ Page 2/2
Test Name: LAC ∇ < > Type: Serum ∇ Level L Level H

ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name LAC
PLASMA/CSF APPLICATION
∇
Test Name: LAC ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 8.1* 12.2*
Point 1: # † 8.1* 12.2*
Point 2 ∇
Point 2:
Point 3 ∇
Point 3:
Point 4 ∇
Point 4:
1-point cal. point
MB type factor
† System Calibrator Cat No: 66300 † System Calibrator Cat No: 66300.
* Values set for working in SI units mmol/L. To work in (mg/dL) multiply by 9.01 * Values set for working in SI units mmol/L. To work in (mg/dL) multiply by 9.01
2009-08
LACTATE, AU2700/AU5400 Plasma/CSF Application LACTATE, AU680/AU480 Plasma/CSF Application
Test Name: LAC ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: LAC ∇ < > Type: Serum ∇ Operation: Yes ∇
Reagents: R1 Volume 160 Dilution 0 Min OD Max OD Pre-Dilution Rate 1 ∇ Min.OD -0.1 Max.OD 2.5
μL μL
Reagent OD limit:
Method FIXED ∇
Measuring Point1 First 1 Last 15 LIH Influence Check #
Measuring Point2 First Last Lipemia +++++
Linearity Limit % Iceterus +++
Specific Test Parameters Lag Time Check ∇ Hemolysis +++++
Test Name: LAC ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # Level L: # Level H: # Test Name: LAC ∇ < > Type: Serum ∇
∇
Normal Ranges: Age L Age H Value/Flag: # Low High
∇
ο 1. # ∇ # # # # # #
ο 3. # ∇ # # # # # # ο 1. # ∇ # # # # # #
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
PLASMA/CSF APPLICATION
General ISE
Test Name: LAC ∇ < > Type Serum ∇ Test Name: LAC ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇ Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # ∇
Point 1: # † 8.1* 12.2* Point 1: # ∇ † 8.1* 12.2*
Point 9 ∇
† System Calibrator Cat No: 66300 Point-1 Reagent Blank 30 Day 0 Hour
∇
* Values set for working in SI units mmol/L. To work in (mg/dL) multiply by 9.01
ф AU680
2009-08
LDL-CHOLESTEROL
OSR6183 4 x 27 mL R1
4x 9 mL R2
OSR6283 4 x 51.3 mL R1
4 x 17.1 mL R2
Intended Use
Enzymatic colour test for the quantitative determination of LDL-cholesterol in human serum and plasma on Beckman Coulter AU analysers. For in vitro
Summary1,2,3
LDL-cholesterol (LDL C) constitutes the largest portion of the LDL molecule, formed via the action of lipoprotein lipase on VLDL. LDL-cholesterol plays a
causal role in the development of coronary heart disease (CHD), with numerous clinical and epidemiological studies demonstrating its atherogenic
properties. LDL-cholesterol has the strongest association with coronary mortality of all lipid and lipoprotein variables (GRIPS study), with a combination of
raised LDL-cholesterol and elevated triglyceride levels constituting an especially high risk. LDL-cholesterol evaluation provides early recognition of
atherosclerosis risk and may be used to determine the response to lipid-lowering drug therapy.
High levels of LDL-cholesterol are associated with increased cardiovascular risk and familial hyperlipidaemia. Reduced levels of LDL-cholesterol may be
found in malabsorption and malnutrition.
In 1988 the National cholesterol Education Program Adult Treatment Panel (NCEP-ATP) developed recommendations for the diagnosis and treatment of
patients with hypercholesterolemia. These recommendations defined LDL-cholesterol as the primary target of therapy.
The 2001 update of these guidelines (NCEP-ATP III) put further emphasis on better risk identification and more aggressive cholesterol-lowering treatment.
Test Principle4
A protecting agent in R1 protects LDL from enzymatic reactions. All non-LDL lipoproteins (HDL, VLDL, CM) are broken down by reaction with cholesterol
esterase (CHE) and cholesterol oxidase (CHO). Hydrogen peroxide (POD) produced by this reaction is decomposed by catalase in R1.
When R2 is added, the protecting reagent is released from LDL and catalase inactivated by sodium azide. LDL can now be quantified by the CHO/PAP
system.
Reaction Principle R2 Phase
CHE and CHO
2 LDL-cholesterol + 2 H2O + 2 O2 2 Cholest-4-en-3-one + 2 Fatty acids + 2 H2O2
Deprotecting Reagent
POD + -
2 H2O2 + 4–AA + HDAOS Blue Dye + OH + 3 H2O
Cholesterol esterase 3.7 IU/mL
Cholesterol oxidase 3.7 IU/mL
Peroxidase 4.9 IU/mL
Sodium azide 0.1 %
Good’s Buffer (pH 6.8) 25 mmol/L
4-aminoantipyrine 0.8 mmol/L
Catalase 743 IU/mL
HDAOS 0.47 mmol/L
Detergents
R2 reagent: Harmful, contains sodium azide. R22 Harmful if swallowed.
Safety Phrases:
S36, S60: Wear suitable protective clothing. This material and its container must be disposed of as hazardous waste.
Reagent Preparation
The colour of R1 may turn to light green when stored on board the analyser. This does not affect the performance of the reagent.
Specimen5
Serum and heparinised plasma: Stable for 7 days when stored at 2…8°C and 1 day when stored at 15…25°C.
Test Procedure
Calibration
LDL-Cholesterol Calibrator ODC0012.
The calibrator is traceable to the US CDC (Centre for Disease Control) LDL-cholesterol reference method.
Recalibrate the assay every 30 days and perform reagent blank every 7 days, or when the following occur:

2009-11
Quality Control
HDL/LDL-Cholesterol Control Serum ODC0005 or other control materials with values determined by this method may be used.
Calculation
The Beckman Coulter analysers automatically compute the LDL-cholesterol concentration of each sample.
National Cholesterol Education Program (NCEP) guidelines6
< 2.6 mmol/L (100 mg/dL) Optimal
2.6 – 3.3 mmol/L (100 – 129 mg/dL) Near Optimal / Above Optimal
3.4 – 4.1 mmol/L (130 – 159 mg/dL) Borderline High
4.1 – 4.9 mmol/L (160 – 189 mg/dL) High
≥ 4.9 mmol/L (190 mg/dL) Very High
values.
Linearity
The test is linear within a concentration range of 0.26 - 10.3 mmol/L (10 - 400 mg/dL).
Precision
1.57 0.04 2.26 0.04 2.71
2.63 0.04 1.36 0.06 2.34
3.70 0.07 1.76 0.10 2.68
Sensitivity
The lowest detectable level represents the lowest measurable level of LDL-cholesterol that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient serum samples were used to compare this OSR6183 LDL-cholesterol assay on the AU600 against another commercially available
LDL-cholesterol assay. Results of linear regression analysis were as follows:
Triglyceride: Interference less than 10% up to 11.3 mmol/L triglyceride
7
Limitations
When triglyceride in a sample exceeds 11.3 mmol/L (1000 mg/dL), dilute the sample with a saline solution, repeat assay and multiply result by dilution
factor.
8
Samples from patients with Type III Hyperlipoproteinemia give discrepant results to the reference method (bias + 30%).
®
Artificial lipid mixtures, as contained in some solutions for intravenous infusions (e.g. Intralipid ), can interfere with this reagent. Samples from patients
receiving these lipid mixtures should not be analysed for LDL-cholesterol using this assay.
Carry over from this LDL-cholesterol reagent to Lipase reagent may result in elevated lipase values. Please refer to Contamination Parameters.
† LDL-Cholesterol Calibrator Cat. No. ODC0012.
‡ Perform reagent blank every 7 days.
BIBLIOGRAPHY
1. Cremer P, Nagel D, Mann H, Labrot B, Muller – Berninger R, Elster H, et al. Ten year follow-up results from the Goettingen Risk, Incidence and prevalance study
(GRIPS). 1. Risk factors for myocardial infarction in a cohort of 5790 men. Atherosclerosis 1997;129(2):221-230.
2. National cholesterol education program. Expert panel on detection, evaluation and treatment of high blood cholesterol in adults (adult treatment panel II). NIH
Publication No. 93-3095,1995.
3. Naito HK, Strong JP, Scott MG et al. Atherogenesis: current topics on etiology and risk factors. Clin Chem 1995; 41(1):132-133.
4. Miki Y. A homogeneous assay for the selective measurement of LDL-cholesterol in serum. Enzymatic selective protection method. Clin Lab 1999;45:398-401.
6. National cholesterol education program expert panel. Executive summary of the third report of the national cholesterol education program (NCEP) expert panel
on detection, evaluation, and treatment of high blood cholesterol in adults (Adult Treatment Panel III). JAMA 2001;285:2486-2497.
8. Esteban-Salan M, Guimon-Bardesi A, de La Viuda-Unzueta JM, Azcarate-Ania MN, Pascual-Usandizaga P, Amoroto-Del-Rio E. Analytical and clinical evaluation
of two homogeneous assays for LDL-cholesterol in hyperlipidemic patients. Clin Chem 2000;46(8 Pt 1):1121-31.

2009-11
LDL CHOLESTEROL, AU400/AU640 Serum/Plasma Application LDL CHOLESTEROL, AU600 Serum/Plasma Application

General LIH ISE Range Test No # Test name LDL-c ∇ Sample type Ser ∇ Page 1/2
Test Name: LDL-c ∇ < > Type: Serum ∇ Operation: Yes ∇ Sample vol. 2 Dil. vol 0 μL Min. OD Max. OD
General LIH ISE Range Test No # Test name LDL-c ∇ Sample type Ser ∇ Page 2/2
Level L Level H
Test Name: LDL-c ∇ < > Type: Serum ∇
Sex Age L Age H L H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic value # #
General ISE Test No # Test name LDL-c ∇
Test Name: LDL-c Type Serum Cal type 8 AB ∇ Count #
∇ < > ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H Point 1 # ∇ † 12* 19*
Point 1: # † 12* 19* Point 2 ∇
Point 7:
1-point cal. point
1-Point Cal. Point: ο With CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ MB type factor
MB Type Factor: Calibration Stability Period: 30‡ Calibrator stability period 30‡

† LDL Cholesterol Calibrator Cat. No.: ODC0012 † LDL Cholesterol Calibrator Cat. No.: ODC0012
* Values set for working in SI units (mmol/L) To work in mg/dL multiply by 38.7 * Values set for working in SI units (mmol/L) To work in mg/dL multiply by 38.7
‡ Perform reagent blank every 7 days ‡ Perform reagent blank every 7 days.
2009-08
LDL CHOLESTEROL, AU2700/AU5400 Serum/Plasma Application LDL CHOLESTEROL, AU680/AU480 Serum/Plasma Application
Test Name: LDL-C ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: LDL-C ∇ < > Type: Serum ∇ Operation: Yes ∇
Reagents: R1 Volume 144 Dilution 0 Min OD Max OD Pre-Dilution Rate 1 ∇ Min.OD Max.OD
μL μL
Reagent OD limit:
Method END ∇
Test Name: LDL-C ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: LDL-C ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # # Specificl Ranges: From To Low High
ο 3. # ∇ # # # # # #
ο 1. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
Test Name: LDL-C ∇ < > ψ Type Serum ∇
Test Name: LDL-C ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † 12* 19*
Point 1: # ∇ † 12* 19*
Point 2:
Point 3:
Point 3: ∇
Point 4:
Point 5:
Point 6:
Point 6: ∇
Point 7:
MB Type Factor: Calibration Stability Period: 30 ‡ Point 9 ∇
† LDL Cholesterol Calibrator Cat. No.: ODC0012 Calibrator OD Conc Low High Stability
* Values set for working in mmol/L. To work in SI units ( mgldL) multiply by 38.7 Point-1 ∇ Reagent Blank 7 Day 0 Hour
‡ Perform reagent blank every 7 days Point-2 ∇ Calibration 30 Day 0 Hour
2009-08
MAGNESIUM
OSR6189 4 x 40 mL R1
Intended Use
Photometric colour test for the quantitative determination of magnesium in human serum, plasma and urine on Beckman Coulter analysers. For
Summary1,2
Magnesium is an essential factor in many important enzymatic reactions, either as an integral part of a metalloenzyme or as an activator, and
plays an important role in glycolysis, cellular respiration and transmembranous calcium transport. Magnesium is mainly regulated by the rate of
renal magnesium excretion, which along with calcium is subject to the effects of parathyroid hormone. Increasing calcium reabsorption thus
leads to competitive inhibition of magnesium absorption.
Magnesium measurements are used in the diagnosis and treatment of hypomagnesemia (abnormally low) and hypermagnesemia (abnormally
high). The best-defined manifestation of magnesium deficiency is impairment of neuromuscular function e.g. hyperirritability, tetany,
convulsions, and electrocardiographic changes. Hypomagnesemia is observed in cases of diabetes, chronic alcoholism, forced diuresis,
hyperthyroidism, hypoparathyroidism, hypocalcemia, malabsorption and acute pancreatitis. Increased serum magnesium levels have been
found in cases of renal failure, dehydration, severe diabetic acidosis and Addison’s Disease.
Test Principle3,4,5
The Magnesium reagent utilises a direct method in which magnesium ions form a coloured complex with xylidyl blue in a strongly basic solution.
The colour produced is measured bichromatically at 520/800 nm and is proportional to the magnesium concentration in the sample. Calcium
interference is eliminated by glycoletherdiamine-N,N,N`,N`-tetraacetic acid (GEDTA).
Reaction Principle
pH 11.4
2+
Mg + Xylidyl blue Purple complex

∈-Amino-n Caproic Acid 450 mmol/L
Tris 100 mmol/L
Glycoletherdiamine-N,N,N’,N’ tetraacetic acid 0.12 mmol/L
Xylidyl blue 0.18 mmol/L
Preservative
Reagent Preparation
The reagent is stable, unopened, up to the stated expiry date when stored at 2…8°C. Once open, reagent stored on board the instrument is
stable for 14 days.
Specimen
6
Serum or heparinised plasma. Do not use EDTA, oxalate or citrate plasma.
Separate serum from red blood cells immediately.
Haemolysed samples should be avoided due to the higher magnesium concentration in erythrocytes.
1
Magnesium in serum is stable for 7 days when stored at 15…25°C.
7
Urine: Acidified with 6M HCl. Collect timed 24 hour specimen using standard laboratory procedures.
Store at 2...8°C.
Test Procedure
statement.
Calibration
The magnesium values of both calibrators are traceable to the National Institute of Standards and Technology (NIST) Standard Reference
Material (SRM) 909b Level 2. Recalibrate the assay every 7 days, or when the following occur:
Quality Control

2009-08
Calculation
The Beckman Coulter analysers automatically compute the magnesium concentration of each sample.
Serum Men 0.73 – 1.06 mmol/L (1.8 – 2.6 mg/dL)
Women 0.77 – 1.03 mmol/L (1.9 – 2.5 mg/dL)
Urine Adults 3 – 5 mmol/24 h (73 – 122 mg/24 h)
findings.
from these values.
Linearity
The test is linear within a concentration range of 0.2 – 3.3 mmol/L (0.5 – 8.0 mg/dL) for serum and plasma.
The test is linear within a concentration range of 0.2 – 9.25 mmol/L (0.5 – 22.5 mg/dL) for urine.
Precision
0.80 0.01 1.02 0.01 1.15
1.09 0.01 0.75 0.01 1.29
3.02 0.03 0.90 0.04 1.24
0.78 0.01 1.15 0.04 4.96
3.76 0.03 0.71 0.10 2.73
7.81 0.06 0.71 0.27 3.40
Sensitivity
The lowest detectable level in urine on the AU2700 was estimated at 0.02 mmol/L.
The lowest detectable level represents the lowest measurable level of magnesium that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this Magnesium OSR6189 assay on the AU640 against another commercially available
magnesium assay. Results of linear regression analysis were as follows:
Patient urine samples were used to compare this Magnesium OSR6189 assay on the AU2700 against another commercially available
magnesium assay. Results of linear regression analysis were as follows:
Calcium: Interference less than 3% up to 30 mg/dL or 7.5 mmol/L calcium
®
Calcium: Interference less than 10% up to 40 mg/dL or 10 mmol/L calcium
8
BIBLIOGRAPHY
1. Dörner K. Magnesium (Mg). In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
Verlagsgesellschaft mbH, 1998:339-340.
2. Jacob RA. Trace Elements. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:521-524.
3. Mann CK, Yoe JH. Spectrophotometric determination of magnesium with sodium 1-Azo-2-hydroxy-3-(2,4-dimethylcarboxanilido) naphthalene-1’-
(2-hydroxybenzene-5-sulfonate). Anal Chem 1956;28:202-205.
4. Mann CK, Yoe JH. Spectrophotometric determination of magnesium with 1-Azo-2-hydroxy-3-(2,4-dimethylcarboxanilido) naphthalene-1’-(2-hydroxybenzene).
Anal Chim Acta 1957;16:155-160.
5. Bohuon C. Microdosage du magnésium dans divers milieux biologiques. Clin Chim Acta 1962;7:811-817.
6. Kazmierczak SC. Magnesium. In: Kaplan LA, Pesce AJ, eds. Clinical chemistry theory, analysis, and correlation. St Louis: Mosby, 1996:551pp.
7. NCCLS. Urinalysis and collection, transportation, and preservation of urine specimens; approved guideline. NCCLS Document GP16-A2, 2nd ed. Pennsylvania:
NCCLS, 2001.

2009-08
MAGNESIUM, AU400/AU640 Serum/Plasma Application MAGNESIUM, AU600 Serum/Plasma Application

Test No # Test name MG ∇ Sample type Ser ∇ Page 1/2
Test Name: MG ∇ < > Type: Serum ∇ Operation: Yes ∇
R2 Volume 0 μL Dilution 0 μL L H Fst. L 0.9 Fst. H 1.5
Wavelength: Pri. 520 ∇ Sec. 800 ∇ First L 0.9 First H 1.5 Method END ∇ Dynamic range
Method: END ∇ Last L 0.9 Last H 1.5 Reaction + ∇ L 0.2* H 3.3*
General LIH ISE Range Test No # Test name MG ∇ Sample type Ser ∇ Page 2/2
Test Name: MG ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name MG ∇
Test Name: MG ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 3* 5*
Point 1: # † 3* 5*
Point 2 ∇
Point 2:
MB type factor
2009-08
MAGNESIUM, AU2700/AU5400 Serum/Plasma Application MAGNESIUM, AU680/AU480 Serum/Plasma Application
Test Name: MG ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: MG ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Method: END ∇ Last L 0.9 Last H 1.5
Specific Test Parameters Lag Time Check ∇ Hemolysis ++ ∇
Test Name: MG ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: MG ∇ < > Type: Serum ∇

ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: MG ∇ < > Type Serum ∇
Test Name: MG ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † 3* 5* Point 1: # ∇ † 3* 5*
Point 9 ∇
2009-08
MAGNESIUM, AU400/AU640 Urine Application MAGNESIUM, AU600 Urine Application

Test No # Test name MG ∇ Sample type Uri ∇ Page 1/2
Test Name: MG ∇ < > Type: Urine ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name MG ∇ Sample type Uri ∇ Page 2/2
Test Name: MG ∇ < > Type: Urine ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
URINE APPLICATION
General ISE Test No # Test name MG ∇
Test Name: MG ∇ < > Type Urine ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 5* 11*
Point 1: # † 5* 11*
Point 2 ∇
Point 2:
MB type factor
2009-08
MAGNESIUM, AU2700/AU5400 Urine Application MAGNESIUM, AU680/AU480 Urine Application
Test Name: MG ∇ < > Type: Urine ∇ Operation Yes ∇
Test Name: MG ∇ < > Type: Urine ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Linearity Limit %
Test Name: MG ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: MG ∇ < > Type: Urine ∇

ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
URINE APPLICATION
General ISE
General ISE
Test Name: MG ∇ < > Type Urine ∇ Test Name: MG ∇ < > Type Urine ∇ ο Use Serum Cal.
Calibration Type: AB ∇ Formula: Y=AX+B Counts: # Process: CONC ∇ Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # ∇
Point 1: # † 5* 11* Point 1: # ∇ † 5* 11*
Point 9 ∇
2009-08
TOTAL PROTEIN
OSR6132 4 x 25 mL R1
4 x 25 mL R2
OSR6232 4 x 48 mL R1
4 x 48 mL R2
Intended Use
Photometric colour test for the quantitative determination of total protein in human serum and plasma on Beckman Coulter analysers. For in vitro
Summary1
The total serum protein is the sum of all circulating proteins and is a major component of blood. Measurements of total protein are used in the
diagnosis and treatment of a variety of diseases involving the liver, kidney, or bone marrow as well as other metabolic and nutritional disorders.
A deviation of serum total protein from the reference interval indicates the presence of dysproteinemia or a disorder in water balance. Both
conditions can be distinguished by additional performance of serum protein electrophoresis and the determination of haematocrit.
It is also useful in interpreting the significance of the total protein concentration to have more specific knowledge of individual fractions such as
albumins and globulins.
Test Principle2
Cupric ions in an alkaline solution react with proteins and polypeptides containing at least two peptide bonds to produce a violet coloured
complex. The absorbance of the complex at 540/660 nm is directly proportional to the concentration of protein in the sample.
Reaction Principle
2+ OH ¯
Protein + Cu Blue violet complex

Sodium hydroxide 200 mmol/L
Potassium sodium tartrate 32 mmol/L
Copper sulphate 18.8 mmol/L
Potassium iodide 30 mmol/L
R1 Reagent: Corrosive, contains sodium hydroxide. R34 Causes burns.
R2 Reagent: Irritant. R36/38, R52/53: Irritating to eyes and skin. Harmful to aquatic organisms, may cause long-term adverse effects in the
aquatic environment.
Safety Phrases:
S26 - S36/37/39, S45, S60, S61: In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable
protective clothing, gloves and eye/face protection. In case of accident or if you feel unwell, seek medical advice immediately (show the label
where possible). This material and its container must be disposed of as hazardous waste. Avoid release to the environment. Refer to special
instructions/safety data sheets.
Reagent Preparation
Specimen
Serum, EDTA or heparinised plasma.
3
Test Procedure
Refer to the appropriate User’s Guide and the accompanying Instrument Setting Sheets for analyser-specific assay instructions.
Calibration
The calibrator total protein value is traceable to the National Institute of Standards and Technology (NIST) Standard Reference Material (SRM)
927c. Recalibrate the assay when the following occur: Change in reagent bottle or significant shift in control values;
Major preventative maintenance was performed on the analyser or a critical part was replaced. Absorption of atmospheric CO2 by the reagent
on board the analyser can impair calibration stability. This effect will vary depending upon the rate of use. Consequently each laboratory should
set a calibration frequency in the instrument parameters appropriate to their usage pattern.
Quality Control
2009-08
Calculation
The Beckman Coulter analysers automatically compute the total protein concentration of each sample.
Serum/plasma
Adults 66 – 83 g/L (6.6 – 8.3 g/dL)
Children (1 - 18 y) 57 – 80 g/L (5.7 – 8.0 g/dL)
New-borns (1 - 30 d) 41 – 63 g/L (4.1 – 6.3 g/dL)
findings.
Data contained within this section is representative of performance on Beckman Coulter Systems. Data obtained in your laboratory may differ
from these values.
Linearity
The test is linear within a concentration range of 30 – 120 g/L (3.0 – 12.0 g/dL).
Precision
35.57 0.18 0.50 0.30 0.84
73.33 0.25 0.34 0.51 0.70
110.80 0.29 0.26 0.71 0.64
Sensitivity
The lowest detectable level represents the lowest measurable level of total protein that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this Total Protein OSR6132 assay on the AU640 against another commercially available total
protein assay. Results of linear regression analysis were as follows:
®
4
‡ Depends on usage pattern in the laboratory
BIBLIOGRAPHY
1. Thomas L. Total Protein. In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
2. Weichselbaum TE. An accurate and rapid method for the determination of proteins in small amounts of blood serum and plasma. Amer J Clin Path 1946;
16:40-48.

2009-08
TOTAL PROTEIN, AU640 Serum/Plasma Application TOTAL PROTEIN, AU600 Serum/Plasma Application

Test No # Test name TP ∇ Sample type Ser ∇ Page ½
Test Name: TP ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name TP ∇ Sample type Ser ∇ Page 2/2
Test Name: TP ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name TP ∇
Test Name: TP ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 147* 237*
Point 1: # † 147* 237*
Point 2 ∇
Point 2:
MB type factor
# User defined
2009-08
TOTAL PROTEIN, AU2700/AU5400 Serum/Plasma Application TOTAL PROTEIN, AU400 Serum/Plasma Application
Specific Test Parameters Specific Test Parameters

General LIH ISE Range General LIH ISE Range
Test Name: TP ∇ < > Type: Serum ∇ Operation: Yes ∇ Test Name: TP ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 5 μL Dilution 0 μL Pre-Dilution Rate: 1 Sample: Volume 6 μL Dilution 0 μL Pre-Dilution Rate: 1
Reagents: R1 Volume 33 μL Dilution 90 μL Min OD Max OD Reagents: R1 Volume 50 μL Dilution 140 μL Min OD Max OD
R2 Volume 33 μL Dilution 10 μL L H R2 Volume 50 μL Dilution 10 μL L H
Reagent OD limit: Reagent OD limit:
Wavelength: Pri. 540 ∇ Sec. 660 ∇ First L -0.5 First H 0.1 Wavelength: Pri. 540 ∇ Sec. 660 ∇ First L -0.5 First H 0.1
Method: END ∇ Last L -0.5 Last H 0.1 Method: END ∇ Last L -0.5 Last H 0.1
Reaction slope: + ∇ Dynamic Range: Reaction slope: + ∇ Dynamic Range:
Measuring Point 1: First 0 Last 27 L 30* H 120* Measuring Point 1: First 0 Last 27 L 30* H 120*
Measuring Point 2: First 0 Last 10 Correlation Factor: Measuring Point 2: First 0 Last 10 Correlation Factor:
Linearity : % A 1 B 0 Linearity : % A 1 B 0
No Lag Time: ∇ On-board stability period: 30 No Lag Time: ∇ On-board stability period: 30

Test Name: TP ∇ < > Type: Serum ∇ Test Name: TP ∇ < > Type: Serum ∇
Value/Flag: # ∇ Level L: # Level H: # Value/Flag: # ∇ Level L: # Level H: #

Normal Ranges: Age L Age H Normal Ranges: Age L Age H
Sex Year Month Year Month L H Sex Year Month Year Month L H
ο 1. # ∇ # # # # # # ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # # ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 6. # ∇ # # # # # #
7. None Selected # # 7. None Selected # #
8. Out of Range L H # # 8. Out of Range L H # #
Panic Value: # # Unit: g/L* Decimal places: # Panic Value: # # Unit: g/L* Decimal places: #
Calibration Specific Calibration Specific

Test Name: TP ∇ < > Type Serum ∇ Test Name: TP ∇ < > Type Serum ∇
Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1: # † 122* 195* Point 1: # † 150* 244*
Point 2: Point 2:
Point 3: Point 3:
Point 4: Point 4:
Point 5: Point 5:
Point 6: Point 6:
Point 7: Point 7:
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-Point Cal. Point: ο With CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇
MB Type Factor: Calibration Stability Period: ‡ MB Type Factor: Calibration Stability Period: ‡
# User defined. # User defined

* Values set for working in SI units (g/L). To work in g/dL divide by 10. * Values set for working in SI units (g/L). To work in g/dL divide by 10.
‡ Depends on usage pattern in the laboratory ‡ Depends on usage pattern in the laboratory.
2009-08
TOTAL PROTEIN, AU680/AU480 Serum/Plasma Application
Test Name: TP ∇ < > Type: Serum ∇ Operation Yes ∇

Method END ∇
Linearity Limit % Icterus +++ ∇
Lag Time Check ∇ Hemolysis +++ ∇

Test Name: TP ∇ < > Type: Serum ∇

ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
General ISE
Test Name: TP ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # ∇ † 122* 195*
Point 3: ∇
Point 6: ∇
Point 9 ∇
# User defined
Point-1 ∇ Reagent Blank ‡ Day ‡ Hour † System Calibrator Cat. No.: 66300
Point-2 ∇ Calibration ‡ Day ‡ Hour * Values set for working in SI units (g/L). To work in g/dL divide by 10.
MB Type Factor: 1-Point Calibration Point ∇ ο with Conc-0 ‡ Depends on usage pattern in the laboratory
ф AU680
2009-08
TRIGLYCERIDE
OSR60118 4 x 20 mL R1
4x 5 mL R2
OSR61118 4 x 50 mL R1
4 x 12.5 mL R2
Intended Use
Enzymatic colour test for the quantitative determination of triglyceride in human serum and plasma on Beckman Coulter analysers. For in vitro
Summary1,2
Measurements of triglyceride are used in the diagnosis and treatment of patients with acute and chronic pancreatitis, diabetes mellitus,
nephrosis, extrahepatic biliary obstruction, and other diseases involving lipid metabolism, or various endocrine disorders.
Clinically, triglyceride assays are used to help classify various genetic and metabolic lipoprotein disorders, and in the assessment of risk factors
for atherosclerosis and coronary artery disease.
Test Principle3,4,5
This Triglyceride procedure is based on a series of coupled enzymatic reactions. The triglycerides in the sample are hydrolysed by a
combination of microbial lipases to give glycerol and fatty acids. The glycerol is phosphorylated by adenosine triphosphate (ATP) in the
presence of glycerol kinase (GK) to produce glycerol-3-phosphate. The glycerol-3-phosphate is oxidised by molecular oxygen in the presence of
GPO (glycerol phosphate oxidase) to produce hydrogen peroxide (H2O2) and dihydroxyacetone phosphate. The formed H2O2 reacts with
4-aminophenazone and N,N-bis(4-sulfobutyl)-3,5-dimethylaniline, disodium salt (MADB) in the presence of peroxidase (POD) to produce a
chromophore, which is read at 660/800nm. The increase in absorbance at 660/800nm is proportional to the triglyceride content of the sample.
Reaction Principle
Lipase
Triglycerides + 3 H2O Glycerol + 3 Fatty acids
2+
GK, Mg
Glycerol + ATP Glycerol-3-phosphate + ADP
GPO
Glycerol-3-phosphate + O2 Dihydroxyacetone phosphate + H2O2
POD
-
H2O2 + 4-AAP + MADB Blue Dye + OH + 3 H2O

PIPES buffer (pH 7.5) 50 mmol/L Lipases 1.5 kU/L (25 µkat/L)
2+
Mg 4.6 mmol/L Glycerol kinase 0.5 kU/L (8.3 µkat/L)
MADB 0.25 mmol/L Peroxidase 0.98 kU/L (16.3 µkat/L)
4-Aminoantipyrine 0.5 mmol/L Ascorbate oxidase 1.48 kU/L (24.6 µkat/L)
ATP 1.4 mmol/L Glycerol-3-phosphate oxidase 1.48 kU/L (24.6 µkat/L)
Preservative
Reagent Preparation
Specimen1
Plasma using anticoagulants such as fluoride, citrate and oxalate should be avoided.
Strongly icteric samples should be avoided.
Avoid using vacuum tubes with glycerol-coated stoppers.
9
Stable in serum and plasma for 7 days when stored at 2...8°C and 2 days when stored at 15…25°C.
Test Procedure
Calibration
The calibrator triglyceride value provided in the calibrator package insert is traceable to the Isotope Dilution Mass Spectrometry Reference
Method.
2009-08
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the triglyceride concentration of each sample.
Normal < 1.70 mmol/L (150 mg/dL)
Borderline high 1.70 – 2.25 mmol/L (150 – 199 mg/dL)
High 2.26 – 5.64 mmol/L (200 – 499 mg/dL)
Very high ≥ 5.65 mmol/L (500 mg/dL)
findings.
from these values.
Linearity
Prozone settings must be applied when using the Triglyceride reagent, refer to Setting Sheet for specific instrument details.
Precision
0.47 0.01 1.06 0.01 1.76
4.28 0.03 0.72 0.04 1.03
10.20 0.08 0.79 0.15 1.46
Sensitivity
The lowest detectable level represents the lowest measurable level of triglyceride that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this Triglyceride OSR61118 assay on the AU640 against another commercially available
triglyceride assay. Results of linear regression analysis were as follows:
7
Limitations
Note: Triglycerides GPO enzymatic methodologies are subject to a strong negative interference from patient samples with extremely elevated
8
triglyceride levels. While these samples are extremely lipemic in appearance and typically have triglyceride levels exceeding 20 mmol/L, results
can be erroneously reported as being within the linear range of the assay. In order to identify grossly lipemic samples exhibiting this
phenomenon, Data Check Parameters are provided. If the reaction kinetics of a test exhibit the characteristics of one of these elevated
triglyceride samples, the analysis result will be flagged (F, Z, @ or &). Grossly lipemic samples under rare circumstances may evade the Data
Check Parameters and should routinely be diluted 1 part sample to 4 parts saline prior to analysis, and the results multiplied by 5.
1
If compensating for free glycerol, subtract 0.11 mmol/L (10 mg/dL) from the triglyceride value obtained. This correction is mainly applicable to
healthy individuals, but for some diseases, for example diabetes or liver disorders, a higher concentration of free glycerol may occur.
BIBLIOGRAPHY
1. Tietz NW, ed. Clinical guide to laboratory tests, 3rd ed. Philadelphia: WB Saunders Company, 1995: 610-611.
2. Riesen WF. Lipid metabolism. In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
3. Jacobs NJ, Van Denmark PJ. Arch Biochem Biophys 1960;88:250-255.
4. Koditschek LK, Umbreit WW. Alpha-glycerophosphate oxidase in streptococcus faecium F 24. J Bacteriol 1969; 98:1063-1068.
5. Trinder P. Ann Clin Biochem, 1969; 6:24-27.
6. National Cholesterol Education Program Expert Panel. Executive summary of the third report of the National Cholesterol Education Program (NCEP) Expert
Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). JAMA 2001;285: 2486-2497.
8. Shephard MD, Whiting MJ. Falsely low estimation of triglycerides in lipemic plasma by the enzymatic triglyceride method with modified Trinder’s chromogen.
Clin Chem 1990; 36:325-329.
9. Guder, WG, Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of anticoagulants in diagnostic laboratory investigations and
stability of blood, plasma and serum samples. WHO/DIL/LAB/99.1 Rev.2: 44pp.

2009-08
TRIGLYCERIDE, AU400/AU640 Serum/Plasma Application TRIGLYCERIDE, AU600 Serum/Plasma Application

Test No # Test name TRIG ∇ Sample type Ser ∇ Page ½
Test Name: TRIG ∇ < > Type: Serum ∇ Operation: Yes ∇
Fst. L -0.1 Fst. H 0.3
Reagent OD limit: Method END ∇ Dynamic range
Wavelength: Pri. 660 ∇ Sec. 800 ∇ First L -0.1 First H 0.3 Reaction + ∇ L 0.1* H 11.3*
Method: END ∇ Last L -0.1 Last H 0.3 Point 1 Fst 0 Lst 27 Correlation factor A 1
Reaction slope: + ∇ Dynamic Range: Point 2 Fst 0 Lst 10 B 0
Measuring Point 1: First 0 Last 27 L 0.1* H 11.3* Sample Pre-dil. Rate ¤
Measuring Point 2: First 0 Last 10 Correlation Factor: Linearity Fst % Sec %
Linearity : % A 1 B 0 No lag time ∇ On-board stability period 30
No Lag Time: ∇ On-board stability period: 30 Select using Space key, or select from list displayed by Guide key
Test No # Test name TRIG ∇

General ISE Cal type 8 AB ∇ Count #
Test Name: TRIG ∇ < > Type Serum ∇ Selection calibrator
Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Point 1 # ∇ † 12* 19*
Point 2 ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H Point 3 ∇
Point 1: # † 12* 19* Point 4 ∇
1-point cal. Point
Point 5: MB type factor
Point 6: Calibrator stability period 30
Point 7:
MB Type Factor: Calibration Stability Period: 30 Data Check Parameters
Test No # Name TRIG ∇ Type Ser ∇

Data Check Parameters
Logic check-1 YES Logic Check-2 NO
Check Point-1 10 Check Point-1 0
Test Name: TRIG ∇ < > Type: Serum ∇
Check Point-2 22 Check Point Interval
Check Point-3 27 Decision value-1 0.0000
√ Logic Check-1 Logic Check -2 Logic Check-3 Decision value-1 1.0500 Decision value-2 0.0000
Check Point-1: 10 Check Point-1: Check Point-1: Decision value-2 9.0000 Limit Point 1 0
Check Point-2: 22 Check Point Interval: Check Point Interval: Decision value-3 0.1000 Limit Point 2 0
Check Point-3: 27 Decision Value-1: Decision Value-1: Limit Point 1 0 Logic Check-3 NO
Decision Value-1: 1.0500 Decision Value-2: Decision Value-2: Limit Point 2 22
Decision Value-2: 9.0000 Limit Point 1: Limit Point 1: Check Point-1 0
Decision Value-3: 0.1000 Limit Point 2: Limit Point 2: Check Point Interval
Limit Point 1: 0 Decision value-1 0.0000
Limit Point 1 0
Limit Point 2 0

2009-08
TRIGLYCERIDE, AU2700/AU5400 Serum/Plasma Application TRIGLYCERIDE, AU680 Serum/Plasma Application
Test Name: TRIG ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: TRIG ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Reaction slope: + ∇ Dynamic Range: Common Rgt. Type None Name Correlation Factor A 1 B 0
General ISE Parameters Calibration Parameters
Test Name: TRIG ∇ < > Type: Serum ∇ General ISE
Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Test Name: TRIG ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # † 10* 17*
Point 2:
Point 1: # ∇ † 10* 17*
Point 3:
Point 4:
Point 3: ∇
Point 5:
MB Type Factor: Calibration Stability Period: 30 Point 8 ∇ Operation # ∇
Point 9 ∇
Data Check Parameters <Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration
Point-1 Reagent Blank 30 Day 0 Hour
Test Name: TRIG ∇ < > Type: Serum ∇ ∇
√ Logic Check-1 Logic Check -2 Logic Check-3
Check Point-1: 11 Check Point-1: Check Point-1:
Check Point-2: 17 Check Point Interval: Check Point Interval: Parameters Misc
Check Point-3: 27 Decision Value-1: Decision Value-1:
CheckedTests Contamination Data Check
Decision Value-1: 0.940 Decision Value-2: Decision Value-2:
Parameters Parameters
Decision Value-2: 9.0000 Limit Point 1: Limit Point 1:
Decision Value-3: 0.100 Limit Point 2: Limit Point 2: Test Name: TRIG ∇ < > Type: Serum ∇
Limit Point 1: 0
Limit Point 2: 22 √ Logic Check-1 Logic Check -2 Logic Check-3
Check Point-2: 17 Check Point Interval: Check Point Interval:
Check Point-3: 27
Decision Value-2: 9.0000 Decision Value-2: Decision Value 2:
# User defined.
Decision Value-3: 0.1000
Limit Point 1: 0 Limit Point 1: Limit Point 1:
Check Pattern: Pattern1 ∇
2009-08
TRIGLYCERIDE, AU480 Serum/Plasma Application
Test Name: TRIG ∇ < > Type: Serum ∇ Operation Yes ∇

Correlation Factor A 1 B 0
Method END ∇

General ISE
Test Name: TRIG ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # ∇ † 10* 17*
Point 3: ∇
Point 6: ∇
Point 9 ∇
Point-1 ∇ Reagent Blank 30 Day 0 Hour
Parameters Misc

Test Name: TRIG ∇ < > Type: Serum ∇

Check Point-3: 27
Limit Point 2: 22 Limit Point 2: Limit Point 2: # User defined.
Check Pattern: Pattern1 ∇ † System Calibrator Cat. No.: 66300
2009-08
TRIGLYCERIDE, AU400/AU640 Paediatric Application TRIGLYCERIDE, AU600 Paediatric Application

Test No # Test name TRIGP ∇ Sample type Ser ∇ Page ½
Test Name: TRIGP ∇ < > Type: Serum ∇ Operation: Yes ∇
Fst. L -0.1 Fst. H 0.3
Test No # Test name TRIGP ∇

General ISE Cal type 8 AB ∇ Count #
Test Name: TRIGP ∇ < > Type Serum ∇ Selection calibrator
Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Point 1 # ∇ † 12* 19*
Point 2 ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H Point 3 ∇
Point 1: # † 12* 19* Point 4 ∇
1-point cal. Point
Point 7:
Test No # Name TRIGP ∇ Type Ser ∇

Test Name: TRIGP ∇ < > Type: Serum ∇
Limit Point 1 0
Limit Point 2 0

2009-08
TRIGLYCERIDE, AU2700/AU5400 Paediatric Application TRIGLYCERIDE, AU680 Paediatric Application
Test Name: TRIGP ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: TRIGP ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: TRIGP ∇ < > Type: Serum ∇ General ISE
Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Test Name: TRIGP ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # † 10* 17*
Point 2:
Point 1: # ∇ † 10* 17*
Point 3:
Point 4:
Point 3: ∇
Point 5:
Point 9 ∇
Test Name: TRIGP ∇ < > Type: Serum ∇ Point-1 ∇ Reagent Blank 30 Day 0 Hour
Decision Value-3: 0.100 Limit Point 2: Limit Point 2: Test Name: TRIGP ∇ < > Type: Serum ∇
Limit Point 1: 0
Check Point-3: 27
# User defined.
2009-08
TRIGLYCERIDE, AU480 Paediatric Application
Test Name: TRIGP ∇ < > Type: Serum ∇ Operation Yes ∇

Method END ∇

General ISE
Test Name: TRIGP ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # ∇ † 10* 17*
Point 3: ∇
Point 6: ∇
Point 9 ∇
Parameters Misc

Test Name: TRIGP ∇ < > Type: Serum ∇

Check Point-3: 27
Check Pattern: Pattern1 ∇ † System Calibrator Cat. No.: 66300
2009-08
UIBC
OSR6124 4 x 27 mL R1
4 x 3 mL R1a
4 x 6 mL R2
4 x 2 mL R2a
Intended Use
Photometric colour test for the quantitative determination of unsaturated iron binding capacity (UIBC) in human serum and plasma on Beckman
Summary1,2,3
Iron participates in a variety of vital processes in the body varying from cellular oxidative mechanisms to the transport and delivery of oxygen to
body cells. It is a constituent of the oxygen-carrying chromoproteins, haemoglobin and myoglobin, as well as various enzymes, such as
cytochrome oxidase and peroxidases. The remaining body iron is present in the flavoproteins, the iron-sulphur proteins, as well as storage iron-
ferritin, and transport iron-transferrin.
Measured serum iron concentration is principally the Fe(III) bound to serum transferrin and does not include the iron contained in serum as free
haemoglobin. Since normally only about one-third of the iron-binding sites of transferrin are occupied by Fe(III), serum transferrin has
considerable reserve iron-binding capacity. This is called the serum unsaturated or latent iron-binding capacity. UIBC measurements can be
used in conjunction with serum iron concentration to obtain the total-iron binding capacity (TIBC) i.e. the maximum concentration of iron that
serum proteins, principally transferrin, can bind.
TIBC is decreased in chronic infections, malignancy, in iron poisoning, renal disease, nephrosis, kwashiorkor and thalassemia. Common causes
for an increase in TIBC include iron deficiency anemia, late pregnancy, oral contraception and viral hepatitis.
Test Principle
2+
Fe from reagent 1 reacts with Nitroso-PSAP from reagent 2 to form an intense green complex. If sample is added a part or all of the iron ions
bind specifically with transferrin at unsaturated iron binding sites at alkaline pH. They are thus not available for the colour reaction with Nitroso-
PSAP. The difference between the resulting changes in the measured absorbances with or without samples is equivalent to the iron quantity
bound to transferrin. This is the unsaturated iron binding capacity (UIBC).

Tris Buffer (pH 8.1) 180 mmol/L
Iron 6.9 µmol/L
Nitroso-PSAP 176 µmol/L
Hydroxylammonium chloride 36 mmol/L
Thiourea 175 mmol/L
Preservative

R1: Harmful, contains thiourea. R40: Limited evidence of a carcinogenic effect.
R1a: Irritant, contains hydroxylammonium chloride. R43: May cause sensitisation by skin contact.
R2: Contains hydroxylammonium chloride. May produce an allergic reaction. R36/R40 Limited evidence of a carcinogenic effect. Irritant to eyes.
Safety Phrases:
S24, S26, S36/37, S60, S61: Avoid contact with skin. Wear suitable protective clothing and gloves. Wear eye/face protection in case of contact
with eyes, rinse immediately with plenty of water and seek medical advise. This material and its container must be disposed of as hazardous
waste. Avoid release to the environment. Refer to special instructions/safety data sheets.
Reagent Preparation
R1: The entire contents of bottle R1a must be transferred into the entire volume of R1. Mix by gentle inversion before placing on board the instrument.

Specimen
4
Haemolysed samples should be avoided. Remove serum from red cells immediately to avoid haemolysis.
5
Test Procedure
Calibration

2009-08
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the UIBC concentration of each sample.
Adults 27.8 – 53.7 µmol/L (155 – 300 µg/dL)
findings.

from these values.
Linearity
The test is linear within a concentration range of 9.8 – 80.6 µmol/L (55 – 450 µg/dL).
Precision
29.17 0.39 1.35 0.46 1.59
35.47 0.38 1.06 0.55 1.56
62.09 0.58 0.93 1.04 1.68
Sensitivity
The lowest detectable level represents the lowest measurable level of UIBC that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient serum samples were used to compare this UIBC OSR6124 assay on the AU2700 against another commercially available UIBC assay.
y = 0.867x + 3.295 r = 0.997 n = 117 Sample range = 14.20-56.66 µmol/L
®
7

BIBLIOGRAPHY
2. Fairbanks VF, Klee GG. Biochemical aspects of hematology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia: WB Saunders
Company, 1999;1698-1703.
3. Woo J, Henry JB. Metabolic intermediates and inorganic ions. In: Henry JB, ed. Clinical diagnosis and management by laboratory methods. Philadelphia:WB
Saunders Company, 1996:188-190.
serum samples.WHO/DIL/LAB/99.1 Rev.2:36pp.
5. Perrotta G. Iron and total iron binding capacity. In: Kaplan LA, Pesce AJ, eds. Clinical chemistry theory, analysis, and correlation. St Louis: Mosby, 1996:714pp.
6. Data on file at Beckman Coulter.

2009-08
UIBC, AU400/AU640 Serum/Plasma Application UIBC, AU600 Serum/Plasma Application

Test No # Test name UIBC ∇ Sample type Ser ∇ Page 1/2
Test Name: UIBC ∇ < > Type: Serum ∇ Operation: Yes ∇
Reagent OD limit: Wave Main 800 Sub ∇ Lst. L -0.1 Lst. H 0.3
Wavelength: Pri. 800 ∇ Sec. None ∇ First L -0.1 First H 0.3 Method END ∇ Dynamic range
Method: END ∇ Last L -0.1 Last H 0.3 Reaction - ∇ L 9.85* H 80.6*
General LIH ISE Range Test No # Test name UIBC ∇ Sample type Ser ∇ Page 2/2
Test Name: UIBC ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name UIBC ∇
Test Name: UIBC ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 340* 510*
Point 1: # † 340* 510*
Point 2 ∇
Point 2:
MB type factor
# User defined
† System Calibrator Cat No: 66300
* Values set for working in SI units (µmol/L). To work In µg/dL multiply by 5.585
2009-08
Test Name: UIBC ∇ < > Type: Serum ∇ Operation Yes ∇

μL μL
Reagent OD limit:
Wavelength: Pri. 800 ∇ Sec. None ∇ First L -0.1 First H 0.3
Reaction slope: - ∇ Dynamic Range: Common Rgt. Type None Name Correlation Factor A 1 B 0
Measuring Point 1: First 0 Last 27 L 9.85* H 80.6* Wavelength Pri 800 ∇nm Sec. None ∇nm Factor for Maker A 1 B 0
Test Name: UIBC ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: UIBC ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: UIBC ∇ < > Type Serum ∇ Test Name: UIBC Type Serum Use Serum Cal.
∇ < > ∇ ο
Point 1: # † 340* 510* Point 1: # ∇ † 340* 510*
Point 9 ∇
† System Calibrator Cat No: 66300 Calibrator OD Conc Low High Stability
* Values set for working in SI units (µmol/L). To work In µg/dL multiply by 5.585 Point-1 ∇ Reagent Blank 14 Day 0 Hour
2009-08
UIBC
OSR61205 4 x 27 mL R1
4 x 3 mL R1a
4 x 6 mL R2
4 x 2 mL R2a
Intended Use
Photometric colour test for the quantitative determination of unsaturated iron binding capacity (UIBC) in human serum and plasma on Beckman
Summary1,2,3
Iron participates in a variety of vital processes in the body varying from cellular oxidative mechanisms to the transport and delivery of oxygen to
body cells. It is a constituent of the oxygen-carrying chromoproteins, haemoglobin and myoglobin, as well as various enzymes, such as
cytochrome oxidase and peroxidases. The remaining body iron is present in the flavoproteins, the iron-sulphur proteins, as well as storage iron-
ferritin, and transport iron-transferrin.
Measured serum iron concentration is principally the Fe(III) bound to serum transferrin and does not include the iron contained in serum as free
haemoglobin. Since normally only about one-third of the iron-binding sites of transferrin are occupied by Fe(III), serum transferrin has
considerable reserve iron-binding capacity. This is called the serum unsaturated or latent iron-binding capacity. UIBC measurements can be
used in conjunction with serum iron concentration to obtain the total-iron binding capacity (TIBC) i.e. the maximum concentration of iron that
serum proteins, principally transferrin, can bind.
TIBC is decreased in chronic infections, malignancy, in iron poisoning, renal disease, nephrosis, kwashiorkor and thalassemia. Common causes
for an increase in TIBC include iron deficiency anemia, late pregnancy, oral contraception and viral hepatitis.
Test Principle
2+
Fe from reagent 1 reacts with Nitroso-PSAP from reagent 2 to form an intense green complex. If sample is added a part or all of the iron ions
bind specifically with transferrin at unsaturated iron binding sites at alkaline pH. They are thus not available for the colour reaction with Nitroso-
PSAP. The difference between the resulting changes in the measured absorbances with or without samples is equivalent to the iron quantity
bound to transferrin. This is the unsaturated iron binding capacity (UIBC).

Iron 6.9 µmol/L
Nitroso-PSAP 176 µmol/L
Hydroxylammonium chloride 36 mmol/L
Preservative

R1a: Irritant, contains hydroxylammonium chloride. R43: May cause sensitisation by skin contact.
R2: Contains hydroxylammonium chloride. May produce an allergic reaction.
Safety Phrases:
S24,S37,S60. Avoid contact with skin. Wear suitable gloves. This material and its container must be disposed of as hazardous waste.
Reagent Preparation

Specimen
4
Haemolysed samples should be avoided. Remove serum from red cells immediately to avoid haemolysis.
5
Test Procedure
statement.
Calibration
The calibrator value is traceable to the National Institute of Standards and Technology (NIST) Standard Reference Material (SRM) 937.

2009-08
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the UIBC concentration of each sample.
Adults 27.8 – 63.6 µmol/L (155 – 355 µg/dL)
findings.

from these values.
Linearity
The test is linear within a concentration range of 10 – 100 µmol/L (55 – 550 µg/dL).
Precision
24.56 0.54 2.19 0.82 3.35
35.57 0.39 1.10 0.68 1.93
72.54 0.47 0.65 0.73 1.01
Sensitivity
The lowest detectable level represents the lowest measurable level of UIBC that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient serum samples were used to compare this UIBC OSR61205 assay on the AU640 against another commercially available UIBC assay.
y = 0.988x + 1.956 r = 0.992 n = 115 Sample range = 11.21 – 75.63 µmol/L
®
7

BIBLIOGRAPHY
Company, 1999;1698-1703.
3. Woo J, Henry JB. Metabolic intermediates and inorganic ions. In: Henry JB, ed. Clinical diagnosis and management by laboratory methods. Philadelphia:WB
Saunders Company, 1996:188-190.
serum samples.WHO/DIL/LAB/99.1 Rev.2:36pp.
5. Perrotta G. Iron and total iron binding capacity. In: Kaplan LA, Pesce AJ, eds. Clinical chemistry theory, analysis, and correlation. St Louis: Mosby, 1996:714pp.
6. Data on file at Beckman Coulter Biomedical Ltd.

2009-08

Test No # Test name UIBC ∇ Sample type Ser ∇ Page 1/2
Method: END ∇ Last L -0.1 Last H 0.3 Reaction - ∇ L 10.0* H 100.0*
General LIH ISE Range Test No # Test name UIBC ∇ Sample type Ser ∇ Page 2/2
Test Name: UIBC ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name UIBC ∇
Test Name: UIBC ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 420* 700*
Point 1: # † 420* 700*
Point 2 ∇
Point 2:
MB type factor
# User defined
2009-08
UIBC, AU2700/AU5400 Serum/Plasma Application UIBC, AU680/AU480 Serum/Plasma Application
Test Name: UIBC ∇ < > Type: Serum ∇ Operation Yes ∇

μL μL
Reagent OD limit:
Measuring Point 1: First 0 Last 27 L 10* H 100* Wavelength Pri 800 ∇nm Sec. None ∇nm Factor for Maker A 1 B 0
Test Name: UIBC ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: UIBC ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: UIBC ∇ < > Type Serum ∇ Test Name: UIBC Type Serum Use Serum Cal.
∇ < > ∇ ο
Point 1: # † 420* 700* Point 1: # ∇ † 420* 700*
Point 9 ∇
† System Calibrator Cat No: 66300 Point-1 ∇ Reagent Blank 14 Day 0 Hour
* Values set for working in SI units (µmol/L). To work In µg/dL multiply by 5.585 Point-2 Calibration 14 Day 0 Hour
∇
ф AU680
2009-08
UREA
OSR6134 4 x 25 mL R1
4 x 25 mL R2
OSR6234 4 x 53 mL R1
4 x 53 mL R2
OSR6534 4 x 103 mL R1
4 x 103 mL R2
Intended Use
Kinetic UV test for the quantitative determination of urea in human serum, plasma and urine on Beckman Coulter analysers. For in vitro
Urea reagent OSR6534 for use on the AU680, AU2700 and AU5400 systems only.
Summary1,2
Urea is synthesised in the liver as the final product of protein and amino acid metabolism. Urea synthesis is therefore dependant on daily protein
intake and endogenous protein metabolism. Most of the urea produced during these metabolic processes is eliminated by glomerular filtration,
with 40 – 60% diffusing back into the blood, irrespective of the flow rate in the proximal tubule. Rediffusion in the distal tubule depends on the
urinary flow and is regulated by antidiuretic hormone. During diuresis, there is minimal rediffusion of urea into the blood; a large quantity of urea
is excreted in the urine and plasma urea concentration is low. During antidiuresis, which may occur in oliguric heart failure, exsiccosis or thirst,
urea rediffuses in the tubules at an increased rate and plasma urea is increased. In pre- and post renal kidney failure, the tubular urine flow is
decreased, resulting in increased rediffusion of urea in the distal tubules and increased creatinine secretion. Prerenal elevation of urea occurs in
cardiac decompensation, increased protein catabolism and water depletion. Urea levels may be elevated due to renal causes such as acute
glomerulonephritis, chronic nephritis, polycystic kidney, tubular necrosis and nephrosclerosis. Post renal elevation of urea may be caused by
obstruction of the urinary tract. Plasma urea concentration is determined by renal perfusion, urea synthesis rate, and glomerular filtration rate
(GFR) and may be increased in acute renal failure, chronic renal failure and prerenal azotaemia. In dialysis patients the urea concentration is
representative of protein degradation and is also an indicator of metabolic status. In end-stage renal failure, the urotoxic signs, in particular
those relating to the gastrointestinal system, correlate well with urea concentration. Serum urea and serum creatinine determinations are
frequently performed together in the differential diagnosis of kidney function.
Test Principle3
Urea is hydrolysed in the presence of water and urease to produce ammonia and carbon dioxide. The ammonia produced in the first reaction
+
combines with 2-oxoglutarate and NADH in the presence of glutamate-dehydrogenase (GLDH) to yield glutamate and NAD . The decrease in
NADH absorbance per unit time is proportional to the urea concentration.
Reaction Principle
Urease
+ 2-
Urea + 2 H2O 2 NH4 + CO3
GLDH
+ +
2-Oxoglutarate + 2 NH4 + 2 NADH 2 L-Glutamate + 2 NAD + 2 H2O

Tris buffer 100 mmol/L
NADH ≥ 0.26 mmol/L
Tetra-Sodium diphosphate 10 mmol/L
EDTA 2.65 mmol/L
2-Oxoglutarate ≥ 9.8 mmol/L
Urease ≥ 17.76 kU/L
ADP ≥ 2.6 mmol/L
GLDH ≥ 0.16 kU/L
Preservative

Reagent Preparation
The reagents are ready for use and can be placed directly on board the instrument

Specimen
1
Serum and EDTA or lithium heparinised plasma. Do not use ammonium heparinised plasma.
4
Stable in serum and plasma for 7 days when stored at 2...25°C.
Haemolysed and strongly icteric samples should be avoided.
5
Urine: 24-hour collection without preservatives is recommended.
4

2009-08
Test Procedure
Calibration
The urea values of both calibrators are traceable to the National Institute of Standards and Technology (NIST) Reference Material (SRM) 909b
Level 1.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the urea concentration of each sample.
Reference Intervals
1
Serum/Plasma Adult (Global) 2.8 – 7.2 mmol/L (17 – 43 mg/dL)
6
Newborn 1.4 – 4.3 mmol/L (8.4 – 25.8 mg/dL)
Infant/child 1.8 – 6.4 mmol/L (10.8 – 38.4 mg/dL)
7
Urine 250 – 570 mmol/day (15,000 – 34,200 mg/day)
findings.

from these values.
Linearity
The test is linear within a concentration range of 0.8 – 50 mmol/L (5 – 300 mg/dL) for serum and plasma.
Precision
1.97 0.05 2.28 0.06 3.25
9.71 0.10 1.03 0.23 2.38
37.40 0.34 0.91 0.91 2.42
78.81 0.81 1.02 2.44 3.09
283.97 2.50 0.88 9.68 3.41
436.70 4.80 1.10 14.01 3.21
Sensitivity
The lowest detectable level using serum settings on an AU600 analyser was calculated as 0.38 mmol/L.
The lowest detectable level represents the lowest measurable level of urea that can be distinguished from zero. It is calculated as three
standard deviations of 20 replicates of an analyte free sample.
Method Comparison
Patient serum samples were used to compare this Urea OSR6134 assay on the AU600 against another commercially available urea assay.
y = 0.928x - 0.206 r = 0.999 n = 116 Sample range = 1.38 – 39.44 mmol/L
Patient urine samples were used to compare this Urea OSR6134 assay on the AU2700 against another commercially available urea assay.
®
8
Limitations
2009-08

‡ Dilute samples and Urine Calibrator Cat. No. ODC0025 1:10 with purified H2 0.
† System Calibrator Cat. No.: 66300/ Urine Calibrator Cat. No.ODC0025
Ω Depends on usage pattern in the laboratory.
BIBLIOGRAPHY
1. Thomas L. Urea and blood urea nitrogen (BUN). In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results.
Frankfurt/Main: TH-Books Verlagsgesellschaft, 1998:374- 377.
2. Newman DJ, Price CP. Renal function and nitrogen metabolites. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:
WB Saunders Company, 1999;1239-1241.
3. Talke H, Schubert GE. Enzymatische harnstoffbestimmung in blut und serum im optischen test nach Warburg. Klin Wochenschr 1965;43:174-75.
plasma and serum samples. WHO/DIL/LAB/99.1 Rev.2:44pp, 49pp.
7. Kazmierczak. Urea. In: Kaplan LA, Pesce AJ, eds. Clinical chemistry theory, analysis and correlation. St. Louis: Mosby; 1996:500pp.

2009-08
UREA, AU400/AU640 Serum/Plasma Application UREA, AU600 Serum/Plasma Application

Test No # Test name UREA ∇ Sample type Ser ∇ Page ½
Test Name: UREA ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 2.5 Dil. Vol 0 µL Min. OD Max. OD
Sample: Volume 2.5 µL Dilution 0 µL Pre-Dilution Rate: 1 Reagent 1 vol 50 Dil. Vol 40 µL L 1.0 H 2.5
Method: RATE ∇ Last L 1.5 Last H 2.5 Reaction - ∇ L 0.8* H 50*
General LIH ISE Range Test No # Test name UREA ∇ Sample type Ser ∇ Page 2/2
Test Name: UREA ∇ < > Type: Serum ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name UREA ∇
Test Name: UREA ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Calibration Type: AB Y=AX+B Counts: # Formula 1 Y=AX+B ∇ Process Conc ∇
∇ Formula: ∇ Process: CONC ∇
Point 1: # † 82* 152* Point 1 # ∇ † 82* 152*
MB type factor
MB Type Factor: Calibration Stability Period: Ω
Calibrator stability period Ω
# User defined
† System Calibrator Cat. No.: 66300 # User defined ¤ Analyser default value Ω Depends on usage pattern in the laboratory
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 6. † System Calibrator Cat. No.: 66300
Ω Depends on usage pattern in the laboratory * Values set for working in SI units (mmol/L). To work in mg/dL multiply by 6
2010-06
UREA, AU2700/AU5400 Serum/Plasma Application UREA, AU680/AU480 Serum/Plasma Application
Test Name: UREA ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: UREA ∇ < > Type: Serum ∇ Operation: Yes ∇

µL µL
Reagent OD limit:
Test Name: UREA ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: UREA ∇ < > Type: Serum ∇
# ∇ # # # # # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # # # # # # #
∇
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: UREA ∇ < > Type Serum ∇ R
Test Name: UREA Type Serum Use Serum Cal.
∇ < > ∇
Point 1: # † 106* 192* Point 1: # ∇ † 82* 152*
Point 9 ∇
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 6. Point-1 ∇ Reagent Blank Ω Day 0 Hour
Ω Depends on usage pattern in the laboratory Point-2 ∇ Calibration Ω Day 0 Hour
Ф AU680 MB Type Factor: 1-Point Calibration Point ∇ R with Conc-0
2010-06
UREA, AU400/AU640 Urine Application UREA, AU600
System Reagent: OSR6134, OSR6234 Reagent ID: 034 Manual Dilution Standard Mode Urine Application
Test Name: UREA ∇ < > Type: Urine ∇ Operation: Yes ∇ Test No # Test name UREA ∇ Sample type Uri ∇ Page 1/2
Sample: Volume 2 µL Dilution 0 µL Pre-Dilution Rate: 10 Sample vol. 2‡ Dil. vol 0 µL Min. OD Max. OD
Reagents: R1 Volume 50 µL Dilution 40 µL Min OD Max OD Reagent 1 vol 50 Dil. vol 40 µL L 1.0 H 2.5
R2 Volume 50 µL Dilution 10 µL L 1.0 H 2.5 Reagent 2 vol 50 Dil. vol 10 µL Reagent OD limit
Reagent OD limit: Fst. L 1.5 Fst. H 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.5 First H 2.5 Wave Main 340 Sub 660 ∇ Lst. L 1.5 Lst. H 2.5
Method: RATE ∇ Last L 1.5 Last H 2.5 Method RATE ∇ Dynamic range
Reaction slope: - ∇ Dynamic Range: Reaction - ∇ L 10 H 750*
Measuring Point 1: First 12 Last 18 L 10 H 750* Point 1 Fst 12 Lst 18 Correlation factor A 1
Linearity : 25 % A 1 B 0 Sample Pre-dil. Rate □
Linearity Fst 25 % Sec %
Test No # Test name UREA ∇ Sample type Uri ∇ Page 2/2
Test Name: UREA ∇ < > Type: Urine ∇ Level L Level H
Normal range
Sex Age L Age H L H
R 1. 1 # ∇ # Y # M # Y # M→ # #
# ∇ # # # # # #
R 2. 2 # ∇ # Y # M # Y # M→ # #
# ∇ # # # # # #
R 3. 3 # ∇ # Y # M # Y # M→ # #
# ∇ # # # # # #
R 4. 4 # ∇ # Y # M # Y # M→ # #
# ∇ # # # # # #
R 5. 5 # ∇ # Y # M # Y # M→ # #
# ∇ # # # # # #
R 6. 6 # ∇ # Y # M # Y # M→ # #
# ∇ # # # # # #
7 Non select # #
8 Out of range # #
L H
URINE APPLICATION

General ISE
Test No # Test name UREA ∇
Test Name: UREA ∇ < > Type Urine ∇
Point 1: # † 1000* 2100* Point 1 # ∇ † 1000* 2100*
Point 7:
Point 7 ∇
MB Type Factor: Calibration Stability Period: Ω MB type factor
# User defined
† Urine Calibrator Cat. No.: ODC0025 ‡ Dilute samples and Urine Calibrator Cat. No. ODC0025 1:10 with purified H2 0.
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 6. # User defined ¤ Analyser default value Ω Depends on usage pattern in the laboratory
Ω Depends on usage pattern in the laboratory † Urine Calibrator Cat. No.: ODC0025
2010-06
UREA, AU2700/AU5400 Urine Application UREA, AU680/AU480 Urine Application
Test Name: UREA ∇ < > Type: Urine ∇ Operation Yes ∇
Test Name: UREA ∇ < > Type: Urine ∇ Operation: Yes ∇
µL µL
Reagent OD limit:
Measuring Point 1: First 12 Last 18 L 10 H 750* Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0
No Lag Time: YES ∇ On-board stability period: 30 Measuring Point1 First 12 Last 18
Specific Test Parameters Lag Time Check YES ∇
Test Name: UREA ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: UREA ∇ < > Type: Urine ∇
# ∇ # # # # # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # # # # # # #
∇
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
URINE APPLICATION
General ISE
General ISE
Test Name: UREA ∇ < > Type Urine ∇ R
Test Name: UREA ∇ < > Type Urine ∇ Use Serum Cal.
Point 1: # † 1000* 2100* Point 1: # ∇ † 1000* 2100*
Point 9 ∇
2010-06
UREA, AU400/AU640 Paediatric Application UREA, AU600 Paediatric Application

Test No # Test name UREAP ∇ Sample type Ser ∇ Page 1/2
Test Name: UREAP ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 2.5 Dil. vol 10 µL Min. OD Max. OD
Sample: Volume 2.5 µL Dilution 10 µL Pre-Dilution Rate: 1 Reagent 1 vol 50 Dil. vol 30 µL L 1.0 H 2.5
General LIH ISE Range Test No # Test name UREAP ∇ Sample type Ser ∇ Page 2/2
Test Name: UREAP ∇ < > Type: Serum ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name UREAP
∇
Test Name: UREAP ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Calibration Type: AB Y=AX+B Counts: # Formula 1 Y=AX+B ∇ Process Conc ∇
∇ Formula: ∇ Process: CONC ∇
Point 1: # † 90* 140* Point 1 # ∇ † 90* 140*
MB type factor
# User defined
† System Calibrator Cat. No.: 66300 # User defined ¤ Analyser default value Ω Depends on usage pattern in the laboratory
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 6. † System Calibrator Cat. No.: 66300
Ω Depends on usage pattern in the laboratory * Values set for working in SI units (mmol/L). To work in mg/dL multiply by 6.
2010-06
UREA, AU2700/AU5400 Paediatric Application UREA, AU680/AU480 Paediatric Application
Test Name: UREAP ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: UREAP ∇ < > Type: Serum ∇ Operation: Yes ∇

µL µL
Reagent OD limit:
Test Name: UREAP ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: UREAP ∇ < > Type: Serum ∇
# ∇ # # # # # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # # # # # # #
∇
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: UREAP ∇ < > Type Serum ∇ R
Test Name: UREAP ∇ < > Type Serum ∇ Use Serum Cal.
Point 1: # † 90* 140* Point 1: # ∇ † 90* 140*
Point 9 ∇
2010-06
UREA STAT
OSR6141 4 x 25 mL R1
4 x 25 mL R1-2
OSR6541 4 x 50 mL R1
4 x 50 mL R1-2
Intended Use
Kinetic UV test for the quantitative determination of urea in human serum, plasma and urine on Beckman Coulter analysers. For in vitro diagnostic use
only.
Urea reagent OSR6541 for use on the AU680, AU2700 and AU5400 systems only.
Summary1,2
Urea is synthesised in the liver as the final product of protein and amino acid metabolism. Urea synthesis is therefore dependant on daily protein intake and
endogenous protein metabolism. Most of the urea produced during these metabolic processes is eliminated by glomerular filtration, with 40-60% diffusing
back into the blood, irrespective of the flow rate in the proximal tubule. Rediffusion in the distal tubule depends on the urinary flow and is regulated by
antidiuretic hormone. During diuresis, there is minimal rediffusion of urea into the blood; a large quantity of urea is excreted in the urine and plasma urea
concentration is low. During antidiuresis, which may occur in oliguric heart failure, exsiccosis or thirst, urea rediffuses in the tubules at an increased rate and
plasma urea is increased. In pre- and post renal kidney failure, the tubular urine flow is decreased, resulting in increased rediffusion of urea in the distal
tubules and increased creatinine secretion. Prerenal elevation of urea occurs in cardiac decompensation, increased protein catabolism and water
depletion. Urea levels may be elevated due to renal causes such as acute glomerulonephritis, chronic nephritis, polycystic kidney, tubular necrosis and
nephrosclerosis. Post renal elevation of urea may be caused by obstruction of the urinary tract. Plasma urea concentration is determined by renal
perfusion, urea synthesis rate, and glomerular filtration rate (GFR) and may be increased in acute renal failure, chronic renal failure and prerenal
azotaemia. In dialysis patients the urea concentration is representative of protein degradation and is also an indicator of metabolic status. In end-stage
renal failure the urotoxic signs, in particular those relating to the gastrointestinal system, correlate well with urea concentration. Serum urea and serum
creatinine determinations are frequently performed together in the differential diagnosis of kidney function.
Test Principle3
Urea is hydrolysed in the presence of water and urease to produce ammonia and carbon dioxide. The ammonia produced in the first reaction combines
+
with 2-oxoglutarate and NADH in the presence of glutamate-dehydrogenase (GLDH) to yield glutamate and NAD . The decrease in NADH absorbance per
unit time is proportional to the urea concentration.
Reaction Principle
Urease
+ 2-
Urea + 2 H2O 2 NH4 + CO3
GLDH
+ +
2-Oxoglutarate + 2 NH4 + 2 NADH 2 L-Glutamate + 2 NAD + 2 H2O

Tris buffer 100 mmol/L
NADH ≥ 0.26mmol/L
Tetra-Sodium diphosphate 10 mmol/L
EDTA 2.65 mmol/L
2-Oxoglutarate ≥ 9.8 mmol/L
Urease ≥ 17.76 kU/L
ADP ≥ 2.6 mmol/L
GLDH ≥ 0.16 kU/L
Preservative

To avoid the possible build-up of azide compounds, flush wastepipes with water after the disposal of undiluted reagent.
Reagent Preparation
To prepare the working reagent, slowly add the contents of the bottle labelled R1-2 to the bottle labelled R1. Mix gently by inversion and place on board the
instrument.

The reagent components are stable, unopened, up to the stated expiry date when stored at 2…8°C. Once prepared, reagent stored on board the
instrument is stable for 21 days.
Specimen
1
Serum and EDTA or lithium heparinised plasma. Do not use ammonium heparinised plasma.
4
5
Urine: 24-hour collection without preservatives is recommended.
4
Stable in urine for 7 days at 2…8°C and 2 days when stored at 15…25°C.
Test Procedure

2010-06
Calibration
The urea values of both calibrators are traceable to the National Institute of Standards and Technology (NIST) Reference Material (SRM) 909b Level 1.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the urea concentration of each sample.
Reference Intervals
1
Serum/Plasma Adult 2.8 – 7.2 mmol/L (17 – 43 mg/dL)
6
Urine 250 – 570 mmol/day (15,000 – 34,200 mg/day)

values.
Linearity
The test is linear within a concentration range of 0.8 – 50 mmol/L (5 – 300 mg/dL) for serum and plasma.
Precision
The following data was obtained on an AU600 using 2 serum controls analysed over 20 days.
4.90 0.09 1.80 0.12 2.50
25.64 0.46 1.80 0.56 2.20
93.7 1.93 2.06 3.46 3.70
276 3.25 1.18 11.02 3.99
402 3.92 0.97 14.65 3.64
Sensitivity
The lowest detectable level using serum settings on an AU600 analyser was calculated as 0.23 mmol/L.
The lowest detectable level represents the lowest measurable level of urea that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison
Patient serum samples were used to compare this Urea OSR6141 assay on the AU600 against another commercially available urea assay. Results of
Patient urine samples were used to compare this Urea OSR6x41 assay on the AU2700 against another commercially available urea assay. Results of
®
7
Limitations

2010-06
± Depends on usage pattern in the laboratory.
BIBLIOGRAPHY
1. Thomas L. Urea and blood urea nitrogen (BUN). In:Thomas L, ed. Clinical laboratory diagnostics.Use and assessment of clinical laboratory results.
2. Newman DJ, Price CP. Renal function and nitrogen metabolites. In: Burtis CA, Ashwood ER, eds.Tietz textbook of clinical chemistry. Philadelphia:WB Saunders
Company, 1999;1239-1241.
3. Talke H, Schubert GE. Enzymatische harnstoffbestimmung in blut und serum im optischen test nach Warburg. Klin Wochenschr 1965;43:174-75.
6. Kazmierczak. Urea. In: Kaplan LA, Pesce AJ, eds. Clinical chemistry theory, analysis and correlation. St. Louis: Mosby; 1996:500pp.
EN.01 LOSR6x41.02 Metabolite

2010-06
UREA - STAT, AU400/AU640 Serum/Plasma Application UREA - STAT, AU600 Serum/Plasma Application

Test No # Test name UREAS ∇ Sample type Ser ∇ Page 1/2
Test Name: UREAS ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 2.5 µL Dilution 0 µL Pre-Dilution Rate: 1 Reagent 1 vol 100 Dil. vol 50 µL L 0.7 H 2.5
General LIH ISE Range Test No # Test name UREAS ∇ Sample type Ser ∇ Page 2/2
Test Name: UREAS ∇ < > Type: Serum ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name UREAS ∇
Test Name: UREAS ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 100* 183*
Point 1: # † 100* 183*
Point 2 ∇
Point 2:
MB type factor
MB Type Factor: Calibration Stability Period: Ω Calibrator stability period Ω
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 6. * Values set for working in SI units (mmol/L). To work in mg/dL multiply by 6.
Ω Depends on usage pattern in the Laboratory Ω Depends on usage pattern in the Laboratory
2010-06
UREA - STAT, AU2700/AU5400 Serum/Plasma Application UREA-STAT, AU680/AU480 Serum/Plasma Application
System Reagent: OSR6141, OSR6541 Reagent ID: 041 System Reagent: OSR6141, OSR6541 (AU680) Reagent ID: 041
Test Name: UREAS ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: UREAS ∇ < > Type: Serum ∇ Operation: Yes ∇

µL µL
Reagent OD limit:
Test Name: UREAS ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: UREAS ∇ < > Type: Serum ∇
# ∇ # # # # # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # # # # # # #
∇
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: UREAS ∇ < > Type Serum ∇ R
Test Name: UREAS Type Serum Use Serum Cal.
∇ < > ∇
Point 1: # † 100* 183* Point 1: # ∇ † 100* 183*
Point 9 ∇
Ф AU680 Point-2 ∇ Calibration Ω Day 0 Hour
Ω Depends on usage pattern in the Laboratory MB Type Factor: 1-Point Calibration Point ∇ R with Conc-0
2010-06
UREA - STAT, AU400/AU640 URINE Application UREA - STAT, AU600
System Reagent: OSR6141 Reagent ID: 041 Manual Dilution Standard Mode URINE Application
System Reagent: OSR6141 Reagent ID: 041
Test No # Test name UREAS ∇ Sample type Uri ∇ Page 1/2
Test Name: UREAS ∇ < > Type: Urine ∇ Operation: Yes ∇
Sample vol. 2‡ Dil. vol 0 µL Min. OD Max. OD
Reagent 1 vol 100 Dil. vol 50 µL L 0.7 H 2.5
Reagent 2 vol 0 Dil. vol 0 µL Reagent OD limit
Fst. L 1.3 Fst. H 2.5
Reagent OD limit:
Wave Main 340 Sub 660 ∇ Lst. L 1.3 Lst. H 2.5
Method RATE ∇ Dynamic range
Reaction - ∇ L 10* H 750*
Point 2 Fst Lst B 0
Linearity Fst 25 % Sec %
No lag time YES ∇ On-board stability period 21
Test No # Test name UREAS ∇ Sample type Uri ∇ Page 2/2
Test Name: UREAS ∇ < > Type: Urine ∇ Level L Level H
Normal range
Sex Age L Age H L H
R 1. 1 # ∇ # Y # M # Y # M→ # #
# ∇ # # # # # #
R 2. 2 # ∇ # Y # M # Y # M→ # #
# ∇ # # # # # #
3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # #
R 4. 4 # ∇ # Y # M # Y # M→ # #
# ∇ # # # # # #
R 5. 5 # ∇ # Y # M # Y # M→ # #
# ∇ # # # # # #
R 6. 6 # ∇ # Y # M # Y # M→ # #
# ∇ # # # # # #
7 Non select # #
8 Out of range # #
L H
URINE APPLICATION
General ISE
Test No # Test name UREAS ∇
Test Name: UREAS ∇ < > Type Urine ∇
Point 1: # † 1000* 2100* Point 1 # ∇ † 1000* 2100*
MB Type Factor: Calibration Stability Period: Ω MB type factor
# User defined ‡ Dilute samples and Urine Calibrator Cat. No. ODC0025 1:10 with purified H2O
† Urine System Calibrator Cat. No.: ODC0025 # User defined ¤ Analyser default value
* Values set for working in SI units (mmol/L). To work in mg/dL multiply by 6. † Urine System Calibrator Cat. No.: ODC0025
Ω Depends on usage pattern in the Laboratory * Values set for working in SI units (mmol/L). To work in mg/dL multiply by 6
2010-06
UREA - STAT, AU2700/AU5400 Urine Application UREA-STAT, AU680/AU480 Urine Application
System Reagent: OSR6141, OSR6541 Reagent ID: 041 System Reagent: OSR6141, OSR6541 (AU680) Reagent ID: 041
Test Name: UREAS ∇ < > Type: Urine ∇ Operation Yes ∇
Test Name: UREAS ∇ < > Type: Urine ∇ Operation: Yes ∇
µL µL
Reagent OD limit:
No Lag Time: YES ∇ On-board stability period: 21 Measuring Point1 First 2 Last 6
Specific Test Parameters Lag Time Check YES ∇
Test Name: UREAS ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: UREAS ∇ < > Type: Urine ∇
# ∇ # # # # # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # # # # # # #
∇
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
URINE APPLICATION
General ISE
General ISE
Test Name: UREAS ∇ < > Type Urine ∇ R
Test Name: UREAS ∇ < > Type Urine ∇ Use Serum Cal.
Point 1: # † 1000* 2100* Point 1: # ∇ † 1000* 2100*
Point 9 ∇
† Urine System Calibrator Cat. No.: ODC0025 Calibrator OD Conc Low High Stability
Ф AU680 Point-2 ∇ Calibration Ω Day 0 Hour
Ω Depends on usage pattern in the Laboratory MB Type Factor: 1-Point Calibration Point ∇ R with Conc-0
2010-06
URIC ACID
OSR6098 4 x 12 mL R1
4 x 5 mL R2
OSR6198 4 x 30 mL R1
4 x 12.5 mL R2
OSR6298 4 x 42.3 mL R1
4 x 17.7 mL R2
Intended Use
Enzymatic colour test for the quantitative determination of uric acid in human serum, plasma and urine on Beckman Coulter analysers. For in vitro
Summary1,2
Uric Acid is the major product of purine catabolism in humans. Most uric acid formation occurs in the liver, and is eliminated via the kidney, with the body
uric acid pool determined by the balance between synthesis and elimination. Hyperuricaemia is divided into primary and secondary classifications,
involving either overproduction or reduced elimination. Primary hyperuricaemia is also known as the idiopathic or familial form. In the vast majority of
affected cases, reduced tubular secretion of uric acid is responsible for the elevation in uric acid levels. Approximately 1% of patients with primary
hyperuricaemia have an enzymatic defect in purine metabolism which results in overproduction of uric acid. Primary hyperuricaemia is associated with
gout, Lesch-Nyhan syndrome, Kelley Seegmiller syndrome and increased phosphoribosyl pyrophosphate synthase activity. Secondary hyperuricaemia
may be caused by increased nutritional purine uptake, associated with increased uric acid excretion in the urine. Secondary hyperuricaemia is associated
with numerous conditions including renal insufficiency, myeloproliferative diseases, haemolytic diseases, psoriasis, polycythemia vera, type I glycogen
storage disease, excess alcohol consumption, lead intoxication, a purine-rich diet, fasting, starvation and chemotherapy.
Hypouricaemia may result from decreased uric acid production, such as occurs in hereditary xanthinuria, hereditary purine nucleoside phosphorylase
deficiency and allopurinol therapy. Hypouricaemia may also be due to increased renal uric acid excretion, which may occur in malignant diseases, AIDS,
Fanconi syndrome, diabetes mellitus, severe burns and hypereosinophilic syndrome. In addition, hypouricaemia may result from treatment with uricosuric
agents and ingestion of X-ray contrast media. Quantitation of urinary uric acid excretion may assist in the selection of appropriate treatment for
hyperuricaemia, providing an indication of whether patients should be treated with uricosuric drugs to enhance renal excretion, or allopurinol to supress
purine synthesis.
Test Principle3
Uric acid is converted by uricase to allantoin and hydrogen peroxide. The Trinder reaction is utilised to measure H2O2. The formed H2O2 reacts with
N,N-bis(4-sulfobutyl)-3,5-dimethylaniline, disodium salt (MADB) and 4-aminophenazone in the presence of peroxidase to produce a chromophore, which is
read biochromatically at 660/800nm. The amount of dye formed is proportional to the uric acid concentration in the sample.
Reaction principle
Uricase
Uric acid + O2 + 2 H2O Allantoin + CO2 + H2O2
Peroxidase
+ -
2 H2O2 + MADB + 4-Aminophenazone Blue dye + OH + 3 H2O

Final concentration of active ingredients:
Phosphate Buffer (pH 7.5) 42 mmol/L
MADB 0.15 mmol/L
4-Aminophenazone 0.30 mmol/L
Peroxidase ≥ 5.9 kU/L (98 μkat/L)
Uricase ≥ 0.25 kU/L (4.15 μkat/L)
Ascorbate Oxidase ≥ 1.56 kU/L (26 μkat/L)
Preservative

To avoid the possible build-up of azide compounds, flush waste-pipes with water after the disposal of undiluted reagent. Dispose of all waste material in
accordance with local guidelines.
Refer to safety data sheet for further information.
Risk Phrases
R43: May cause sensitisation by skin contact.
Safety Phrases
S24,S26,S37: Avoid contact with skin. In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable gloves.
S60: This material and its container must be disposed of as hazardous waste.
Reagent Preparation

30 days.

2009-08
Specimen4,5
Stable in serum and plasma for 7 days when stored at 2…8°C and 3 days when stored at 15…25°C.
Urine: Stable in urine for 4 days when stored at 15…25°C. To prevent urate precipitation in urine specimens after collection, add a sufficient volume of
sodium hydroxide to bring the pH between 8 and 9.
EDTA plasma will give a 5 – 10% lower recovery compared to serum or heparinised plasma.
Test Procedure
Refer to the appropriate User’s Guide and Setting Sheet for analyser-specific assay instructions for the sample type as listed in the Intended Use
statement.
Calibration
The uric acid values of both calibrators are traceable to the Isotope Dilution Mass Spectrometry Reference Method (IDMS).
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the uric acid concentration of each sample.
Serum Male 208.3 – 428.4 µmol/L (3.5 – 7.2 mg/dL)
Female 154.7 – 357.0 µmol/L (2.6 – 6.0 mg/dL)
Urine, 24h Average diet 1488 – 4463 µmol/d (250 – 750 mg/d)

Data contained within this section are representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ from these
values.
Linearity
The test is linear within a concentration range of 89 – 1785 µmol/L (1.5 – 30 mg/dL) for serum and plasma. The test is linear within a concentration range of
119 – 23,800 µmol/L (2 – 400 mg/dL) for urine.
Precision
171.84 1.91 1.11 2.94 1.71
388.23 6.02 1.55 9.48 2.44
1362.06 9.90 0.73 28.86 2.12
1360.16 14.39 1.06 22.63 1.66
3660.61 57.57 1.57 71.24 1.95
5604.48 98.50 1.76 93.44 1.67
Sensitivity
The lowest detectable level using serum settings on an AU2700 analyser was estimated at 2 µmol/L.
The lowest detectable level using urine settings on an AU640 analyser was estimated at 10 µmol/L.
The lowest detectable level represents the lowest measurable level of uric acid that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient samples were used to compare this uric acid assay on the AU640 against another commercially available uric acid assay. Results of linear
Serum Samples:
y = 0.964x – 12.498 r = 0.999 n = 116 Sample range = 94 – 1531 µmol/L
Urine Samples:
y = 0.982x + 80.76 r = 1.000 n = 151 Sample range = 142 – 20,653 µmol/L
Results of studies conducted on serum samples to evaluate the susceptibility of the method to interference were as follows:
Ascorbate: Interference less than 5% up to 20 mg/dL ascorbate.
Icterus: Interference less than 5% up to 40 mg/dL or 684 µmol/L unconjugated bilirubin.
Interference less than 10% up to 20 mg/dL or 342 µmol/L conjugated bilirubin.
Haemolysis: Interference less than 5% up to 5 g/L haemoglobin.
® ®
Lipemia: Interference less than 5% up to 1000 mg/dL Intralipid. (AU480 Interference less than 10% up to 1000 mg/dL Intralipid. )
2009-08
Results of studies conducted on urine samples to evaluate the susceptibility of the method to interference were as follows:
Ascorbate: Interference less than 5% up to 50 mg/dL ascorbate.
In very rare cases gammopathy, especially IgM (Waldenström’s macroglobulinemia), may cause unreliable results.
7

BIBLIOGRAPHY
1. Thomas L. Uric acid. In: Thomas L, Hrsg. Labor und Diagnose. Indikation und Bewertung von Laborbefunden für die medizinische Diagnostik.
2. Newman DJ, Price CP. Renal function and nitrogen metabolites. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:
WB Saunders Company, 1999;1245-50.
3. Barham D, Trinder P. An improved colour reagent for the determination of blood glucose by the oxidase system. Analyst 1972;97:142-5.
plasma and serum samples. WHO/DIL/LAB/99.1 Rev.2: 44pp & 49pp.
5. Data on file at OLMEI.
Philadelphia: WB Saunders Company, 1999;1838pp.
7. Young DS, Effects of Drugs on CLINICAL Laboratory Tests, AACC, 5th ed. CCPress, 2000.

2009-08
URIC ACID, AU400/AU640 Serum/Plasma Application URIC ACID, AU600 Serum/Plasma Application

Test No # Test name UA ∇ Sample type Ser ∇ Page ½
Test Name: UA ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name UA ∇ Sample type Ser ∇ Page 2/2
Test Name: UA ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name UA ∇
Test Name: UA ∇ < > Type Serum ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 2800* 5200*
Point 1: # † 2800* 5200*
Point 2 ∇
Point 2:
MB type factor
# User defined
* Values set for working in SI units (µmol/L) . To work in mg/dL divide by 59.5
2009-08
URIC ACID, AU2700/AU5400 Serum/Plasma Application URIC ACID, AU680/AU480 Serum/Plasma Application
Test Name: UA ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: UA ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: UA ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: UA ∇ < > Type: Serum ∇

ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: UA ∇ < > Type Serum ∇
Test Name: UA ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † 2800* 5200* Point 1: # ∇ † 2800* 5200*
Point 9 ∇
† System Calibrator Cat. No.: 66300. Calibrator OD Conc Low High Stability
* Values set for working in SI units (µmol/L) . To work in mg/dL divide by 59.5 Point-1 ∇ Reagent Blank 30 Day 0 Hour
2009-08
URIC ACID, AU400/AU640 Urine Application URIC ACID, AU600 Urine Application
General LIH ISE Range Test No # Test name UA ∇ Sample type Uri ∇ Page 1/2
Test Name: UA ∇ < > Type: Urine ∇ Operation: Yes ∇ Sample vol. 1.5 Dil. vol 0 μL Min. OD Max. OD
Sample: Volume 1.5 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 2 vol 25 Dil. vol 10 Reagent OD limit
μL
Reagents: R1 Volume 60 μL Dilution 90 μL Min OD Max OD Fst. L -0.1 Fst. H 0.3
R2 Volume 25 μL Dilution 10 μL L H Wave Main 660 Sub 800 ∇ Lst. L -0.1 Lst. H 0.3
Wavelength: Pri. 660 ∇ Sec. 800 ∇ First L -0.1 First H 0.3 Reaction + L 119* H 23800*
∇
Measuring Point 1: First 0 Last 27 L 119* H 23800* Sample Pre-dil. Rate ¤
Linearity : % A 1 B 0 Linearity Fst % Sec %
No Lag Time: ∇ On-board stability period: 30 No lag time ∇ On-board stability period 30
Specific Test Parameters Test No # Test name UA ∇ Sample type Uri ∇ Page 2/2
Level L Level H
Test Name: UA ∇ < > Type: Urine ∇ Value/flag # ∇ # #
Normal range
Value/Flag: # ∇ Level L: # Level H: # Sex Age L Age H L H
Normal Ranges: Age L Age H 1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 7 Non select # #
ο 6. # ∇ # # # # # # 8 Out of range # #
7. None Selected # # L H
8. Out of Range L H # # Panic value # #
Panic Value: # # Unit: µmol/L* Decimal places: # Select the function using the Function key or the Mouse
URINE APPLICATION
Calibration Specific Test No # Test name UA ∇
General ISE
Test Name: UA ∇ < > Type Urine ∇ Formula 1 Y=AX+B Process Conc
∇ ∇
Point 1: # † 12250* 22750* Point 2 ∇
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ MB type factor
# User defined † Urine Calibrator Cat. No.: ODC0025
† Urine Calibrator Cat. No.: ODC0025 * Values set for working in SI units (µmol/L). To work in mg/dL divide by 59.5
* Values set for working in SI units (µmol/L) . To work in mg/dL divide by 59.5
2009-08
URIC ACID, AU2700/AU5400 Urine Application URIC ACID, AU680/AU480 Urine Application
Test Name: UA ∇ < > Type: Urine ∇ Operation Yes ∇
Test Name: UA ∇ < > Type: Urine ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Linearity Limit %
Test Name: UA ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: UA ∇ < > Type: Urine ∇

ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
URINE APPLICATION
General ISE
General ISE
Test Name: UA ∇ < > Type Urine ∇ Test Name: UA ∇ < > Type Urine ∇ ο Use Serum Cal.
Point 1: # † 12250* 22750* Point 1: # ∇ † 12250* 22750*
Point 9 ∇
* Values set for working in SI units (µmol/L) . To work in mg/dL divide by 59.5 Point-1 ∇ Reagent Blank 30 Day 0 Hour
2009-08
URINARY/CSF PROTEIN
OSR6170 4 x 19 mL R1
1 x 3 mL Calibrator
Intended Use
Photometric colour test for the quantitative determination of total protein in human urine and cerebrospinal fluid (CSF) on Beckman Coulter
Summary1
Measurement of total protein in urine is important in the diagnosis and treatment of diseases associated with renal, cardiac and thyroid function.
These diseases are often characterised by proteinuria of which there are four main types: (a) increased glomerular permeability (glomerular
proteinuria) (b) defective tubular reabsorption (tubular proteinuria) (c) increased concentration of low molecular weight protein (overload
proteinuria) (d) abnormal secretion of protein into the urinary tract (postrenal proteinuria). Increased levels of urinary protein may also be
present following strenuous exercise or in the following conditions: monoclonal gammopathies, nephritis, diabetic nephropathy or urinary tract
infections.
The measurement of total protein in CSF is important in detecting increased permeability of the blood/brain barrier to plasma proteins or to
detect increased intrathecal production of immunoglobulins. Increased permeability of the blood brain barrier may result from conditions such as
brain tumour, intracerebral haemmorhage or by inflammation caused by bacterial or viral meningitis, encephalitis or poliomyelitis. Determination
of increased intrathecal synthesis of immunoglobulins is important in the diagnosis of demyelinating diseases such as multiple sclerosis.
Test Principle1,2,3
Pyrogallol red is combined with molybdate to form a red complex with a maximum absorbance at 470nm. The assay is based on the shift in
absorption that occurs when the pyrogallol red-molybdate complex binds basic amino groups of protein molecules. A blue-purple complex is
formed with a maximum absorbance at 600 nm. The absorbance of this complex is directly proportional to the protein concentration in the
sample.

R1 Calibrator
Pyrogallol Red 47 µmol/L Human Serum Albumin 0.50 g/L
Sodium Molybdate 320 µmol/L Also contains preservatives
Succinic Acid 50 mmol/L
Sodium Benzoate 3.5 mmol/L
Sodium Oxalate 1.0 mmol/L
Methanol 0.8 % w/v

Calibrator: Irritant, contains a mixture of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one. R43; May cause sensitisation
by skin contact. Biological materials of human origin contained in the calibrator were tested for anti-HCV, HbsAg and Anti-HIV 1/2 on a single
donor basis using FDA approved methods and were found to be non-reactive. As there is no known test method that can offer complete
assurance that products derived from human blood will not transmit infectious agents, this product should be handled as a potentially infectious
material.
Safety Phrases:
S24, S37, S60. Avoid contact with skin. Wear suitable protective clothing and gloves. This material and its container must be disposed of as
hazardous waste.
To avoid the possible build-up of azide compounds, flush waste-pipes with water after the disposal of calibrator.
Reagent Preparation
R1 is ready for use and can be placed directly on board the instrument. Protect R1 from direct sunlight. The Calibrator is ready for use.

1. The unopened reagent and calibrator are stable until the expiration date printed on the label when stored at 2…8°C.
2. Once open, reagent stored on board the instrument is stable for 90 days. The opened calibrator is stable until the expiration date on the
label, provided that the stopper and cap are replaced immediately after each use to avoid contamination and the calibrator is stored at
2…8°C.
Specimen
Urine or cerebrospinal fluid.
4,5,6
Urine: A 24 hour or 12 hour urine specimen with no preservative is preferred.
7
Sample Stability: Analyse fresh otherwise stable stored at 2...8°C for up to 48 hours.
Urine samples contaminated by haemoglobin will result in a falsely elevated value.
CSF: Beckman Coulter recommends that CSF samples be collected in plain collection devices.
In the case of CSF samples care should be taken to avoid blood contamination during collection.
4
Sample Stability: Analyse fresh, otherwise stable stored at 4°C for up to 72 hours.
As with all dye based methods, analysis of urine samples containing immunoglobulin light chains (i.e. Bence-Jones Protein) may result in the
underestimation of protein. Where such samples are suspected it is recommended that the sample be concentrated and further analysed via
1
electrophoresis.
Discrepancies may arise when analysing total urine protein in samples from patients who have been treated with polypeptide-based plasma
8
substitutes. The polypeptides from the plasma substitute may be excreted into the urine and result in an elevated total urine protein result.
Where such samples are suspected it is recommended that the sample be concentrated and further analysed via electrophoresis.

2009-08
Test Procedure
Calibration
Use Calibrator provided in the kit. For value assigned to the calibrator please refer to bottle label. The calibrator is traceable to a primary
standard which is prepared gravimetrically using reagent grade human serum albumin.
Quality Control
Control material with values determined by this Beckman Coulter system may be used.
Calculation
The Beckman Coulter analysers automatically compute the total protein concentration of each sample.
Reference Intervals
1
Urine 0.05 – 0.08 g/day at rest
Value may increase to up to 0.30 g/day following exercise.
1
CSF (Adults) 0.15 – 0.45 g/L
1
CSF (newborn <1month) 0.15 – 1.30 g/L
findings.

from these values.
Linearity
The test is linear within a concentration range of 0.01 – 2.00 g/L.
Precision
9
Estimates of precision based on NCCLS recommendations are consistent with typical performance. The following data was obtained on an
AU640 using 3 urine pools analysed over 20 days.
n=80 Within run Total
0.15 0.003 1.7 0.007 4.8
0.53 0.006 1.2 0.010 1.9
1.52 0.011 0.7 0.026 1.7
Sensitivity
The lowest detectable level on an AU640 analyser was estimated at 0.007 g/L.
The lowest detectable level represents the lowest measurable level of total protein that can be distinguished from zero. It is calculated as the
absolute mean plus three standard deviations of 20 replicates of an analyte-free sample.
Method Comparison
Patient urine samples were used to compare this Urinary/CSF Protein OSR6170 assay on the AU640 against another commercially available
urinary/CSF protein assay. Results of linear regression analysis were as follows:
y = 0.957x + 0.009 r = 0.998 n = 108 Sample range = 0.01 – 1.99 g/L
Results of studies conducted on the AU400, AU600/AU640, and AU2700/AU5400 show that the following substances interfere with this
Urinary/CSF Protein procedure by < 10%:
Substance Level Tested (mmol/L) Level Tested (g/L)
Ammonia 139
Ascorbate 1.1
Bilirubin 0.3
Citric Acid 10
Creatinine 26
2+
Cu 1.6
3+
Fe 1.1
Gentamycin 0.04
Glucose 277
Oxalic Acid 7.8
Tartaric Acid 13
Tobramycin 0.04
Uric Acid 18
10

† Urinary/CSF Protein Calibrator supplied with the kit
* Values set for working in SI units (g/L). To work in traditional units (mg/dL) multiply by 100.

2009-08
BIBLIOGRAPHY
nd
1. Tietz NW. Textbook of clinical chemistry, 2 ed. Philadelphia: WB Saunders Company, 1994.
2. Fujita Y, Mori I, Kitano S. Color reaction between pyrogallol red molybdenum (VI) complex and protein. Bunseki Kagaku 1983;32:379-386.
3. Watanabe N, Kamei S, Ohkubo A, Yamanaka M, Ohsawa S,Makino K, Tokuda K. Urinary protein as measured with pyrogallol
red-molybdate complex manually and in Hitachi 726 automated analyser. Clin Chem 1986;32:1551-1544.
rd
4. Tietz NW. Clinical guide to laboratory tests, 3 ed. Philadelphia: WB Saunders Company, 1995:518-520.
5. NCCLS. Urinalysis and collection, transportation and preservation of urine specimens; approved guideline - second edition. NCCLS
document GP16-A2; 2001.
nd
6. Young DS. Effects of preanalytical variables on clinical laboratory tests, 2 ed. AACC Press, 1997.
rd
7. First MR. Renal Function. In: Kaplan LA, Pesce, AJ, eds. Clinical chemistry: theory, analysis and correlation, 3 ed. St. Louis: Mosby-Year
Book, 1996:484-504.
8. Pena C, Martinez-Bru C, Homs R, Planella T, Cortes M. Effect of plasma replacement therapy on determinations of urine protein
concentration [Letter]. Clin Chem 1998;44:359-360.
9. NCCLS. Evaluation of precision performance of clinical chemistry devices; approved guideline. NCCLS document EP5-A;1999.
th
10. Young DS. Effects of drugs on clinical laboratory tests, AACC, 5 ed. AACC Press, 2000.

2009-08
URINARY/CSF PROTEIN, AU400/AU640 Urine/CSF Application URINARY/CSF PROTEIN, AU600 Urine/CSF Application

Test No # Test name UCSFP ∇ Sample type Uri ∇ Page 1/2
Test Name: UCSFP ∇ < > Type: Urine ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name UCSFP ∇ Sample type Uri ∇ Page 2/2
Test Name: UCSFP ∇ < > Type: Urine ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic Value: # # Unit: g/L Decimal places: # Panic value # #
URINE/CSF APPLICATION
General ISE Test No # Test name UCSFP ∇
Test Name: UCSFP ∇ < > Type Urine ∇ Cal type 8 AB ∇ Count #
Point 1 # ∇ † 4* 9*
Point 1: # † 4* 9*
Point 2 ∇
Point 2:
MB type factor
# User defined
† Urinary/CSF Protein Calibrator supplied with kit
* Values set for working in g/L. To work in traditional units (mg/dL) multiply by 100
2009-08
URINARY/CSF PROTEIN, AU600 Automated Pre-Dilution URINARY/CSF PROTEIN, AU2700/AU5400 Urine/CSF Application
Urine/CSF Application System Reagent: OSR6170 Reagent ID: 070
Specific test parameters General LIH ISE Range
Test Name: UCSFP ∇ < > Type: Urine ∇ Operation: Yes ∇

Test No # Test name UCSFP ∇ Sample type Uri ∇ Page 1/2
Reagent 2 vol 0 Dil. vol 0 μL Reagent OD limit
Reagent OD limit:
Fst. L 0.05 Fst. H 0.2
Wave Main 600 Sub 800 ∇ Lst. L 0.05 Lst. H 0.2
Method END ∇ Dynamic range
Reaction + ∇ L 0.01* H 2.00*
Point 2 Fst Lst B 0
Sample Pre-dil. Rate 5
Test No # Test name UCSFP ∇ Sample type Uri ∇ Page 2/2 Test Name: UCSFP ∇ < > Type: Urine ∇
Level L Level H
Normal range
Sex Age L Age H L H
ο 1. # ∇ # # # # # #
1 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # #
2 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # #
3 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # #
4 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # #
5 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # #
6 # ∇ # Y # M # Y # M→ # #
7 Non select # #
8 Out of range # #
L H Panic Value: # # Unit: g/L* Decimal places: #
Panic value # #
Calibration specific General ISE
URINE/CSF APPLICATION
Test No # Test name UCSFP ∇ Test Name: UCSFP ∇ < > Type Urine ∇
Cal type 8 AB ∇ Count # Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇
Selection calibrator Cal. No. OD CONC Factor/OD-L Factor/OD-H
Cal No OD Conc Factor/OD-L Factor/OD-H Point 1: # † 4* 9*
Point 1 # ∇ † 4* 9* Point 2:
Point 2 ∇ Point 3:
Point 7 ∇ 1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇
1-point cal. point MB Type Factor: Calibration Stability Period: 999
MB type factor
Select the function using the Function key or the Mouse # User defined.
# User defined ¤ Analyser default value * Values set for working in g/L. To work in traditional units (mg/dL) multiply by 100
2009-08
URINARY/CSF PROTEIN, AU680/AU480 Urine/CSF Application
Test Name: UCSFP ∇ < > Type: Urine ∇ Operation Yes ∇

Method END ∇
Linearity Limit %
Lag Time Check ∇

Test Name: UCSFP ∇ < > Type: Urine ∇
URINE/CSF
ο 1. # ∇ # # # # # #
URINE/CSF
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
APPLICATION
General ISE
Test Name: UCSFP ∇ < > Type Urine ∇ ο Use Serum Cal.
APPLICATION
Point 1: # ∇ † 4* 9*
Point 3: ∇
Point 6: ∇
Point 9 ∇
Point-1 ∇ Reagent Blank 999 Day 0 Hour # User defined.
Point-2 Calibration 999 Day 0 Hour † Urinary/CSF Protein Calibrator supplied with kit
∇
MB Type Factor: 1-Point Calibration Point ∇ ο with Conc-0 ф AU680
2009-08
α-1 ACIDGLYCOPROTEIN
OSR6162 4 x 20 mL R1
4 x 4.5 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of α-1 acidglycoprotein in human serum and plasma on Beckman Coulter analysers. For
Summary1
α-1 acidglycoprotein (also known as orosomucoid) contains a high percentage of carbohydrate and a large number of sialic acid residues giving it a very
high net negative charge and high solubility in water. α-1 acidglycoprotein is synthesised primarily by the hepatic parenchymal cells, but granulocytes and
monocytes may also contribute significantly to plasma levels in sepsis. It binds and inactivates a large number of basic and lipophilic compounds including
progesterone and related hormones as well as the progesterone antagonist RU486. It also binds and reduces the bioavailability of many drugs including
cocaine and benzodiazepine.
α-1 acidglycoprotein is an acute phase reactant, plasma levels show a 3-4 fold increase in most conditions associated with inflammation and tissue
necrosis. It may be one of the most reliable indicators of clinical activity of ulcerative colitis. Levels are also increased by glucocorticoid effects, either
endogenous (e.g. Cushing's syndrome) or exogenous as in prednisolone or dexamethasone therapy.
Plasma levels are decreased by oestrogens and due to nephrotic syndrome and protein losing enteropathies.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, human α-1 acidglycoprotein reacts specifically with the anti-human
α-1 acidglycoprotein antibodies to yield insoluble aggregates. The absorbance of these aggregates is proportional to the α-1 acidglycoprotein

Tris buffer (pH 7.6) 81 mmol/L
Goat anti-human α-1 acidglycoprotein antiserum Variable
Preservative

Harmful, contains sodium azide. R22; Harmful if swallowed.
Safety Phrases:
S36, S60. Wear suitable protective clothing. This material and its container must be disposed of as hazardous waste.
Reagent Preparation

90 days.
Specimen
2
Stable in serum and plasma for 5 months when stored at 2…25°C.
Test Procedure
Calibration
Serum Protein Multi-Calibrator 2 Cat. No. ODR3023.
The calibrator α-1 acidglycoprotein values are traceable to IFCC (International Federation of Clinical Chemistry) standard CRM 470.
Change in reagent lot or significant shift in control values,
Following calibration, the resulting curve should be visually reviewed, on the Beckman Coulter analyser, for acceptability using the software options -
Routine, Calibration Monitor, Calibration Curve. Quality control procedures should be undertaken immediately following calibration in accordance with good
laboratory practice.
Quality Control
ITA Control Sera ODC0014, ODC0015 and ODC0016 or other control materials with values determined by this Beckman Coulter system may be used.
Calculation
The Beckman Coulter analysers automatically compute the α-1 acidglycoprotein concentration of each sample.
EN.01 BLOSR6x62.01 Specific Protein

2009-08
Serum Adult 0.5 – 1.2 g/L (50 – 120 mg/dL)

values.
Linearity
The test is linear within a concentration range of 0.2 – 2.0 g/L (20 – 200 mg/dL).
Precision
0.27 0.01 2.23 0.01 4.18
1.05 0.02 2.10 0.03 2.92
1.57 0.03 1.63 0.03 2.11
Sensitivity
The lowest detectable level on an AU600 analyser was calculated as 0.047 g/L.
The lowest detectable level represents the lowest measurable level of α-1 acidglycoprotein that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this α-1 Acidglycoprotein OSR6162 assay on the AU640 against another commercially available α-1
acidglycoprotein assay. Results of linear regression analysis were as follows:
®
4
Limitations
Samples with extremely abnormal optical characteristics, especially turbidity, may produce atypical results.
Samples with very high α-1 acidglycoprotein concentrations (> 5 g/L) can generate false low results without appropriate "Z" flags due to excess antigen in
the sample.

† Serum Protein Multi-Calibrator 2 Cat. No.: ODR3023
* Values set for working in SI units (g/L). To work in mg/dL multiply by 100.
BIBLIOGRAPHY
1. Johnson AM, Rohlfs EM, Silverman LM. Proteins. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders Company,
1999; 485-486.
2. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of Anticoagulants in Diagnostic Laborator Investigations and Stability of Blood, Plasma and
3. Baudner S, Dati F. Standardization of the measurement of 14 proteins in human serum based on the new IFCC/BCR/CAP International reference material CRM
470. J Lab Med 1996;20:145-152.
Specific Protein BLOSR6x62.01 EN.01

2009-08
α-1-ACIDGLYCOPROTEIN, AU400/AU640 Serum/Plasma Application α-1-ACIDGLYCOPROTEIN, AU600 Serum/Plasma Application

Test No # Test name AAG ∇ Sample type Ser ∇ Page ½
Test Name: AAG ∇ < > Type: Serum ∇ Operation: Yes ∇
Reagent OD limit: Wave Main 660 Sub NONE ∇ Lst. L -0.1 Lst. H 1.5
General LIH ISE Range Test No # Test name AAG ∇ Sample type Ser ∇ Page 2/2
Test Name: AAG ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name AAG ∇
Test Name: AAG ∇ < > Type Serum ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
1-Point Cal. Point: ο with CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ 1-point cal. Point
MB type factor
# User defined
Specific Protein BSOSR6x62.01

2009-08
α-1-ACIDGLYCOPROTEIN, AU2700/AU5400 α-1-ACIDGLYCOPROTEIN, AU680/AU480 Serum/Plasma Application
Serum/Plasma Application Parameters Specific Test Parameters
System Reagent: OSR6162 Reagent ID: 062 General LIH ISE HbA1c Calculated Test Range
Specific Test Parameters Test Name: AAG ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: AAG ∇ < > Type: Serum ∇ Operation: Yes ∇ Sample Volume 1.6 μL Dilution 0 μL OD Limit
Sample: Volume 1.6 μL Dilution 0 μL Pre-Dilution Rate: 1 Rgt. Volume R1(R1-1) 160 μL Dilution 0 μL Reagent OD Limit
Reagents: R1 Volume 160 μL Dilution 0 μL Min OD Max OD First Low -0.1 High 1.5
R2 Volume 36 μL Dilution 10 μL L H Last Low -0.1 High 1.5
Reagent OD limit: R2(R2-1) 36 μL Dilution 10 μL
Wavelength: Pri. 660 ∇ Sec. None ∇ First L -0.1 First H 1.5 Dynamic Range Low 0.2* High 2*
Method: END ∇ Last L -0.1 Last H 1.5 Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Reaction slope: + ∇ Dynamic Range: Wavelength Pri 660 ∇nm Sec. None ∇nm Factor for Maker A 1 B 0
Measuring Point 1: First 0 Last 27 L 0.2* H 2* Method END ∇
Measuring Point 2: First 0 Last 10 Correlation Factor: Reaction Slope + ∇ Onboard Stability Period 90 Day 0 Hour
Linearity : % A 1 B 0 Measuring Point1 First 0 Last 27 LIH Influence Check # ∇
No Lag Time: ∇ On-board stability period: 90 Measuring Point2 First 0 Last 10 Lipemia +++++ ∇

Test Name: AAG ∇ < > Type: Serum ∇ Test Name: AAG ∇ < > Type: Serum ∇
Value/Flag: # ∇ Level L: # Level H: # Value/Flag: # ∇ Low High

Normal Ranges: Age L Age H Level # # Panic Value
Sex Year Month Year Month L H Specificl Ranges: From To Low High
ο 2. # ∇ # # # # # # ο 1. # ∇ # # # # # #
ο 3. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 4. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 4. # ∇ # # # # # #
6. # # # # # # # ο 5. # ∇ # # # # # #
ο ∇
ο 6. # ∇ # # # # # #
Panic Value: # # Unit: g/L* Decimal places: #
Calibration Specific General ISE
General ISE
Test Name: AAG ∇ < > Type Serum ∇ ο Use Serum Cal.
Test Name: AAG Type Serum
∇ < > ∇
Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: # ∇
Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇ <Calibrator Parameters> OD Range
Calibrator OD Conc Low High Slope Check + ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H Point 1: # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5 Point 2: # ∇ † -0.1 2.5 Allowance Range Check
Point 2: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 4: # ∇ † -0.1 2.5 ο Reagent Blank
Point 4: # † -0.1 2.5 Point 5: # ∇ † -0.1 2.5 ο Calibration
Point 5: # † -0.1 2.5 Point 6: ∇
Point 9 ∇
1-Point Cal. Point: ο with CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇
MB Type Factor: Calibration Stability Period: 999 <Point Cal. For No. of Correction Points Use Master Curve
# User defined. Point-1 ∇ Reagent Blank 999 Day 0 Hour
† Serum Protein Multi-Calibrator 2 Cat. No.: ODR3023 Point-2 ∇ Calibration 999 Day 0 Hour
MB Type Factor: 1-Point Calibration Point None ∇ ο with Conc-0
Ф AU680

2009-08
α-1 ANTITRYPSIN
OSR6163 4 x 20 mL R1
4 x 6.5 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of α-1 antitrypsin in human serum and plasma on Beckman Coulter analysers. For in vitro
Summary1,2
α-1 antitrypsin, a glycoprotein with a molecular mass of 51 kDa is present in approximately equal concentrations in plasma and interstitial fluid. Although
there is some local tissue synthesis (e.g. in monocytes and macrophages) nearly all plasma α-1 antitrypsin is synthesised by the hepatic parenchymal
cells. Also referred to as serine protease inhibitor (α1- proteinase inhibitor or α1-Pi) it inhibits the serine proteases trypsin, chymotrypsin as well as
pancreatric and especially granulocytic elastase. It forms tetrahedral complexes with the active sites of serine proteases thus blocking their enzyme activity.
Physiologically it is most important as an inhibitor of leukocyte elastase, which is released in the process of phagocytosis by polymorphonuclear
leukocytes.
Increased levels of α-1 antitrypsin are common as it is an acute phase reactant whose plasma concentrations rise several fold in the case of acute or
chronic inflammation. Elevated levels are also seen in late pregnancy and during oestrogen therapy because the synthesis of α-1 antitrypsin is stimulated
by oestrogens.
Low levels of α-1 antitrypsin are found in neonatal respiratory distress syndrome, severe neonatal hepatitis, preterminal disease of the pancreas and in
severe protein losing enteropathies. A hereditary deficiency of α-1 antitrypsin is associated with pulmonary emphysema and diseases of the liver including
neonatal cholestasis (hepatitis), cirrhosis and hepatocellular carcinoma.
In α-1 antitrypsin deficiency, levels can fall within the reference interval because of its acute phase function in cases of inflammation. CRP can be
measured to distinguish such cases.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, human α-1 antitrypsin reacts specifically with the anti-human α-1 antitrypsin antibodies
to yield insoluble aggregates. The absorbance of these aggregates is proportional to the α-1 antitrypsin concentration in the sample.
Goat anti-human α-1 antitrypsin antibodies Variable
Preservative
Harmful, contains sodium azide. R22; Harmful if swallowed.
Safety Phrases:
Reagent Preparation
90 days.
Specimen
3
Stable in serum and plasma for 5 months when stored at 2…8°C and 3 months when stored at 15…25°C.
Test Procedure
Calibration
The calibrator α-1 antitrypsin values are traceable to IFCC (International Federation of Clinical Chemistry) standard CRM 470.
Quality Control

2009-08
Calculation
The Beckman Coulter analysers automatically compute the α-1 antitrypsin concentration of each sample.
Serum Adult 0.9 – 2.0 g/L (90 – 200 mg/dL)
values.
Linearity
Precision
0.40 0.006 1.42 0.009 2.24
2.98 0.029 0.96 0.059 1.98
4.43 0.036 0.81 0.094 2.12
Sensitivity
The lowest detectable level on an AU600 analyser was calculated as 0.01 g/L.
The lowest detectable level represents the lowest measurable level of α-1 antitrypsin that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient serum samples were used to compare this α-1 Antitrypsin OSR6163 assay on the AU640 against another commercially available
α-1 antitrypsin assay. Results of linear regression analysis were as follows:
®
5
Limitations
Samples with very high α-1 antitrypsin concentrations (> 16 g/L) can generate false low results without appropriate "Z" flags due to excess antigen in the
sample.
BIBLIOGRAPHY
1. Johnson AM, Rohlfs EM, Silverman LM. Proteins. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders
Company, 1999;486-490.
2. Thomas L. a1-Antitrypsin (a1-AT). In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
4. Baudner S, Dati F. Standardization of the measurement of 14 proteins in human serum based on the new IFCC/BCR/CAP International reference
material CRM 470. J Lab Med 1996;20:145-152.

2009-08
α-1-ANTITRYPSIN, AU400/AU640 Serum/Plasma Application α-1-ANTITRYPSIN, AU600 Serum/Plasma Application

Test No # Test name AAT ∇ Sample type Ser ∇ Page ½
Test Name: AAT ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name AAT ∇ Sample type Ser ∇ Page 2/2
Test Name: AAT ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name AAT ∇
Test Name: AAT ∇ < > Type Serum ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
MB type factor
# User defined

2009-08
α-1-ANTITRYPSIN, AU2700/AU5400 Serum/Plasma Application α-1-ANTITRYPSIN, AU680/AU480 Serum/Plasma Application
Test Name: AAT ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: AAT ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Measuring Point 1: First 0 Last 27 L 0.3* H 5* Wavelength Pri 600 ∇nm Sec. None ∇nm Factor for Maker A 1 B 0
Test Name: AAT ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: AAT ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: AAT ∇ < > Type Serum ∇ Test Name: AAT Type Serum Use Serum Cal.
∇ < > ∇ ο
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇
† Serum Protein Multi-Calibrator 2 Cat. No.: ODR3023 Calibrator OD Conc Low High Stability
* Values set for working in SI units (g/L). To work in mg/dL multiply by 100. Point-1 ∇ Reagent Blank 999 Day 0 Hour

2009-08
APO A1
OSR6142 4 x 13 mL R1
4 x 13 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of Apo A1 (Apolipoprotein A1) in human serum on Beckman Coulter analysers. For
Summary1,2,3
Lipids are transported throughout the body by complex structures called lipoproteins. Lipoproteins are classified into five major density classes:
chylomicrons, very low-density lipoprotein (VLDL), intermediate density lipoprotein (IDL), low-density lipoprotein (LDL) and high-density
lipoprotein (HDL). Over the past several decades, decreased serum levels of high-density lipoprotein (HDL) and increased levels of low-density
lipoprotein (LDL) have been associated with increased risk of coronary artery disease.
Associated with these lipoproteins, at least five major apolipoproteins have been described and have been labelled A through E. The principle
apolipoproteins of HDL are the A apolipoproteins, constituting nearly 90% of the protein mass. Apo A1 has a role in the removal of excess
cholesterol from the tissues and reduced levels of Apo A1 have been observed in patients with coronary heart disease. Apo A1 measurements
are frequently used in characterising patients with genetic disorders that lead to low HDL cholesterol concentrations. Apo B plays an essential
role in the delivery of cholesterol to the tissues, the most abundant form of Apo B, Apo B100 is present in all atherogenic lipoprotein fractions
VLDL, IDL and LDL. Elevated levels of ApoB100 are associated with an increased risk of coronary artery disease.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, Apo A1 reacts specifically with anti-human Apo A1 antibodies to yield
insoluble aggregates. The absorbance of these aggregates is proportional to the Apo A1 concentration in the sample.

TRIS buffer (pH 7.4) 8 mmol/L
Polyethylene glycol 6000 3.5 % w/v
Goat anti-Apo A1 antibodies ≈ 0.14 g/L
Preservative

Reagent Preparation

Specimen4
Serum: Stable for 8 days when stored at 2...8°C and 1 day when stored at 15…25°C.
Test Procedure
statement.
Calibration
Apo A1 & B Calibrators Cat. No. ODR3005 for 3 point calibration.
The calibrator Apo A1 values are traceable to the WHO International Reference Material, SP1-01.
Following calibration, the resulting curve should be visually reviewed, on the Beckman Coulter analyser, for acceptability using the software
options - Routine, Calibration Monitor, Calibration Curve. Quality control procedures should be undertaken immediately following calibration in
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the Apo A1 concentration of each sample.

2009-08
Male 1.05 – 1.75 g/L (105 – 175 mg/dL)
Female 1.05 – 2.05 g/L (105 – 205 mg/dL)
findings.

from these values.
Linearity
Precision
0.87 0.01 0.78 0.01 1.45
1.61 0.02 1.01 0.04 2.34
1.96 0.02 0.80 0.04 2.02
Sensitivity
The lowest detectable level represents the lowest measurable level of Apo A1 that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this Apo A1 OSR6142 assay on the AU600 against another commercially available Apo A1
®
5
Limitations

† Apo A1 & B Calibrator Cat. No.: ODR3005 for 3 point Calibration
‡ When using 5 point calibration choose Apo A1 & B Calibrator Cat No. ODR3022 and set Cal Type as 13 - 5AB
‡ When using 5 point calibration choose Apo A1 & B Calibrator Cat No. ODR3022 and set Cal Type as 5AB
BIBLIOGRAPHY
1. Stein EA. Lipids, lipoproteins, and apolipoproteins. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:454-456.
3. Bhatnagar D, Durrington PN. Measurement and clinical significance of apolipoproteins A-1 and B. In: Rifai N, Warnick GR, Dominiczak MH, eds. Handbook of
lipoprotein testing. Washington: AACC Press, 1997:177-198.

2009-08
Apo A1, AU400/AU640 Serum Application Apo A1, AU600 Serum Application

Test No # Test name Apo A1 ∇ Sample type Ser ∇ Page 1/2
Test Name: Apo A1 ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name Apo A1 ∇ Sample type Ser ∇ Page 2/2
Test Name: Apo A1 ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
SERUM APPLICATION
General ISE Test No # Test name Apo A1 ∇
Test Name: Apo A1 ∇ < > Type Serum ∇ Cal type 11‡ 3AB‡ ∇ Count #
Calibration Type: 3AB‡ ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
1-Point Cal. Point: with CONC-0 Slope Check: + # 1-point cal. point
MB type factor
† APO A1 & B Calibrator Cat. No.: ODR3005 for 3 point Calibration † APO A1 & B Calibrator Cat. No.: ODR3005 for 3 point Calibration
‡ When using 5 point calibration choose Apo A1 & B Calibrator Cat. No. ODR3022 and set Cal Type as 5AB ‡ When using 5 point calibration choose Apo A1 & B Calibrator Cat. No. ODR3022 and set Cal Type as 13-5AB
* Values set for working in SI units (g/L). To work in mg/dL multiply by 100 * Values set for working in SI units (g/L). To work in mg/dL multiply by 100

2009-08
APO AI, AU2700/AU5400 Serum Application APO A1, AU680/AU480 Serum Application
Test Name: APO A1 ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: APO A1 ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: APO A1 ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: APO A1 ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
SERUM APPLICATION
General ISE
General ISE
Test Name: APO A1 ∇ < > Type Serum ∇ Test Name: APO A1 ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: 3AB‡ ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇ Calibration Type: 3AB‡ ∇ Formula: POLYGONAL ∇ Counts: # ∇
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇
† APO A1 & B Calibrator Cat. No.: ODR3005 for 3 point Calibration Calibrator OD Conc Low High Stability
‡ When using 5 point calibration choose Apo A1 & B Calibrator Cat. No. ODR3022 and set Cal Type as 5AB Point-1 ∇ Reagent Blank 999 Day 0 Hour
* Values set for working in SI units (g/L). To work in mg/dL multiply by 100 Point-2 ∇ Calibration 999 Day 0 Hour

2009-08
Apo B
OSR6143 4 x 13 mL R1
4 x 7 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of Apo B (Apolipoprotein B) in human serum on Beckman Coulter analysers. For in vitro
Summary1,2,3
Lipids are transported throughout the body by complex structures called lipoproteins. Lipoproteins are classified into five major density classes:
chylomicrons, very low-density lipoprotein (VLDL), intermediate density lipoprotein (IDL), low-density lipoprotein (LDL) and high-density lipoprotein (HDL).
Over the past several decades, decreased serum levels of high-density lipoprotein (HDL) and increased levels of low-density lipoprotein (LDL) have been
associated with increased risk of coronary artery disease.
Associated with these lipoproteins, at least five major apolipoproteins have been described and have been labelled A through E. The principle
apolipoproteins of HDL are the A apolipoproteins, constituting nearly 90% of the protein mass. Apo A1 has a role in the removal of excess cholesterol from
the tissues and reduced levels of Apo A1 have been observed in patients with coronary heart disease. Apo A1 measurements are frequently used in
characterising patients with genetic disorders that lead to low HDL cholesterol concentrations. Apo B plays an essential role in the delivery of cholesterol to
the tissues, the most abundant form of Apo B, Apo B100 is present in all atherogenic lipoprotein fractions VLDL, IDL and LDL. Elevated levels of Apo B100
are associated with an increased risk of coronary artery disease.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, Apo B reacts specifically with anti-human Apo B antibodies to yield insoluble
aggregates. The absorbance of these aggregates is proportional to the Apo B concentration in the sample.
TRIS buffer (pH 7.4) 8.6 mmol/L
Polyethylene glycol 6000 4 % w/v
Goat anti-Apo B antibodies ≈ 1.93 g/L
Preservative
Reagent Preparation
90 days.
Specimen
4
Serum: Stable for 8 days when stored at 2...8°C and 1 day when stored at 15…25°C.
Lipemic samples should be avoided.
Test Procedure
Calibration
The calibrator Apo B values are traceable to the WHO International Reference Material, SP3-07.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the Apo B concentration of each sample.

2009-08
Male 0.60 – 1.40 g/L (60 – 140 mg/dL)
Female 0.55 – 1.30 g/L (55 – 130 mg/dL)
values.
Linearity
Precision
0.61 0.01 1.16 0.01 1.92
1.49 0.01 0.89 0.02 1.27
1.98 0.01 0.71 0.03 1.33
Sensitivity
The lowest detectable level represents the lowest measurable level of Apo B that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison
Patient serum samples were used to compare this Apo B OSR6143 assay on the AU600 against another commercially available Apo B assay. Results of
y = 0.951x - 0.01 r = 0.992 n = 80 Sample range = 0.48 – 1.68 g/L
®
5
Limitations
† Apo A1 & B Calibrator Calibrator Cat. No.: ODR3005 for 3 point Calibration
‡ When using 5 point calibration choose Apo A1 & B Calibrator Cat No. ODR3022 and set Cal Type as 13 - 5AB
‡ When using 5 point calibration choose Apo A1 & B Calibrator Cat No. ODR3022 and set Cal Type as 5AB
BIBLIOGRAPHY
1. Stein EA. Lipids, lipoproteins, and apolipoproteins. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:454-456.
3. Bhatnagar D, Durrington PN. Measurement and clinical significance of apolipoproteins A-1 and B. In: Rifai N, Warnick GR, Dominiczak MH, eds. Handbook of
lipoprotein testing. Washington: AACC Press, 1997:177-198.

2009-08
Apo B, AU400/AU640 Serum Application Apo B, AU600 Serum Application

Test No # Test name Apo B ∇ Sample type Ser ∇ Page 1/2
Test Name: Apo B ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name Apo B ∇ Sample type Ser ∇ Page 2/2
Test Name: Apo B ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
SERUM APPLICATION
General ISE Test No # Test name Apo B ∇
Test Name: Apo B ∇ < > Type Serum ∇ Cal type 11‡ 3AB‡ ∇ Count #
Calibration Type: 3AB‡ ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
1-Point Cal. Point: ο with CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor
† APO A1 & B Calibrator Cat. No.: ODR3005 for 3 point Calibration † APO A1 & B Calibrator Cat. No.: ODR3005 for 3 point Calibrator
‡ When using 5 point calibration choose Apo A1 & B Calibrator Cat No. ODR3022 and set Cal Type as 5AB ‡ When using 5 point calibration choose Apo A1 & B Calibrator Cat No. ODR3022 and set Cal Type as13-5AB
* Values set for working in SI units (g/L). To work in mg/dL multiply by 100. * Values set for working in SI units (g/L). To work in mg/dL multiply by 100.

2009-08
APO B, AU2700/AU5400 Serum Application APO B, AU680/AU480 Serum Application
Test Name: APO B ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: APO B ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Measuring Point2 First 0 Last 10 Lipemia +++ ∇
Test Name: APO B ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: APO B ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
SERUM APPLICATION
General ISE
General ISE
Test Name: APO B ∇ < > Type Serum ∇ Test Name: APO B ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: 3AB‡ ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇ Calibration Type: 3AB‡ ∇ Formula: POLYGONAL ∇ Counts: # ∇
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇
† APO A1 & B Calibrator Cat. No.: ODR3005 for 3 point Calibration Calibrator OD Conc Low High Stability
‡ When using 5 point calibration choose Apo A1 & B Calibrator Cat No. ODR3022 and set Cal Type as 5AB Point-1 ∇ Reagent Blank 999 Day 0 Hour
* Values set for working in SI units (g/L). To work in mg/dL multiply by 100. Point-2 ∇ Calibration 999 Day 0 Hour
Ф AU680 MB Type Factor: 1-Point Calibration Point None ∇ ο with Conc-0

2009-08
ASO
OSR6194 4 x 51 mL R1
4 x 7 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of ASO (Anti-Streptolysin O) antibodies in human serum on Beckman Coulter analysers. For
Summary1,2
Group A streptococcus is one of the most common causes of human bacterial infections, causing diseases such as acute rheumatic fever, acute
glomerulonephritis, acute pharyngitis, sinusitis, pneumonia, septic scarlet fever, gangrene, and lymphangitis. Streptolysin O is a haemolysin produced by
group A streptococci. In an infected individual streptolysin O acts as a protein antigen to which the patient mounts an antibody response. Titres rise as early
as 1 week and peak 3 – 6 weeks after infection. In the absence of complications or re-infection, the ASO titre will usually fall to pre-infection levels within
6 – 12 months.
Test Principle
When a sample is mixed with R1 buffer and R2 latex solution, Anti-Streptolysin O antibodies react specifically with Streptolysin-O coated latex to yield
insoluble aggregates. The absorbance of these aggregates is proportional to the ASO concentration in the sample.
Streptolysin-O coated latex < 0.2 % w/v
Preservative
Reagent Preparation
R1 is ready for use and can be placed directly on board the instrument. R2 should be mixed by inversion 5 – 10 times before placing on board the
instrument and at weekly intervals thereafter.
The reagents are stable, unopened, up to the stated expiry date when stored at 2…8 °C. Once open, reagents stored on board the instrument are stable
for 60 days.
Do not use if there are visible signs of microbial growth, turbidity of precipitate, or any change in reagent colour.
Specimen
3
Serum: Stable for 8 days when stored at 2…8 °C and 2 days when stored at 15…25 °C.
Test Procedure
Calibration
This assay may be calibrated using multi-point calibration or single point calibration assay. For multi-point calibration use Serum Protein
Multi-Calibrator Cat. No.: ODR3021. For single point calibration Serum Protein Multi-Calibrator Cat. No.: ODR3021, Calibrator 4 may be used. MC Cal A
Cat No. ODR30037 for Mastercurve enabled systems only. Please refer to User Guide for further instructions.
st 4,5
Calibrator ASO values are traceable to the WHO NIBSC 1 International standard for Anti- Streptolysin O, AST.
Following calibration, the resulting curve should be visually reviewed for acceptability, on the Beckman Coulter analyser, using the software options to
access the Calibration Monitor. Quality control procedures should be undertaken immediately following calibration in accordance with good laboratory
practice.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the ASO concentration of each sample.
Adults ≤ 200 IU/mL
Children ≤ 150 IU/mL
2009-08
values.
Linearity
The test is linear within a concentration range of 100 – 1000 lU/mL.
Precision
Mean IU/mL SD CV% SD CV%
152 2.50 1.65 3.77 2.49
318 3.84 1.21 7.74 2.43
644 7.61 1.18 16.90 2.63
Sensitivity
The lowest detectable level in serum on an AU400 analyser was calculated as 7 IU/mL.
The lowest detectable level represents the lowest measurable level of Anti-Streptolysin O that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this ASO OSR6194 assay on the AU2700 against another commercially available ASO method. Results of
y = 1.031x - 16.575 r = 0.992 n = 102 Sample range = 106 – 941 IU/mL
®
6
Limitations
† For multi-point calibration use Serum Protein Multi-Calibrator Cat. No.: ODR3021.
For single-point calibration use Serum Protein Multi-Calibrator ODR3021 (Cal. No. 4), with calibration type “AB”. Set the factor range at
0 to 99999.9 when using calibration type “AB”.
₪ MC Cal A Cat. No. ODR30037
* Values set for working in IU/mL
BIBLIOGRAPHY
1. Thomas L. Streptococcus pyogenes infection. In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
4. Blirup-Jensen S, Johnson AM, Larsen M. Protein standardization IV: value transfer procedure for the assignment of serum protein values from a reference
preparation to a target material.Clin Chem Lab Med 2001;39:1110-1122.
5. Spaun J, Bentzon MW, Olesen Larsen S, Hewitt LF. International standard for anti-streptolysin-O. Bull Wld Hlth Org 1961;24:271-79.

2009-08
ASO, AU400/AU640 Serum Application ASO, AU600 Serum Application
Test No # Test name ASO ∇ Sample type Ser ∇ Page 1/2
Test Name: ASO ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 2.0 µL Dilution 0 µL Pre-Dilution Rate: 1 Reagent 1 vol 182 Dil. vol 0 µL L -0.1 H 2.5
R2 Volume 25 µL Dilution 0 µL L -0.1 H 2.5 Fst. L -0.1 Fst. H 1.5
Wavelength: Pri. 600 ∇ Sec. None ∇ First L -0.1 First H 1.5 Method FIXED ∇ Dynamic range
Method: FIXED ∇ Last L -0.1 Last H 1.5 Reaction + ∇ L 100* H 1000*
General LIH ISE Range Test No # Test name ASO ∇ Sample type Ser ∇ Page 2/2
Test Name: ASO ∇ < > Type: Serum ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
Panic Value: # # Unit: IU/mL* Decimal places: # Panic value # #
SERUM APPLICATION
General ISE Test No # Test name ASO ∇
Test Name: ASO ∇ < > Type Serum ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
1-Point Cal. Point: R With CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor
# User defined ₪ MC Cal A ODC30037
† For multi-point calibration use Serum Protein Multi-Calibrator Cat. No.: ODR3021
† For multi-point calibration use Serum Protein Multi-Calibrator Cat. No.: ODR3021
For single-point calibration use Serum Protein Multi-Calibrator ODR3021 (Cal. No. 4), with calibration type “AB” * Set
For single-point calibration use Serum Protein Multi-Calibrator ODR3021 (Cal. No. 4), with calibration type “AB” *Set the
the factor range at 0 to 99999.9 when using calibration type “AB”
factor range at 0 to 99999.9 when using calibration type “AB”

2009-11
ASO, AU2700/AU5400 Serum Application ASO, AU680/AU480 Serum Application
Test Name: ASO ∇ < > Type: Serum ∇ Operation: Yes ∇ Test Name: ASO ∇ < > Type: Serum ∇ Operation Yes ∇
Sample: Volume 1.6 µL Dilution 0 µL Pre-Dilution Rate: 1

Sample Volume 1.6 µL Dilution 0 µL OD Limit
R2 Volume 20 µL Dilution µL L -0.1 H 2.5
Reagent OD limit:
Method: FIXED ∇ Last L -0.1 Last H 1.5 R2(R2-1) 20 Dilution 0
µL µL
Reaction slope: + ∇ Dynamic Range: Dynamic Range Low 100* High 1000*
Measuring Point 2: First Last Correlation Factor: Wavelength Pri 600 ∇nm Sec. None ∇nm Factor for Maker A 1 B 0
Linearity : % A 1 B 0 Method FIXED ∇
Test Name: ASO ∇ < > Type: Serum ∇ Parameters Specific Test Parameters
Normal Ranges: Age L Age H Test Name: ASO ∇ < > Type: Serum ∇
R 1. Value/Flag: # ∇ Low High
# ∇ # # # # # #
# ∇ # # # # # #
R 4. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 5. # ∇ # # # # # # R 2. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 3. # ∇ # # # # # #
7. None Selected # # R 4. # ∇ # # # # # #
8. Out of Range L H # # R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # #
Panic Value: # # Unit: IU/mL* Decimal places: #
SERUM APPLICATION
Unit IU/mL* Decimal Places #
General ISE
Test Name: ASO ∇ < > Type Serum ∇ General ISE
Test Name: ASO ∇ < > Type Serum ∇ R Use Serum Cal.
Cal. No. OD CONC Factor/OD-L Factor/OD-H <Calibrator Parameters> OD Range
Point 1: # † -0.1 2.5 Calibrator OD Conc Low High Slope Check + ∇
Point 2: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 4: # ∇ † -0.1 2.5 R Reagent Blank
Point 6: Point 5: # ∇ † -0.1 2.5 R Calibration
1-Point Cal. Point: R with CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ Point 7 ∇ Advanced Calibration
Point 9 ∇
# User defined <Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
† For multi-point calibration use Serum Protein Multi-Calibrator Cat. No.: ODR3021 Master Curve> OD Range
For single-point calibration use Serum Protein Multi-Calibrator ODR3021 (Cal. No. 4), with calibration type “AB” * Set Calibrator OD Conc Low High Stability
the factor range at 0 to 99999.9 when using calibration type “AB” Point-1 ∇ Reagent Blank 999 Day 0 Hour
* Values set for working in IU/mL Point-2 Calibration 999 Day 0 Hour
∇
ф AU680
MB Type Factor: 1-Point Calibration Point None ∇ R with Conc-0

2009-11
ASO, AU680/AU480 Mastercurve Serum Application
Test Name: ASO ∇ < > Type: Serum ∇ Operation Yes ∇

R2(R2-1) 20 µL Dilution 0µL
Wavelength Pri 600 ∇nm Sec. None ∇nm Factor for Maker A 1 B 0
Method FIXED ∇

Test Name: ASO ∇ < > Type: Serum ∇

R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # #
Unit IU/mL* Decimal Places #
SERUM APPLICATION
General ISE
Test Name: ASO ∇ < > Type Serum ∇ R Use Serum Cal.
Calibration Type: 5MC ∇ Formula: POLYGONAL ∇ Counts: # ∇

Calibrator OD Conc Low High Slope Check ∇
Point 1: ∇ 126*
Point 2: ∇ 233* Allowance Range Check
Point 3: ∇ 353*
Point 4: ∇ 465* R Reagent Blank
Point 5: ∇ 570* R Calibration
Point 6: ∇
Point 8 ∇ Operation Yes ∇
Point 9 ∇
<Point Cal. For No. of Correction Points 1 ∇ Use Master Curve MC1 ∇ R Lot Calibration
Point-1 # ∇ ₪ 0.0799 0.1865 Reagent Blank 999 Day 0 Hour
# User defined
MB Type Factor: 1-Point Calibration Point None ∇ R with Conc-0 * Values set for working in IU/mL
₪ MC Cal A ODR30037
ф AU680
2009-11
β-2 MICROGLOBULIN
OSR6151 4 x 10 mL R1
4 x 8 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of β-2 microglobulin in human serum and plasma on Beckman Coulter analysers. For in vitro
Summary1,2
β-2 microglobulin is a low molecular weight (11.8 kDa) protein found on the cell membrane of all nucleated cells including lymphocytes. The protein is the
light or β-chain of the class I human leukocyte antigen (HLA) and consists of a single polypeptide chain with one intrachain disulfide bridge. Changes in
serum β-2 microglobulin levels or β-2 microglobulin excretion are caused by an increase in production, a change in glomerular filtration rate (GFR) or
tubular reabsorption.
Since the lymphatic system is the main synthesis site of β-2 microglobulin, all conditions with an increased proliferation rate of lymphocytic cells are
associated with elevated serum concentrations e.g. multiple myelomas, Hodgkin's lymphomas, chronic lymphocytic leukemias and other malignant non-
Hodgkin's lymphomas. Production of β-2 microglobulin is also increased in all diseases that are associated with activation of the immune system e.g.
certain autoimmune diseases; bacterial and viral infections (HIV,CMV,EBV); infectious mononucleosis and transplant rejection.
In healthy people, β-2 microglobulin is synthesised at a relatively constant rate and released into the body fluids during the process of natural cell
regeneration, it is subject to free glomerular filtration and tubular reabsorption. A reduction in GFR prolongs the half-life of β-2 microglobulin and serum
concentrations increase exponentially. Excretion of β-2 microglobulin is increased in the case of tubular damage e.g. due to bacterially induced interstitial
nephritis and cadmium nephropathy. β-2 microglobulin is frequently used to test renal tubular function, particularly in kidney transplant patients in whom
rejection of the allograft will manifest as an increase in serum β-2 microglobulin due to tubulopathy.
Test Principle
When a sample is mixed with R1 buffer and R2 latex solution, human β-2 microglobulin reacts specifically with anti-human β-2 microglobulin antibodies
coated on the latex particles, to yield insoluble aggregates. The absorbance of these aggregates is proportional to the β-2 microglobulin concentration in
the sample.

Latex particles coated with anti-human β-2 microglobulin Variable
Preservative

Reagent Preparation

90 days.
Specimen
3
Test Procedure
Calibration
st
The calibrator β-2 microglobulin values provided in the calibrator package insert are traceable to the WHO 1 International Standard 1985.
Quality Control

2009-08
Calculation
The Beckman Coulter analysers automatically compute the β-2 microglobulin concentration of each sample.
Serum, plasma (< 60 years) 0.8 – 2.4 mg/L (0.08 – 0.24 mg/dL)
(> 60 years) ≤ 3.0 mg/L (≤ 0.3 mg/dL)

values.
Linearity
The test is linear within a concentration range of 0.5 – 16.0 mg/L (0.05 – 1.6 mg/dL).
Precision
Mean mg/L SD CV% SD CV%
0.63 0.02 2.46 0.01 2.36
6.37 0.07 1.09 0.12 1.89
9.92 0.10 1.02 0.16 1.64
Sensitivity
The lowest detectable level on an AU600 analyser was estimated at 0.064 mg/L.
The lowest detectable level represents the lowest measurable level of β-2 microglobulin that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient serum samples were used to compare this β-2 Microglobulin OSR6151 assay on the AU600 against another commercially available
β-2 microglobulin assay.
Log (y) = 0.998log(x) – 0.072 r = 0.980 n = 62 Sample range = 11.0 – 56.5 mg/L
®
4
Limitations
Samples from patients with paraproteinemia may occasionally give spuriously elevated results, such samples should be diluted prior to analysis.
Samples containing elevated levels of rheumatoid factor may cause positive interference in this assay. Results from such patients should be interpreted
with care.
Samples with very high β-2 microglobulin concentrations (> 2000 mg/L) can generate false low results without appropriate “Z” flags due to excess antigen
in the sample.

* Values set for working in mg/L. To work in mg/dL divide by 10.
BIBLIOGRAPHY
1. Thomas L. β2-microglobulin (β2-M). In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
2. Grant GH, Silverman LM, Christenson RH. Amino Acids and proteins. In: Teitz NW, ed. Fundamentals of clinical chemistry. Philadelphia: WB Saunders

2009-08
β-2-MICROGLOBULIN, AU400/AU640 Serum/Plasma Application β-2-MICROGLOBULIN, AU600 Serum/Plasma Application

Test No # Test name B2M ∇ Sample type Ser ∇ Page 1/2
Test Name: B2M ∇ < > Type: Serum ∇ Operation: Yes ∇
Method: FIXED ∇ Last L -0.1 Last H 1.5 Reaction + ∇ L 0.5* H 16.0*
General LIH ISE Range Test No # Test name B2M ∇ Sample type Ser ∇ Page 2/2
Test Name: B2M ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic Value: # # Unit: mg/L* Decimal places: # Panic value # #
General ISE Test No # Test name B2M ∇
Test Name: B2M ∇ < > Type Serum ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
1-Point Cal. Point: with CONC-0 Slope Check: + Advanced Calibration: # 1-point cal. point
ο ∇ ∇
MB type factor
† Serum Protein Multi-Calibrator 2 Cat. No.: ODR3023 † Serum Protein Multi-Calibrator 2 Cat. No.: ODR3023
* Values set for working in mg/L. To work in mg/dL divide by 10. * Values set for working in mg/L. To work in mg/dL divide by 10.

2009-08
β-2-MICROGLOBULIN, AU2700/AU5400 Serum/Plasma Application β-2-MICROGLOBULIN, AU680/AU480 Serum/Plasma Application
Test Name: B2M ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: B2M ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: B2M ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: B2M ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: mg/L* Decimal places: # 7. No demographics # #
Unit mg/L* Decimal Places #
General ISE
General ISE
Test Name: B2M ∇ < > Type Serum ∇
Test Name: B2M ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
1-Point Cal. Point: ο With CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
* Values set for working in mg/L. To work in mg/dL divide by 10. Point-1 ∇ Reagent Blank 999 Day 0 Hour

2009-08
C3
OSR6159 4 x 10 mL R1
4 x 8 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of C3 (Complement 3) in human serum and plasma on Beckman Coulter analysers. For in vitro
Summary1,2
The Complement system consists of about 20 plasma proteins as well as receptors on blood cells that play an important role in inflammation by facilitating
phagocytosis through opsonisation, lysing foreign cells, increasing vascular permeability and attracting phagocytes. Activation or consumption of
complement occurs in a number of disorders, particularly those involving immune complex deposition e.g. SLE, mixed cryoglobulinaemia and some forms
of vasculitis, however this may be compensated for in part by the synthesis of acute phase reactants. C3 comprises about 30% of the total plasma
concentration of complement components and is consumed by activation of both the classical and alternative pathways. C4 levels fall only as a
consequence of classical pathway activation, therefore if hypocomplementemia is present, measurement of both C3 and C4 can determine whether the
classical or alternative pathway has been activated.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, human C3 reacts specifically with anti-human C3 antibodies to yield insoluble
aggregates. The absorbance of these aggregates is proportional to the C3 concentration in the sample.

Polyethylene glycol 6000 1.6% w/v
Goat anti-C3 antibodies Variable
Preservative

Reagent Preparation

for 90 days.
Specimen
3
Stable in serum and plasma for 8 days when stored at 2...8 °C and 4 days when stored at 15…25 °C.
Test Procedure
Calibration
Serum Protein Multi-Calibrator Cat. No. ODR3021. MC Cal A Cat No. ODR30037 for Mastercurve enabled systems only. Please refer to User Guide for
further instructions.
®
The calibrator values are traceable to ERM – DA470 for C3c.
practice.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the C3 concentration of each sample.

2009-08
Adults and children 0.9 – 1.8 g/L (90 – 180 mg/dL)

values.
Linearity
Precision
0.68 0.01 1.24 0.01 1.38
1.39 0.01 0.84 0.02 1.37
4.75 0.03 0.64 0.05 1.01
Sensitivity
The lowest detectable level represents the lowest measurable level of C3 that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison
Patient serum samples were used to compare this C3 OSR6159 assay on the AU600 against another commercially available C3 assay. Results of linear
®
6
Limitations

† Serum Protein Multi-Calibrator Cat. No.: ODR3021
BIBLIOGRAPHY
1. Ismail AA, Snowden N. Autoantibodies and specific serum proteins in the diagnosis of rheumatological disorders. Ann Clin Biochem 1999;36:565-578.
2. Thomas L. The complement system. In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
470. J Lab Med 1996;20:145-152.
Specific Protien BLOSR6x59.01 EN.01

2009-08
C3, AU400/AU640 Serum/Plasma Application C3, AU600 Serum/Plasma Application

Test No # Test name C3 ∇ Sample type Ser ∇ Page ½
Test Name: C3 ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name C3 ∇ Sample type Ser ∇ Page 2/2
Test Name: C3 ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name C3 ∇
Test Name: C3 ∇ < > Type Serum ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
1-Point Cal. Point: with CONC-0 Slope Check: + # 1-point cal. Point
MB type factor
† Serum Protein Multi-Calibrator Cat. No.: ODR3021 † Serum Protein Multi-Calibrator Cat. No.: ODR3021

2009-08
C3, AU2700/AU5400 Serum/Plasma Application C3, AU680/AU480 Serum/Plasma Application
Test Name: C3 ∇ < > Type: Serum ∇ Operation Yes ∇
μL μL
Reagent OD limit:
Test Name: C3 ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: C3 ∇ < > Type: Serum ∇

ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: C3 ∇ < > Type Serum ∇
Test Name: C3 ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: Process: CONC ∇ Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: # ∇
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇
† Serum Protein Multi-Calibrator Cat. No.: ODR3021 Point-1 ∇ Reagent Blank 999 Day 0 Hour
* Values set for working in SI units (g/L). To work in mg/dL multiply by 100. Point-2 Calibration 999 Day 0 Hour
∇
Ф AU680

2009-08
C3, AU680/AU480 Mastercurve Serum/Plasma Application

Method END ∇

Test Name: C3 ∇ < > Type: Serum ∇

ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
General ISE
Point 1: ∇ 0.41*
Point 2: ∇ 1.55* Allowance Range Check
Point 3: ∇ 2.69*
Point 4: ∇ 3.86* ο Reagent Blank
Point 5: ∇ 4.93* ο Calibration
Point 6: ∇
Point 9 ∇
<Point Cal. For No. of Correction Points 1 ∇ Use Master Curve MC1 ∇ ο Lot Calibration
Point-2 Calibration 999 Day 0 Hour
# User defined
∇
MB Type Factor: 1-Point Calibration Point None ∇ ο with Conc-0 ₪ MC Cal A ODR30037
ф AU680

2009-08
C3, AU400/AU640 Paediatric Application C3, AU600 Paediatric Application

Test No # Test name C3P ∇ Sample type Ser ∇ Page 1/2
Test Name: C3P ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name C3P ∇ Sample type Ser ∇ Page 2/2
Test Name: C3P ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name C3P ∇
Test Name: C3P ∇ < > Type Serum ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
MB type factor

2009-08
C3, AU2700/AU5400 Paediatric Application C3, AU680/AU480 Paediatric Application
Test Name: C3P ∇ < > Type: Serum ∇ Operation Yes ∇
μL μL
Reagent OD limit:
Test Name: C3P ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: C3P ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: C3P ∇ < > Type Serum ∇ Test Name: C3P ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: Process: CONC ∇ Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: # ∇
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇
† Serum Protein Multi-Calibrator Cat. No.: ODR3021 Calibrator OD Conc Low High Stability

2009-08
C4
OSR6160 4 x 10 mL R1
4 x 8 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of C4 (Complement 4) in human serum and plasma on Beckman Coulter analysers. For in vitro
Summary1,2
The Complement system consists of about 20 plasma proteins as well as receptors on blood cells that play an important role in inflammation by facilitating
phagocytosis through opsonisation, lysing foreign cells, increasing vascular permeability and attracting phagocytes. Activation or consumption of
complement occurs in a number of disorders, particularly those involving immune complex deposition e.g. SLE, mixed cryoglobulinaemia and some forms
of vasculitis, however this may be compensated for in part by the synthesis of acute phase reactants. C3 comprises about 30% of the total plasma
concentration of complement components and is consumed by activation of both the classical and alternative pathways. C4 levels fall only as a
consequence of classical pathway activation, therefore if hypocomplementemia is present, measurement of both C3 and C4 can determine whether the
classical or alternative pathway has been activated.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, human C4 reacts specifically with anti-human C4 antibodies to yield insoluble
aggregates. The absorbance of these aggregates is proportional to the C4 concentration in the sample.

Polyethylene glycol 6000 1.6% w/v
Goat anti-C4 antibodies Variable
Preservative

Reagent Preparation

for 90 days.
Specimen
3
Stable in serum and plasma for 8 days when stored at 2...8 °C and 2 days when stored at 15…25 °C.
Test Procedure
Calibration
®
The calibrator values are traceable to ERM – DA470.
practice.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the C4 concentration of each sample.

2009-08
Adults and Children 0.1 – 0.4 g/L (10 – 40 mg/dL)
values.
Linearity
Precision
0.16 0.00 2.31 0.00 2.48
0.33 0.00 0.84 0.00 1.19
1.43 0.01 0.94 0.02 1.36
Sensitivity
The lowest detectable level represents the lowest measurable level of C4 that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison
Patient serum samples were used to compare this C4 OSR6160 assay on the AU600 against another commercially available C4 assay. Results of linear
®
6
Limitations

BIBLIOGRAPHY
2. Thomas L. The complement system. In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
470. J Lab Med 1996;20:145-152.

2009-08
C4, AU400/AU640 Serum/Plasma Application C4, AU600 Serum/Plasma Application

Test No # Test name C4 ∇ Sample type Ser ∇ Page ½
General LIH ISE Range Test No # Test name C4 ∇ Sample type Ser ∇ Page 2/2
Test Name: C4 ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name C4 ∇
Test Name: C4 ∇ < > Type Serum ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
MB type factor
# User defined

2009-08
C4, AU2700/AU5400 Serum/Plasma Application C4, AU680/AU480 Serum/Plasma Application
μL μL
Reagent OD limit:
Test Name: C4 ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: C4 ∇ < > Type: Serum ∇

ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: C4 ∇ < > Type Serum ∇
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇

2009-08
C4, AU680/AU480 Mastercurve Serum/Plasma Application

Method END ∇

Test Name: C4 ∇ < > Type: Serum ∇

ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
General ISE
Test Name: C4 Type Serum Use Serum Cal.
∇ < > ∇ ο

Point 1: # ∇ 0.15*
Point 2: # ∇ 0.48* Allowance Range Check
Point 3: # ∇ 0.78*
Point 4: # ∇ 1.11* ο Reagent Blank
Point 5: # ∇ 1.40* ο Calibration
Point 6: ∇
Point 9 ∇
Point-2 Calibration 999 Day 0 Hour
# User defined
∇
ф AU680

2009-08
C4, AU400/AU640 Paediatric Application C4, AU600 Paediatric Application

Test No # Test name C4P ∇ Sample type Ser ∇ Page ½
General LIH ISE Range Test No # Test name C4P ∇ Sample type Ser ∇ Page 2/2
Test Name: C4P ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE
Test No # Test name C4P ∇
Test Name: C4P ∇ < > Type Serum ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
MB type factor
# User defined

2009-08
C4, AU2700/AU5400 Paediatric Application C4, AU680/AU480 Paediatric Application
Test Name: C4P ∇ < > Type: Serum ∇ Operation Yes ∇
μL μL
Reagent OD limit:
Test Name: C4P ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: C4P ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: C4P ∇ < > Type Serum ∇ Test Name: C4P ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇

2009-08
CERULOPLASMIN
OSR6164 4 x 18 mL R1
4 x 5 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of ceruloplasmin in human serum and plasma on Beckman Coulter analysers. For in vitro
Summary1,2
Ceruloplasmin is the primary copper containing protein in plasma. It is a late acute phase reactant synthesised by the liver. Acute phase reactant refers to
proteins whose serum concentrations rise significantly during acute inflammation due to causes including surgery, myocardial infarction, infections and
tumours. Ceruloplasmin’s main clinical importance is in the diagnosis of Wilson’s disease. Here plasma ceruloplasmin concentration is reduced while
dialysable copper concentration is increased. Increased ceruloplasmin levels are particularly notable in diseases of the reticuloendothelial system such as
Hodgkin’s disease as well as during pregnancy or the use of contraceptive pills. Low plasma levels of ceruloplasmin are found in malnutrition,
malabsorption, nephrosis and severe liver disease, particularly biliary cirrhosis.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, human ceruloplasmin reacts specifically with the anti-human ceruloplasmin antibodies
to yield insoluble aggregates. The absorbance of these aggregates is proportional to the ceruloplasmin concentration in the sample.

Solution of polymers in phosphate buffered saline (pH 7.4 – 7.6)
Polyethylene glycol 6000 5% w/v
Rabbit anti-human ceruloplasmin antiserum Variable
Preservative

Reagent Preparation

90 days.
Specimen
3
Test Procedure
Calibration
The calibrator values are traceable to the IFCC (International Federation of Clinical Chemistry) standard CRM 470.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the ceruloplasmin concentration of each sample.
Adults 200 – 600 mg/L (20 – 60 mg/dL)

2009-08
values.
Linearity
The test is linear within a concentration range of 60 – 2000 mg/L (6 – 200 mg/dL).
Precision
120.30 1.10 0.94 2.90 2.44
934.10 11.80 1.27 22.10 2.36
1810.50 17.10 0.94 84.80 4.68
Sensitivity
The lowest detectable level on an AU640 analyser was calculated at 4 mg/L.
The lowest detectable level represents the lowest measurable level of ceruloplasmin that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient serum samples were used to compare this Ceruloplasmin OSR6164 assay on the AU640 against another commercially available ceruloplasmin
assay. Results of Deming linear regression analysis were as follows:
y = 1.020x – 3.5 r = 0.990 n = 50 Sample range = 90 – 510 mg/L
Lipemia: Interference less than 3% up to 1000 mg/dL Intralipid®
5
Limitations
Samples from patients with abnormal lipoprotein metabolism such as those seen in cholecystitis or obstructive liver disease may give artificially negative
Ceruloplasmin results. These samples are characterised by having extremely elevated Cholesterol values (>10 mmol/L) and elevated Bilirubin. Such
samples should be diluted 1 part sample to 4 parts deionised water prior to analysis and the result multiplied by 5.

† System Calibrator Cat. No.: ODR3023
* Values set for working in SI units (mg/L). To work in mg/dL divide by 10.
BIBLIOGRAPHY
1. Johnson AM, Rohlfs EM, Silverman LM. Proteins. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders Company,
1999;490-492.
2. Kazmierczak SC. Ceruloplasmin. In: Kaplan LA, Pesce AJ, eds. Clinical chemistry theory, analysis, correlation. St Louis: Mosby, 1996:966pp.
3. Tietz NW, ed. Clinical guide to laboratory tests, 3rd ed. Philadelphia: WB Saunders Company, 1995:122pp.
470. J Lab Med 1996;20:145-152.

2009-08
CERULOPLASMIN, AU400/AU640 Serum/Plasma Application CERULOPLASMIN, AU600 Serum/Plasma Application

Test No # Test name CER ∇ Sample type Ser ∇ Page 1/2
Test Name: CER ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name CER ∇ Sample type Ser ∇ Page 2/2
Test Name: CER ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name CER ∇
Test Name: CER ∇ < > Type Serum ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
1-Point Cal. Point: ο with CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor
# User defined

2008-08
CERULOPLASMIN, AU2700/AU5400 Serum/Plasma Application CERULOPLASMIN, AU680/AU480 Serum/Plasma Application
Test Name: CER ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CER ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Test Name: CER ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CER ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: CER ∇ < > Type Serum ∇
Test Name: CER ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇
† Serum Protein Multi-Calibrator 2 Cat. No.: ODR3023 Calibrator OD Conc Low High Stability
* Values set for working in SI units (mg/L). To work in mg/dL divide by 10. Point-1 ∇ Reagent Blank 999 Day 0 Hour

2008-08
CRP
OSR6147 4 x 14 mL R1
4 x 6 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of C-reactive protein (CRP) in human serum and plasma on Beckman Coulter analysers. For in
vitro diagnostic use only.
Summary1,2
C-reactive protein (CRP) is an acute phase protein synthesised by the liver in response to the release of inflammatory cytokines such as interleukin-6. CRP
exhibits dramatic increases in concentration following acute or chronic inflammation that may accompany bacterial infections - the most potent stimulus to
CRP production; autoimmune or immune complex disease; tissue necrosis and malignancy; myocardial infarction; and trauma. The increase occurs within
24 – 48 hours and the level may be 2000 times normal. In many cases the changes in plasma CRP level precede changes in the clinical symptoms.
The degree of elevation of CRP reflects the mass or activity of the inflammed tissue and in acute inflammation or infection correlates well with disease
activity. Because the increase is non-specific, it cannot be interpretated without a complete clinical history, and even then only by comparison with previous
values. A persistently raised CRP level generally indicates that therapy is ineffective.
Normal CRP levels do not exclude the presence of minor degrees of acute, localised inflammation or some chronic diseases such as SLE and ulcerative
colitis.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, CRP reacts specifically with anti-human CRP antibodies to yield insoluble aggregates.
The absorbance of these aggregates is proportional to the CRP concentration in the sample.

Goat anti-CRP antibodies ≈ 0.6 g/L
Preservative

Warning – this preparation contains a substance not yet tested completely.
Reagent Preparation

for 90 days.
Specimen
3
Serum and heparinised plasma: Stable in serum and plasma for 2 months when stored at 2...8 °C and 11 days when stored at 15…25 °C.
Test Procedure
Calibration
Serum Protein Multi-Calibrator Cat. No. ODR3021.
The calibrator CRP values are traceable to IFCC (International Federation of Clinical Chemistry) standard CRM 470.
Following calibration, the resulting curve should be visually reviewed, on the Beckman Coulter analyser, for acceptability using the software options to
access Calibration Monitor. Quality control procedures should be undertaken immediately following calibration in accordance with good laboratory practice.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the CRP concentration of each sample.

2009-08
Serum-adult Upper limit is 5 mg/L

values.
Linearity
The test is linear within a concentration range of 5 – 300 mg/L
Prozone settings must be applied when using the CRP reagent, refer to Setting Sheet for specific instrument details.
Precision
10.90 0.31 2.81 0.65 5.99
89.56 2.24 2.49 2.29 2.55
168.07 1.71 1.02 2.17 1.29
Sensitivity
The lowest detectable level represents the lowest measurable level of CRP that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison
Patient serum samples were used to compare this CRP OSR6147 assay on the AU600 against another commercially available CRP assay. Results of
y = 0.939x – 0.817 r = 0.994 n = 79 Sample range = 10 – 212 mg/L
®
5
Limitations
This product is not suitable for cardiovascular risk assessment or diagnostic evaluation of neonates. The CRP Latex reagent Cat. No.: OSR6199 is
available for this purpose.
Samples from patients with abnormal lipoprotein metabolism such as those seen in cholecystitis or obstructive liver disease may give artificially elevated
CRP results. These samples are characterised by having extremely elevated Cholesterol values (> 10 mmol/L) and elevated Bilirubin. Such samples
should be diluted 1 part sample to 4 parts deionised water prior to analysis and the result multiplied by 5.
Samples with very high CRP concentrations (> 750 mg/L) can generate false low results without appropriate "Z" flags due to excess antigen in the sample.
In very rare cases gammopathy, especially monoclonal IgM (Waldenström’s Macroglobulinemia), may cause unreliable results.

* Values set for working in mg/L
BIBLIOGRAPHY
1. Whicher J. C-reactive protein (CRP). In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
1987:334pp.
470. J Lab Med 1996;20:145-152.

2009-08
CRP, AU400/AU640 Serum/Plasma Application CRP, AU600 Serum/Plasma Application

Test No # Test name CRP ∇ Sample type Ser ∇ Page 1/2
Test Name: CRP ∇ < > Type: Serum ∇ Operation: Yes ∇
Fst. L -0.1 Fst. H 1.5
Reagent OD limit: Method FIXED ∇ Dynamic range
Wavelength: Pri. 340 ∇ Sec. 800 ∇ First L -0.1 First H 1.5 Reaction + ∇ L 5* H 300*
Method: FIXED ∇ Last L -0.1 Last H 1.5 Point 1 Fst 11 Lst 27 Correlation factor A 1
Reaction slope: + ∇ Dynamic Range: Point 2 Fst Lst B 1*
Measuring Point 2: First Last Correlation Factor: Linearity Fst % Sec %
Linearity : % A 1 B 1* No lag time ∇ On-board stability period 90
Test No # Test name CRP ∇

General ISE Cal type 14 6AB ∇ Count #
Test Name: CRP ∇ < > Type Serum ∇ Selection calibrator
Calibration Type: 6AB ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇ Point 1 # ∇ † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Cal. No. OD CONC Factor/OD-L Factor/OD-H Point 3 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 6 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 7 ∇
1-point cal. Point
Point 5: # † -0.1 2.5 MB type factor
Point 6: # † -0.1 2.5 Calibrator stability period 999
Point 7:
Test No # Name CRP ∇ Type Ser ∇

Test Name: CRP ∇ < > Type: Serum ∇ Check Point-2 14 Check Point Interval
Decision value-1 0.4 Decision value-2 0.0000
√ Logic Check-1 Logic Check –2 Logic Check -3
Decision value-2 2.0 Limit Point 1 0
Decision value-3 0.3 Limit Point 2 0
Limit Point 1 10 Logic Check-3 NO
Limit Point 2 11
Limit Point 1 0
Limit Point 2 0
# User defined

2009-08
CRP, AU2700/AU5400 Serum/Plasma Application CRP, AU680/AU480 Serum/Plasma Application
Test Name: CRP ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CRP ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Measuring Point 1: First 11 Last 27 L 5* H 300* Wavelength Pri 340 ∇nm Sec. 800 ∇nm Factor for Maker A 1 B 1*
Linearity : % A 1 B 1* Reaction Slope + ∇ Onboard Stability Period 90 Day 0 Hour
Test Name: CRP ∇ < > Type: Serum ∇ General ISE
Calibration Type: 6AB ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇ Test Name: CRP ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # † -0.1 2.5
Point 2: # † -0.1 2.5
Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5
Point 2: # ∇ † -0.1 2.5 Allowance Range Check
Point 4: # † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5
Point 4: # ∇ † -0.1 2.5 ο Reagent Blank
Point 6: # † -0.1 2.5 Point 5: # † -0.1 2.5
∇ ο Calibration
Point 7: Point 6: # † -0.1 2.5
∇
1-Point Cal. Point: ο with CONC-0 Slope Check: + ∇ Advanced Calibration: # Point 7 ∇ Advanced Calibration
Point 9 ∇
Test Name: CRP ∇ < > Type: Serum ∇
√ Logic Check-1 Logic Check-2 Logic Check-3
Check Point-1: 11 Check Point-1: Check Point-1: Parameters Misc
Check Point-3: 18 Decision Value-1: Decision Value-1: CheckedTests Contamination Data Check
Decision Value-1: 0.4 Decision Value-2: Decision Value-2: Parameters Parameters
Test Name: CRP ∇ < > Type: Serum ∇
Limit Point 1: 10
Limit Point 2: 11
Check Point-3: 18
# User defined. Decision Value-1: 0.4000 Decision Value-1: Decision Value-1:
† Serum Protein Multi-Calibrator Cat. No.: ODR3021 Decision Value-2: 2.0000 Decision Value-2: Decision Value 2:
ф AU680

2009-08
CRP Latex
OSR6199 4 x 30 mL R1
4 x 30 mL R2
OSR6299 4 x 50 mL R1
4 x 50 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of C-reactive protein (CRP) in human serum and plasma on Beckman Coulter analysers. For
in vitro diagnostic use only. The normal application is to be used for the detection and evaluation of infection, tissue injury, inflammatory disorders and
associated diseases. The Highly Sensitive application is not a general screening test for infection/inflammation.
Summary
C-reactive protein (CRP) is one of the most sensitive acute-phase reactants. With the Beckman Coulter System CRP Latex reagent, CRP can be
measured down to very low concentrations, however due to the non specificity of CRP and the wide inter-individual variation, interpretation of CRP levels
must be undertaken with care, usually in comparison with previous CRP values or other markers.
Depending on the application used (different instrument settings) two measuring ranges are available:
1,2,3,4
1. Normal Application (0.2-480 mg/L): C-reactive protein levels in serum can rise dramatically after myocardial infarction, trauma, infection,
inflammation, surgery, or neoplastic proliferation. The increase occurs within 24 to 48 hours, and the level may be 2000 times normal.
Studies have shown that the detection of much lower CRP levels can indicate an increased risk for coronary heart disease in asymptomatic patients. CRP
concentrations above 3 mg/L at the time of hospital admission can predict subsequent cardiac events.
5,6
2. Highly Sensitive Application (0.08 – 80 mg/L): Cord blood normally has very low CRP concentrations (median 0.12 mg/L). In the diagnostic evaluation
of neonates with suspected infection, measurements of serial CRP levels are useful. Two low CRP levels obtained 24 hours apart, 8-48 hours after
presentation, indicate that bacterial infection is unlikely. Thus CRP Latex reagent is providing a valuable tool for the early diagnosis of infection in preterm
infants and neonates. It assesses both the need for and the effectiveness of antibiotic treatment however CRP values alone cannot be used as a basis for
early discontinuance of antibiotic therapy.
Test Principle
When a sample is mixed with R1 buffer and R2 latex suspension, CRP reacts specifically with anti-human CRP antibodies coated on the latex particles to
yield insoluble aggregates. The absorbance of these aggregates is proportional to the CRP concentration in the sample.

Glycine buffer 100 mmol/L
Latex, coated with anti-CRP antibodies < 0.5 % w/v
Preservative < 0.1 % w/v

Antisera was produced in healthy animals in facilities free from rinderpest, foot and mouth disease, peste des petits ruminants, Rhift Valley fever, bovine
spongiform encephalopathy and blue tongue disease.
Reagent Preparation
R1 is ready for use and can be placed directly on board the instrument. R2 should be mixed by inversion 5 - 10 times before placing on board the

90 days.
Specimen
7
Serum, EDTA and lithium heparinised plasma: Stable in serum and plasma for 2 months when stored at 2...8°C and 11 days when stored at 15…25°C.
Test Procedure
statement.
Data check parameters must be applied when using Highly Sensitive application on AU2700/AU5400, see setting sheets for specific instrument
details.
Calibration
Application Calibrator Cat. No.
Normal CRPLatex Calibrator Normal Set ODC0026
Highly sensitive CRP Latex Calibrator Highly Sensitive Set ODC0027
The calibrator CRP values are traceable to IFCC (International Federation of Clinical Chemistry) standard CRM 470.
Recalibrate when the following occur:

2009-08
Quality Control
ITA Control Sera ODC0014, ODC0015 and ODC0016 should be used for the Normal and Highly Sensitive applications, and CRP (Latex) Control Sera
ODC0013 (two levels) should be used for the Highly Sensitive application only. Other control materials with values determined by this Beckman Coulter
system may also be used.
Calculation
The Beckman Coulter analysers automatically compute the CRP concentration of each sample.
Reference Intervals
8
Normal application < 5 mg/L
9
Highly sensitive application < 1 mg/L

values.
Linearity
The test is linear within a concentration range of 0.2 - 480mg/L for the Normal Application (AU400/640, AU600, AU2700/5400), 0.08 - 80 mg/L
(AU400/640, AU600, AU2700/5400), for the Highly Sensitive Application.
Precision
The following data was obtained for CRP (Latex) normal and highly sensitive assays on an AU640, AU400 and AU2700 using 3 serum pools analysed
over 20 days.
AU640:
Application Mean mg/L SD CV% SD CV%
Normal 6.56 0.07 1.09 0.12 1.85
64.79 0.78 1.20 2.16 3.34
137.71 0.96 0.70 2.26 1.64
Highly Sensitive 0.21 0.01 4.08 0.01 4.87
59.38 0.54 0.91 1.00 1.68
146.95 1.12 0.76 2.34 1.59
AU400:
Normal 6.44 0.05 0.80 0.20 3.12
64.02 0.56 0.87 1.83 2.86
137.70 1.02 0.74 3.63 2.63
60.12 0.33 0.55 0.76 1.27
144.18 1.08 0.75 2.78 1.93
AU2700:
Normal 6.59 0.21 3.22 0.25 3.79
62.84 0.64 1.02 1.21 1.92
137.52 1.28 0.93 2.27 1.65
9.70 0.09 0.92 0.19 1.94
48.28 0.35 0.73 0.73 1.50
Sensitivity
The lowest detectable level was estimated as follows:
Application Lowest Detectable Level (mg/L)
AU640 AU400 AU2700
Normal 0.09 0.10 0.14
Highly Sensitive 0.02 0.02 0.07
The lowest detectable level represents the lowest measurable level of CRP that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison
Patient serum samples were used to compare the normal and highly sensitive CRP Latex OSR6199 assays on the AU640 against another commercially
available CRP assay (Method 2). Results of linear regression analysis were as follows:
Normal Highly Sensitive
Y Method AU640 AU640
X Method Method 2 Method 2
Slope 1.024 0.993
Intercept 0.546 - 0.825
Correlation Coeff (r) 0.998 0.997
No. of samples 119 118
Range (mg/L) 0.20 – 167.38 0.20 – 155.86

2009-08
Results of studies conducted to evaluate the susceptibility of CRP Latex normal,and highly sensitive assays to interference were as follows:
Normal Application
®
Highly Sensitive Application
®
10
Limitations
Samples containing heterophilic antibodies can cause falsely elevated results.
Samples with very high CRP concentrations (> 750 mgmL) can generate false low results without appropriate "Z" flags due to excess antigen in
the sample.
In very rare cases Gammopathy, especially monoclonal IgM (Waldenström’s macroglobinemia), may cause unreliable results.

† CRP Latex Calibrator Normal Set ODC0026
† CRP Latex Calibrator Highly Sensitive Set ODC0027
* Values set for working in mg/L.
BIBLIOGRAPHY
1987:334pp.
2. Morrow DA, Ridker PM. C-reactive protein, inflammation, and coronary risk. Med Clin North Am 2000;84:149-161.
3. Ridker PM. Novel risk factors and markers for coronary disease. Adv Intern Med 2000;45:391-419.
4. Liuzzo G, Biasucci LM, Gallimore JR, Grillo RL, Rebuzzi AG, Pepys MB, Maseri A. The prognostic value of C-reactive protein and serum amyloid A protein in
severe unstable angina. N Engl J Med 1994;331:417-424.
5. Wasunna A, Whitelaw A, Gallimore R, Hawkins PN, Pepys MB. C-reactive protein and bacterial infection in preterm infants. EurJ Pediatr 1990;149:424-427.
6. Benitz WE, Han MY, Madan A, Ramachandra P. Serial serum C-reactive protein levels in the diagnosis of neonatal infection. Pediatrics 1998;102: 4:E41.
470. J Lab Med 1996;20:145-152.
9. Pearson TA, Mensah GA, Alexander RW, Anderson JL, Cannon RO, Criqui M et al. Markers of inflammation and cardiovascular disease. Application to clinical
and public health practice. A statement for healthcare professionals from the Centers for Disease Control and Prevention and the American Heart Association.
Circulation. 2003;107:499-511.

2009-08
CRP Latex, AU400/AU640 Normal Serum/Plasma Application CRP Latex, AU680/AU480 Normal Serum/Plasma Application
Test Name: CRPN ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CRPN ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 2 Dilution 0 Pre-Dilution Rate: 1 Sample Volume 1.6 μL Dilution 0 μL OD Limit
μL μL
Reagent OD limit:
Test Name: CRPN ∇ < > Type Serum ∇ General ISE
Calibration Type: 6AB ∇ Formula: SPLINE ∇ Counts: # Process: CONC ∇ Test Name: CRPN ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: 6AB ∇ Formula: SPLINE ∇ Counts: # ∇

Point 1: # 0§ -0.1 2.5
Point 2: # 10† -0.1 2.5
Point 1: # ∇ 0§ -0.1 2.5
Point 3: # 40† -0.1 2.5
Point 2: # ∇ 10† -0.1 2.5 Allowance Range Check
Point 4: # 160† -0.1 2.5
Point 3: # ∇ 40† -0.1 2.5
Point 5: # 320† -0.1 2.5
Point 4: # ∇ 160† -0.1 2.5 ο Reagent Blank
Point 6: # 480† -0.1 2.5 Point 5: # 320† -0.1 2.5
∇ ο Calibration
Point 7: Point 6: # 480† -0.1 2.5
∇
1-Point Cal. Point: ο with CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ Point 7 ∇ Advanced Calibration
Point 9 ∇
Test Name: CRPN ∇ < > Type: Serum ∇
Logic Check-1 Logic Check –2 Logic Check -3
Check Point-1: Check Point-1: Check Point-1:
Parameters Misc
Check Point-2: Check Point Interval: Check Point Interval:
Check Point-3: Decision Value-1: Decision Value-1: CheckedTests Contamination Data Check
Decision Value-1: Decision Value-2: Decision Value-2: Parameters Parameters
Decision Value-2: Limit Point 1: Limit Point 1:
Decision Value-3: Limit Point 2: Limit Point 2: Test Name: CRPN ∇ < > Type: Serum ∇
Limit Point 1:
Limit Point 2: Logic Check-1 Logic Check -2 Logic Check-3
# User defined Check Point-3:
§ For the zero calibrator saline should be used Decision Value-1: Decision Value-1: Decision Value-1:
† CRP Latex calibrator Normal Set Cat. No.: ODC0026
Decision Value-2: Decision Value-2: Decision Value 2:
ф AU680 Decision Value-3:
Limit Point 1: Limit Point 1: Limit Point 1:
Check Pattern: ∇

2009-08
CRP Latex, AU400/AU640 Highly Sensitive Serum/Plasma Application CRP Latex, AU680/AU480 Highly Sensitive Serum/Plasma Application
Test Name: CRPHS ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CRPHS ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Test Name: CRPHS ∇ < > Type Serum ∇ General ISE
Calibration Type: 6AB ∇ Formula: SPLINE ∇ Counts: # Process: CONC ∇ Test Name: CRPHS ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # 0§ -0.1 2.5
Point 2: # 2.5† -0.1 2.5
Point 1: # ∇ 0§ -0.1 2.5
Point 3: # 10† -0.1 2.5
Point 2: # ∇ 2.5† -0.1 2.5 Allowance Range Check
Point 4: # 20† -0.1 2.5
Point 3: # ∇ 10† -0.1 2.5
Point 5: # 80† -0.1 2.5
Point 4: # ∇ 20† -0.1 2.5 ο Reagent Blank
Point 6: # 160† -0.1 2.5 Point 5: # 80† -0.1 2.5
∇ ο Calibration
Point 7: Point 6: # 160† -0.1 2.5
∇
Point 9 ∇
Test Name: CRPHS Type: Serum Point-1 ∇ Reagent Blank 999 Day 0 Hour
∇ < > ∇
Logic Check-1 Logic Check –2 Logic Check -3
Parameters Misc
Check Point-3: Decision Value-1: Decision Value-1: CheckedTests Contamination Data Check
Decision Value-1: Decision Value-2: Decision Value-2: Parameters Parameters
Decision Value-2: Limit Point 1: Limit Point 1:
Decision Value-3: Limit Point 2: Limit Point 2: Test Name: CRPHS ∇ < > Type: Serum ∇
Limit Point 1:
Limit Point 2: Logic Check-1 Logic Check -2 Logic Check-3
# User defined Check Point-3:
§ For the zero calibrator saline should be used Decision Value-1: Decision Value-1: Decision Value-1:
† CRP Latex calibrator Highly Sensitive Set Cat. No.: ODC0027
Decision Value-2: Decision Value-2: Decision Value 2:
ф AU680 Decision Value-3:
Check Pattern: ∇

2009-08
CRP Latex, AU600 Normal Serum/Plasma Application CRP Latex, AU600 Highly Sensitive Serum/Plasma Application
Test No # Test name CRPN ∇ Sample type Ser ∇ Page ½ Test No # Test name CRPHS ∇ Sample type Ser ∇ Page ½
Sample vol. 2 Dil. Vol 0 μL Min. OD Max. OD Sample vol. 3 Dil. Vol 0 μL Min. OD Max. OD
Reagent 1 vol 150 Dil. Vol 0 μL L 0.6 H 2.5 Reagent 1 vol 75 Dil. Vol 0 μL L -0.1 H 2.5
Reagent 2 vol 150 Dil. Vol 10 μL Reagent OD limit Reagent 2 vol 75 Dil. Vol 10 μL Reagent OD limit
Fst. L -0.1 Fst. H 2.5 Fst. L -0.1 Fst. H 2.5
Wave Main 570 Sub NONE ∇ Lst. L -0.1 Lst. H 2.5 Wave Main 570 Sub NONE ∇ Lst. L -0.1 Lst. H 2.5
Method FIXED ∇ Dynamic range Method FIXED ∇ Dynamic range
Reaction + ∇ L 0.2* H 480* Reaction + ∇ L 0.08* H 80*
Point 1 Fst 12 Lst 22 Correlation factor A 1 Point 1 Fst 12 Lst 22 Correlation factor A 1
Point 2 Fst Lst B 0 Point 2 Fst Lst B 0
Sample Pre-dil. Rate ¤ Sample Pre-dil. Rate ¤
Linearity Fst % Sec % Linearity Fst % Sec %
No lag time ∇ On-board stability period 90 No lag time ∇ On-board stability period 90
Select using Space key, or select from list displayed by Guide key Select using Space key, or select from list displayed by Guide key
Test No # Test name CRPN ∇ Test No # Test name CRPHS ∇
Cal type 14 6AB ∇ Count # Cal type 14 6AB ∇ Count #

Formula 10 SPLINE ∇ Process Conc ∇ Formula 10 SPLINE ∇ Process Conc ∇
Selection calibrator Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H Cal No OD Conc Factor/OD-L Factor/OD-H
Point 1 # ∇ 0§ -0.1 2.5 Point 1 # ∇ 0§ -0.1 2.5
Point 2 # ∇ 10† -0.1 2.5 Point 2 # ∇ 2.5† -0.1 2.5
Point 3 # ∇ 40† -0.1 2.5 Point 3 # ∇ 10† -0.1 2.5
Point 4 # ∇ 160† -0.1 2.5 Point 4 # ∇ 20† -0.1 2.5
Point 5 # ∇ 320† -0.1 2.5 Point 5 # ∇ 80† -0.1 2.5
Point 6 # ∇ 480† -0.1 2.5 Point 6 # ∇ 160† -0.1 2.5
1-point cal. Point 1-point cal. Point
MB type factor MB type factor
Calibrator stability period 999 Calibrator stability period 999
Select the function using the Function key or the Mouse Select the function using the Function key or the Mouse
Data Check Parameters Data Check Parameters
Test No # Name CRPN ∇ Type Ser ∇ Test No # Name CRPHS ∇ Type Ser ∇
Logic check-1 NO Logic Check-2 NO Logic check-1 NO Logic Check-2 NO

Check Point-1 0 Check Point-1 0 Check Point-1 0 Check Point-1 0
Check Point-2 0 Check Point Interval Check Point-2 0 Check Point Interval
Check Point-3 0 Decision value-1 0.0000 Check Point-3 0 Decision value-1 0.0000
Decision value-1 0.0000 Decision value-2 0.0000 Decision value-1 0.0000 Decision value-2 0.0000
Decision value-2 0.0000 Limit Point 1 0 Decision value-2 0.0000 Limit Point 1 0
Decision value-3 0.0000 Limit Point 2 0 Decision value-3 0.0000 Limit Point 2 0
Limit Point 1 0 Logic Check-3 NO Limit Point 1 0 Logic Check-3 NO
Limit Point 2 0 Limit Point 2 0
Check Point Interval Check Point Interval
# User defined ¤ Analyser default value # User defined ¤ Analyser default value
§ For the zero calibrator saline should be used § For the zero calibrator saline should be used
† CRP Latex calibrator Normal Set Cat. No.: ODC0026 † CRP Latex calibrator Highly Sensitive Set Cat. No.: ODC0027
* Values set for working in mg/L * Values set for working in mg/L

2009-08
CRP Latex, AU2700/AU5400 Normal Serum/Plasma Application CRP Latex, AU2700/AU5400 Highly Sensitive Serum/Plasma
Application
Specific Test Parameters System Reagent: OSR6199, OSR6299 Reagent ID: 099
General LIH ISE Range Specific Test Parameters
Test Name: CRPN ∇ < > Type: Serum ∇ Operation: Yes ∇
Test Name: CRPHS ∇ < > Type: Serum ∇ Operation: Yes ∇
Reagents: R1 Volume 120 μL Dilution 0 μL Min OD Max OD Sample: Volume 3.5 μL Dilution 0 μL Pre-Dilution Rate: 1
R2 Volume 120 μL Dilution 10 μL L 0.6 H 2.5 Reagents: R1 Volume 70 μL Dilution 0 μL Min OD Max OD
Reagent OD limit: R2 Volume 70 μL Dilution 10 μL L -0.1 H 2.5
Wavelength: Pri. 570 ∇ Sec. None ∇ First L -0.1 First H 2.5 Reagent OD limit:
Method: FIXED ∇ Last L -0.1 Last H 2.5 Wavelength: Pri. 570 ∇ Sec. None ∇ First L -0.1 First H 2.5
Reaction slope: + ∇ Dynamic Range: Method: FIXED ∇ Last L -0.1 Last H 2.5
Measuring Point 1: First 12 Last 22 L 0.2* H 480* Reaction slope: + ∇ Dynamic Range:
Measuring Point 2: First Last Correlation Factor: Measuring Point 1: First 12 Last 22 L 0.08* H 80*
Linearity : % A 1 B 0 Measuring Point 2: First Last Correlation Factor:
No Lag Time: ∇ On-board stability period: 90 Linearity : % A 1 B 0
General ISE Calibration Specific
General ISE
Test Name: CRPHS ∇ < > Type: Serum ∇
Calibration Type: 6AB ∇ Formula: SPLINE ∇ Counts: # Process: CONC ∇
Calibration Type: 6AB ∇ Formula: SPLINE ∇ Counts: # Process: CONC ∇
Point 1: # 0§ -0.1 2.5 Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 2: # 10† -0.1 2.5 Point 1: # 0§ -0.1 2.5
Point 3: # 40† -0.1 2.5 Point 2: # 2.5† -0.1 2.5
Point 4: # 160† -0.1 2.5 Point 3: # 10† -0.1 2.5
Point 5: # 320† -0.1 2.5 Point 4: # 20† -0.1 2.5
Point 6: # 480† -0.1 2.5 Point 5: # 80† -0.1 2.5
Point 7: Point 6: # 160† -0.1 2.5
Point 7:

Test Name: CRPHS ∇ < > Type: Serum ∇
Logic Check-1 Logic Check-2 Logic Check-3

Check Point-1: Check Point-1: Check Point-1: √ Logic Check-1 Logic Check-2 Logic Check-3
Check Point-2: Check Point Interval: Check Point Interval: Check Point-1: 11 Check Point-1: Check Point-1:
Check Point-3: Decision Value-1: Decision Value-1: Check Point-2: 15 Check Point Interval: Check Point Interval:
Decision Value-1: Decision Value-2: Decision Value-2: Check Point-3: 27 Decision Value-1: Decision Value-1:
Decision Value-2: Limit Point 1: Limit Point 1: Decision Value-1: 0.0 Decision Value-2: Decision Value-2:
Decision Value-3: Limit Point 2: Limit Point 2: Decision Value-2: 2.0 Limit Point 1: Limit Point 1:
Limit Point 1: Decision Value-3: 0.35 Limit Point 2: Limit Point 2:
Limit Point 2: Limit Point 1: 17
Limit Point 2: 25
# User defined.
§ For the zero calibrator saline should be used # User defined.
† CRP Latex calibrator Normal Set Cat. No.: ODC0026 § For the zero calibrator saline should be used
* Values set for working in mg/L † CRP Latex calibrator Highly Sensitive Set Cat. No.: ODC0027

2009-08
D-DIMER
OSR60135 2 x 12.5 mL R1
2 x 12.5 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of D-Dimer in human plasma on Beckman Coulter analysers. For in vitro diagnostic
use only.
Summary1,2,3
Plasmin degradation of cross-linked fibrin results in the formation of specific degradation products including D-Dimer. As D-Dimer is released
into the circulation during the fibrinolytic process, the measurement of D-Dimer and higher molecular weight oligomers containing D-Dimer
epitopes is considered to reflect the overall activity of clot formation and lysis. Elevated levels of D-Dimer can occur in a variety of clinical
conditions associated with fibrin breakdown, including Deep Vein Thrombosis (DVT), Pulmonary Embolism (PE) and Disseminated Intravascular
Coagulation (DIC). D-Dimer is generally used for its negative predictive value.
Test Principle
When a sample is mixed with R1 buffer and R2 latex suspension, D-Dimer reacts specifically with anti-human D-Dimer antibodies coated on the
latex particles to yield insoluble aggregates. The absorbance of these aggregates is proportional to the D-Dimer concentration in the sample.
Contents
Tris/HCl
NaCl
Bovine Serum Albumin
Latex coated monoclonal anti human D-Dimer antibodies (mouse)
Preservative
R28 Very toxic if swallowed.
R32 Contact with acids liberates very toxic gas.
R36/37/38 Irritating to eyes, respiratory system and skin.
R50/53 Very toxic to aquatic organisms, may cause long-term adverse effects in the aquatic environment.
To avoid the possible build-up of azide compounds, flush waste-pipes with water after the disposal of material.
Safety data sheet available for professional user on request
Biological materials of human origin contained in this product were tested for Anti-HCV, HbsAg and Anti-HIV 1/2 on a single donor basis using
FDA approved methods and were found to be non-reactive. As there is no known test method that can offer complete assurance that products
Reagent Preparation
The reagents are ready for use and can be placed directly on board the instrument. Mix the R2 gently before use and on a weekly basis
thereafter.
Specimen4
Citrated plasma. Stable in plasma for 4 days when stored at 2…8°C and 6 months when stored at -20°C. Lithium Heparin plasma may also be
used. Unlike when using citrated plasma, there is no sample dilution with heparin tubes. Therefore the D-Dimer values in heparin plasma are on
average 16% higher over the entire measuring range.
Use samples undiluted.
Test Procedure
statement. Data check parameters are required. See Setting Sheet for specific instrument details.
Calibration
D-Dimer Calibrator Cat. No. ODR3033
The D-Dimer assay has been calibrated against another commercially available immuno-turbidimetric assay which reports results in µg FEU/mL.
Recalibrate the assay every 30 days or when the following occur:
Following calibration, the resulting curve should be visually reviewed on the Beckman Coulter analyser for acceptability using the software
options - Routine, Calibration Monitor, Calibration Curve.
Quality control procedures should be undertaken immediately following calibration in accordance with good laboratory practice.
Quality Control
D-Dimer Controls Cat. No. ODC0029 or other control materials with values determined by this Beckman Coulter system may be used.
Each laboratory should establish its own control frequency, however good laboratory practice suggests that controls be tested each day patient
2009-08
Calculation
The Beckman Coulter analysers automatically compute the D-Dimer concentration of each sample.
Reference Interval < 0.5 µg FEU/mL. Expected values may vary with age, sex, sample type, diet and geographical location. Each laboratory
should verify the transferability of the expected values to its own population, and if necessary determine its own reference interval according to
good laboratory practice. For diagnostic purposes, results should always be assessed in conjunction with the patient's medical history, clinical
examinations and other findings.
Note: D-Dimer analysis is system specific and values generated from different manufacturers may differ significantly. Each laboratory must
determine reference intervals for their individual test populations, reagents and instruments, accordingly.
Clinical Performance in the Evaluation of Venous Thromboembolism using the D-Dimer assay
The diagnostic utility of the D-Dimer assay for exclusion of Venous Thromboembolism (VTE) was evaluated in a clinical study conducted on 248
patients. Samples with suspected Venous Thromboembolism (VTE) were analysed for D-Dimer using the D-Dimer assay. A negative diagnosis
was confirmed using established clinical procedures in 85 out of 86 patients with D-Dimer concentration
< 0.5 µg FEU/mL. Of the 162 patients with D-Dimer concentration > 0.5 µg FEU/mL, 48 were confirmed as positive for VTE using established
clinical procedures.
Results were analysed using a clinical cut-off for D-Dimer concentration of 0.5 µg FEU/mL, whereby samples with D-Dimer concentration
< 0.5 µg FEU/mL were considered negative, and samples with D-Dimer concentration > 0.5 µg FEU/mL were considered positive. Results are
summarised in the table below:
n % Sensitivity (95% CI) % Specificity (95% CI) % Negative Predictive Value (NPV) (95% CI)
248 98.0 (87.8 – 99.9) 42.7 (35.8 – 49.9) 98.8 (92.8 – 99.9)
This data is representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ from these values.
CI = Confidence Interval
The diagnostic performance of the method was evaluated and compared to the BioMerieux Vidas D-Dimer assay using ROC (Receiver
Operating Characteristics) analysis. Results are summarised in the table below.
Diagnostic (n)
n = 248
Absent 199 / Present 49
Test Area 95% CI SE

Vidas µg FEU/mL 0.87 0.82 to 0.92 0.025
Beckman Coulter µg 0.87 0.82 to 0.92 0.025
FEU/mL
SE = Systematic Error
In a paper by Altman and Bland in the British Medical Journal in 1994, it was remarked that “Predictive values observed in one study do not
7
apply universally”. This make direct comparison of NPV results between studies difficult.
Data contained within this section are representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ
from these values.
Linearity
The test is linear within a concentration range of 0.25 – 8 µg FEU/mL. Samples exceeding the upper limit of linearity should not be diluted, but
instead should be reported as > 8 µg FEU/mL.
Prozone settings must be applied when using the D-Dimer OSR60135 assay.
Precision
The following data were obtained on an AU640, AU400 and AU2700 using 3 plasma pools analysed over 20 days.
AU640:
Mean µg FEU/mL SD CV% SD CV%
0.28 0.01 4.60 0.03 9.14
0.55 0.02 4.22 0.04 7.95
5.66 0.04 0.69 0.17 3.02
AU400:
0.28 0.02 6.12 0.03 9.44
0.57 0.02 3.21 0.05 7.99
5.86 0.03 0.55 0.15 2.48
AU2700:
0.28 0.01 4.42 0.02 8.17
0.54 0.01 2.06 0.02 4.44
6.18 0.03 0.42 0.16 2.52
Sensitivity
Analyser Lowest Detectable Level (µg FEU/mL)
AU640 0.08
AU400 0.05
AU2700 0.03
The lowest detectable level represents the lowest measurable level of D-Dimer that can be distinguished from zero. It is calculated as the

2009-08
Method Comparison
Patient plasma samples were used to compare this D-Dimer (OSR60135) assay on the AU640 against another commercially available D-Dimer
y = 1.010x + 0.079 r = 0.996 n = 104 Sample range = 0.28- 7.53 µg FEU/mL
®
Rheumatoid Factor: Interference less than 10% up to 100 IU/mL
Heparin: Interference less than 10% up to 1.5 IU/mL
6
Limitations8
The D-Dimer assay has been optimised to reduce the risk of prozone occurrence in the presence of abnormally high D-Dimer concentrations.
Samples with very elevated D-Dimer concentrations (>200 µg FEU/mL) can generate false low results without appropriate “Z” flags due to
excess antigen in the sample, such as can occur during lysis therapy. In very rare cases gammopathy, especially monoclonal IgM
(Waldenström’s macroglobulinemia), may cause unreliable results.
Samples containing heterophilic antibodies may cause falsely elevated results.
† D-Dimer calibrator ODR3033
* Values set for working in µg FEU/mL
BIBLIOGRAPHY
1. Budzynski AJ, Marder VJ, Parker ME, Shames P, Brizuela BS, Olexa SA. Antigenic markers of fragment DD, a unique plasmic derivative of
human crosslinked fibrin. Blood 1979:54:794-804.
2. Gaffney PJ, Brasher M. Subunit structure of the plasmin-induced degradation products of crosslinked fibrin. Biochem Biophys Acta
1973;295:308-313.
3. Wakai A, Gleeson A, Winter D. Role of D-Dimer Testing in Emergency Medicine. Emerg. Med J. 2003; 20:319-325.
4. Guder, WG, Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of anticoagulants in diagnostic laboratory investigations
and stability of blood, plasma and serum samples. WHO/DIL/LAB/99.1 Rev.2, 2002.
5. Data on file at Beckman Coulter Biomedical Ltd..
th
6. Young DS. Effects of Drugs on Clinical Laboratory Tests, AACC, 5 ed. AACC Press, 2000.
7. Altman DG, Bland JM. Diagnostic tests 2: Predictive values. BMJ. 1994; 309: 102.
8. Selby C. Interference in Immunoassay. Ann Clin Biochem 1999; 36:704-721.

2009-08
D-DIMER, AU400/AU640 Citrated Plasma Application D-DIMER, AU600 Citrated Plasma Application
System Reagent: OSR60135 Reagent ID: 135 System Reagent: OSR60135
Reagent ID: 135
Test No # Test name DDIM ∇ Sample type Ser ∇ Page ½
Test Name: DDIM ∇ < > Type: Serum ∇ Operation: Yes ∇
Reagent 2 vol 125 Dil. Vol 0 μL Reagent OD limit
Fst. L -2.0 Fst. H 2.5
R2 Volume 125 μL Dilution 0 μL L -2.0 H 2.5 Wave Main 800 Sub NONE ∇ Lst. L -2.0 Lst. H 2.5
Wavelength: Pri. 800 ∇ Sec. None ∇ First L -2.0 First H 2.5 Reaction + ∇ L 0.25* H 8*
Reaction slope: + Dynamic Range: Point 2 Fst Lst B 0
∇
Measuring Point 1: First 13 Last 27 L 0.25* H 8*
Test No # Test name DDIM ∇

Formula 10 SPLINE ∇ Process Conc ∇
Test Name: D
DDIM ∇ < > Type Serum ∇ Selection calibrator
Calibration Type: 6AB ∇ Formula: SPLINE ∇ Counts: # Process: CONC ∇ Point 1 # ∇ † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
Point 6 # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5
Point 7 ∇
Point 4: # † -0.1 2.5 1-point cal. Point
Point 7:
Test No # Name DDIM ∇ Type Ser ∇

Logic check-1 NO Logic Check-2 NO
Test Name: DDIM ∇ < > Type: Serum ∇ Check Point-2 0 Check Point Interval
CITRATED PLASMA APPLICATION

Logic Check-1 Logic Check -2 √ Logic Check-3 Decision value-1 0.0000 Decision value-2 0.0000
Check Point-1: Check Point-1: Check Point-1: 16 Decision value-2 0.0000 Limit Point 1 0
Check Point-2: Check Point Interval: Check Point Interval: 2 Decision value-3 0.0000 Limit Point 2 0
Check Point-3: Decision Value-1: Decision Value-1: 0.9 Limit Point 1 0 Logic Check-3 YES
Decision Value-1: Decision Value-2: Decision Value-2: 0.2 Limit Point 2 0
Decision Value-2: Limit Point 1: Limit Point 1: 12 Check Point-1 16
Decision Value-3: Limit Point 2: Limit Point 2: 24 Check Point Interval 2
Limit Point 1: Decision value-1 0.9
Limit Point 1 12
Limit Point 2 24

† D-Dimer Calibrator Cat. No.: ODR3033 † D-Dimer Calibrator Cat. No.: ODR3033
* Values set for working in µg FEU/mL. * Values set for working in µg FEU/mL.

2009-08
D-DIMER, AU2700/AU5400 Citrated Plasma Application D-DIMER, AU680/AU480 Citrated Plasma Application
Test Name: DDIM ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: DDIM ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Test Name: DDIM ∇ < > Type: Serum ∇ General ISE
Calibration Type: 6AB ∇ Formula: SPLINE ∇ Counts: # Process: CONC ∇ Test Name: DDIM ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # † -0.1 2.5
Point 2: # † -0.1 2.5
CITRATED SERUM
Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5
PLASMA
Point 4: # † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5
Point 6: # † -0.1 2.5 Point 5: # † -0.1 2.5
∇ ο Calibration
Point 7: Point 6: # † -0.1 2.5
∇
Point 9 ∇
APPLICATION
Test Name: DDIM ∇ < > Type: Serum ∇ Point-1 ∇ Reagent Blank 30 Day 0 Hour
APPLICATION
Logic Check-1 Logic Check -2 √ Logic Check-3
Check Point-1: Check Point-1: Check Point-1: 16
Check Point-2: Check Point Interval: Check Point Interval: 2 Parameters Misc
Check Point-3: Decision Value-1: Decision Value-1: 0.9
Decision Value-1: Decision Value-2: Decision Value-2: 0.2
Decision Value-2: Limit Point 1: Limit Point 1: 12
Decision Value-3: Limit Point 2: Limit Point 2: 24 Test Name: DDIM ∇ < > Type: Serum ∇
Limit Point 1:
Limit Point 2: Logic Check-1 Logic Check -2 √ Logic Check-3
Check Point-2: Check Point Interval: Check Point Interval: 2
Check Point-3:
# User defined. Decision Value-2: Decision Value-2: Decision Value 2: 0.2
† D-Dimer Calibrator Cat. No.: ODR3033
Decision Value-3:
* Values set for working in µg FEU/mL.
Limit Point 1: Limit Point 1: Limit Point 1: 12
ф AU680
Check Pattern: ∇

2009-08
FERRITIN
OSR6150 4 x 24 mL R1
4 x 5 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of ferritin in human serum on Beckman Coulter analysers. For in vitro diagnostic use only.
Summary1
1
Serum ferritin is a sensitive indicator of body iron stores; it has been shown to correlate with stainable bone-marrow iron. Serum ferritin is especially useful
in distinguishing iron deficiency from the anemia of chronic disorders because in the latter ferritin levels are increased. A serum ferritin level of less than
10 µg/L almost always indicates iron deficiency. Serum ferritin is also increased in other anemias including aplastic anemia, sideroblastic anemia, and
chronic haemolytic anemias. In idiopathic hemochromatosis and, in multiple transfused patients, the serum ferritin may be extremely high.
Test Principle2,3
Latex agglutination reactions occur as a result of antibody-coated latex beads aggregating if antigen is present in sufficient quantity. Immune complexes
formed in solution scatter light in proportion to their size, shape and concentration. Under conditions of antibody excess, increasing amounts of antigen
result in higher scatter. Turbidimeters measure the reduction of incident light due to reflection, absorption, or scatter.
In the procedure, the measurement of the decrease in light intensity transmitted (increase in absorbance) through particles suspended in solution as a
result of complexes formed during the antigen-antibody reaction, is the basis of this assay. The Ferritin reagent is a suspension of polystyrene latex
particles, of uniform size, coated with polyclonal rabbit anti-ferritin antibody. When serum, containing ferritin, is mixed with the Ferritin reagent, an
agglutination mixture occurs. This is measured spectrophotometrically on Beckman Coulter Chemistry Analysers.
Latex particles coated with rabbit anti-human ferritin
Preservative.
To avoid the possible build up of azide compounds, flush waste-pipes with water after the disposal of undiluted reagent.
Reagent Preparation
instrument.
The reagents are stable unopened, up to the stated expiry date when stored at 2…8°C. Once open, reagents stored on board the instrument are stable for
30 days. Once on board the R2 must be mixed at weekly intervals (see Reagent Preparation).
Specimen
Serum samples only are the recommended specimens.
4,5
Stability in serum: at 2…8°C up to 7 days
at - 20°C up to 6 months

Test Procedure
Data check parameters are required, see setting sheets for specific instrument details.
Calibration
Serum Protein Multi-Calibrator Cat No. ODR3021.
The calibrator ferritin values assigned to the calibrators are traceable to the 3rd International Standard for ferritin, Recombinant NIBSC code: 94/572.
Quality Control
ITA Control Sera ODC0014, ODC0015, ODC0016 or other control materials with values determined by this Beckman Coulter system may be used.
Calculation
The Beckman Coulter analysers automatically compute the ferritin concentration of each sample.
2009-08
Serum
New-born Infants: 25 – 200 µg/L
6 months to 15 years: 7 – 142 µg/L
Adult male: 20 – 300 µg/L
Adult female: 10 – 120 µg/L
values.
Linearity
The test is linear within a concentration range of 8.0 – 450 µg/L.
Precision
Mean μg/L SD CV% SD CV%
40 0.91 2.25 1.40 3.43
101 2.03 2.00 2.86 2.81
383 4.79 1.25 8.14 2.12
Sensitivity
The lowest detectable level in serum on an AU2700 was estimated at 6.4 µg/L.
The lowest detectable level represents the lowest measurable level of ferritin that can be distinguished from zero. It is calculated as the absolute mean plus
Method Comparison
Patient samples were used to compare this Ferritin OSR6150 assay on an AU640 against another commercially available ferritin assay. Results of linear
regression analysis were as follows.
y = 0.964x – 2.549 r = 0.995 n = 100 Sample Range = 20 – 408 µg/L
Lipemic: Interference less than 20% up to 400 mg/dL Intralipid®
Triglyceride: Interference less than 10% up to 1500 mg/dL triglyceride
Rheumatoid Factor: Interference less than 5% up to 500 IU/mL RF
6
Limitations
7
Certain specimens may contain heterophilic antibodies. Such specimens may result in falsely elevated values when tested with the Ferritin assay.
Samples with extremely abnormal optical characteristics, especially turbidity, may produce atypical results. Such samples must be serially diluted and
results compared to ensure that no such interference exists.
For diagnostic purposes the Ferritin results should always be assessed in conjunction with other available information e.g., patients medical history, clinical
impressions and results of other tests.
Samples with very high Ferritin concentrations (> 20,000 IU/mL) can generate false low results without appropriate "Z" flags due to excess antigen in the
sample.
† Serum Protein Multi-Calibrator Cat No.: ODR3021.
* Values set for working in SI units µg/L equivalent to ng/mL.
Bibliography
1. Fairbanks VF, Klee GG. Biochemical aspects of hematology.In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia: WB Saunders Company,
1987:823pp.
2. Bernard A, Lauwerys R. Turbidimetric latex immunoassay for serum ferritin. Journal of Immunological methods 1984;71:141 147.
3. Thompson SG, Principles for competitive-binding assays. In: Kaplan LA, Pesce AJ ,eds, Clinical Chemistry theory, analysis and correlation. St Louis: Mosby,
1996:265pp.
4. Tietz NW, ed. Clinical guide to laboratory tests. 3rd ed. Philadelphia: WB Saunders Company, 1995:234pp.
5. Young DS, ed. Effects of preanalytical variables on clinical laboratory tests, 2nd ed. Washington : AACC Press, 1997:3-220pp.
7. Selby C. Interference in Immunoassay. Ann Clin Biochem 1999;36:704-721.

2009-08
FERRITIN, AU400/AU640 Serum Application FERRITIN, AU600 Serum Application

Test No # Test name FERR ∇ Sample type Ser ∇ Page ½
Test Name: FERR ∇ < > Type: Serum ∇ Operation: Yes ∇
Fst. L 0.8 Fst. H 1.8
Wave Main 600 Sub NONE ∇ Lst. L 0.8 Lst. H 1.8
Wavelength: Pri. 600 ∇ Sec. None ∇ First L 0.8 First H 1.8 Reaction + ∇ L 8* H 450*
Method: FIXED ∇ Last L 0.8 Last H 1.8 Point 1 Fst 12 Lst 18 Correlation factor A 1
Reaction slope: + ∇ Dynamic Range: Point 2 Fst Lst B 0
Calibration Specific Test No # Test name FERR ∇

General ISE
Cal type 13 5AB ∇ Count #
Test Name: FERR ∇ < > Type Serum ∇ Formula 3 POLYGONAL ∇ Process Conc ∇
Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇ Cal No OD Conc Factor/OD-L Factor/OD-H
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 6 ∇
Point 7 ∇
Point 5: # † -0.1 2.5
1-point cal. Point
1-Point Cal. Point: ο With CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ Select the function using the Function key or the Mouse
SERUM APPLICATION
Test No # Name FERR ∇ Type Ser ∇

Test Name: FERR ∇ < > Type: Serum ∇ Check Point-1 0 Check Point-1 0
Logic Check-1 Logic Check -2 √ Logic Check-3 Check Point-3 0 Decision value-1 0.0000
Check Point-1: Check Point-1: Check Point-1: 11 Decision value-1 0.0000 Decision value-2 0.0000
Check Point-3: Decision Value-1: Decision Value-1: 1.2000 Decision value-3 0.0000 Limit Point 2 0
Decision Value-1: Decision Value-2: Decision Value-2: 0.0400 Limit Point 1 0 Logic Check-3 YES
Decision Value-2: Limit Point 1: Limit Point 1: 11 Limit Point 2 0
Limit Point 1: Check Point Interval 2
Decision value-2 0.0400
Limit Point 1 11
Limit Point 2 15
# User defined
† Serum Protein Multi-Calibrator ODR3021 # User defined ¤ Analyser default value
* Values set for working in SI units μg/L equivalent to ng/mL † Serum Protein Multi-Calibrator ODR3021
* Values set for working in SI units μg/L equivalent to ng/mL

2009-08
FERRITIN, AU2700/AU5400 Serum Application FERRITIN, AU680 Serum Application
Test Name: FERR ∇ < > Type: Serum ∇ Operation Yes ∇

μL μL
Reagent OD limit:
Wavelength: Pri. 600 ∇ Sec. None ∇ First L 0.8 First H 1.8
Method: FIXED ∇ Last L 0.8 Last H 1.8
Measuring Point2 First Last Lipemia ++++ ∇
Calibration Specific Lag Time Check ∇ Hemolysis +++++ ∇
General ISE
Test Name: FERR ∇ < > Type: Serum ∇ Calibrators Calibration Specific STAT Table Calibration
General ISE
Test Name: FERR ∇ < > Type Serum ∇ ο Use Serum Cal.
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: # ∇
Point 1: # † -0.1 2.5 <Calibrator Parameters> OD Range
Point 2: # † -0.1 2.5 Calibrator OD Conc Low High Slope Check + ∇
Point 3: # † -0.1 2.5 Point 1: # † -0.1 2.5
∇
Point 4: # † -0.1 2.5 Point 2: # † -0.1 2.5 Allowance Range Check
∇
Point 5: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 6: Point 4: # ∇ † -0.1 2.5 ο Reagent Blank
Point 7: Point 5: # ∇ † -0.1 2.5 ο Calibration
1-Point Cal. Point: ο with CONC-0 Slope Check: + ∇ Advanced Calibration # ∇ Point 6: ∇
MB Type Factor: Calibration Stability Period: 14 Point 7 ∇ Advanced Calibration
Point 9 ∇
SERUM APPLICATION
Test Name: FERR ∇ < > Type: Serum ∇ Calibrator OD Conc Low High Stability
Logic Check-1 Logic Check -2 √ Logic Check-3 MB Type Factor: 1-Point Calibration Point None ∇ ο with Conc-0
Parameters Misc
Limit Point 1: Test Name: FERR ∇ < > Type: Serum ∇
Limit Point 2:
Check Point-3:
# User defined. Decision Value-1: Decision Value-1: Decision Value-1: 0.70
† Serum Protein Multi-Calibrator ODR3021 Decision Value-2: Decision Value-2: Decision Value 2: 0.04
* Values set for working in SI units μg/L equivalent to ng/mL Decision Value-3:
Check Pattern: ∇

2009-08
FERRITIN, AU480 Serum Application

Method FIXED ∇

General ISE
Test Name: FERR ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # ∇ † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 5: # ∇ † -0.1 2.5 ο Calibration
Point 6: ∇
Point 9 ∇
Parameters Misc

Test Name: FERR ∇ < > Type: Serum ∇

Check Point-3:
Decision Value-2: Decision Value-2: Decision Value 2: 0.0400
Decision Value-3:
Limit Point 2: Limit Point 2: Limit Point 2: 15 # User defined.
Check Pattern: ∇ † Serum Protein Multi-Calibrator ODR3021
* Values set for working in SI units μg/L equivalent to ng/mL

2009-08
FERRITIN
OSR61138 4 x 24 mL R1
4 x 5 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of ferritin in human serum and plasma on Beckman Coulter analysers. For in vitro diagnostic
use only.
Summary1,2
Measurement of ferritin in serum or plasma is a useful indicator of body iron stores in normal persons, and in individuals with iron deficiency. A ferritin level
of less than 10 µg/L usually indicates iron deficiency anemia. An increased ferritin concentration is observed in a large number of chronic diseases. These
include chronic infections; chronic inflammatory disorders; heart disease; and malignancies, especially lymphomas, leukemias, breast cancer and
neuroblastoma. In patients who have any of these disorders together with iron deficiency, ferritin concentration is often normal. Ferritin concentrations may
be extremely high in patients suffering from hemochromatosis and certain liver diseases.
Test Principle3,4
In the Beckman Coulter procedure, the measurement of the decrease in light intensity transmitted (increase in absorbance) through particles suspended in
solution as a result of complexes formed during the antigen-antibody reaction, is the basis of this assay. The Ferritin reagent is a suspension of polystyrene
latex particles, of uniform size, coated with polyclonal rabbit anti-ferritin antibody. When a sample containing ferritin is mixed with the Ferritin reagent, an
Preservative.
R1 reagent:
Irritant, contains a mixture of 5-chloro-2-methyl-2H-isothiazol-3-one [EC No 247-500-7] and 2-methyl-2H-isothiazol-3-one [EC No 220-239-6] (3:1).
R43: may cause sensitization by skin contact.
Safety Phrases: S24, S37, S60. Avoid contact with skin. Wear suitable gloves. This material and its container must be disposed of as hazardous waste.
R1 and R2 reagents:
Reagent Preparation
R1 is ready for use and can be placed directly on board the instrument. R2 should be mixed by inversion 5-10 times before placing on board the
instrument.
Specimen
Serum, Li-heparin plasma and EDTA plasma samples are the recommended specimens.
5,6
Stable in serum and plasma for 7 days when stored at 2…8°C and for 12 months when stored at -20°C.
Test Procedure
Calibration
rd
The calibrator ferritin values assigned to the calibrators are traceable to the 3 International Standard for ferritin, Recombinant NIBSC code: 94/572.
Quality Control
2009-08
Calculation
Reference Intervals
1
Serum Adults
7
Serum/Plasma Children
Up to 1 month: 6 – 400 µg/L
1 to 6 months: 6 – 410 µg/L
1 to 5 years: 6 – 60 µg/L
6 to 19 years: 6 – 320 µg/L
values.
Linearity
Precision
Mean µg/L SD CV% SD CV%
40 0.84 2.1 1.31 3.3
144 2.73 1.9 3.17 2.2
424 3.94 0.9 7.49 1.8
Sensitivity
Method Comparison
Patient samples were used to compare this Ferritin OSR61138 assay on an AU640 against another commercially available ferritin assay.
y = 0.98x + 2 r = 0.998 n = 109 Sample Range = 11 – 407 µg/L
Triglyceride: Interference less than 10% up to 1500 mg/dL triglyceride
8
Limitations
9
Certain specimens may contain heterophilic antibodies. Such specimens may result in falsely elevated values when tested with the Ferritin assay.
For diagnostic purposes the Ferritin results should always be assessed in conjunction with other available information e.g., patients medical history, clinical
impressions and results of other tests.
Samples with very high Ferritin concentrations (> 20,000 IU/mL) can generate false low results without appropriate "Z" flags due to excess antigen in the
sample.
Bibliography
2. Tietz NW, ed. Clinical guide to laboratory tests. 3rd ed. Philadelphia: WB Saunders Company, 1995:234-5.
3. Bernard A, Lauwerys R. Turbidimetric latex immunoassay for serum ferritin. Journal of Immunological methods 1984;71:141-147.
1996:265pp.
5. WHO. Use of anticoagulants in diagnostic laboratory investigations and stability of blood, plasma and serum samples. WHO/DIL/LAB/99.1 rev.2, 2002:31.
6. Young DS, ed. Effects of preanalytical variables on clinical laboratory tests, 2nd ed. Washington : AACC Press, 1997:3 -220pp.
8. Young DS. Effects of Drugs on Clinical Laboratory Tests, 5th ed. AACC Press, 2000.

2009-08
FERRITIN, AU400/AU640 Serum/Plasma Application FERRITIN, AU600 Serum/Plasma Application

Fst. L 0.8 Fst. H 1.8
Wave Main 600 Sub ∇ Lst. L 0.8 Lst. H 1.8
Test No # Test name FERR ∇

Test Name: FERR ∇ < > Type Serum ∇ Selection calibrator
Point 2 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 6 ∇
Point 4: # † -0.1 2.5 Point 7 ∇
1-point cal. Point
Point 7:

Logic Check-1 Logic Check -2 √ Logic Check-3 Decision value-1 0.0000 Decision value-2 0.0000
Check Point-1: Check Point-1: Check Point-1: 11 Decision value-2 0.0000 Limit Point 1 0
Check Point-3: Decision Value-1: Decision Value-1: 1.2000 Limit Point 1 0 Logic Check-3 YES
Decision Value-1: Decision Value-2: Decision Value-2: 0.0400 Limit Point 2 0
Decision Value-3: Limit Point 2: Limit Point 2: 15 Check Point Interval 2
Limit Point 1 11
Limit Point 2 15

* Values set for working in SI units µg/L, equivalent to ng/mL. * Values set for working in SI units µg/L, equivalent to ng/mL

2009-08

μL μL
Reagent OD limit:
Linearity : % A 1 B 0 Reaction Slope + ∇ Onboard Stability Period 30 Day Hour
Test Name: FERR ∇ < > Type: Serum ∇ General ISE
Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇ Test Name: FERR ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # † -0.1 2.5
Point 2: # † -0.1 2.5
SERUM/PLASMA
Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5
Point 4: # † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5
Point 6: Point 5: # † -0.1 2.5
∇ ο Calibration
Point 7: Point 6:
Point 9 ∇
SERUM APPLICATION
APPLICATION
Test Name: FERR ∇ < > Type: Serum ∇ ∇
Parameters Misc
Decision Value-3: Limit Point 2: Limit Point 2: 15 Test Name: FERR ∇ < > Type: Serum ∇
Limit Point 1:
Limit Point 2: Logic Check-1 Logic Check -2 √ Logic Check-3
Check Point-3:
# User defined. Decision Value-2: Decision Value-2: Decision Value 2: 0.04
† Serum Protein Multi-Calibrator Cat. No.: ODR3021 Decision Value-3:
* Values set for working in SI units µg/L, equivalent to ng/mL.
Check Pattern: ∇

2009-08
FERRITIN
OSR61203 4 x 24 mL R1
4 x 12 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of ferritin in human serum and plasma on Beckman Coulter AU analysers. For in vitro
Summary1,2
Measurement of ferritin in serum or plasma is a useful indicator of body iron stores in normal persons, and in individuals with iron deficiency. A ferritin level
of less than 10 µg/L usually indicates iron deficiency anemia. An increased ferritin concentration is observed in a large number of chronic diseases. These
include chronic infections; chronic inflammatory disorders; heart disease; and malignancies, especially lymphomas, leukemias, breast cancer and
neuroblastoma. In patients who have any of these disorders together with iron deficiency, ferritin concentration is often normal. Ferritin concentrations may
be extremely high in patients suffering from hemochromatosis and certain liver diseases.
Test Principle3,4
In the Beckman Coulter procedure, the measurement of the decrease in light intensity transmitted (increase in absorbance) through particles suspended in
solution as a result of complexes formed during the antigen-antibody reaction, is the basis of this assay. The Ferritin reagent is a suspension of polystyrene
latex particles, of uniform size, coated with polyclonal rabbit anti-ferritin antibody. When a sample containing ferritin is mixed with the Ferritin reagent, an

Glycine buffer (R1: pH 8.3, R2: pH 7.3 ) 170 mmol/L
Preservative.

Reagent Preparation
instrument.

Specimen
Serum, Li-heparin plasma and EDTA plasma samples are the recommended specimens.
5,6
Stable in serum and plasma for 7 days when stored at 2…8°C and for 12 months when stored at -20°C.
Test Procedure
Calibration
rd
The calibrator ferritin values assigned to the calibrators are traceable to the 3 International Standard for ferritin, Recombinant NIBSC code: 94/572.
Quality Control

2009-11
Calculation
Reference Intervals
1
Serum Adults
7
Serum/Plasma Children
Up to 1 month: 6 – 400 µg/L
1 to 5 years: 6 – 60 µg/L
6 to 19 years: 6 – 320 µg/L

values.
Linearity
Precision
25 0.55 2.24 0.92 3.71
148 0.91 0.61 2.21 1.49
438 4.18 0.95 4.95 1.13
Sensitivity
Method Comparison
Patient samples were used to compare this Ferritin OSR61203 assay on an AU400 against another commercially available ferritin assay.
y = 1.09x + 0.25 r = 0.994 n = 462 Sample Range = 14 – 434 µg/L
Bilirubin: Interference less than 5% up to 40 mg/dL
8
Limitations
This assay has been specifically designed to substantially reduce the risk of interference from HAMA or Heterophilic antibodies, however as with all
immuno-assays there is always a small risk from such interferences and therefore for diagnostic purposes the Ferritin results should always be assessed in
9,10
conjunction with other available information e.g., patients medical history, clinical impressions and results of other tests.
To investigate samples which are believed to contain interference, a number of approaches can be adopted. Serial dilution may reveal incorrect recovery,
PEG precipitation, pre-treatment with non immune serum or assay of the sample in an alternate assay system are all useful in identifying whether the
10
sample contains an interferent. The results of such samples should be interpreted with extreme care.
Samples with very high Ferritin concentrations (> 20,000 µg/L) can generate false low results without appropriate "Z" flags due to excess antigen in the
sample.

Bibliography
2. Tietz NW, ed. Clinical guide to laboratory tests. 3rd ed. Philadelphia: WB Saunders Company, 1995:234-5.
3. Bernard A, Lauwerys R. Turbidimetric latex immunoassay for serum ferritin. Journal of Immunological methods 1984;71:141-147.
1996:265pp.
5. WHO. Use of anticoagulants in diagnostic laboratory investigations and stability of blood, plasma and serum samples. WHO/DIL/LAB/99.1 rev.2, 2002:31.
6. Young DS, ed. Effects of preanalytical variables on clinical laboratory tests, 2nd ed. Washington : AACC Press, 1997:3 -220pp.
8. Young DS. Effects of Drugs on Clinical Laboratory Tests, 5th ed. AACC Press, 2000.
10. Ismail AA. On the interpretation of affirmative follow-up tests in immunoassays: what must not be done? Ann Clin Biochem 2006;43(4):249-251.

2009-11

Fst. L 0.5 Fst. H 2.5
Wave Main 660 Sub ∇ Lst. L 0.5 Lst. H 2.5
Test No # Test name FERR ∇

Test Name: FERR ∇ < > Type Serum ∇ Selection calibrator
Calibration Type: 5AB ∇ Formula: SPLINE ∇ Counts: # Process: CONC ∇ Point 1 # ∇ † -2.0 2.5
Point 2 # ∇ † -2.0 2.5
Point 1: # † -2.0 2.5 Point 4 # ∇ † -2.0 2.5
Point 2: # † -2.0 2.5 Point 5 # ∇ † -2.0 2.5
Point 3: # † -2.0 2.5 Point 6 ∇
Point 4: # † -2.0 2.5 Point 7 ∇
1-point cal. Point
Point 7:
1-Point Cal. Point: R with CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇

Check Point-1 11 Check Point-1
Decision Value-2: 2.0000 Limit Point 1: Limit Point 1: Check Point-1
Limit Point 1
Limit Point 2

† System Calibrator Cat. No.: ODR3021 † System Calibrator Cat. No.: ODR3021
* Values set for working in SI units µg/L, equivalent to ng/mL. * Values set for working in SI units µg/L, equivalent to ng/mL

2010-07
FERRITIN, AU2700/AU5400 Serum/Plasma Application FERRITIN, AU680/AU480 Serum/Plasma Application

µL µL
Reagent OD limit:
Test Name: FERR ∇ < > Type: Serum ∇ General ISE
Calibration Type: 5AB ∇ Formula: SPLINE ∇ Counts: # Process: CONC ∇ Test Name: FERR ∇ < > Type Serum ∇ R Use Serum Cal.

Point 1: # † -2.0 2.5
Point 2: # † -2.0 2.5
SERUM/PLASMA
Point 1: # ∇ † -2.0 2.5
Point 3: # † -2.0 2.5
Point 4: # † -2.0 2.5
Point 3: # ∇ † -2.0 2.5
Point 5: # † -2.0 2.5
Point 4: # ∇ † -2.0 2.5 R Reagent Blank
Point 6: Point 5: # † -2.0 2.5 R Calibration
∇
Point 7: Point 6:
Point 9 ∇
Data Check Parameters <Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
SERUM APPLICATION
APPLICATION
Parameters Misc
Decision Value-3: 0.0500 Limit Point 2: Limit Point 2: Test Name: FERR ∇ < > Type: Serum ∇
Limit Point 1: 11
Check Point-3: 27
# User defined. Decision Value-2: 2.0000 Decision Value-2: Decision Value 2:
† System Calibrator Cat. No.: ODR3021 Decision Value-3: 0.0500
* Values set for working in SI units µg/L, equivalent to ng/mL.
ф AU680

2010-07
HAPTOGLOBIN
OSR6165 4 x 16.5 mL R1
4 x 4.5 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of haptoglobin in human serum and plasma on Beckman Coulter analysers. For in vitro
Summary1
Haptoglobin is an α2-glycoprotein that binds haemoglobin irreversibly. Each haptoglobin monomer can bind up to 2 haemoglobin αβ dimers, the equivalent
of one intact haemoglobin molecule. The complexes are rapidly removed by the hepatic Kupffer cells, where the proteins are degraded and the amino
acids and iron reutilised. Haptoglobin is therefore central to the preservation of iron and prevents possible damage to the renal tubules by preventing
excretion of haemoglobin. Haptoglobin assays are used primarily to screen for or to follow the course of haemolytic disorders. Plasma levels are increased
in the presence of acute inflammatory processes, tissue necrosis or malignancy; protein losing syndromes (e.g. nephrotic syndrome or protein losing
enteropathies); and in corticosteroid therapy. Plasma levels are decreased in haemolytic disease and ineffective erythropoiesis, genetic deficiency; with
either endogenous or exogenous oestrogens, and in hepatocellular diseases.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, human haptoglobin reacts specifically with the anti-human haptoglobin antibodies to
yield insoluble aggregates. The absorbance of these aggregates is proportional to the haptoglobin concentration in the sample.
Goat anti-human haptoglobin antibodies Variable
Preservative
Harmful, contains sodium azide. R22 Harmful if swallowed.
Safety Phrases: S36, S60. Wear suitable protective clothing. This material and its container must be disposed of as hazardous waste.
Reagent Preparation
90 days.
Specimen
Serum and EDTA or heparinised plasma. Haemolysed samples should be avoided.
2
Stable in serum and plasma for 8 months when stored at 2…8°C and 3 months when stored at 15…25°C.
Test Procedure
Calibration
The calibrator haptoglobin values are traceable to IFCC (International Federation of Clinical Chemistry) standard CRM 470.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the haptoglobin concentration of each sample.
Adults 0.3 – 2.0 g/L (30 – 200 mg/dL)

2009-08
values.
Linearity
Precision
0.41 0.01 1.12 0.01 2.57
2.05 0.03 1.20 0.05 2.61
3.72 0.05 1.22 0.10 2.64
Sensitivity
The lowest detectable level represents the lowest measurable level of haptoglobin that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient serum samples were used to compare this Haptoglobin OSR6165 assay on the AU2700 against another commercially available haptoglobin
®
4
Limitations
BIBLIOGRAPHY
1. Johnson AM, Rohlfs EM, Silverman LM. Proteins. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia: WB Saunders Company,
1999;494-497.
470. J Lab Med 1996;20:145-152.

2009-08
HAPTOGLOBIN, AU400/AU640 Serum/Plasma Application HAPTOGLOBIN, AU600 Serum/Plasma Application

Test No # Test name HAPT ∇ Sample type Ser ∇ Page ½
Test Name: HAPT ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name HAPT ∇ Sample type Ser ∇ Page 2/2
Test Name: HAPT ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name HAPT ∇
Test Name: HAPT ∇ < > Type Serum ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
MB type factor
# User defined

2009-08
HAPTOGLOBIN, AU2700/AU5400 Serum/Plasma Application HAPTOGLOBIN, AU680/AU480 Serum/Plasma Application
Test Name: HAPT ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: HAPT ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: HAPT ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: HAPT ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: HAPT ∇ < > Type Serum ∇ Test Name: HAPT Type Serum Use Serum Cal.
∇ < > ∇ ο
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇

2009-08
HbA1c (Hemoglobin A1c)
OSR6192 HbA1c 2 x 19 mL R1
2 x 19 mL R2
Total Hemoglobin 2 x 37.5 mL R1
Intended Use
Immuno-inhibition test for the quantitative determination of HbA1c (Hemoglobin A1c), in human blood, on Beckman Coulter AU analysers. For in vitro
The absolute HbA1c and Total Hemoglobin (THb) values generated as part of the HbA1c assay are intended for use in the calculation of the HbA1c/Total
Hemoglobin ratio, and must not be used individually for diagnostic purposes.
Summary1
HbA1c is formed by the non-enzymatic glycation of free amino groups at the N-terminus of the β-chain of hemoglobin A0. The level of HbA1c is proportional
to the level of glucose in the blood. As the glucose remains bound to the red cell throughout its life cycle, measurement of HbA1c provides an indication of
the mean daily blood glucose concentration over the preceding two months. Measurement of HbA1c is, therefore, considered to be an important diagnostic
tool in the monitoring of dietary control and therapeutic regimes during the treatment of diabetes. Effective control of blood glucose levels is important in the
prevention of ketosis and hyperglycaemia, and may reduce the prevalence and severity of late diabetic complications such as retinopathy, neuropathy,
nephropathy and cardiac disease.
Test Principle
The concentrations of both HbA1c and Total Hemoglobin are determined. The HbA1c/Total Hemoglobin ratio is expressed as percentage HbA1c
(%HbA1c). The assay for percent HbA1c, involves the use of four reagents: Total Hemoglobin R1, HbA1c R1 antibody reagent, HbA1c R2 agglutinator
reagent, and Hemoglobin Denaturant (sold separately as Cat No.OSR0004).
In a pre-treatment step the whole blood is mixed with Hemoglobin Denaturant (OSR0004) in a 1 : 41 dilution and incubated for a minimum of five minutes
at room temperature. The red blood cells are lysed and the hemoglobin chain is hydrolysed by the protease present in the reagent.
Total Hemoglobin is measured via the conversion of all hemoglobin derivatives into alkaline haematin in the alkaline solution of a non-ionic detergent.
Addition of the pre-treated blood sample to the Total Hemoglobin reagent results in a green solution, which is measured at 600nm.
HbA1c is measured in a latex agglutination inhibition assay. An agglutinator, consisting of a synthetic polymer containing multiple copies of the
immunoreactive portion of HbA1c, causes agglutination of latex coated with HbA1c specific mouse monoclonal antibodies. In the absence of HbA1c in the
sample, the antibody-coated microparticles in the HbA1c R1 and the agglutinator in the HbA1c R2 will agglutinate. Agglutination leads to an increase in the
absorbance of the suspension. The presence of HbA1c in the sample results in a decrease in the rate of agglutination of the HbA1c R1 and the
agglutinator in the HbA1c reagent R2. The increase in absorbance is therefore inversely proportional to the concentration of HbA1c in the sample.
The increase in absorbance due to agglutination is measured at 700nm.
Contents, Reagent Composition
HbA1c
HbA1c R1 HbA1c R2
HbA1c Antibody (mouse) coupled particles HbA1c Hapten
Bovine Serum Albumin Bovine Serum Albumin
Buffer pH 8.1 Buffer pH 2.0
Surfactant: 0.6% Non-ionic detergent Surfactant
Preservative 0.1% Proclin Preservative 0.1% Proclin
Total Hemoglobin
Total Hemoglobin R1
Sodium hydroxide 0.4%; pH 13
Surfactant: 0.7% Non-ionic detergent

HbA1c R1 and R2:
R43: May cause sensitisation by skin contact. Product contains a mixture of 5-chloro-2-methyl-4-isothiazolin -3-one [EC No 247-500-7] and 2-methyl-2 H –
isothiazol-3-one [EC No 220-239-6] (3:1).
Reagent Preparation
Total Hemoglobin R1 and HbA1c R2 are ready for use, and can be placed directly on board the instrument. HbA1c R1 should be mixed by inversion
5 – 10 times before placing on board the instrument and at weekly intervals thereafter.

30 days.
Specimen
K2-EDTA or NH4-heparinised whole blood. Before use, pretreat samples and controls by dilution of whole blood with Hemoglobin denaturant 1 : 41 (for
example 25 µLof sample or control plus 1000 µL of Hemoglobin Denaturant Cat No. OSR0004). Due to the viscosity of whole blood, a volume of less than
25 µL is not recommended. Mix thoroughly, avoid foaming and incubate for 5 minutes at room temperature before use. Calibrators do not require pre-
treatment. Please note that only Hemoglobin Denaturant OSR0004 can be used with this method.
Samples (non-pretreated) are stable up to 1 week when stored at 25°C, 2 weeks when stored at 2…8°C and up to 6 months when frozen at ≤ -70°C.
Hemolysed (pretreated) samples are stable up to 8 hours when stored at room temperature, up to 48 hours when stored at 2…8°C, if stored in a sealed
container.
Note: All pretreated samples should be mixed thoroughly immediately prior to assay.
2009-12
Test Procedure
Total Hemoglobin and HbA1c tests must be performed on each pre-treated sample and control. Refer to the appropriate User’s Guide and the
accompanying Instrument Setting Sheets for analyser-specific assay instructions.
Calibration
HbA1c Calibrator (ODR3032).
Calibrator 1 is used for calibration of the Total Hemoglobin assay only.
Calibrators 1 to 6 are used for calibration of the HbA1c assay.
Note: Calibrators do not require pre-treatment with Hemoglobin Denaturant.
For calibration procedure, please refer to HbA1c Calibrator ODR3032 Instructions for Use.
Following calibration, the resulting curve should be visually reviewed, on the Beckman Coulter analyser, for acceptability. Quality control procedures should
be undertaken immediately following calibration in accordance with good laboratory practice.
Traceability2,3
2
The calibrator HbA1c values are traceable to the IFCC HbA1c reference method via IFCC HbA1c reference material. Total haemoglobin values assigned
to the THb calibrator are traceable to the IRMM haemoglobin cyanide standard (BCR-522).The relationship between results from the NGSP network
(DCCT aligned) and the IFCC network has been evaluated, and a Master Equation has been developed for interconversion of results from IFCC to
DCCT/NGSP units.
MASTER EQUATION
DCCT/NGSP = (0.915 x IFCC) + 2.15
Definition of the relationship between the two networks links IFCC traceable results to clinically meaningful HbA1c results from the DCCT and the United
Kingdom Prospective Diabetes Study (UKPDS). The Master Equation also provides these DCCT results with traceability to a higher order reference
method.
% HbA1c results generated during this assay are automatically recalculated to DCCT aligned units by the instrument, using the IFCC approved Master
Equation (NGSP = (0.915 x IFCC) + 2.15). Results are therefore expressed in DCCT aligned units as recommended by the IFCC.
Quality Control
HbA1c Control material ODC0022 or other control materials with values determined by this Beckman Coulter system may be used.
Additional pre-treatment/QC recommendations for manual method:
1. Strictly follow the pre-treatment directions. Incubate controls with the denaturant at room temperature for exactly 5 minutes before placing onto the
system. It is imperative that this step is controlled as accurately as possible. Once the effectiveness of the denaturant has been demonstrated in this
manner for the minimum defined pre-treatment time, patient samples can be processed with the minimum pre-treatment time of 5 minutes up to the
maximum defined stability of the pre-treated sample of 8 hours at room temperature.
2. Once a control has been assayed – discard it – never rerun pre-treated HbA1c controls: instead repeat the pre-treatment step. This will ensure that any
issues with the pre-treatment process or denaturant solution are identified and the performance of the HbA1c assay (OSR6192) is effectively
monitored.
3. If at any time HbA1c controls are out of range, discontinue the use of the current open bottle of OSR0004 Hemoglobin Denaturant.
Reference Interval2
Adults: 4.0 – 6.2% (DCCT)
2.0 – 4.4% HbA1c (IFCC units; recalculated using Master Equation)
should always be assessed in conjunction with the patient’s medical history, clinical examinations and other findings.
values.
Analytical Range
Total Hemoglobin
The analytical range for Total Hemoglobin is 7-23 g/dL (4.4-14.3 mmol/L). When the result for Total Hemoglobin is outside the analytical range the
calculated % HbA1c should not be reported. Settling of the red cells before the aliquot is taken for pre-treatment may cause an elevated Total Hemoglobin
result. Samples exceeding the upper limit of Total Hemoglobin may be mixed well and the analyses repeated on a freshly denatured sample.
HbA1c
The analytical range of this assay extends from the concentration of calibrator 1 to calibrator 6. Samples exceeding the upper limit of the analytical range
for HbA1c should not be diluted, but instead should be reported as “% HbA1c > 14.5%”.
% HbA1c
The reportable range for the calculated % HbA1c is approximately 3.2% to 14.5% for a sample with a total hemoglobin of 14.5 g/dL (9 mmol/L).
Note: a calculated result may be outside the reportable range based on either the Total Hemoglobin or the HbA1c result being outside of their respective
analytical ranges.
Precision
The following data was obtained on an AU640 analyser using 3 whole blood pools analysed over 20 days.
AU640:
n=80 Within run Total
Mean % SD CV% SD CV%
4.98 0.04 0.74 0.07 1.32
6.56 0.08 1.25 0.12 1.78
10.01 0.08 0.77 0.12 1.18

2009-12
Sensitivity
The lowest detectable level on an AU640 analyser was calculated as:
HbA1c 0.02mmol/L (0.03g/dL); THb 0.05mmol/L (0.08 g/dL).
The lowest detectable level represents the lowest measurable level of HbA1c that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient samples were used to compare this HbA1c OSR6192 assay on the AU640 against another commercially available immunoturbidimetric HbA1c
y = 1.092x - 0.147 r = 0.995 n = 97 Sample range = 5.1 – 11.5 %HbA1c
Icterus: Interference less than 10% up to 513µmol/L (30 mg/dL) Bilirubin
®
Triglyceride: Interference less than 10% up to 18mmol/L (1600 mg/dL) Triglyceride
Rheumatoid factor (RF) up to 2000 IU/mL, acetylsalicylic acid (60 mg/dL), sodium cyanate (50 mg/dL) and urea (83 mmol/L) do not interfere with this
assay.
4
Limitations5
Shortened red cell survival time will reduce the exposure of red cells to glucose, with a resultant decrease in % HbA1c values. Percentage HbA1c results
are therefore not reliable where red cell survival time is reduced. Causes of reduced red cell survival time include hemolytic anaemia, or other hemolytic
diseases, significant blood loss and pregnancy.
Samples containing hemoglobin variants S and C may produce up to a 40% elevation of the expected HbA1c value in this assay. Samples containing
> 10% of haemoglobin F may yield a lower than expected result with this test. HbA1c results obtained by this test method for blood samples containing Hb
variants S, C and F (>10%) should not be compared to published normal or abnormal values. A sample containing hemoglobin E was shown not to
interfere with this test.
The labile fraction of glycated hemoglobin (Schiff base attachment of glucose to HbA or HbA1c) does not affect the assay result due to the specificity of the
antibody for the stable ketoamine.
As with any chemical reaction, users should be aware of the possible effect on results due to unknown interferences from medication or endogenous
substances. For diagnostic purposes, the HbA1c results should always be assessed in conjunction with other available information e.g. patient’s medical
history, clinical status and results of other tests.

† HbA1c Calibrator Cat. No.: ODR3032.
* Values set for working in mmol/L. To work in g/dL, multiply by 1.6125.
** Concentration of 0.1 mmol/L and highest calibrators.
¥ For determination of %HbA1c the above tests (THb and HbA1c) are used. A third test, % HbA1c, must also be entered in the general tests and the
calculated tests of the test section menu (no settings required). Set this test as CALCULATED TEST in the INTER TEST menu. Enter the formula
(A/B)*a + b, where A=HbA1c, B=THb, a=91.5, b=02.15.
ж Set this test as COMMON TEST PARAMETER TEST NAME in CALCULATED TEST. Enter the formula (A/B)*a+b, where A = HbA1c , B = THb,
a = 91.5 and b = 2.15 in Parameters Specific Test Parameters CALCULATED TEST
BIBLIOGRAPHY
1. Niederau CM, Reinauer H Glycohemoglobins In: Thomas L, ed. Clinical Laboratory Diagnostics. Use and assessment of clinical laboratory results.
Frankfurt/Main: TH-Books Verlagsgesellschaft mbH, 1998: 142-148
2. Jeppsson J-O, Kobold U, Barr J, Finke A, Hoezel W, Hoshino T, Miedema K, Mosca A, Mauri P, Paroni R, Thienpont L, Umemoto M, Weykamp C. Approved
IFCC reference method for the Measurement of HbA1c in Human Blood. Clin Chem Lab Med 2002; 40(1): 78-89.
3. Hoelzel W, Weykamp C, Jeppsson J-O, Miedema, Barr JR, Goodall I, Hoshino T, John, WG, Kobold U, Little R, Mosca A, Mauri P, Paroni R, Susanto F, Takei I,
Theinpoint L, Umemoto M, Wiedmeyer H-M. IFCC Reference System for Measurement of Hemoglobin A1c in Human Blood and the National Standardization
Schemes in the United States, Japan, and Sweden: A Method-Comparison Study. Clin Chem 2004; 50(1): 166-174

2009-12
TOTAL Hb, AU400/AU640 Whole Blood Application HbA1c, AU400/AU640 Whole Blood Application
System Reagent: OSR6192 Reagent ID: 092 System Reagent : OSR6192 Reagent ID : 035
Test Name: THb¥ ∇ < > Type: Serum ∇ Operation: Yes ∇ Test Name: HbA1c¥ ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 11 µL Dilution 0 µL Pre-Dilution Rate: 1 Sample: Volume 3.5 µL Dilution 0 µL Pre-Dilution Rate: 1
Reagents: R1 Volume 150 µL Dilution 0 µL Min OD Max OD Reagents: R1 Volume 76 µL Dilution 0 µL Min OD Max OD
R2 Volume 0 µL Dilution 0 µL L H R2 Volume 76 µL Dilution 0 µL L -2.0 H 2.5
Wavelength: Pri. 600 ∇ Sec. 700 ∇ First L -2.0 First H 2.5 Wavelength: Pri. 700 ∇ Sec. None ∇ First L -2.0 First H 2.5
Method: END ∇ Last L -2.0 Last H 2.5 Method: FIXED ∇ Last L -2.0 Last H 2.5
Measuring Point 1: First 0 Last 27 L 4.4* H 14.3* Measuring Point 1: First 13 Last 22 L ** H **
Measuring Point 2: First Last Correlation Factor: Measuring Point 2: First Last Correlation Factor:

Test Name: THb¥ ∇ < > Type: Serum ∇ Test Name: HbA1c¥ ∇ < > Type: Serum ∇

R 1. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # # R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # # R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 6. # ∇ # # # # # #
Panic Value: # # Unit: mmol/L* Decimal places: # Panic Value: # # Unit: mmol/L* Decimal places: #

Test Name: THb¥ ∇ < > Type Serum ∇ Test Name: HbA1c¥ ∇ < > Type Serum ∇
WHOLE BLOOD APPLICATION

Calibration Type: AB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Calibration Type: 6AB ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1: # † -999999 999999 Point 1: # † -2.0 2.5
Point 2: Point 2: # † -2.0 2.5
Point 3: Point 3: # † -2.0 2.5
Point 4: Point 4: # † -2.0 2.5
Point 5: Point 5: # † -2.0 2.5
Point 6: Point 6: # † -2.0 2.5
Point 7: Point 7:
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-Point Cal. Point: R with CONC-0 Slope Check: - ∇ Advanced Calibration: # ∇
MB Type Factor: Calibration Stability Period: 14 MB Type Factor: Calibration Stability Period: 14

† HbA1c Calibrator ODR3032 † HbA1c Calibrator ODR3032
* Values set for working in mmol/L . To work in g/dL multiply by 1.6125. * Values set for working in mmol/L. To work in g/dL multiply by 1.6125.
¥ For determination of %HbA1c the above tests (THb and HbA1c) are used. A third test %HbA1c must also be entered in the general ** Concentration of 0.1 mmol/L and highest calibrator.
tests and the calculated tests of the test section menu (no settings required). Set this test as CALCULATED TEST in the INTER ¥ For determination of %HbA1c the above tests (THb and HbA1c) are used. A third test %HbA1c must also be entered in the general
TEST menu. Enter the formula (A/B)*a+b, , where A=HbA1c , B=THb and a=91.5 and b=2.15. tests and the calculated tests of the test section menu (no settings required). Set this test as CALCULATED TEST in the INTER
TEST menu. Enter the formula (A/B)*a+b, where A=HbA1c , B=THb and a=91.5 and b=2.15.
2010-06
TOTAL Hb, AU600 Whole Blood Application HbA1c, AU600 Whole Blood Application
Specific test parameters Specific test parameters
Test No # Test name THb¥ ∇ Sample type Ser ∇ Page 1/2 Test No # Test name HbA1c¥ ∇ Sample type Ser ∇ Page 1/2
Sample vol. 11 Dil. vol 0 µL Min. OD Max. OD Sample vol. 3.5 Dil. vol 0 µL Min. OD Max. OD
Reagent 1 vol 150 Dil. vol 0 µL L H Reagent 1 vol 76 Dil. vol 0 µL L -2.0 H 2.5
Reagent 2 vol 0 Dil. vol 0 µL Reagent OD limit Reagent 2 vol 76 Dil. vol 0 µL Reagent OD limit
Fst. L -2.0 Fst. H 2.5 Fst. L -2.0 Fst. H 2.5
Wave Main 600 Sub 700 ∇ Lst. L -2.0 Lst. H 2.5 Wave Main 700 Sub NONE ∇ Lst. L -2.0 Lst. H 2.5
Method END ∇ Dynamic range Method FIXED ∇ Dynamic range
Reaction + ∇ L 4.4* H 14.3* Reaction + ∇ L ** H **
Point 1 Fst 0 Lst 27 Correlation factor A 1 Point 1 Fst 13 Lst 22 Correlation factor A 1
Point 2 Fst Lst B 0 Point 2 Fst Lst B 0
Sample Pre-dil. Rate ¤ Sample Pre-dil. Rate ¤
Linearity Fst % Sec % Linearity Fst % Sec %

No lag time ∇ On-board stability period 30 No lag time ∇ On-board stability period 30
Select using Space key, or select from list displayed by Guide key Select using Space key, or select from list displayed by Guide key
Test No # Test name THb¥ ∇ Sample type Ser ∇ Page 2/2 Test No # Test name HbA1c¥ ∇ Sample type Ser ∇ Page 2/2
Level L Level H Level L Level H
Value/flag # ∇ # # Value/flag # ∇ # #
Normal range Normal range
Sex Age L Age H L H Sex Age L Age H L H
1 # ∇ # Y # M # Y # M→ # # 1 # ∇ # Y # M # Y # M→ # #
2 # ∇ # Y # M # Y # M→ # # 2 # ∇ # Y # M # Y # M→ # #
3 # ∇ # Y # M # Y # M→ # # 3 # ∇ # Y # M # Y # M→ # #
4 # ∇ # Y # M # Y # M→ # # 4 # ∇ # Y # M # Y # M→ # #
5 # ∇ # Y # M # Y # M→ # # 5 # ∇ # Y # M # Y # M→ # #
6 # ∇ # Y # M # Y # M→ # # 6 # ∇ # Y # M # Y # M→ # #
7 Non select # # 7 Non select # #
8 Out of range # # 8 Out of range # #
L H L H
Panic value # # Panic value # #
Calibration specific Calibration specific
Test No # Test name THb¥ ∇ Test No # Test name HbA1c¥ ∇

Cal type 8 AB ∇ Count # Cal type 14 6AB ∇ Count #
Formula 1 Y=AX+B ∇ Process Conc ∇ Formula 3 POLYGONAL ∇ Process Conc ∇
Selection calibrator Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H Cal No OD Conc Factor/OD-L Factor/OD-H
Point 1 # ∇ † -999999 999999 Point 1 # ∇ † -2.0 2.5
Point 2 Point 2 # ∇ † -2.0 2.5
Point 3 Point 3 # ∇ † -2.0 2.5
Point 4 Point 4 # ∇ † -2.0 2.5
Point 5 Point 5 # ∇ † -2.0 2.5
Point 6 ∇ Point 6 # ∇ † -2.0 2.5
1-point cal. point 1-point cal. point
MB type factor MB type factor
Calibrator stability period 14 Calibrator stability period 14
# User defined ¤ Analyser default value # User defined ¤ Analyser default value
† HbA1c Calibrator ODR3032. † HbA1c Calibrator ODR3032
* Values set for working in mmol/L. To work in g/dL multiply by 1.6125. * Values set for working in mmol/L. To work in g/dL multiply by 1.6125.
¥ For determination of %HbA1c the above tests (THb and HbA1c) are used. A third test %HbA1c must also be entered in ** Concentration of 0.1 mmol/L and highest calibrator.
the general tests and the calculated tests of the test section menu (no settings required). Set this test as CALCULATED ¥ For determination of %HbA1c the above tests (THb and HbA1c) are used. A third test %HbA1c must also be entered in the general
TEST in the INTER TEST menu. Enter the formula (A/B)*a+b, where A=HbA1c , B=THb and a=91.5 and b=2.15 tests and the calculated tests of the test section menu (no settings required). Set this test as CALCULATED TEST in the
INTER TEST menu. Enter the formula (A/B)*a+b, where A=HbA1c , B=THb and a=91.5 and b=2.15.

2010-06
TOTAL Hb, AU2700/AU5400 Whole Blood Application HbA1C, AU2700/AU5400 Whole Blood Application
System Reagent: OSR6192 Reagent ID: 092 System Reagent : OSR6192 Reagent ID : 035
Test Name: THb¥ ∇ < > Type: Serum ∇ Operation: Yes ∇ Test Name: HbA1c¥ ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample: Volume 9.5 µL Dilution 0 µL Pre-Dilution Rate: 1 Sample: Volume 3 µL Dilution 0 µL Pre-Dilution Rate: 1
Reagents: R1 Volume 130 µL Dilution 0 µL Min OD Max OD Reagents: R1 Volume 65 µL Dilution 0 µL Min OD Max OD
R2 Volume 0 µL Dilution 0 µL L H R2 Volume 65 µL Dilution 0 µL L -0.1 H 2.5
Wavelength: Pri. 600 ∇ Sec. 700 ∇ First L -2.0 First H 2.5 Wavelength: Pri. 700 ∇ Sec. None ∇ First L -2.0 First H 2.5
Method: END ∇ Last L -2.0 Last H 2.5 Method: FIXED ∇ Last L -2.0 Last H 2.5
Measuring Point 1: First 0 Last 27 L 4.4* H 14.3* Measuring Point 1: First 13 Last 22 L ** H **
Measuring Point 2: First Last Correlation Factor: Measuring Point 2: First Last Correlation Factor:

Test Name: THb¥ ∇ < > Type: Serum ∇ Test Name: HbA1c¥ ∇ < > Type: Serum ∇

R 1. R 1. # ∇ # # # # # #
# ∇ # # # # # #
R 2. R 2. # ∇ # # # # # #
# ∇ # # # # # #
R 3. R 3. # ∇ # # # # # #
# ∇ # # # # # #
R 4. R 4. # ∇ # # # # # #
# ∇ # # # # # #
R 5. R 5. # ∇ # # # # # #
# ∇ # # # # # #
R 6. R 6. # ∇ # # # # # #
# ∇ # # # # # #
Panic Value: # # Unit: mmol/L* Decimal places: # Panic Value: # # Unit: mmol/L* Decimal places: #
General ISE
General ISE
Test Name: HbA1c¥ ∇ < > Type Serum ∇

Test Name: THb¥ ∇ < > Type Serum ∇
Point 1: # † -2.0 2.5
Point 1: # † -999999 999999
Point 2: # † -2.0 2.5
Point 2:
Point 3: # † -2.0 2.5
Point 3:
Point 4: # † -2.0 2.5
Point 4:
Point 5: # † -2.0 2.5
Point 5:
Point 6: # † -2.0 2.5
Point 6:
Point 7:
Point 7:
1-Point Cal. Point: R with CONC-0 Slope Check: - ∇ Advanced Calibration: # ∇
# User defined
# User defined † HbA1c Calibrator ODR3032.
† HbA1c Calibrator ODR3032. * Values set for working in mmol/L. To work in g/dL multiply by 1.6125.
* Values set for working in mmol/L. To work in g/dL multiply by 1.6125. ** Concentration of 0.1 mmol/L and highest calibrator.
¥ For determination of %HbA1c the above tests (THb and HbA1c) are used. A third test %HbA1c must also be entered in ¥ For determination of %HbA1c the above tests (THb and HbA1c) are used. A third test %HbA1c must also be entered in the general
the general tests and the calculated tests of the test section menu (no settings required). Set this test as CALCULATED TEST in tests and the calculated tests of the test section menu (no settings required). Set this test as CALCULATED TEST in the INTER
the INTER TEST menu. Enter the formula (A/B)*a+b, where A=HbA1c , B=THb and a=91.5 and b=2.15. TEST menu. Enter the formula (A/B)*a+b, where A=HbA1c , B=THb and a=91.5 and b=2.15.

2010-06
TOTAL Hb, AU680/AU480 Whole Blood Application
Test Name: THbж ∇ < > Type: Serum ∇ Operation Yes ∇

Method END ∇
WHOLE
WHOLE
Test Name: THbж ∇ < > Type: Serum ∇

BLOOD
R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # #
BLOOD
R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # #
General ISE
Test Name: THbж ∇ < > Type Serum ∇ R Use Serum Cal.
APPLICATION
Y=AX+B
APPLICATION
Calibration Type: AB ∇ Formula: ∇ Counts: # ∇
Point 1: # ∇ † -999999 999999
Point 3: ∇
Point 4: ∇ R Reagent Blank
Point 5: ∇ R Calibration
Point 6: ∇
Point 9 ∇
Master Curve> OD Range # User defined
Calibrator OD Conc Low High Stability † HbA1c Calibrator ODR3032.
Point-1 ∇ Reagent Blank 14 Day 0 Hour * Values set for working in mmol/L. To work in g/dL multiply by 1.6125.
Point-2 ∇ Calibration 14 Day 0 Hour ж Set this test in Common Test Parameter test name as CALCULATED TEST. Enter the formula (A/B)*a+b, where A=HbA1c ,
MB Type Factor: 1-Point Calibration Point R with Conc-0 B=THb and a=91.5 and b=2.15 in Parameters Specific Test Parameters CALCULATED TEST
∇
ф AU680

2010-06
HbA1C, AU680/AU480 Whole Blood Application
Test Name: HbA1cж ∇ < > Type: Serum ∇ Operation Yes ∇

Dynamic Range Low ** High **
Method FIXED ∇

Test Name: HbA1cж ∇ < > Type: Serum ∇

R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # #
General ISE
Test Name: HbA1cж ∇ < > Type Serum ∇ R Use Serum Cal.

Calibrator OD Conc Low High Slope Check - ∇
Point 1: # ∇ † -2.0 2.5
Point 3: # ∇ † -2.0 2.5
Point 5: # ∇ † -2.0 2.5 R Calibration
Point 6: # ∇ † -2.0 2.5
Point 9 ∇
Calibrator OD Conc Low High Stability † HbA1c Calibrator ODR3032.
Point-1 ∇ Reagent Blank 14 Day 0 Hour * Values set for working in mmol/L. To work in g/dL multiply by 1.6125.
Point-2 ∇ Calibration 14 Day 0 Hour **Concentration of 0.1 mmol/L and highest calibrator.
MB Type Factor: 1-Point Calibration Point R with Conc-0 ж Set this test in Common Test Parameter test name as CALCULATED TEST. Enter the formula (A/B)*a+b, where A=HbA1c ,
∇
B=THb and a=91.5 and b=2.15 in Parameters Specific Test Parameters CALCULATED TEST
ф AU680

2010-06
HbA1c APT (Hemoglobin A1c, Whole Blood Application)
OSR61177 HbA1c APT 2 x 19 mL R1
2 x 19 mL R2
Total Hemoglobin APT 2 x 38 mL R1
HbA1c APT Denaturant 2 x 55 mL R1
Intended Use
Immuno-inhibition test for the quantitative determination of HbA1c (Hemoglobin A1c) in human whole blood on Beckman Coulter AU680 with whole blood
automated pre-treatment (APT) capability only.
For in vitro diagnostic use only.
The absolute HbA1c and Total Hemoglobin (THb) values generated as part of the HbA1c assay are intended for use in the calculation of the HbA1c/Total
Hemoglobin ratio, and must not be used individually for diagnostic purposes.
Summary1
HbA1c is formed by the non-enzymatic glycation of free amino groups at the N-terminus of the β-chain of hemoglobin A0. The level of HbA1c is proportional
to the level of glucose in the blood. As the glucose remains bound to hemoglobin in the red cell throughout the life cycle of the cell, measurement of HbA1c
provides an indication of the mean daily blood glucose concentration over the preceding two months. Measurement of HbA1c is, therefore, considered to
be an important diagnostic tool in the monitoring of dietary control and therapeutic regimes during the treatment of diabetes. Effective control of blood
glucose levels is important in the prevention of ketosis and hyperglycaemia, and may reduce the prevalence and severity of late diabetic complications
such as retinopathy, neuropathy, nephropathy and cardiac disease.
Test Principle
The concentrations of both HbA1c and Total Hemoglobin are determined. The HbA1c/Total Hemoglobin ratio is expressed as percentage HbA1c
(%HbA1c). The assay for percent HbA1c involves the use of four reagents: Total Hemoglobin APT R1, HbA1c APT R1 antibody reagent, HbA1c APT R2
agglutinator reagent, and HbA1c APT Denaturant R1.
Whole blood is pretreated automatically on board the AU680. The red blood cells are lysed and the hemoglobin chain is hydrolysed by the protease
present in the reagent.
Total Hemoglobin is measured via the conversion of all hemoglobin derivatives into alkaline haematin in the alkaline solution of a non-ionic detergent.
Addition of the pre-treated blood sample to the Total Hemoglobin reagent results in a green solution, which is measured at 600 nm.
HbA1c is measured in a latex agglutination inhibition assay. An agglutinator, consisting of a synthetic polymer containing multiple copies of the
immunoreactive portion of HbA1c, causes agglutination of latex coated with HbA1c specific mouse monoclonal antibodies. In the absence of HbA1c in the
sample, the antibody-coated microparticles in the HbA1c APT R1 and the agglutinator in the HbA1c APT R2 will agglutinate. Agglutination leads to an
increase in the absorbance of the suspension. The presence of HbA1c in the sample results in a decrease in the rate of agglutination of the HbA1c APT
R1 and the agglutinator in the HbA1c APT R2. The increase in absorbance is therefore inversely proportional to the concentration of HbA1c in the sample.
The increase in absorbance due to agglutination is measured at 700 nm.
Contents, Reagent Composition
HbA1c
HbA1c APT R1 HbA1c APT R2
HbA1c Antibody (mouse) coupled particles HbA1c Hapten
Bovine Serum Albumin Bovine Serum Albumin
Buffer pH 8.1 Buffer pH 2.0
Surfactant 0.6% Non-ionic detergent Surfactant
Preservative 0.1% Proclin Preservative 0.1% Proclin
Total Hemoglobin
Total Hemoglobin APT R1
Sodium hydroxide 0.4%; pH 13
Surfactant 0.7% Non-ionic detergent
HbA1c APT Denaturant
HbA1c APT Denaturant R1
Porcine Pepsin
Buffer pH 2.4
Preservative

Hazard Warnings and Risk Phrases
HbA1c APT R1 and HbA1c APT R2
R43: May cause sensitisation by skin contact. Product contains a mixture of 5-chloro-2-methyl-4-isothiazolin -3-one [EC No 247-500-7] and
2-methyl-2 H –isothiazol-3-one [EC No 220-239-6] (3:1).
Reagent Preparation
Total Hemoglobin APT R1 and HbA1c APT R2 are ready for use, and can be placed directly on board the instrument.
HbA1c APT R1 and HbA1c APT Denaturant R1 should be mixed by inversion 5 – 10 times before placing on board the instrument and at weekly intervals
thereafter.
Do not mix reagents between kit lots. Do not top up reagent vials.

The reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C. Once open, Total Hemoglobin APT R1, HbA1c APT R1, HbA1c
APT R2 and HbA1c APT Denaturant reagents stored on board the instrument are stable for 30 days.

2009-08
Specimen
K2-EDTA or NH4-heparinised whole blood. Calibrators do not require pre-treatment. Please note that only Beckman Coulter HbA1c APT Denaturant
can be used with this method.
Whole blood samples should be mixed by inverting 5 – 10 times before placing on board the instrument.
Samples with any hematocrit disorders may result in instrument flags. They have to be mixed thoroughly by inversion and analyzed subsequently by
making the test requisition and placing them in a rack on the rack feeder unit close to the sampling lane. However, the rack should not be placed closer
than 3 positions from the head of the queue. Do not remove the racks which are about to be transferred onto the conveyor belt for processing at the
sampling position. If there are more than 3 racks in the queue on the rack feeder, then manually move those racks backwards to allow the HbA1c test
sample to be placed close to the front for priority sampling.
Sedimentation of erythrocytes can lead to a "Tx" flagged result. A valid final HbA1c value can be obtained by mixing the sample thoroughly and analysing
immediately as described above for samples with haematocrit disorders.
Please confirm that the dynamic ranges are set appropriately according to the Setting Sheets to ensure correct flagging.
Samples (non-pretreated) are stable up to 1 week when stored at 25°C, 2 weeks when stored at 2…8°C and up to 6 months when frozen at ≤ -70°C.
Reconstituted control material should not be pre-treated with HbA1c APT Denaturant, but should be placed on-board the instrument for on-board
denaturation.
Test Procedure
Total Hemoglobin and HbA1c tests must be performed on each pre-treated sample and control. Refer to the appropriate User’s Guide and the
accompanying Instrument Setting Sheets for analyser-specific assay instructions.
Calibration
HbA1c Calibrator Cat. No.: ODR3032.
Calibrator 1 is used for calibration of the Total Hemoglobin APT assay only.
Calibrators 1 to 6 are used for calibration of the HbA1c APT assay.
Note: Calibrators do not require pre-treatment with HbA1c APT Denaturant.
For calibration procedure, please refer to HbA1c Calibrator ODR3032 Instructions for Use.
Following calibration, the resulting curve should be visually reviewed, on the Beckman Coulter analyser for acceptability. Quality control procedures should
be undertaken immediately following calibration in accordance with good laboratory practice.
Traceability2,3
2
The calibrator HbA1c values are traceable to the IFCC HbA1c reference method via IFCC HbA1c reference material. Total hemoglobin values assigned to
the THb calibrator are traceable to the IRMM hemoglobin cyanide standard (BCR-522). The relationship between results from the NGSP network
(DCCT aligned) and the IFCC network has been evaluated, and a Master Equation has been developed for interconversion of results from IFCC to
DCCT/NGSP units.
MASTER EQUATION
DCCT/NGSP = (0.915 x IFCC) + 2.15
Definition of the relationship between the two networks links IFCC traceable results to clinically meaningful HbA1c results from the DCCT and the United
Kingdom Prospective Diabetes Study (UKPDS). The Master Equation also provides these DCCT results with traceability to a higher order reference
method.
%HbA1c results generated during this assay are automatically recalculated to DCCT aligned units by the instrument, using the IFCC approved Master
Equation (NGSP = (0.915 x IFCC) + 2.15). Results are therefore expressed in DCCT aligned units as recommended by the IFCC.
Quality Control
HbA1c Control material ODC0022 or other control materials with values determined by this Beckman Coulter system may be used.
Each laboratory should establish its own control frequency, however good laboratory practice suggests that controls be tested each day patient samples
Ensure that controls are run at every calibration event.
Reference Interval2
Adults: 4.0 – 6.2% (DCCT)
2.0 – 4.4 %HbA1c (IFCC units; recalculated using Master Equation)
should always be assessed in conjunction with the patient’s medical history, clinical examinations and other findings.
values.
Analytical Range
Total Hemoglobin
The analytical range for Total Hemoglobin is 4.4 – 14.3 mmol/L (7 – 23 g/dL).
HbA1c
DCCT
The analytical range of this assay extends from 3.2 %HbA1c to the concentration of Calibrator 6 which approximately corresponds to 14.5 %HbA1c at a
total hemoglobin level of 9 mmol/L (14.5 g/dL). Samples exceeding the upper limit of linearity should not be diluted, but instead should be reported as
>14.5%.
Precision
The following data was obtained on an AU680 analyser using 3 whole blood pools analysed over 20 days.
AU680:
Mean % SD CV% SD CV%
5.20 0.02 0.43 0.05 0.95
7.34 0.05 0.73 0.07 0.92
11.17 0.06 0.56 0.10 0.88
2009-08
Sensitivity
The lowest detectable level on an AU680 analyser was calculated as:
HbA1c 0.01 mmol/L (0.02 g/dL)
THb 0.05 mmol/L (0.08 g/dL)
The lowest detectable level represents the lowest measurable level of HbA1c that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient samples were used to compare this HbA1c APT OSR61177 assay on the AU680 against the HbA1c OSR6192 on the AU640 assay. Results of
y = 1.026x + 0.437 r = 0.966 n = 105 Sample range = 4.80 – 13.45 %HbA1c
Patient samples were used to compare this HbA1c APT OSR61177 assay on the AU680 against an HPLC method. Results of linear regression analysis
were as follows:
y = 1.039x + 0.512 r = 0.960 n = 127 Sample range = 4.60 – 12.90 %HbA1c
Icterus: Interference less than 10% up to 513 µmol/L (30 mg/dL) Bilirubin
®
Triglyceride: Interference less than 10% up to 18 mmol/L (1600 mg/dL) Triglyceride
Rheumatoid factor (RF) up to 2000 IU/mL, acetylsalicylic acid (60 mg/dL), sodium cyanate (50 mg/dL) and urea (83 mmol/L) do not interfere with this
assay.
4
Limitations5
Shortened red cell survival time will reduce the exposure of red cells to glucose, with a resultant decrease in %HbA1c values. Percentage HbA1c results
are therefore not reliable where red cell survival time is reduced. Causes of reduced red cell survival time include hemolytic anaemia, or other hemolytic
diseases, significant blood loss and pregnancy.
Samples containing hemoglobin variants S and C may produce up to a 40% elevation of the expected HbA1c value in this assay. Samples containing
> 10% of haemoglobin F may yield a lower than expected result with this test. HbA1c results obtained by this test method for blood samples containing
Hb variants S, C and F (> 10%) should not be compared to published normal or abnormal values. A sample containing hemoglobin E was shown not to
interfere with this test.
The labile fraction of glycated hemoglobin (Schiff base attachment of glucose to HbA or HbA1c) does not affect the assay result due to the specificity of the
antibody for the stable ketoamine.
As with any chemical reaction, users should be aware of the possible effect on results due to unknown interferences from medication or endogenous
substances. For diagnostic purposes, the HbA1c results should always be assessed in conjunction with other available information e.g. patient’s medical
history, clinical status and results of other tests.

# User defined
† HbA1c Calibrator Cat. No.: ODR3032
** Concentration of 0.1 mmol/L and highest calibrator.
§ For determination of %HbA1c the above tests (THb and HbA1c) are used. %HbA1c is calculated automatically by the analyser using the following
formula : (A/B)* a + b, where A = HbA1c, B = THb, a = 91.5, b = 2.15.
BIBLIOGRAPHY
1. Niederau CM, Reinauer H Glycohemoglobins In: Thomas L, ed. Clinical Laboratory Diagnostics. Use and assessment of clinical laboratory results.
Frankfurt/Main: TH-Books Verlagsgesellschaft mbH, 1998: 142-148.
2. Jeppsson J-O, Kobold U, Barr J, Finke A, Hoezel W, Hoshino T, Miedema K, Mosca A, Mauri P, Paroni R, Thienpont L, Umemoto M, Weykamp C. Approved
IFCC reference method for the Measurement of HbA1c in Human Blood. Clin Chem Lab Med 2002; 40(1): 78-89.
3. Hoelzel W, Weykamp C, Jeppsson J-O, Miedema, Barr JR, Goodall I, Hoshino T, John, WG, Kobold U, Little R, Mosca A, Mauri P, Paroni R, Susanto F, Takei I,
Theinpoint L, Umemoto M, Wiedmeyer H-M. IFCC Reference System for Measurement of Hemoglobin A1c in Human Blood and the National Standardization
Schemes in the United States, Japan, and Sweden: A Method-Comparison Study. Clin Chem 2004; 50(1): 166-174.

2009-08
TOTAL Hb, AU680 HbA1c Automated APT Application
Operation: Yes ∇
100.HbA1c% 101.T-Hb 102.HbA1c 100.HbA1c 101.T-Hb 102.HbA1c
OD Limit Min OD -2.0000 -2.0000
Sample Volume 5 μL 11.0 µL 3.5 µL Max OD 3.0000 3.0000
Rgt. Volume: R1(R1-1) 200 µL 150 µL 76 µL Reagent OD Limit
R2(R2-1) 0 μL 0 µL 76 µL First Low -2.0000 -2.0000
Wavelength: Pri. 600 ∇nm 700 ∇nm High 3.0000 3.0000
Sec 700 ∇nm None ∇nm Last Low -2.0000 -2.0000
Method: END ∇ FIXED ∇ High 3.0000 3.0000
Reaction Slope: + ∇ + ∇ Dynamic Range
Low 4.4000* **
Measuring Point-1 First 0 13 High 14.300* **
Last 27 22 Correlation Factor A 1 1
Measuring Point-2 First B 0 0
Last Factor for Maker A 1 1
Linearity Limit: 0 % 0 % B 0 0
Lag Time Check: No ∇ No ∇ Onboard Stability Period 30 Day
Hour

Test Name: THb§ ∇ < > Type: Whole Blood ∇

Normal Ranges: From To Low High
ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
General ISE
Test Name: THb§ ∇ < > Type Whole Blood ∇ ο Use Serum Cal.

HbA1c AUTOMATED APT APPLICATION

Point 1: # ∇ † -999999 999999
Point 3: ∇
Point 6: ∇
Point 9 ∇
Calibrator OD Conc Low High Stability † HbA1c Calibrator ODR3032
Point-1 ∇ Reagent Blank 14 Day 0 Hour * Values set for working in mmol/L. To work in g/dL, multiply by 1.6125.
Point-2 ∇ Calibration 14 Day 0 Hour ** Concentration of 0.1 mmol/L and highest calibrator.
§ For determination of %HbA1c the above tests (THb and HbA1c) are used. %HbA1c is calculated automatically by the analyser using
MB Type Factor: 1-Point Calibration Point None ∇ ο with Conc-0 the following formula: (A/B)*a+b, where A=HbA1c , B=THb and a=91.5 and b=2.15.

2009-08
HbA1c, AU680 HbA1c Automated APT Application
System Reagent: OSR61177 HbA1C Reagent ID: 177
Denaturant Reagent ID: 200
Operation: Yes ∇
100.HbA1c% 101.T-Hb 102.HbA1c 100.HbA1c 101.T-Hb 102.HbA1c
OD Limit Min OD -2.0000 -2.0000
Sample Volume 5 μL 11.0 µL 3.5 µL Max OD 3.0000 3.0000
Rgt. Volume: R1(R1-1) 200 µL 150 µL 76 µL Reagent OD Limit
R2(R2-1) 0 μL 0 µL 76 µL First Low -2.0000 -2.0000
Wavelength: Pri. 600 ∇nm 700 ∇nm High 3.0000 3.0000
Sec 700 ∇nm None ∇nm Last Low -2.0000 -2.0000
Method: END ∇ FIXED ∇ High 3.0000 3.0000
Reaction Slope: + ∇ + ∇ Dynamic Range
Low 4.4000* **
Measuring Point-1 First 0 13 High 14.300* **
Last 27 22 Correlation Factor A 1 1
Measuring Point-2 First B 0 0
Last Factor for Maker A 1 1
Linearity Limit: 0 % 0 % B 0 0
Lag Time Check: No ∇ No ∇ Onboard Stability Period 30 Day
Hour

Test Name: HbA1c§ ∇ < > Type: Whole Blood ∇

Normal Ranges: From To Low High
ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
General ISE
Test Name: HbA1c§ ∇ < > Type Whole Blood ∇ ο Use Serum Cal.

Calibrator OD Conc Low High Slope Check - ∇
HbA1c AUTOMATED APT APPLICATION
Point 1: # ∇ † -2.0 3.0
Point 3: # ∇ † -2.0 3.0
Point 6: # ∇ † -2.0 3.0
Point 9 ∇
<Point Cal. For No. of Correction Points Use Master Curve # User defined
† HbA1c Calibrator ODR3032
** Concentration of 0.1 mmol/L, and highest calibrator.
Point-1 ∇ Reagent Blank 14 Day 0 Hour § For determination of %HbA1c the above tests (THb and HbA1c) are used. %HbA1c is calculated automatically by the analyser using
Point-2 ∇ Calibration 14 Day 0 Hour the following formula: (A/B)*a+b, where A=HbA1c, B=THb and a=91.5 and b=2.15.

2009-08
IgA
OSR61171 4 x 14 mL R1
4 x 11 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of immunoglobulin A (IgA) in human serum and plasma on Beckman Coulter analysers. For
Summary1,2
Immunoglobulin classes IgG, IgA, IgM, IgD and IgE are present in descending order of concentration in the serum of healthy people. IgA antibodies occur
as serum IgA and as secretory IgA. Unlike secretory IgA the specific role of serum IgA is unclear. IgA does not cross the placenta, therefore it is not present
in fetal blood. Secretory IgA consists of a dimer connected by a J-chain and has a secretory component that protects the molecule from proteolytic
enzymes. Secretory IgA is the predominant immunoglobulin of body secretions such as saliva, tears, colostrum, nasal secretions, tracheobronchial mucus
and gastrointestinal secretions. Essential functions of secretory IgA are the binding of microorganisms on mucous membranes, the activation of the
alternative complement pathway and activation of inflammatory reactions, it particularly plays a major role in the protection of the respiratory, genitourinary,
and gastrointestinal tracts against infection.
Changes in serum immunoglobulin concentrations can be classified as follows:
- Hypogammaglobulinemias, individuals with secretory IgA deficiency are found to suffer more commonly from mucosal infections, atopy, and autoimmune
diseases. Individuals with absent IgA have a higher than expected incidence of rheumatic disorders and lymphoma.
- Polyclonal gammopathies, increased levels occur in chronic liver disease; chronic infections, especially of the GI and respiratory tracts; neoplasia of the
lower GI tract; inflammatory bowel disease; some immunodeficiency states such as Wiskott-Aldrich syndrome and rheumatoid arthritis.
- Monoclonal gammopathies, e.g. in IgA type multiple myeloma.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, human IgA reacts specifically with anti-human IgA antibodies to yield insoluble
aggregates. The absorbance of these aggregates is proportional to the IgA concentration in the sample.
Polyethylene glycol 6000 3.5 %
Goat anti-IgA antibodies Dependent on titre
Preservative
Reagent Preparation
for 90 days.
Specimen
3
Stable in serum and plasma for 8 months when stored at 2…25 °C.
Test Procedure
Calibration
Serum Protein Multi-Calibrator ODR3021.
The calibrator IgA values are traceable to IFCC (International Federation of Clinical Chemistry) standard CRM 470.
Quality Control

2009-08
Calculation
The Beckman Coulter analysers automatically compute the IgA concentration of each sample.
Adults 0.7 – 4.0 g/L (70 – 400 mg/dL)
values.
Linearity
Precision
The following data was obtained on an AU640 analyser using 3 serum pools analysed over 20 days.
1.02 0.01 1.41 0.03 3.39
2.40 0.04 1.52 0.09 3.85
4.79 0.10 2.18 0.19 4.01
Sensitivity
The lowest detectable level represents the lowest measurable level of IgA immunoglobulins that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this IgA assay on the AU2700 against another commercially available IgA assay. Results of linear
®
RF: Interference less than 10% up to 600 IU/mL
5
Limitations
The IgA assay has been optimised to reduce the risk of prozone occurrence in the presence of abnormally high immunoglobulin concentrations.
However, as a precaution samples from patients with suspected paraproteinaemia should also be tested by electrophoresis.
Samples with very high IgA concentrations (> 100 g/L polyclonal) can generate false low results without appropriate "Z" flags due to excess antigen in the
sample.
When running elevated samples on any of the AU system analysers, “F” flags or a combination of “F” and “Z” flags may be obtained. Such samples should
be diluted using physiological saline so as to recover close to the middle of the measuring range.
† Serum Protein Multicalibrator Cat. No.: ODR3021
BIBLIOGRAPHY
1. Thomas L. Immunoglobulins (Ig). In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books
470. J Lab Med 1996;20:145-152.

2009-08
IgA, AU400/AU640 Serum/Plasma Application IgA, AU600 Serum/Plasma Application
General LIH ISE Range Specific Test Parameters
Test Name: IgA ∇ < > Type: Serum ∇ Operation: Yes ∇ Test No # Test name IgA ∇ Sample type Ser ∇ Page 1/2
Sample: Volume 2 μL Dilution 0 μL Pre-Dilution Rate: 1 Sample vol. 2 Dil. Vol 0 μL Min. OD Max. OD
Reagents: R1 Volume 70 μL Dilution 115 μL Min OD Max OD Reagent 1 vol 70 Dil. Vol 115 μL L H
Reagent 2 vol 55 Dil. Vol 10 μL Reagent OD limit
Fst. L -0.1 Fst. H 1.5
Linearity : % A 1 B 0 Linearity Fst % Sec %
No Lag Time: ∇ On-board stability period: 90 No lag time ∇ On-board stability period 90
Calibration Specific Test No # Test name IgA ∇

General ISE
Cal type 13 5AB ∇ Count #
Test Name: IgA ∇ < > Type Serum ∇ Formula 3 POLYGONAL ∇ Process Conc ∇
Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇ Cal No OD Conc Factor/OD-L Factor/OD-H
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 6 ∇
Point 7 ∇
Point 5: # † -0.1 2.5
1-point cal. Point
1-Point Cal. Point: ο with CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ Select the function using the Function key or the Mouse
Test No # Name IgA ∇ Type Ser ∇


Test Name: IgA ∇ < > Type: Serum ∇
√ Logic Check-1 Logic Check -2 Logic Check-3 Check Point-3 27 Decision value-1 0.0000
Check Point-1: 11 Check Point-1: Check Point-1: Decision value-1 0.0000 Decision value-2 0.0000
Check Point-3: 27 Decision Value-1: Decision Value-1: Decision value-3 0.0150 Limit Point 2 0
Decision Value-2: 0.7700 Limit Point 1: Limit Point 1: Limit Point 2 27 Logic Check-3 NO
Limit Point 1: 16 Check Point Interval
Decision value-2 0.0000
Limit Point 1 0
Limit Point 2 0
# User defined
† Serum Protein Multicalibrator Cat. No.: ODR3021 # User defined ¤ Analyser default value
* Values set for working in SI units (g/L). To work in mg/dL multiply by 100. † Serum Protein Multicalibrator Cat. No.: ODR3021

2009-08
IgA, AU2700/AU5400 Serum/Plasma Application IgA, AU680 Serum/Plasma Application
Test Name: IgA ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: IgA ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: IgA ∇ < > Type: Serum ∇ General ISE
Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇ Test Name: IgA ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # † -0.1 2.5
Point 2: # † -0.1 2.5
Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5
Point 4: # † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5
Point 6: Point 5: # † -0.1 2.5
∇ ο Calibration
Point 7: Point 6:
Point 9 ∇
Test Name: IgA ∇ < > Type: Serum ∇ ∇
Parameters Misc
Decision Value-3: 0.0230 Limit Point 2: Limit Point 2: Test Name: IgA ∇ < > Type: Serum ∇
Limit Point 1: 16
Check Point-3: 27
# User defined Decision Value-2: 0.7700 Decision Value-2: Decision Value 2:
† Serum Protein Multicalibrator Cat. No.: ODR3021 Decision Value-3: 0.0160

2009-08
IgA, AU480 Serum/Plasma Application
Test Name: IgA ∇ < > Type: Serum ∇ Operation Yes ∇

Method END ∇

General ISE
Test Name: IgA ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # ∇ † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 6:
Point 9 ∇
Parameters Misc

Test Name: IgA ∇ < > Type: Serum ∇

Check Point-3: 27
Limit Point 1: 16 Limit Point 1: Limit Point 1: # User defined
Limit Point 2: 27 Limit Point 2: Limit Point 2: † Serum Protein Multicalibrator Cat. No.: ODR3021
Check Pattern: Pattern1 ∇ * Values set for working in SI units (g/L). To work in mg/dL multiply by 100.

2009-08
IgA, AU400/AU640 Paediatric Application IgA, AU600 Paediatric Application

Test No # Test name IgAP ∇ Sample type Ser ∇ Page 1/2
Test Name: IgAP ∇ < > Type: Serum ∇ Operation: Yes ∇
Fst. L -0.1 Fst. H 1.5
Wavelength: Pri. 600 ∇ Sec. 800 ∇ First L -0.1 First H 1.5 Reaction + ∇ L 0.1* H 7*
Test No # Test name IgAP ∇

Test Name: IgAP ∇ < > Type Serum ∇ Selection calibrator
Point 2 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 6 ∇
Point 4: # † -0.1 2.5 Point 7 ∇
1-point cal. Point
Point 7:
Test No # Name IgAP ∇ Type Ser ∇

Test Name: IgAP ∇ < > Type: Serum ∇
Check Point-3: 27 Decision Value-1: Decision Value-1: Limit Point 1 16
Decision Value-1: 0.0000 Decision Value-2: Decision Value-2: Limit Point 2 27 Logic Check-3 NO
Limit Point 1 0
Limit Point 2 0

† Serum Protein Multicalibrator Cat. No.: ODR3021 † Serum Protein Multicalibrator Cat. No.: ODR3021
* Values set for working in SI units (g/L). To work in mg/dL multiply by 100. * Values set for working in SI units (g/L). To work in mg/dL multiply by 100

2009-08
IgA, AU2700/AU5400 Paediatric Application IgA, AU680 Paediatric Application
Test Name: IgAP ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: IgAP ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: IgAP ∇ < > Type: Serum ∇ General ISE
Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇ Test Name: IgAP ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # † -0.1 2.5
Point 2: # † -0.1 2.5
Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5
Point 4: # † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5
Point 6: Point 5: # † -0.1 2.5
∇ ο Calibration
Point 7: Point 6:
1-Point Cal. Point: ο With CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ Point 7 ∇ Advanced Calibration
Point 9 ∇
Test Name: IgAP ∇ < > Type: Serum ∇ Point-1 ∇ Reagent Blank 999 Day 0 Hour
Parameters Misc
Decision Value-3: 0.0230 Limit Point 2: Limit Point 2: Test Name: IgAP ∇ < > Type: Serum ∇
Limit Point 1: 16
Check Point-3: 27
* Values set for working in SI units (g/L). To work in mg/dL multiply by 100

2009-08
IgA, AU480 Paediatric Application
Test Name: IgAP ∇ < > Type: Serum ∇ Operation Yes ∇

Method END ∇

General ISE
Test Name: IgAP ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # ∇ † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 6:
Point 9 ∇
Parameters Misc

Test Name: IgAP ∇ < > Type: Serum ∇

Check Point-3: 27
Check Pattern: Pattern1 ∇ * Values set for working in SI units (g/L). To work in mg/dL multiply by 100

2009-08
IgG
OSR61172 4 x 22 mL R1
4 x 20 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of immunoglobulin G (IgG) in human serum, plasma and cerebrospinal fluid on Beckman
Coulter AU analysers. For in vitro diagnostic use only.
Summary1,2,3,4
Immunoglobulin classes IgG, IgA, IgM, IgD and IgE are present in descending order of concentration in the serum of healthy people. IgG which consists of
four subclasses, comprises approximately 75% of total immunoglobulins and 10 – 20% of total serum protein. Approximately one half of total body IgG is
present in plasma while the other half, because of IgG's low molecular weight (150 kDa), is distributed in the interstitial fluid to act against tissue infection.
IgG is particularly important in the body's long-term defence against infection as it presents a slower but more sustained response than IgM to primary
antigenic stimulus; however, the levels of IgG rise rapidly and early on re-exposure to the same antigenic stimulus. IgG promotes phagocytosis and
activates complement.
- Hypogammaglobulinemias, IgG deficiency may be genetic as in severe combined immunodeficiency or acquired as in AIDS. Definitive diagnosis of the
clinical syndrome requires extensive evaluation of humoral and cellular functions in the immune response. A decrease in IgG also occurs as a result of
thermal burns, nephrotic syndrome, protein losing enteropathies and non-IgG myelomas.
- Polyclonal gammopathies, levels of IgG are increased in autoimmune diseases (systemic lupus erythematosus, rheumatoid arthritis, Sjögren's
syndrome), sarcoidosis, chronic liver disease, some parasitic diseases and chronic or recurrent infections.
- Monoclonal gammopathies, e.g. in IgG type multilple myeloma, lymphomas, leukemia, and other malignancies.
IgG is the only immunoglobulin that crosses the placenta and is therefore of special importance in the infant’s defence against infection.
IgG determination in cerebrospinal fluid (CSF) is used for evaluation of infections involving the central nervous system, neoplasms or primary neurological
diseases. CSF IgG measurements may be used to determine the IgG CSF/albumin CSF ratio which is an important factor in differentiating between
intrathecal and localised synthesis of IgG.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, human IgG reacts specifically with anti-human IgG antibodies to yield insoluble
aggregates. The absorbance of these aggregates is proportional to the IgG concentration in the sample.

Goat anti-IgG antibodies Dependent on titre
Preservative

Reagent Preparation

for 90 days.
Specimen
Serum and EDTA or heparinised plasma
5
Stable in serum and plasma for 8 months when stored at 2…8°C and 4 months when stored at 15…25 °C.
6
Cerebrospinal fluid: Stable for 72 hours when stored at 4°C. Stable for 6 months when stored frozen at -20 °C.
For CSF determinations, strongly icteric or turbid samples should be avoided.
Test Procedure
Calibration
Serum Protein Multi-Calibrator ODR3021
The calibrator IgG values are traceable to IFCC (International Federation of Clinical Chemistry) standard CRM 470.
For CSF application, recalibrate every 2 days.

2010-04
Quality Control
CSF: Control materials with values determined by this Beckman Coulter method may be used.
Calculation
The Beckman Coulter analysers automatically compute the IgG concentration of each sample.
Adults (Serum) 7 – 16 g/L (700 – 1600 mg/dL)
8
CSF 15 – 20 y 35 mg/L ± 20 mg/L
21 – 40 y 42 mg/L ± 14 mg/L
41 – 60 y 47 mg/L ± 10 mg/L
IgG Index 0.3 – 0.6

values.
Linearity
Serum: the test is linear within a concentration range of 0.75 – 30.0 g/L (75 – 3000 mg/dL).
CSF: the test is linear within a concentration range of 20 – 500 mg/L (2.0 – 50 mg/dL)
Precision
The following data was obtained on an AU640 analyser using 3 serum pools analysed over 20 days:
4.31 0.05 1.14 0.14 3.29
10.88 0.16 1.45 0.38 3.49
21.73 0.49 2.24 1.01 4.66
The following data was obtained on an AU640 analyser using 3 CSF pools analysed over 20 days:
34.86 0.93 2.68 3.32 9.53
101.34 0.90 0.89 3.72 3.67
355.74 3.29 0.93 10.01 2.81
Sensitivity
The lowest detectable level in CSF on an AU640 analyser was estimated at 4.20 mg/L.
The lowest detectable level represents the lowest measurable level of IgG immunoglobulins that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this IgG assay on the AU2700 against another commercially available IgG assay. Results of linear
Patient CSF samples were used to compare this IgG assay on the AU2700 against another commercially available IgG assay. Results of linear regression
were as follows.
y = 0.962x - 3.451 r = 0.997 n = 86 Sample range = 20.59 – 441.85 mg/L
Serum Icterus: Interference less than 3% up to 40 mg/dL or 684 µmol/L bilirubin
®
RF: Interference less than 10% up to 1200 IU/mL
CSF Icterus: Interference less than 10% up to 36 mg/dL or 205 µmol/L bilirubin
9
Limitations
The IgG assay has been optimised to reduce the risk of prozone occurrence in the presence of abnormally high immunoglobulin concentrations.
Samples with very high IgG concentrations (> 300 g/L polyclonal) can generate false low results without appropriate "Z" flags due to excess antigen in the
sample.
Please note that there is a requirement to dedicate a separate test channel specifically for CSF sample settings when assigning this IgG application on
Beckman Coulter AU analysers.
The result of IgG CSF samples may be elevated when immediately following a serum sample. In order to eliminate this effect, it is recommended to:
(a) Calibrate CSF IgG separately to other calibrators
(b) Avoid freely alternating serum and CSF samples
2010-04
(c) When changing from serum to CSF samples, place a sample cup containing 2% Wash Solution Cat. No.: OSR0001 in the first position of the rack and
requisition a test for this sample.

† Serum Protein Multicalibrator Cat. No.: ODR3021.
CSF: Prepare a 1 + 44 dilution of calibrator No. 4 in 0.9% saline. Then make four subsequent 1 + 1 dilutions of this to prepare a 5 point calibration
curve. Insert the adjusted values in the calibration curve.
∂ Separate CSF channel from other types (see IFU for further instruction).
CSF: Values set for working in SI units (mg/L). To work in mg/dL divide by 10.
CSF: note: Refer to IFU for limitations
BIBLIOGRAPHY
3. Colbert D, ed. Fundamentals of clinical physiology, 1st ed. Hertfordshire; Prentice Hall, 1993:296pp.
4. Mayne PD, ed. Clinical chemistry in diagnosis and treatment, 6th ed. Glasgow:Arnold, 1994:322-326, 323pp.
6. Tietz NW, ed Clinical guide to laboratory tests, 3rd ed. Philadelphia WB Saunders Company, 1995: 360pp
470. J Lab Med 1996;20:145-152.
8. Painter PC, Cope JY, Smith JL Reference information for the clinical laboratory. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry,Philadelphia:
WB Saunders Company,1999; 1820pp.

2010-04
IgG, AU400/AU640 Serum/Plasma Application IgG, AU600 Serum/Plasma Application

Test No # Test name IgG ∇ Sample type Ser ∇ Page 1/2
Test Name: IgG ∇ < > Type: Serum ∇ Operation: Yes ∇
Fst. L -0.1 Fst. H 1.5
Wave Main 600 Sub ∇ Lst. L -0.1 Lst. H 1.5
Test No # Test name IgG ∇

Formula 6 EIA Type 1 ∇ Process Conc ∇
Test Name: IgG ∇ < > Type Serum ∇ Selection calibrator
Calibration Type: 5AB ∇ Formula: EIA Type 1 ∇ Counts: # Process: CONC ∇ Point 1 # ∇ † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 6 ∇
Point 4: # † -0.1 2.5 Point 7 ∇
1-point cal. Point
Point 7:
Test No # Name IgG ∇ Type Ser ∇

Test Name: IgG ∇ < > Type: Serum ∇
Limit Point 1 0
Limit Point 2 0


2010-06
IgG, AU2700/AU5400 Serum/Plasma Application IgG, AU680 Serum/Plasma Application
Test Name: IgG ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: IgG ∇ < > Type: Serum ∇ Operation: Yes ∇
µL µL
Reagent OD limit:
Test Name: IgG ∇ < > Type: Serum ∇ General ISE
Calibration Type: 5AB ∇ Formula: EIA Type 1 ∇ Counts: # Process: CONC ∇ Test Name: IgG ∇ < > Type Serum ∇ R Use Serum Cal.
Calibration Type: 5AB ∇ Formula: EIA Type 1 ∇ Counts: # ∇

Point 1: # † -0.1 2.5
Point 2: # † -0.1 2.5
Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5
Point 4: # † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5
∇
Point 9 ∇
Decision Value-3: 0.3000 Limit Point 2: Limit Point 2: Test Name: IgG ∇ < > Type: Serum ∇
Limit Point 1: 11
Check Point-3: 20

2010-06
IgG, AU480 Serum/Plasma Application
Test Name: IgG ∇ < > Type: Serum ∇ Operation Yes ∇

Method END ∇

General ISE
Test Name: IgG ∇ < > Type Serum ∇ R Use Serum Cal.

Point 1: # ∇ † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 6: ∇
Point 9 ∇
Parameters Misc


Check Point-3: 20
Check Pattern: Pattern1 ∇ † Serum Protein Multicalibrator Cat. No.: ODR3021

2010-06
IgG, AU400/AU640 CSF Application∂ IgG, AU600 CSF Application∂
Test No # Test name IgG ∇ Sample type Others ∇ Page 1/2
Test Name: IgG ∇ < > Type: Others ∇ Operation: Yes ∇
Wavelength: Pri. 340 ∇ Sec. None ∇ First L -0.1 First H 2.5 Method END Dynamic range
∇
General LIH ISE Range Test No # Test name IgG ∇ Sample type Others ∇ Page 2/2
Test Name: IgG ∇ < > Type: Others ∇ Level L Level H
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
CSF APPLICATION
General ISE Test No # Test name IgG ∇
Test Name: IgG ∇ < > Type Others ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
1-Point Cal. Point: R with CONC-0 Slope Check: + # 1-point cal. point
MB type factor
† Serum Protein Multicalibrator Cat. No.: ODR3021. Prepare a 1+44 dilution of calibrator No. 4 in 0.9% † Serum Protein Multicalibrator Cat. No.: ODR3021. Prepare a 1+44 dilution of calibrator No. 4 in 0.9%
saline. Then make four subsequent 1+1 dilutions of this to prepare a 5 point calibration curve. Insert the adjusted saline. Then make four subsequent 1+1 dilutions of this to prepare a 5 point calibration curve. Insert the adjusted
values in the calibration curve. ∂ Separate CSF channel from other types (see IFU for further instruction) values in the calibration curve. ∂ Separate CSF channel from other types (see IFU for further instruction)
* Values set for working in SI Units (mg/L). To work in mg/dL divide by 10. * Values set for working in SI Units (mg/L). To work in mg/dL divide by 10.
Note: Refer to IFU for limitations. Note: Refer to IFU for limitations.
2010-06
IgG, AU2700/AU5400 CSF Application∂ IgG, AU680/AU480 CSF Application∂
Test Name: IgG ∇ < > Type: Others ∇ Operation Yes ∇
Test Name: IgG ∇ < > Type: Others ∇ Operation: Yes ∇

µL µL
Reagent OD limit:
Linearity Limit %
Test Name: IgG ∇ < > Type: Others ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: IgG ∇ < > Type: Others ∇
# ∇ # # # # # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # # # # # # #
∇
R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
CSF APPLICATION
General ISE
General ISE
Test Name: IgG ∇ < > Type Others ∇ R
Test Name: IgG ∇ < > Type Others ∇ Use Serum Cal.
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4: # ∇ † -0.1 2.5 R Reagent Blank
Point 5: # † -0.1 2.5 Point 5: # ∇ † -0.1 2.5 R Calibration
1-Point Cal. Point: R with CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
† Serum Protein Multicalibrator Cat. No.: ODR3021. Prepare a 1+44 dilution of calibrator No. 4 in 0.9%
saline. Then make four subsequent 1+1 dilutions of this to prepare a 5 point calibration curve. Insert the adjusted Point-1 ∇ Reagent Blank 2 Day 0 Hour
values in the calibration curve. ∂ Separate CSF channel from other types (see IFU for further instruction) Point-2 ∇ Calibration 2 Day 0 Hour
* Values set for working in SI Units (mg/L). To work in mg/dL divide by 10. MB Type Factor: 1-Point Calibration Point None ∇ R with Conc-0
Note: Refer to IFU for limitations.
ф U680

2010-06
IgG, AU400/AU640 Paediatric Application IgG, AU600 Paediatric Application

Test No # Test name IgGP ∇ Sample type Ser ∇ Page 1/2
Test Name: IgGP ∇ < > Type: Serum ∇ Operation: Yes ∇
Fst. L -0.1 Fst. H 1.5
Wave Main 600 Sub ∇ Lst. L -0.1 Lst. H 1.5
Test No # Test name IgGP ∇

Formula 6 EIA Type 1 ∇ Process Conc ∇
Test Name: IgGP ∇ < > Type Serum ∇ Selection calibrator
Calibration Type: 5AB ∇ Formula: EIA Type 1 ∇ Counts: # Process: CONC ∇ Point 1 # ∇ † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 6 ∇
Point 4: # † -0.1 2.5 Point 7 ∇
1-point cal. Point
Point 7:
Test No # Name IgGP ∇ Type Ser ∇

Test Name: IgGP ∇ < > Type: Serum ∇
Limit Point 1 0
Limit Point 2 0


2010-06
IgG, AU2700/AU5400 Paediatric Application IgG, AU680 Paediatric Application
Test Name: IgGP ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: IgGP ∇ < > Type: Serum ∇ Operation: Yes ∇
µL µL
Reagent OD limit:
Test Name: IgGP ∇ < > Type: Serum ∇ General ISE
Calibration Type: 5AB ∇ Formula: EIA Type 1 ∇ Counts: # Process: CONC ∇ Test Name: IgGP ∇ < > Type Serum ∇ R Use Serum Cal.

Point 1: # † -0.1 2.5
Point 2: # † -0.1 2.5
Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5
Point 4: # † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5
∇
1-Point Cal. Point: R With CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ Point 7 ∇ Advanced Calibration
Point 9 ∇
Test Name: IgGP ∇ < > Type: Serum ∇ Point-1 ∇ Reagent Blank 999 Day 0 Hour
Decision Value-3: 0.3000 Limit Point 2: Limit Point 2: Test Name: IgGP ∇ < > Type: Serum ∇
Limit Point 1: 11
Check Point-3: 20

2010-06
IgG, AU480 Paediatric Application
Test Name: IgGP ∇ < > Type: Serum ∇ Operation Yes ∇

Method END ∇

General ISE
Test Name: IgGP ∇ < > Type Serum ∇ R Use Serum Cal.

Point 1: # ∇ † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 6: ∇
Point 9 ∇
Parameters Misc

Test Name: IgGP ∇ < > Type: Serum ∇

Check Point-3: 20
Check Pattern: Pattern1 ∇ * Values set for working in SI units (g/L). To work in mg/dL multiply by 100

2010-06
IgM
OSR61173 4 x 14 mL R1
4 x 11 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of immunoglobulin M (IgM) in human serum and plasma on Beckman Coulter analysers. For
Summary1,2,3
Immunoglobulin classes IgG, IgA, IgM, IgD and IgE are present in descending order of concentration in the serum of healthy people. IgM composes
approximately 7% of the total plasma immunoglobulins and is the first immunoglobulin to respond to an antigenic stimulus. IgM normally circulates in
plasma in a pentameric form and because of its relatively large molecular mass (971 kDa), 75-80% of IgM is located intravascularly. The IgM class
includes the natural antibodies, e.g. the ABO blood group isohaemagglutinins, saline Rh and antibodies to IgG e.g. rheumatoid factors.
The essential functions of IgM in the immune response are the agglutination of pathogens and the activation of the classical complement pathway.
- Hypogammaglobulinemias, IgM deficiency is rare and is associated with recurrent pyrogenic infections.
- Polyclonal gammopathies, levels of IgM are increased in primary biliary cirrhosis, haemoprotozoan infections such as malaria, viral or bacterial infections
and rheumatoid arthritis.
- Monoclonal gammopathies, e.g. in Waldenström’s macroglobulinemia and malignant lymphoma.
Elevated levels of IgM in cord serum or during the first four weeks of life may indicate intrauterine or neonatal infections such as rubella, cytomegalovirus,
toxoplasmosis or syphilis.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, human IgM reacts specifically with anti-human IgM antibodies to yield insoluble
aggregates. The absorbance of these aggregates is proportional to the IgM concentration in the sample.
Goat anti-IgM antibodies Dependent on titre
Preservative
Reagent Preparation
for 90 days.
Specimen
4
Stable in serum and plasma for 4 months when stored at 2…8 °C and 2 months when stored at 15…25 °C.
Lipemic samples should be avoided.
Test Procedure
Calibration
Serum Protein Multi-Calibrator ODR3021.
The calibrator IgM values are traceable to IFCC (International Federation of Clinical Chemistry) standard CRM 470.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the IgM concentration of each sample.
2009-08
Adults 0.4 – 2.3 g/L (40 – 230 mg/dL)
Children 0.2 – 2.0 g/L (20 – 200 mg/dL)
values.
Linearity
Precision
The following data was obtained on an AU640 analyser using 3 serum pools analysed over 10 days.
0.48 0.01 1.69 0.02 3.44
1.14 0.02 1.36 0.04 3.29
2.17 0.05 2.19 0.09 4.08
Sensitivity
The lowest detectable level represents the lowest measurable level of IgM immunoglobulins that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this IgM assay on the AU2700 against another commercially available IgM assay. Results of linear
®
7
Limitations
The IgM assay has been optimised to reduce the risk of prozone occurrence in the presence of abnormally high immunoglobulin concentrations.
Samples with very high IgM concentrations (> 100 g/L polyclonal) can generate false low results without appropriate "Z" flags due to excess antigen in the
sample.
† Serum Protein Multicalibrator Cat. No.: ODR3021
BIBLIOGRAPHY
3. Mayne PD, ed. Clinical chemistry in diagnosis and treatment, 6th ed. Glasgow: Arnold, 1994:322-326,346-347.
470. Lab Med 1996;20:145-152.

2009-08
IgM, AU400/AU640 Serum/Plasma Application IgM, AU600 Serum/Plasma Application

Test No # Test name IgM ∇ Sample type Ser ∇ Page 1/2
Test Name: IgM ∇ < > Type: Serum ∇ Operation: Yes ∇
Fst. L -0.1 Fst. H 1.5
Test No # Test name IgM ∇

Test Name: IgM ∇ < > Type Serum ∇ Selection calibrator
Point 2 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 6 ∇
Point 4: # † -0.1 2.5 Point 7 ∇
1-point cal. point
Point 7:
Test No # Name IgM ∇ Type Ser ∇

Test Name: IgM ∇ < > Type: Serum ∇
Limit Point 1 0
Limit Point 2 0


2009-08
IgM, AU2700/AU5400 Serum/Plasma Application IgM, AU680/AU480 Serum/Plasma Application
Test Name: IgM ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: IgM ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: IgM ∇ < > Type: Serum ∇ General ISE
Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇ Test Name: IgM ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # † -0.1 2.5
Point 2: # † -0.1 2.5
Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5
Point 4: # † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5
Point 6: Point 5: # † -0.1 2.5
∇ ο Calibration
Point 9 ∇
Test Name: IgM Type: Serum Point-1 ∇ Reagent Blank 999 Day 0 Hour
∇ < > ∇
Decision Value-3: 0.3000 Limit Point 2: Limit Point 2: Test Name: IgM ∇ < > Type: Serum ∇
Limit Point 1: 11
Check Point-3: 23
ф AU680

2009-08
IgM, AU400/AU640 Paediatric Application IgM, AU600 Paediatric Application

Test No # Test name IgMP ∇ Sample type Ser ∇ Page 1/2
Test Name: IgMP ∇ < > Type: Serum ∇ Operation: Yes ∇
Fst. L -0.1 Fst. H 1.5
Test No # Test name IgMP ∇

Test Name: IgMP ∇ < > Type Serum ∇ Selection calibrator
Point 2 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 6 ∇
Point 4: # † -0.1 2.5 Point 7 ∇
1-point cal. Point
Point 7:
Test No # Name IgMP ∇ Type Ser ∇

Test Name: IgMP ∇ < > Type: Serum ∇
Limit Point 1 0
Limit Point 2 0


2009-08
IgM, AU2700/AU5400 Paediatric Application IgM, AU680/AU480 Paediatric Application
Test Name: IgMP ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: IgMP ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: IgMP ∇ < > Type: Serum ∇ General ISE
Calibration Type: 5AB ∇ Formula: POLYGONAL ∇ Counts: # Process: CONC ∇ Test Name: IgMP ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # † -0.1 2.5
Point 2: # † -0.1 2.5
Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5
Point 4: # † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5
Point 6: Point 5: # † -0.1 2.5
∇ ο Calibration
Point 9 ∇
Test Name: IgMP ∇ < > Type: Serum ∇ Point-1 ∇ Reagent Blank 999 Day 0 Hour
Decision Value-3: 0.3000 Limit Point 2: Limit Point 2: Test Name: IgMP ∇ < > Type: Serum ∇
Limit Point 1: 11
Check Point-3: 23
ф AU680

2009-08
MICROALBUMIN
OSR6167 4 x 15 mL R1
4 x 5 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of albumin in human urine and cerebrospinal fluid on Beckman Coulter analysers. For in vitro
Summary1,2,3,4
The earliest clinical evidence of nephropathy is the appearance of low but abnormal levels (> 30 mg/day or 20 µg/min) of albumin in the urine, referred to
as microalbuminuria, and patients with microalbuminuria are referred to as having incipient nephropathy. Conventional qualitative tests (chemical strips or
dipsticks) for albuminuria do not detect the small increases in urinary albumin excretion seen in early stages of nephropathy. For this purpose, tests for
microalbuminuria are used. Microalbuminuria is defined as an albumin excretion rate between 30-300 mg/24h on 2 of 3 urine collections.
Microalbuminuria is considered a clinically important indicator of deteriorating renal function in diabetic subjects and regular screening is valuable in
monitoring both type I and type II diabetes.
Prospective studies have demonstrated that increased urinary albumin excretion precedes and is highly predictive of diabetic nephropathy, end stage renal
disease, and proliferative retinopathy in type I diabetes. In patients with type II diabetes increased urinary albumin excretion is an independent predictor of
progressive renal disease, atherosclerotic disease and cardiovascular mortality. Increased urinary albumin excretion, both independently and in
conjunction with hyperinsulinemia, identifies a group of nondiabetic subjects at increased risk of coronary artery disease.
The degree of permeability of the blood-CSF barrier may be evaluated by the simultaneous measurement of serum and CSF albumin. CSF albumin
measurements may also be used to determine the IgG CSF/albumin CSF ratio which is an important factor in differentiating between intrathecal and
localised synthesis of IgG.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, human albumin reacts specifically with anti-human albumin antibodies, to yield
insoluble aggregates. The absorbance of these aggregates is proportional to the albumin concentration in the sample.
Goat anti-human albumin antibodies Variable
Preservative
Harmful, contains sodium azide. R22 Harmful if swallowed.
Safety Phrases:
S36, S60: Wear suitable protective clothing. This material and its container must be disposed of as hazardous waste.
Antisera was produced in healthy animals in facilities free from rinderpest, foot and mouth disease, peste des petits ruminants, Rhift Valley fever, bovine
spongiform encephalopathy and blue tongue disease.
Reagent Preparation
90 days.
Specimen5
Urine: Stable in urine, without preservative, for 1 month when stored at 2…8°C and 7 days when stored at 15…25°C.
6
Cerebrospinal fluid. Stable for 72 hours when stored at 4°C. Stable for 6 months when stored frozen at -20°C.
Test Procedure
Calibration
Microalbumin Calibrator Cat. No. ODR3024.
The calibrator Microalbumin values are traceable to a primary albumin standard.
Quality Control
Control materials, of human origin, with values determined by this Beckman Coulter system may be used.
2009-08
Calculation
The Beckman Coulter analysers automatically compute the albumin concentration of each sample.
Urine:
24-h collection Timed collection Spot collection
(mg/24h) (µg /min) (µg /mg creatinine)
Normal < 30 < 20 < 30
Microalbuminuria 30 – 299 20 – 199 30 – 299
Clinical albuminuria ≥ 300 ≥ 200 ≥ 300
7
CSF 3 mo – 4y 0 – 450mg/L
>4y 100 – 300mg/L
values.
Linearity
The test is linear within a concentration range of 5 – 300 mg/L (0.5 – 30 mg/dL). The test has a prozone tolerance of 6000 mg/L (600 mg/dL).
Microalbumin OSR6167 can exhibit depressed recovery of analyte when urine and CSF proteins are at a level exceeding 6000 mg/L due to antigen
excess effect. Urine and CSF samples should be initially screened by an alternative method for grossly abnormal total protein. Samples with extremely
high levels of protein should not be assayed for Microalbumin. Urine and CSF samples with total protein levels greater than 300 mg/L (the upper limit of the
OSR6167 linearity range) should be diluted to allow measurement within the range of the assay.
Precision
5.43 0.17 3.06 0.27 4.90
107.93 1.43 1.33 2.14 1.98
269.28 2.10 0.78 4.89 1.82
Sensitivity
The lowest detectable level represents the lowest measurable level of albumin that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient urine samples were used to compare this Albumin OSR6167 assay on the AU400 against another commercially available albumin assay. Results
y = 1.109x – 1.99 r = 993 n = 58 Sample range = 2.3 – 304 mg/L
Patient CSF samples were used to compare this Albumin OSR6167 assay on the AU640 against another commercially available albumin assay. Results
of linear regression analysis were as follows.
y = 1.07x – 1.24 r = 0.994 n = 100 Sample range = 9.7 – 91.0 mg/dL
Glucose: Interference less than 5% up to 3000 mg/dL or 166.7 mmol/L glucose
Creatinine: Interference less than 5% up to 300 mg/dL or 26.52 mmol/L creatinine
8
Limitations
The Microalbumin result of a urine or CSF sample may be elevated when it immediately follows a serum sample. In order to eliminate this effect, it is
recommended to:
a) Process microalbumin calibrators separately to other serum calibrators.
b) Avoid freely alternating serum and urine racks.
c) When changing from serum to urine or CSF samples, place a sample cup containing 2% AU detergent in the first position of the urine rack and
requisition a test for this sample.
Prozone or hook effect may occur with very elevated microalbumin samples (>6000 mg/L).
† System Calibrator Cat. No.: ODR3024
* Values set for working in mg/L. To work in (mg/dL), divide by 10.
CSF only Note: Refer to leaflet for limitations
BIBLIOGRAPHY
1. American Diabetes Association. Diabetic Nephropathy. Diabetes Care 25:(Suppl. 1):S85-S89.
2. Sacks DB, Bruns DE, Goldstein DE, MacLaren NK, McDonald JM, Parrot M. Guidelines and recommendations for laboratory analysis in the diagnosis and management of
diabetes mellitus. Clin Chem 2002;48:436-472.
3. Sacks DB. Carbohydrates. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia: WB Saunders Company, 1999;798pp.
4. Newman DJ, Price CP. Renal function and nitrogen metabolites. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia: WB Saunders Company,
1999;1228pp.
5. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of anticoagulants in diagnostic laboratory investigations and stability of blood, plasma and serum
samples. WHO/DIL/LAB/99.1 Rev.2:46pp.
6. Tietz NW, ed Clinical guide to laboratory tests, 3rd ed. Philadelphia WB Saunders Company, 1995:24pp
7. Painter PC, Cope JY, Smith JL Reference information for the clinical laboratory. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry, Philadelphia: WB
Saunders Company,1999;1800pp.

2009-08
MICROALBUMIN, AU400/AU640 Urine Application MICROALBUMIN, AU600 Urine Application

Test No # Test name MALB ∇ Sample type Uri ∇ Page ½
Test Name: MALB ∇ < > Type: Urine ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name MALB ∇ Sample type Uri ∇ Page 2/2
Test Name: MALB ∇ < > Type: Urine ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
URINE APPLICATION
General ISE Test No # Test name MALB ∇
Test Name: MALB ∇ < > Type Urine ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
MB type factor
# User defined
† Microalbumin Calibrator Cat. No.: ODR3024

2009-08
MICROALBUMIN, AU2700 / AU5400 Urine Application MICROALBUMIN, AU680/AU480 Urine Application
Test Name: MALB ∇ < > Type: Urine ∇ Operation Yes ∇
Test Name: MALB ∇ < > Type: Urine ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Linearity Limit %
Test Name: MALB ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: MALB ∇ < > Type: Urine ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
URINE APPLICATION
General ISE
General ISE
Test Name: MALB ∇ < > Type Urine ∇ Test Name: MALB ∇ < > Type Urine ∇ ο Use Serum Cal.
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇
† Microalbumin Calibrator Cat. No.: ODR3024 Calibrator OD Conc Low High Stability

2009-08
MICROALBUMIN, AU400/AU640 CSF Application MICROALBUMIN, AU600 CSF Application

Test No # Test name MALB ∇ Sample type Others ∇ Page 1/2
Test Name: MALB ∇ < > Type: Others ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name MALB ∇ Sample type Others ∇ Page 2/2
Test Name: MALB ∇ < > Type: Others ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
CSF APPLICATION
General ISE Test No # Test name MALB ∇
Test Name: MALB ∇ < > Type Others ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
MB type factor
† Microalbumin Calibrator Cat. No.: ODR3024 † Microalbumin Calibrator Cat. No.: ODR3024
* Values set for working in mg/L. To work in mg/dL divide by 10. * Values set for working in mg/L. To work in mg/dL divide by 10.
Note: Refer to leaflet for limitations. Note: Refer to leaflet for limitations.
2009-08
MICROALBUMIN, AU2700/AU5400 CSF Application MICROALBUMIN, AU680/AU480 CSF Application
Test Name: MALB ∇ < > Type: Others ∇ Operation Yes ∇
Test Name: MALB ∇ < > Type: Others ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Linearity Limit %
Test Name: MALB ∇ < > Type: Others ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: MALB ∇ < > Type: Others ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
CSF APPLICATION
General ISE
General ISE
Test Name: MALB ∇ < > Type Others ∇ Test Name: MALB ∇ < > Type Others ∇ ο Use Serum Cal.
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇
Note: Refer to leaflet for limitations. Point-2 ∇ Calibration 999 Day 0 Hour
Ф AU680 MB Type Factor: 1-Point Calibration Point None ∇ ο with Conc-0

2009-08
MYOGLOBIN
OSR6168 4 x 12 mL R1
4 x 9 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of myoglobin in human serum on Beckman Coulter analysers. For in vitro diagnostic use only.
Summary1,2,3,4
Myoglobin is the O2-binding protein of striated i.e. cardiac and skeletal muscle. Unlike haemoglobin, myoglobin is unable to release O2 except at an
extremely low pO2, therefore may serve as an oxygen reservoir, accessible only under conditions of extreme hypoxia. Myoglobinemia occurs after trauma
to either skeletal or cardiac muscle, e.g. in crush injury, uremia, seizures, inflammatory myopathies, intramuscular injections, exercise, burns and
myocardial infarction. A decrease in myoglobin may be observed in the presence of circulating antibodies to myoglobin, as in many patients with
polymyositis, and rheumatoid arthritis.
A clinical history is required to establish if an increase in serum myoglobin is due to a myocardial infarction or to a release of myoglobin from skeletal
muscle. The serum myoglobin concentration increases before a rise in other biochemical cardiac markers, such as troponin I or T, and CK-MB after a
myocardial infarction. The increase in myoglobin occurs 2-4 hours after the onset of symptoms and returns to reference range within 24 hours of onset.
Myoglobin elevation due to skeletal muscle damage shows a pattern essentially identical to that of total CK. Myoglobin is very useful for the exclusion of an
acute myocardial infarction and for monitoring thrombolytic therapy however it is a non-specific marker for myocardial infarction and must be corroborated
by other cardiac-specific markers.
Test Principle
When a sample is mixed with R1 buffer and R2 latex solution, human myoglobin reacts specifically with anti-human myoglobin antibodies coated on the
latex particles, to yield insoluble aggregates. The absorbance of these aggregates is proportional to the myoglobin concentration in the sample.
Glycine Buffer (pH 9.0) 92 mmol/L
Latex particles coated with anti-human myoglobin (pH 7.3) Variable
Preservative
Reagent Preparation
R1 is ready for use and can be placed directly on board the instrument. R2 should be mixed by inversion 5 - 10 times before placing on board the
90 days.
Specimen
5
Serum: Stable in serum for 1 week when stored at 2…8°C and 2 days when stored at 15…25°C.
Test Procedure
Calibration
Myoglobin Calibrator Cat. No. ODR3025.
Calibrators are traceable to highly purified human cardiac myoglobin and values assigned using a commercially available RIA method.
Quality Control
Control materials of human origin with values determined by this Beckman Coulter system may be used. Due to the lack of international standardisation for
myoglobin, values obtained may differ significantly from other assays. Laboratories should establish their own mean and ranges for any QC prior to routine
use.
Calculation
The Beckman Coulter analysers automatically compute the myoglobin concentration of each sample.
Serum Male 19 – 92 µg/L
Female 12 – 76 µg/L
EN.01 BLOSR6x68.01 Specific Protien

2009-08
values.
Linearity
The test is linear within a concentration range of 30 – 800 µg/L.
Precision
45.15 1.30 2.89 1.88 4.17
377.15 2.57 0.68 9.46 2.51
693.76 5.70 0.82 18.16 2.62
Sensitivity
The lowest detectable level on an AU600 analyser was estimated at 3.14 µg/L.
The lowest detectable level represents the lowest measurable level of myoglobin that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient serum samples were used to compare this Myoglobin OSR6168 assay on the AU600 against another commercially available myoglobin assay.
y = 1.388x - 2.114 r = 0.997 n = 37 Sample range = 3.31 – 300.96 µg/L
®
7
Limitations
Samples containing heterophilic antibodies can cause falsely elevated results.
This assay has a prozone tolerance of 20,000 µg/L. Samples suspected of myoglobin concentrations greater than 20,000 µg/L should be diluted prior to
analysis.
† Myoglobin Calibrator Cat. No.: ODR3025
* Values set for working in µg/L
BIBLIOGRAPHY
1. Fairbanks VF, Klee GG. Biochemical aspects of hematology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders
Company, 1999;1689pp.
3. Puschendorf B, Mair J. Cardiac diseases. In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
4. Thygesen K, Alpert JS et al. Myocardial infarction redefined-A consensus document of the Joint European Society of Cardiology/American College of Cardiology
Committee for the redefinition of myocardial infarction. JACC 2000;36:959-969.

2009-08
MYOGLOBIN, AU400/AU640 Serum Application MYOGLOBIN, AU600 Serum Application

Test No # Test name MYO ∇ Sample type Ser ∇ Page ½
Test Name: MYO ∇ < > Type: Serum ∇ Operation: Yes ∇
Reagent OD limit: 1.5 Wave Main 570 Sub ∇ Lst. L -0.1 Lst. H 1.5
Method: FIXED ∇ Last L -0.1 Last H 1.5 Reaction + ∇ L 30* H 800*
General LIH ISE Range Test No # Test name MYO ∇ Sample type Ser ∇ Page 2/2
Test Name: MYO ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic Value: # # Unit: µg/L* Decimal places: # Panic value # #
SERUM APPLICATION
General ISE Test No # Test name MYO ∇
Test Name: MYO ∇ < > Type Serum ∇ Cal type 12 4AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
MB type factor
† Myoglobin Calibrator Cat. No.: ODR3025 † Myoglobin Calibrator Cat. No.: ODR3025
* Values set for working in µg/L. * Values set for working in µg/L.

2009-08
MYOGLOBIN, AU2700/AU5400 Serum Application MYOGLOBIN, AU680/AU480 Serum Application
Test Name: MYO ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: MYO ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Test Name: MYO ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: MYO ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: µg/L* Decimal places: # 7. No demographics # #
Unit µg/L* Decimal Places #
SERUM APPLICATION
General ISE
General ISE
Test Name: MYO ∇ < > Type Serum ∇ Test Name: MYO ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇
† Myoglobin Calibrator Cat. No.: ODR3025
* Values set for working in µg/L Point-1 ∇ Reagent Blank 999 Day 0 Hour

2009-08
PREALBUMIN
OSR6175 4 x 15 mL R1
4 x 6.5 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of prealbumin in human serum on Beckman Coulter analysers. For in vitro diagnostic use only.
Summary1,2
Prealbumin (transthyretin), a nonglycosylated tetrameric protein composed of four identical subunits is primarily synthesised in the liver. It binds and
transports approximately 10% of both serum thyroxine and triiodothyronine and also plays a significant role in the metabolism of vitamin A by complexing
with retinol-binding protein. Because of its relatively short half life, a high tryptophan content, a high proportion of essential to non essential amino acids,
and small pool size, prealbumin is an excellent indicator of protein status (levels fall during periods of protein malnutrition).It is a negative acute phase
reactant whose levels also fall in inflammation and malignancy as well as in cirrhosis of the liver and protein wasting diseases of the gut or kidneys,
because of decreased synthesis and to a lesser extent increased degradation. Other acute phase proteins may be assayed to differentiate whether a
decrease in prealbumin is due to malnutrition or inflammation. A number of genetic variants have been described of which most are associated with
extracelluar deposition of amyloid fibrils in various tissues. Levels of prealbumin increase in Hodgkin's disease.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, human prealbumin reacts specifically with the anti-human prealbumin antibodies to
yield insoluble aggregates. The absorbance of these aggregates is proportional to the prealbumin concentration in the sample.

Solution of polymers in phosphate buffered saline (pH 7.1 – 7.3)
Rabbit anti-human prealbumin antibodies Variable
Preservative

Reagent Preparation

90 days.
Specimen
3
Serum: Stable for 6 months when stored at 2…8°C and 3 days when stored at 15…25°C.
Test Procedure
Calibration
Prealbumin Calibrator Cat. No. ODR3029.
The calibrator prealbumin values are traceable to IFCC (International Federation of Clinical Chemistry) standard CRM 470.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the prealbumin concentration of each sample.
Serum (Adults) 0.2 – 0.4 g/L (20 – 40 mg/dL)
2009-08
values.
Linearity
Precision
0.06 0.001 2.06 0.001 3.10
0.45 0.01 1.12 0.01 1.86
0.70 0.01 1.90 0.02 2.76
Sensitivity
The lowest detectable level on an AU600 analyser was estimated at 0.004 g/L.
The lowest detectable level represents the lowest measurable level of prealbumin that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient serum samples were used to compare this Prealbumin OSR6175 assay on the AU600 against another commercially available prealbumin assay.
®
5
Limitations

† Prealbumin Calibrator Cat. No.: ODR3029
BIBLIOGRAPHY
1. Johnson AM, Rohlfs EM, Silverman LM. Proteins. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders
Company, 1999;500-501.
rd
2. Tietz NW, ed. Clinical guide to laboratory tests, 3 ed. Philadelphia: WB Saunders Company,1995:608-609.
4. Baudner S, Dati F. Standardization of the measurement of 14 proteins in human serum based on the new IFCC/BCR/CAP international reference
material CRM 470. J Lab Med 1996;20:145-152.
th

2009-08
PREALBUMIN, AU400/AU640 Serum Application PREALBUMIN, AU600 Serum Application

Test No # Test name PALB ∇ Sample type Ser ∇ Page ½
Test Name: PALB ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name PALB ∇ Sample type Ser ∇ Page 2/2
Test Name: PALB ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
SERUM APPLICATION
General ISE Test No # Test name PALB ∇
Test Name: PALB ∇ < > Type Serum ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
1-Point Cal. Point: ο With CONC-0 Slope Check: + ∇ Advanced Calibration: # ∇ 1-point cal. Point
MB type factor
# User defined
* Values set for working in SI units (g/L) . To work in mg/dL multiply by 100
* Values set for working in SI units (g/L) . To work in mg/dL multiply by 100

2009-08
PREALBUMIN, AU2700/AU5400 Serum Application PREALBUMIN, AU680/AU480 Serum Application
Test Name: PALB ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: PALB ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: PALB ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: PALB ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
SERUM APPLICATION
General ISE
General ISE
Test Name: PALB ∇ < > Type Serum ∇ Test Name: PALB ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇
† Prealbumin Calibrator Cat. No.: ODR3029 Calibrator OD Conc Low High Stability
* Values set for working in SI units (g/L) . To work in mg/dL multiply by 100 Point-1 ∇ Reagent Blank 999 Day 0 Hour

2009-08
RF Latex
OSR61105 4 x 24 mL R1
4 x 8 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of RF (rheumatoid factor) antibodies in human serum and plasma on Beckman
Summary1,2
Rheumatoid factors (RF) are antibodies directed against antigenic determinants on the Fc fragment of IgG. These are usually IgM antibodies,
but may be IgG, IgA or IgE. Rheumatoid factor sensitivity in rheumatoid arthritis varies from 30% in population-based studies to 70 – 80% in
hospital-based studies, where the disease tends to be more severe. Higher titres of RF are more specific for the diagnosis of RA and are more
common in patients with rapidly progressive joint destruction and in those with extraarticular manifestations such as subcutaneous rheumatic
nodules. However, RF is a non-specific test and a positive RF is observed in 1 – 5% of the healthy population at low titres and in 15 – 20% of
elderly subjects with other chronic disease states. A positive RF is also seen in autoimmune rheumatic diseases and in non-rheumatic
conditions with variable frequency e.g. SLE, Sjögren’s syndrome, subacute bacterial endocarditis and other bacterial infections, infectious
hepatitis, chronic liver diseases, chronic active pulmonary diseases, parasitic infections and viral infections.
Test Principle
When a sample is mixed with R1 buffer and R2 IgG latex suspension, RF reacts specifically with IgG coated on the latex particles to yield
insoluble aggregates. The absorbance of these aggregates is proportional to the RF concentration in the sample.
Glycine buffer (pH 8.0) 170 mmol/L
Latex coated with human IgG < 0.5%
Preservative 0.09%
Biological materials of human origin contained in R2 were tested for Anti-HCV, HbsAg and Anti-HIV 1/2 on a single donor basis using FDA
approved methods and were found to be non-reactive. As there is no known test method that can offer complete assurance that products
Reagent Preparation
R1 is ready for use and can be placed directly on board the instrument. R2 should be mixed by inversion 5 – 10 times before placing on board
the instrument and at weekly intervals thereafter.
Specimen
Serum and Li-/Na-heparin, Na-/K-EDTA & citric acid plasma.
3
Stable in serum and plasma for:
1 day at 20…25°C
8 days at 2…8°C
3 months at -20°C (avoid repeated freezing and thawing)
Test Procedure
statement. Data check parameters are required, see setting sheets for specific instrument details.
Calibration
RF Latex Calibrator Cat. No.: ODC0028.
4
The calibrator RF value is traceable to WHO International reference material, NIBSC 64/2.
Quality Control
ITA Control Sera ODC0014, ODC0015 and ODC0016 or other control materials with values determined by this Beckman Coulter system may
be used.

2009-08
Calculation
The Beckman Coulter analysers automatically compute the RF concentration of each sample.
Adult ≤ 14 IU/mL
th
This value is based on serum samples from 144 test subjects (97.5 percentile).
findings.
from these values.
Linearity
The test is linear within a concentration range of 10 – 120 lU/mL.
Precision
The following data was obtained for the RF (Latex) assay on an AU640, AU400 and AU2700 using 4 serum pools analysed over 20 days.
AU640:
10.26 0.38 3.72 0.77 7.51
20.06 0.45 2.24 0.79 3.96
76.87 0.75 0.97 2.04 2.66
114.81 0.93 0.81 2.97 2.59
AU400:
9.90 0.30 3.07 0.51 5.11
20.21 0.27 1.35 0.60 2.96
78.71 0.54 0.69 1.41 1.79
117.65 0.61 0.52 1.30 1.11
AU2700:
9.99 0.46 4.63 0.79 7.89
19.74 0.47 2.39 0.64 3.25
75.37 0.47 0.62 0.88 1.16
112.93 0.98 0.87 1.24 1.10
Sensitivity
Analyzer Lowest Detectable Level (IU/mL)
AU2700 1.38
AU640 1.42
AU400 2.22
The lowest detectable level represents the lowest measurable level of RF that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient serum samples were used to compare this RF Latex OSR61105 assay on the AU640 against other commercially available RF Latex
assay (Method 2). Results of linear regression analysis were as follows:
y = 0.996x - 1.217 r = 0.996 n = 55 Sample range = 6.30 – 103.20 IU/mL
®
In very rare cases gammopathy, in particular type IgM (Waldenstrom’s macroglobulinemia), may cause unreliable results.
7
Limitations
Samples with very high RF concentrations (> 1500 IU/mL) can generate false low results without appropriate "Z" flags due to excess antigen in the sample.
† RF Latex Calibrator Cat. No.: ODC0028
§ For the zero calibrator saline should be used
BIBLIOGRAPHY
2. Mierau R, Genth E. Autoantibodies in rheumatoid arthritis. In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results.
3. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of Anticoagulants in Diagnostic Laboratory Investigations and Stability of Blood, Plasma and Serum
Samples. WHO/DIL/LAB/99.1 Rev.2:41pp.
4. Anderson SG, Bentzon MW, Houba V, Krag P. International reference preparation of rheumatoid arthritis serum. Bull Wld Hlth Org 1970;42: 311-318.
5. In-house data on file
6. Tietz NW, ed. Clinical guide to laboratory tests, 2nd ed. Philadelphia: WB Saunders Company,1990.

2009-08
RF Latex, AU400/AU640 Serum/Plasma Application RF Latex, AU600 Serum/Plasma Application

Test No # Test name RF ∇ Sample type Ser ∇ Page ½
Test Name: RF ∇ < > Type: Serum ∇ Operation: Yes ∇
Fst. L -2.0 Fst. H 2.5
Wave Main 660 Sub None ∇ Lst. L -2.0 Lst. H 2.5
Wavelength: Pri. 660 ∇ Sec. NONE ∇ First L -2.0 First H 2.5 Reaction + ∇ L 10* H 120*
Test No # Test name RF ∇

Test Name: RF ∇ < > Type Serum ∇ Selection calibrator
Calibration Type: 6AB ∇ Formula: SPLINE ∇ Counts: # Process: CONC ∇ Point 1 # ∇ 0§ -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 1: # 0§ -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 6 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 7 ∇
1-point cal. Point
Point 7:
Test No # Name RF ∇ Type Ser ∇

Test Name: RF ∇ < > Type: Serum ∇
Limit Point 1 0
Limit Point 2 0

† Latex Calibrator Cat. No.: ODC0028 † Latex Calibrator Cat. No.: ODC0028
* Values set for working in SI IU/mL * Values set for working in IU/mL
§ For the zero calibrator saline should be used § For the zero calibrator saline should be used

2009-08
RF Latex, AU2700/AU5400 Serum/Plasma Application RF Latex, AU680/AU480 Serum/Plasma Application
Test Name: RF ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: RF ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Wavelength: Pri. 660 ∇ Sec. NONE ∇ First L -2.0 First H 2.5
Test Name: RF ∇ < > Type: Serum ∇ General ISE
Calibration Type: 6AB ∇ Formula: SPLINE ∇ Counts: # Process: CONC ∇ Test Name: RF ∇ < > Type Serum ∇ ο Use Serum Cal.

Point 1: # 0§ -0.1 2.5
Point 2: # † -0.1 2.5
Point 1: # ∇ 0§ -0.1 2.5
Point 3: # † -0.1 2.5
Point 4: # † -0.1 2.5
Point 3: # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5
Point 6: # † -0.1 2.5 Point 5: # † -0.1 2.5
∇ ο Calibration
Point 7: Point 6: # † -0.1 2.5
∇
1-Point Cal. Point: ο with CONC-0 Slope Check: + ∇ Advanced Calibration # ∇ Point 7 ∇ Advanced Calibration
Point 9 ∇
Test Name: RF ∇ < > Type: Serum ∇ ∇
Decision Value-3: 0.1500 Limit Point 2: Limit Point 2: Test Name: RF ∇ < > Type: Serum ∇
Limit Point 1: 11
Check Point-3: 27
# User defined Decision Value-2: 2.0000 Decision Value-2: Decision Value 2:
† RF Latex Calibrator Cat. No.: ODC0028
§ For the zero calibrator saline should be used.
Ф AU680 Limit Point 2: 14 Limit Point 2: Limit Point 2:

2009-08
TRANSFERRIN
OSR6152 4 x 7 mL R1
4 x 8 mL R2
Intended Use
Immuno-turbidimetric test for the quantitative determination of transferrin in human serum and plasma on Beckman Coulter analysers. For in vitro
Summary1,2
Transferrin is the principle plasma protein for the transport of iron. Transferrin has two binding sites for iron; they are strong at physiological pH but weaken
with decreasing pH. Transferrin is largely but not exclusively synthesised by the liver.
Measurement of plasma transferrin levels is useful in the differential diagnosis of anemia, and will rise with iron deficiency anemia. In congenital
atransferrinemia, a very low level of transferrin is accompanied by iron overload and a severe hypochromic anemia resistant to iron therapy. High levels of
transferrin occur in pregnancy and during oestrogen administration. It is decreased in conditions, which are associated with increased protein loss such as
nephrotic syndrome, protein-deficiency states, and in chronic liver disease. Transferrin is a negative acute phase reactant and will decrease during any
inflammatory state or malignancy.
Transferrin is often used in conjunction with iron to calculate transferrin saturation.
Test Principle
When a sample is mixed with R1 buffer and R2 antiserum solution, human transferrin reacts specifically with anti-human transferrin antibodies to yield
insoluble aggregates. The absorbance of these aggregates is proportional to the transferrin concentration in the sample.

Goat anti-transferrin antibodies Variable
Preservative

Reagent Preparation

for 90 days.
Specimen
3
Stable in serum and plasma for 8 months when stored at 2…8 °C and 4 months when stored at 15…25 °C.
Test Procedure
Calibration
®
The calibrator values are traceable to ERM – DA470.
practice.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the transferrin concentration of each sample.

2009-08
Serum (Adults) 2.0 – 3.6 g/L (200 – 360 mg/dL)
Serum (Children)
0 − 4 day 1.30 – 2.75 g/L (130 – 275 mg/dL)
3 month – 10 year 2.03 – 3.6 g/L (203 – 360 mg/dL)
values.
Linearity
The test is linear within a concentration range of 0.75 − 7.50 g/L (75 − 750 mg/dL).
Precision
1.47 0.01 0.62 0.01 0.96
2.84 0.02 0.64 0.02 0.86
6.51 0.06 0.88 0.08 1.28
Sensitivity
The lowest detectable level represents the lowest measurable level of transferrin that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient serum samples were used to compare this Transferrin OSR6152 assay on the AU400 against another commercially available transferrin assay.
®
6
Limitations

BIBLIOGRAPHY
1. Grant GH, Silverman LM, Christenson RH. Amino Acids and proteins. In: Teitz NW, ed. Fundamentals of clinical chemistry. Philadelphia: WB Saunders
Company, 1987:333-334pp.
2. Thomas L. Transferrin saturation (TfS). In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
TH-Books Verlagsgesellschaft, 1998:275-277pp.
4. Baudner S, Dati F. Standardization of the measurement of 14 proteins in human serum based on the new IFCC/BCR/CAP International reference material CRM
470. J Lab Med 1996;20:145-152pp.

2009-08
TRANSFERRIN, AU400/AU640 Serum/Plasma Application TRANSFERRIN, AU600 Serum/Plasma Application

Test No # Test name TRF ∇ Sample type Ser ∇ Page ½
Test Name: TRF ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name TRF ∇ Sample type Ser ∇ Page 2/2
Test Name: TRF ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name TRF ∇
Test Name: TRF ∇ < > Type Serum ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
1-Point Cal. Point: with CONC-0 Slope Check: + Advanced Calibration: # 1-point cal. Point
ο ∇ ∇
MB type factor

2009-08
TRANSFERRIN, AU2700/AU5400 Serum/Plasma Application TRANSFERRIN, AU680/AU480 Serum/Plasma Application
Test Name: TRF ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: TRF ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: TRF ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: TRF ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: TRF ∇ < > Type Serum ∇ Test Name: TRF ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇

2009-08
TRANSFERRIN, AU680/AU480 Mastercurve Serum/Plasma Application
Test Name: TRF ∇ < > Type: Serum ∇ Operation Yes ∇

Method END ∇

Test Name: TRF ∇ < > Type: Serum ∇

ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
General ISE
Test Name: TRF ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: ∇ 0.95*
Point 2: ∇ 2.63* Allowance Range Check
Point 3: ∇ 4.13*
Point 4: ∇ 5.75* ο Reagent Blank
Point 5: ∇ 7.21* ο Calibration
Point 6: ∇
Point 9 ∇
Point-2 ∇ Calibration 999 Day 0 Hour # User defined
ф AU680

2009-08
TRANSFERRIN, AU400/AU640 Paediatric Application TRANSFERRIN, AU600 Paediatric Application

Test No # Test name TRFP ∇ Sample type Ser ∇ Page ½
Test Name: TRFP ∇ < > Type: Serum ∇ Operation: Yes ∇
General LIH ISE Range Test No # Test name TRFP ∇ Sample type Ser ∇ Page 2/2
Test Name: TRFP ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
General ISE Test No # Test name TRFP
∇
Test Name: TRFP ∇ < > Type Serum ∇ Cal type 13 5AB ∇ Count #
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
MB type factor

2009-08
TRANSFERRIN, AU2700/AU5400 Paediatric Application TRANSFERRIN, AU680/AU480 Paediatric Application
Test Name: TRFP ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: TRFP ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: TRFP ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: TRFP ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: TRFP ∇ < > Type Serum ∇ Test Name: TRFP ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 9 ∇

2009-08
Carbamazepine
2x R1 Lyo
2 x 16 mL R2 Buffer
2x R2 Lyo
Intended Use
Homogeneous enzyme immunoassay for the quantitative determination of carbamazepine in human serum and plasma on Beckman Coulter analysers.
Summary
1
Carbamazepine is an anticonvulsant drug, used in particular for the treatment of trigeminal neuralgia , all forms of partial epilepsy, generalised tonic-clonic
2-4
seizures, and simple and complex partial seizures. Effective serum concentration of carbamazepine is essential for seizure control. However, serum
5,6
carbamazepine concentrations show only a moderate correlation to dose, due to individual differences in absorption, metabolism and clearance.
6
Moreover, co-administration of other antiepileptic agents can significantly increase serum carbamazepine levels.
2-4
Toxicity of carbamazepine associated with therapy may or may not be dose related. However, the central nervous system symptoms of vertigo,
dizziness, and diplopia are dose-related with chronic therapy. In conjunction with other clinical information, monitoring carbamazepine levels will provide
physicians with an effective tool to aid in adjusting dosage and achieving optimal therapeutic effect while avoiding both subtherapeutic and toxic drug
levels.
Test Principle7
This assay is based on the bacterial enzyme β-galactosidase, which has been genetically engineered into two inactive fragments. These fragments
spontaneously reassociate to form fully active enzyme that, in the assay format, cleaves a substrate, generating a colour change that can be measured
spectrophotometrically.
In the assay, analyte in the sample competes with analyte conjugated to one inactive fragment of β-galactosidase for antibody binding site. If analyte is
present in the sample, it binds to antibody, leaving the inactive enzyme fragments free to form active enzyme. If analyte is not present in the sample,
antibody binds to analyte conjugated on the inactive fragment, inhibiting the reassociation of inactive β-galactosidase fragments, and no active enzyme will
be formed.
The amount of active enzyme formed and resultant absorbance change are directly proportional to the amount of drug present in the sample.
Reagent Composition
R1 Buffer MOPS (3-(N-morpholino)propanesulfonic acid buffer), mouse monoclonal anti-carbamazepine antibodies, stabiliser and preservative.
R1 Lyo Enzyme acceptor, stabiliser, buffer salts and preservative.
R2 Buffer MES (2-(N-morpholino)ethanesulfonic acid buffer) and preservative.
R2 Lyo Enzyme donor conjugated to carbamazepine, chlorophenol red-β-D-galactopyranoside, buffer salts, stabiliser and preservative.

Harmful. Contains sodium azide. R22; Harmful if swallowed.
Safety Phrases:

Reagent Preparation
Remove the kit from refrigerated storage (2…8°C) immediately prior to preparation of the solutions.
Prepare the R2 solution before the R1 solution to minimise possible contamination.

R2 (Enzyme donor solution):
Connect the R2 lyo bottle to the R2 buffer bottle using one of the enclosed adapters. Mix by gentle inversion, ensuring that all the lyophilised material from
the R2 lyo bottle is transferred into the R2 buffer bottle. Avoid the formation of foam. Detach the R2 lyo bottle and adapter from the R2 buffer bottle and
discard. Cap the R2 buffer bottle and let stand for approximately 5 minutes at 15…25°C. Mix again. Record the reconstitution date on the bottle label. Place
the bottle directly into the reagent compartment of the analyser or into refrigerated storage (2…8°C) and let stand for at least 30 minutes. Ensure the
reagent is homogeneous before use.
R1 (Enzyme acceptor solution):

NOTE 1: The components supplied in this kit are intended for use as an integral unit. Do not mix components from different lots.
NOTE 2: Avoid cross-contamination of reagents by matching reagent caps to the appropriate reagent bottle. The R2 working solution (enzyme donor)
should be yellow-orange in colour. A dark red or purple-red colour indicates that the reagent has been contaminated and must be discarded.
NOTE 3: The R1 and R2 solutions must be at the reagent compartment temperature of the analyser before performing the assay.
NOTE 4: To ensure reconstituted R1 stability, protect from prolonged, continuous exposure to bright light.
NOTE 5: Do not freeze reconstituted reagents.
EN.01 BLOSR6414.01 TDM

2009-08
The reagents are stable, unopened, up to the stated expiry date when stored at 2...8°C.
Once reconstituted, reagents stored on board the instrument are stable for 60 days.
Specimen
Serum or plasma (Na or Li heparin; Na EDTA) may be used.
Stable in serum and plasma for 7 days when stored at 2…8°C and 4 weeks when stored at -20˚C.
Avoid repeated freezing and thawing.
Test Procedure
Calibration
Core TDM Multi-calibrator, Cat. No. ODC6411.
The calibrator values provided in the calibrator package insert are traceable to a primary gravimetric standard.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the carbamazepine concentration of each sample.
Expected Values
4,8
The effective serum/plasma carbamazepine concentrations for seizure control have been reported as 8 – 12 µg/mL for adults receiving carbamazepine
5,9-12
as the sole antiepileptic agent. Other therapeutic ranges for carbamazepine have also been suggested
The expected range provided is only a guide for drug dosage. Prior to the adjustment of dose, results should always be assessed in conjunction with other
available information such as patients medical history, mode of drug administration, timing of sample collection, other drug treatments and clinical
13
symptoms.

values.
Linearity
The test is linear within a concentration range of 0.5 µg/mL and the value of the highest calibrator (approximately 20 µg/mL or 84.6 µmol/L).
Specimen results greater than the highest calibrator can be reported as greater than the value of the highest calibrator or diluted one part sample with one
part Core TDM Multi-Calibrator 1 and reassayed. The value obtained on reassay should be derived as follows:
Actual Value = (2 x diluted value) – Concentration of Core TDM Multi-Calibrator 1.
Samples reported as less than the analytical range can be confirmed by diluting one part sample of known value with one part of the original patient
sample. The assayed result of this dilution, when multiplied by 2, should approximate the original value of the known sample to confirm the low patient
result. The confirmed result should be reported as < 0.28 µg/mL. If the assayed result of the first dilution, when multiplied by 2, does not approximate the
original result of the known sample, further dilutions utilising saline are needed.
Precision
The following data was obtained on an AU2700 using serum pools and control sera. Two replicates of each sample were analysed twice daily over
20 days.
Mean µg/mL SD CV% SD CV%
4.2 0.14 3.3 0.28 6.7
7.1 0.14 2.0 0.34 4.8
12.1 0.22 1.8 0.50 4.1
Method Comparison
Patient serum samples were used to compare this Carbamazepine OSR6414 assay on the AU640 against another commercially available carbamazepine
y = 0.942x – 0.07 r = 0.997 n = 135 Sample range = 0.5 – 26.4 µg/mL
Sensitivity
The lowest detectable level on an AU2700 analyser was estimated at 0.28 µg/mL.
The lowest detectable level (LDL) represents the lowest measurable level of carbamazepine that can be distinguished from zero. It is calculated as the
Interference
No significant interference (<10%) was observed with the following substances:
Substance Concentration Substance Concentration
Bilirubin ≤ 1026 µmol/L Total Protein ≤ 120 g/L
Haemoglobin ≤ 10 g/L Triglyceride ≤ 11.3 mmol/L
Rheumatoid Factor ≤ 180 IU/mL
TDM BLOSR6414.01 EN.01
2009-08
Specificity
The following compounds when tested with the Carbamazepine assay, yielded the following percent cross-reactivity results:
Compound Concentration Tested (µg/mL) % Cross reactivity
Amitriptyline 100 18.6
Carbamazepine-10,11-epoxide 250 7.4
Diazepam 250 4.8
Imipramine 200 5.6
Methsuximide 1000 1.0
Nortriptyline 50 17.2
Phenothiazine 200 8.6
Probenecid 500 2.0
Limitations
• Samples containing antibodies to E. coli β-galactosidase may result in artificially high results which will not fit the clinical profile. The incidence of
patients having such antibodies is extremely low.
• As with any assay employing mouse antibodies, the possibility exists for interference by human anti-mouse antibodies (HAMA) in the sample, which
could cause falsely elevated results.

† Core TDM Multi-calibrator Cat. No.: ODC6411
* Values set for working in µg/mL. To work in SI units (µmol/L) multiply by 4.23
BIBLIOGRAPHY
1. Blom S.: Trigeminal neuralgia: its treatment with a new anticonvulsant drug (G-32883) Lancet, Is. 1962; 839-840.
2. Eadie MJ, Tyler JH Anticonvulsant Therapy: Pharmacological Basis and Practice. In: Churchhill Livingstone, Edinburgh. Great Britain, 1974: Chapter 7.
3. Penry JK, Newmark ME. The use of antiepileptic drugs. Annals of Internal Medicine, 1979; 90: 207-218.
4. Scheuer ML, Pedley TA. The evaluation and treatment of seizures. N. Engl. J. Med., 1990; 322 (21):1468-1474.
5. Larkin JG, Herrick AL, McGuire GM, Percy-Robb IW, Brodie MJ. Antiepileptic Drug Monitoring at the Epilepsy Clinic: A Prospective Evaluation. Epilepsia, 1991;
32: 89-95.
6. Altafullah I, Talwar D, Loewenson R, Olson K, Lockman LA. Factors Influencing Serum Levels of Carbamazepine and Carbamazepine-10, 11-epoxide in
children. Epilepsy 1989; Res. 4: 72-80.
7. Henderson DR, Friedman SB, Harris JD, et al. CEDIA™, a new homogeneous immunoassay system. Clin Chem. 1986; 32:1637-1641.
8. Troupin A, Ojemann LM, Halpern L, Dodrill C, Wikus R, Friel P. Carbamazepine - a double blind comparison with phenytoin. Neurology 1977; 27: 511-519.
9. Strandjord RE, Johannessen SI. Single-drug therapy with carbamazepine in patients with epilepsy: serum levels and clinical effects. Epilepsia 1980; 21: 655-
662.
10. Simonsen H, Olsen PZ, Khul V, Lund M, Wendelboe J. A comparative controlled study between carbamazepine and diphenylhydantoin in psychomotor
epilepsy. Epilepsia 1976; 17: 169-176.
11. Shorvon SD, Chadwick D, Galbraith AW, Reynolds EH. One drug for epilepsy. Br. Med. J. 1978;1:474-476.
12. MacKichan JJ, Kutt H. Carbamazepine: Therapeutic use and serum concentration monitoring. In: Taylor WJ, Finn AL eds. Individualizing Drug Therapy:
Practical Applications of Drug Monitoring. New York: Gross, Townsend, Frank, Inc., 1981:1-25.
13. Hallworth M, Capps N, eds. Therapeutic drug monitoring and clinical biochemistry, London:ACB Venture Publications, 1993.

2009-08
CARBAMAZEPINE, AU400/AU640 Serum/Plasma Application CARBAMAZEPINE, AU600 Serum/Plasma Application

Test No # Test name CBZ ∇ Sample type Ser ∇ Page 1/2
Test Name: CBZ ∇ < > Type: Serum ∇ Operation: Yes ∇
Wavelength: Pri. 570 ∇ Sec. 660 ∇ First L -2.00 First H 2.50 Method RATE1 ∇ Dynamic range
Method: RATE1 ∇ Last L -2.00 Last H 2.50 Reaction + ∇ L 0.5* H 20*
Reaction slope: + ∇ Dynamic Range: Point 1 Fst 24 Lst 27 Correlation factor A 1.04
Linearity : % A 1.04 B 0
No Lag Time: NO ∇ On-board stability period: 60 Linearity Fst % Sec %
General LIH ISE Range Test No # Test name CBZ ∇ Sample type Ser ∇ Page 2/2
Test Name: CBZ ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic Value: # # Unit: µg/mL* Decimal places: # Panic value # #
General ISE Test No # Test name CBZ ∇
Test Name: CBZ ∇ < > Type Serum ∇ Cal type 9 AA Count #
∇
Calibration Type: AA ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇
Point 1 # ∇ † -999999 9999999
Point 1: # † -999999 9999999
Point 2 # ∇ † -999999 9999999
Point 2: # † -999999 9999999
MB type factor
# User defined
† Core TDM Multi-Calibrator Cat. No.: ODC6411
† Core TDM Multi-Calibrator Cat. No.: ODC6411
TDM BSOSR6414.01
2009-08
CARBAMAZEPINE, AU2700/AU5400 Serum/Plasma Application CARBAMAZEPINE, AU680/AU480 Serum/Plasma Application
Test Name: CBZ ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: CBZ ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Method: RATE1 ∇ Last L -2.00 Last H 2.50
Measuring Point 1: First 24 Last 27 L 0.5* H 20* Wavelength Pri 570 ∇nm Sec. 660 ∇nm Factor for Maker A 1.04 B 0
Measuring Point 2: First Last Correlation Factor: Method RATE1 ∇
Linearity : % A 1.04 B 0 Reaction Slope + ∇ Onboard Stability Period 60 Day 0 Hour
Test Name: CBZ ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: CBZ ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: µg/mL* Decimal places: # 7. No demographics # #
Unit µg/mL* Decimal Places #
General ISE
General ISE
Test Name: CBZ Type Serum
∇ < > ∇ Test Name: CBZ Type Serum Use Serum Cal.
∇ < > ∇ ο
Calibration Type: AA ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Calibration Type: AA ∇ Formula: Y=AX+B ∇ Counts: # ∇
Point 1: # † -999999 9999999 Point 1: # ∇ † -999999 999999
Point 2: # † -999999 9999999 Point 2: # ∇ † Allowance Range Check
Point 9 ∇
† Core TDM Multi-Calibrator Cat. No.: ODC6411 Calibrator OD Conc Low High Stability
* Values set for working in µg/mL. To work in SI units (µmol/L) multiply by 4.23 Point-1 ∇ Reagent Blank 999 Day 0 Hour
TDM BSOSR6414.01
2009-08
DIGITOXIN
OSR6403 2 x 21 mL R1
2 x 12.8 mL R2
Intended Use
Immuno-inhibition test for the quantitative determination of digitoxin in human serum on Beckman Coulter analysers. For in vitro diagnostic use
only.
Summary
Digitoxin is a naturally occurring cardiac glycoside that is used to treat congestive heart failure and atrial fibrillation. Its structure is the same as
digoxin except for the loss of the C-12 hydroxyl group. This difference results in digitoxin being more lipid soluble and highly bound to protein
with the effect that it has a relatively long half life of 5-7 days and requires higher concentrations to achieve therapeutic effectiveness. Cardiac
1
glycosides have a low therapeutic ratio (a very small difference between therapeutic and tissue toxic levels). There is a marked patient
2
variability in response to the same dose of drug, often resulting in unpredictable serum drug levels. Intoxication symptoms are often
indistinguishable from the original condition for which the drug was prescribed. It may not be immediately apparent whether the patient has been
under or overdosed. Monitoring serum digitoxin levels combined with other clinical data can provide the physician with useful information to aid
3,4
in adjusting patient dosage to achieve optimal therapeutic effect while avoiding useless subtherapeutic or harmful toxic dosage levels.
Test Principle
R1 contains a monoclonal antibody specific for digitoxin that binds to microparticles covalently coated with a digitoxin derivative in R2. The
resulting agglutination reaction causes an increase in turbidity, which can be measured spectrophotometrically at 700 nm. Digitoxin from the
sample competes with the microparticle-bound digitoxin for the antibody, thus inhibiting the agglutination reaction.
Monoclonal anti-digitoxin antibody Variable
Digitoxin – derivative coated microspheres
Reagent Preparation
R1 is ready for use and can be placed directly on board the instrument. R2 should be mixed by inversion 5 – 10 times before placing on board
the instrument and at fortnightly intervals thereafter.
Specimen5
Serum: Stable in serum for 3 months when stored at 2…8°C and 2 weeks when stored at 15…25°C.
Test Procedure
Calibration
Digitoxin Calibrator Cat. No. ODC6403.
These calibrators are gravimetrically prepared then verified by a commercially available process.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the digitoxin concentration of each sample.
Therapeutic Range6
Effective levels 13 – 39 nmol/L (10 – 30 ng/mL)
Toxic Levels >39 nmol/L (30 ng/mL)
Consider these ranges as guidelines only.
from these values.
2009-08
Linearity
The test is linear within a concentration range of 0 – 118 nmol/L (0 - 90 ng/mL).
Samples reported as less than the analytical range can be confirmed by diluting one part sample of known value with one part of the original
patient sample. The assayed result of this dilution, when multiplied by 2, should approximate the original value of the known sample to confirm
the low patient result. The confirmed result should be reported as < 1.2 nmol/L. If the assayed result of the first dilution, when multiplied by 2,
does not approximate the original result of the known sample, further dilutions utilising saline are needed.
Precision
Mean nmol/L SD CV% SD CV%
15.62 0.50 3.21 0.66 4.22
63.71 0.61 0.96 1.36 2.14
111.94 1.23 1.09 1.82 1.63
Sensitivity
The lowest detectable level on an AU640 analyser was estimated at 1.179 nmol/L.
The lowest detectable level represents the lowest measurable level of digitoxin that can be distinguished from zero. It is calculated as the
Method Comparison
Patient serum samples were used to compare this Digitoxin OSR6403 assay on the AU640 against another commercially available digitoxin
y = 1.055x – 1.470 r = 0.961 n = 108 Sample range = 4.59 – 62.36 nmol/L
®
RF: Interference less than 10% up to 505 IU/mL RF
7
Specificity
The following compounds have been tested for cross reactivity in the assay.
Substance Conc. Tested ng/mL Cross-reaction (%)
Digitoxigenin 12.5 332.0
Digitoxigenin-bis-digitoxide 25 144.4
Digitoxigenin-mono-digitoxide 25 168.5
Dihydrodigitoxigenin 25 19.5
Digoxin 25 17.8
Digoxigenin-mono-digitoxide 25 18.4
Digoxigenin-bis-digitoxide 25 11.4
Digoxigenin 25 17.5
A cross-reaction of less than 10% was found for the following: Dihydrodigoxin, 20,22-Dihydrodigoxigenin, Prednisone, Testosterone, and
Oubain.
Limitations
As with any assay employing mouse antibodies, the possibility exists for interference by human anti-mouse antibodies (HAMA) in the sample,
which could cause falsely elevated results.
®
Digoxin Immune Fab, such as Digibind, will interfere with digitalis immunoassay measurements. Thus, the standard serum digitoxin
concentration measurement can be clinically misleading until the Fab fragment is eliminated from the body. Serum digitoxin concentration
® 8
should be obtained before administration of Digibind if at all possible.
In rare cases, samples from digitalised patients may yield clinically implausible values of 0.0 ng/mL digitoxin. In such cases the analysis must be
repeated following ultrafiltration of the samples (exclusion threshold: MW 100,000).
† System Calibrator Cat. No.: ODC6403
* Values set for working in SI units (nmol/L). To work in ng/mL divide by 1.31.
BIBLIOGRAPHY
1. Bresnahan JF, Vliesta RE. Series on pharmacology in practice 3. Digitalis glycosides. Mayo Clin Proc 1979;54:675-84.
2. Surawicz B, Mortelmans S. Factors affecting individual tolerance to digitalis. In: Fisch C, Surawiicz B, eds. Digitalis. New York: Grune and Stratton, 1969;127-47.
3. McNeely MDD. Making digitalis safer. Drug Ther 1975;5:222-4.
4. Taggart AJ, McDevitt DG. Digitalis: Its place in modern therapy. Drugs 1980;20:389-404.
6. Drug Monitoring Data (Pocket Guide II). Therapeutic Drug Monitoring and Toxicology Laboratory Improvement Program. American Association for Clinical
Chemistry, Inc., 2nd Ed. 1994 pp.61.
8. GlaxoSmithKline Product Information, August 2001.

2009-08
DIGITOXIN, AU400/AU640 Serum Application DIGITOXIN, AU600 Serum Application

Test No # Test name DGT ∇ Sample type Ser ∇ Page ½
Test Name: DGT ∇ < > Type: Serum ∇ Operation: Yes ∇
Wavelength: Pri. 700 ∇ Sec. None ∇ First L -0.1 First H 2.5 Method FIXED1 ∇ Dynamic range
Method: FIXED1 ∇ Last L -0.1 Last H 2.5 Reaction + ∇ L 0* H 118*
General LIH ISE Range Test No # Test name DGT ∇ Sample type Ser ∇ Page 2/2
Test Name: DGT ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic Value: # # Unit: nmol/L* Decimal places: # Panic value # #
SERUM APPLICATION
General ISE Test No # Test name DGT ∇
Test Name: DGT ∇ < > Type Serum ∇ Cal type 14 6AB ∇ Count #
Formula 6 EIA TYPE 1 ∇ Process Conc ∇
Calibration Type: 6AB ∇ Formula: EIA TYPE 1 ∇ Counts: # Process: CONC ∇
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
Point 6: # † -0.1 2.5 Point 6 # ∇ † -0.1 2.5
1-Point Cal. Point: ο with CONC-0 Slope Check: - ∇ Advanced Calibration: # ∇ 1-point cal. Point
MB type factor
# User defined
† Digitoxin Calibrator Cat. No.: ODC6403
† Digitoxin Calibrator Cat. No.: ODC6403
* Values set for working in SI units (nmol/L). To work in ng/mL divide by 1.31
TDM BSOSR6403.01
2009-08
DIGITOXIN, AU2700/AU5400 Serum Application DIGITOXIN, AU680/AU480 Serum Application
Test Name: DGT ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: DGT ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Method: FIXED1 ∇ Last L -0.1 Last H 2.5
Measuring Point 2: First Last Correlation Factor: Method FIXED1 ∇
Test Name: DGT ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: DGT ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: nmol/L* Decimal places: # 7. No demographics # #
SERUM APPLICATION
Unit nmol/L* Decimal Places #
General ISE
General ISE
Test Name: DGT ∇ < > Type Serum ∇ Test Name: DGT ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: 6AB ∇ Formula: EIA TYPE 1 ∇ Counts: # Process: CONC ∇ Calibration Type: 6AB ∇ Formula: EIA TYPE 1 ∇ Counts: # ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibrator OD Conc Low High Slope Check - ∇
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 6: # † -0.1 2.5 Point 6: # ∇ † -0.1 2.5
1-Point Cal. Point: ο with CONC-0 Slope Check: - ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
† Digitoxin Calibrator Cat. No.: ODC6403 Calibrator OD Conc Low High Stability
* Values set for working in SI units (nmol/L). To work in ng/mL divide by 1.31 Point-1 ∇ Reagent Blank 999 Day 0 Hour
TDM BSOSR6403.01
2009-08
DIGOXIN
OSR6404 2 x 17.6 mL R1
2 x 12.8 mL R2
Intended Use
Immuno-inhibition test for the quantitative determination of digoxin in human serum on Beckman Coulter analysers. For in vitro diagnostic use only.
Summary
Digoxin is a potent cardiac glycoside widely prescribed for the treatment of patients suffering from congestive heart failure, as well as some types of cardiac
arrhythmias. Digoxin intoxication is a common and serious problem in the clinical setting, which can result from a combination of factors. Cardiac
1
glycosides have a low therapeutic ratio (a very small difference between therapeutic and tissue toxic levels). There is a marked patient variability in
2
response to the same dose of drug, often resulting in unpredictable serum drug levels. Intoxication symptoms are often indistinguishable from the original
condition for which the drug was prescribed. It may not be immediately apparent whether the patient has been under or overdosed. Monitoring serum
digoxin levels combined with other clinical data can provide the physician with useful information to aid in adjusting patient dosage to achieve optimal
3,4
therapeutic effect while avoiding useless subtherapeutic or harmful toxic dosage levels.
Test Principle
R1 contains a monoclonal antibody specific for digoxin that binds to microparticles covalently coated with a digoxin derivative in R2. The resulting
agglutination reaction causes an increase in turbidity, which can be measured spectrophotometrically at 700 nm. Digoxin from the sample competes with
the microparticle-bound digoxin for the antibody, thus inhibiting the agglutination reaction.
Monoclonal anti-digoxin antibody ≤ 1.0 mg/L
BIS-TRIS buffer, pH 6.4 0.25 mol/L
Digoxin - derivative coated microspheres
Reagent Preparation
instrument and at fortnightly intervals thereafter.
30 days.
Specimen5
Serum: Stable in serum for 3 months when stored at 2…8°C and 2 weeks when stored at 15…25°C.
Test Procedure
Calibration
Digoxin Calibrator Cat. No. ODC6404.
These calibrators are gravimetrically prepared then verified by a commercially available process.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the digoxin concentration of each sample.
Therapeutic Range6
Effective levels 1.0 – 2.6 nmol/L (0.8 – 2.0 ng/mL)
Toxic levels > 3.1 nmol/L (2.4 ng/mL)
Consider these ranges as guidelines only.
values.
Linearity
The test is linear within a concentration range of 0 – 6.4 nmol/L (0 - 5 ng/mL).
2009-08
result. The confirmed result should be reported as < 0.2 nmol/L. If the assayed result of the first dilution, when multiplied by 2, does not approximate the
Precision
The following data was obtained on an AU2700 using 3 control sera analysed over 20 days.
Mean nmol/L SD CV% SD CV%
0.98 0.03 3.48 0.05 4.95
2.42 0.05 1.87 0.06 2.46
4.77 0.05 1.10 0.09 1.90
Sensitivity
The lowest detectable level on an AU400 analyser was estimated at 0.19 nmol/L.
The lowest detectable level represents the lowest measurable level of digoxin that can be distinguished from zero. It is calculated as the absolute mean
Method Comparison
Patient serum samples were used to compare this Digoxin OSR6404 assay on the AU400 against another commercially available digoxin assay. Results
y = 1.008x + 0.023 r = 0.976 n = 100 Sample range = 0.35 – 3.35 nmol/L
®
RF: Interference less than 5% up to 490 IU/mL RF
7
Specificity
Cross reactivity for the following substances was determined in accordance with the equation:
Cross-reactivity (%) =
Conc of analyte at 50% signal strength x 100
Conc of cross reactant at same signal level
Substance Cross-reaction (%)
Lanocide C 54
Digoxigenin Bisdigitoxoside 100
Digoxigenin Monodigitoxoside 38
Digoxigenin 13
Digitoxin 11.5
Dihydrodigoxin 2.5
Gitoxin 2
A cross-reaction of less than 1% was found for the following: Dehydroisoandrosterone, Digitonin, Digitoxigenin, D-Digitoxose,
11α-Hydroxyprogesterone, 17α-Hydroxyprogesterone, β-Estradiol, Estriol, Oubain, Prednisone, Prednisolone, Progesterone, Spironolacetone.
Limitations
As with any assay employing mouse antibodies, the possibility exists for interference by human anti-mouse antibodies (HAMA) in the sample, which could
cause falsely elevated results.
The sera from patients in specific patient populations (i.e. patients with renal and/or hepatic failure, newborn infants and pregnant women) have been
8-14
reported to contain an unidentified component that gives positive results with a number of immunoassays. This component has been termed digoxin-like
immunoreactive factor (DLIF). The presence of DLIF in a sample can result in falsely elevated digoxin assay results. The amount of DLIF in these patient
9-12
samples is extremely variable, but in some cases these levels have been shown to approach concentrations that are in the therapeutic range of digoxin.
®
Digoxin Immune Fab, such as Digibind , will interfere with digitalis immunoassay measurements. Thus, the standard serum digoxin concentration
measurement can be clinically misleading until the Fab fragment is eliminated from the body. Serum digoxin concentration should be obtained before
® 15
administration of Digibind if at all possible.
In rare cases, samples from digitalised patients may yield clinically implausible values of 0.0 ng/mL digoxin. In such cases the analysis must be repeated
following ultrafiltration of the samples (exclusion threshold: MW 100,000).
† Calibrator Cat. No.: ODC6404
* Values set for working in SI units (nmol/L). To work in ng/mL divide by 1.28.
BIBLIOGRAPHY
1. Bresnahan JF, Vliesta RE. Series on pharmacology in practice 3. Digitalis glycosides. Mayo Clin Proc 1979;54: 675-84.
2. Surawicz B, Mortelmans S. Factors affecting individual tolerance to digitalis. In: Fisch C, Surawiicz B, eds. Digitalis. New York: Grune and Stratton,
1969;127-47.
3. McNeely MDD. Making digitalis safer. Drug Ther 1975;5:222-4.
4. Taggart AJ, McDevitt DG. Digitalis: Its place in modern therapy. Drugs 1980; 20:389-404.
6. Drug Monitoring Data (Pocket Guide II). Therapeutic drug monitoring and toxicology laboratory improvement program. American Association for Clinical
Chemistry, Inc., 2nd ed. 1994:65pp.
8. Yatscoff RW, Desjardins PRE, Dalton JG. Digoxin-like immunoreactivity in the serum of neonates and uremic patients, as measured in the Abbott
TDx. Clin Chem 1984;30:588.
9. Greenway DC, Nanjii AA. Falsely increased results for digoxin in sera from patients with liver disease: Ten immunoassay kits compared. Clin Chem
1985;31:1078-9.
10. Rosenkranz B. Frolich JC. Falsely elevated digoxin concentrations in patients with liver disease. The Drug Monit 1985;7:202-6.
11. Pudek MR, Seccombe DW, Jacobsen BE, Whitfield MF. Seven different digoxin immunoassay kits compared with respect to interference by a
digoxin-like immunoreactive substance in serum from premature and full-term infants. Clin Chem 1983;29:1972-4.
12. Hick JM, Brett EM. Falsely increased digoxin concentrations in samples from neonates and infants. The Drug Monit 1984;6:461-4.
13. Soldin SJ, Papanastasiou-Diamandi A, Heyes J. Lindwood C, Olley P. Are immunoassays for digoxin reliable? Clin Biochem 1984;17:317-20.
14. Valdes, Jr. R. Endogenous digoxin-like immunoreactive factors: Impact on digoxin measurements and potential physiological implications. Clin Chem
1985;31:1525-32.
15. GlaxoSmithKline Product Information, August 2001.

2009-08
DIGOXIN, AU400/AU640 Serum Application DIGOXIN, AU600 Serum Application

Test No # Test name DIG ∇ Sample type Ser ∇ Page 1/2
Test Name: DIG ∇ < > Type: Serum ∇ Operation: Yes ∇
Wavelength: Pri. 700 ∇ Sec. None ∇ First L -0.1 First H 2.5 Method FIXED1 ∇ Dynamic range
Method: FIXED1 ∇ Last L -0.1 Last H 2.5 Reaction + ∇ L 0* H 6.4*
Measuring Point 1: First 12 Last 27 L 0* H 6.4* Point 2 Fst Lst B 0
General LIH ISE Range Test No # Test name DIG ∇ Sample type Ser ∇ Page 2/2
Test Name: DIG ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic Value: # # Unit: nmol/L* Decimal places: # Panic value # #
SERUM APPLICATION
General ISE Test No # Test name DIG ∇
Test Name: DIG ∇ < > Type Serum ∇ Cal type 14 6AB ∇ Count #
Formula 6 EIA TYPE 1 ∇ Process Conc ∇
Calibration Type: 6AB ∇ Formula: EIA TYPE 1 ∇ Counts: # Process: CONC ∇
Point 1 # ∇ † -0.1 2.5
Point 1: # † -0.1 2.5
Point 2 # ∇ † -0.1 2.5
Point 2: # † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3 # ∇ † -0.1 2.5
Point 4: # † -0.1 2.5 Point 4 # ∇ † -0.1 2.5
Point 5: # † -0.1 2.5 Point 5 # ∇ † -0.1 2.5
Point 6: # † -0.1 2.5 Point 6 # ∇ † -0.1 2.5
1-Point Cal. Point: ο with CONC-0 Slope Check: - ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor
# User defined
† Digoxin Calibrator Cat. No.: ODC6404
† Digoxin Calibrator Cat. No.: ODC6404
* Values set for working in SI units (nmol/L) . To work in ng/mL divide by 1.28
TDM BSOSR6404.01
2009-08
DIGOXIN, AU2700/AU5400 Serum Application DIGOXIN, AU680/AU480 Serum Application
Test Name: DIG ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: DIG ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
Method: FIXED1 ∇ Last L -0.1 Last H 2.5
Dynamic Range Low 0* High 6.4*
Measuring Point 1: First 15 Last 27 L 0* H 6.4* Wavelength Pri 700 ∇nm Sec. None ∇nm Factor for Maker A 1 B 0
Measuring Point 2: First Last Correlation Factor: Method FIXED1 ∇
Test Name: DIG ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: DIG ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: nmol/L* Decimal places: # 7. No demographics # #
Unit nmol/L* Decimal Places #
SERUM APPLICATION
General ISE
General ISE
Test Name: DIG ∇ < > Type Serum ∇ Test Name: DIG ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: 6AB ∇ Formula: EIA TYPE 1 ∇ Counts: # Process: CONC ∇ Calibration Type: 6AB ∇ Formula: EIA TYPE 1 ∇ Counts: # ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibrator OD Conc Low High Slope Check - ∇
Point 1: # † -0.1 2.5 Point 1: # ∇ † -0.1 2.5
Point 3: # † -0.1 2.5 Point 3: # ∇ † -0.1 2.5
Point 6: # † -0.1 2.5 Point 6: # ∇ † -0.1 2.5
1-Point Cal. Point: ο with CONC-0 Slope Check: - ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
† Digoxin Calibrator Cat. No.: ODC6404 Calibrator OD Conc Low High Stability
* Values set for working in SI units (nmol/L) . To work in ng/mL divide by 1.28 Point-1 ∇ Reagent Blank 999 Day 0 Hour
TDM BSOSR6404.01
2009-08
Gentamycin
2x R1 Lyo
2 x 9 mL R2 Buffer
2x R2 Lyo
Intended Use
Homogeneous enzyme immunoassay for the quantitative determination of gentamycin in human serum and plasma on Beckman Coulter analysers. For
Summary
Gentamycin is an aminoglycoside antibiotic used in the treatment of infections caused by E. coli, Klebsiella, Enterobacter, Proteus mirabilis, Pseudomonas
aeruginosa, Serratia, Staphylococcus aureus, Staphylococcus epidermidis and other microorganisms. Gentamycin’s toxic effect is produced by interfering
1,2
with ribosomal protein synthesis. Gentamycin undergoes very little, if any, metabolisation and is, therefore, eliminated as the parent drug by glomerular
filtration.
The therapeutic range should be measured at peak as well as trough concentrations. Peak serum or plasma concentrations of gentamycin are suggested
to ensure that adequate antimicrobial activity is obtained. Trough gentamycin concentrations usually ensure that drug elimination is adequate and the drug
concentration is above minimum inhibitory concentration. Serum or plasma gentamycin concentration is impacted by the mode of administration, the
volume of extracellular fluid, the duration of the treatment and physiological changes during the illness and therapy. Therefore, monitoring of peak and
3,4
trough gentamycin serum or plasma levels is critical in the prevention of serious complications with the adjustment of dosage administration as indicated.
Test Principle5
be formed.
Reagent Composition
R1 Buffer MOPS (3-(N-morpholino)propanesulfonic acid buffer), buffer salts, surfactant and preservative.
R1 Lyo Enzyme acceptor, mouse monoclonal anti-gentamycin antibody, buffer salts, stabiliser and preservative
R2 Buffer MOPS (3-(N-morpholino)propanesulfonic acid buffer), buffer salts, surfactant and preservative
R2 Lyo Enzyme donor conjugated to gentamycin, chlorophenol red-β-D-galactopyranoside, goat anti-mouse antibodies, buffer salts, stabiliser and
preservative.

Safety Phrases:
Reagent Preparation

2009-08
Specimen
Stable in serum and plasma for 7 days when stored at 2…8°C and 4 weeks when stored at -20°C.
Test Procedure
Calibration
Antibiotic TDM Multi-Calibrator, Cat. No. ODC6413.
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the gentamycin concentration of each sample.
Expected Values
In most adults a peak therapeutic response is achieved with gentamycin concentrations between 5-8 µg/mL. Trough concentrations between 1-2 µg/mL
1,4 2-3,6-8
usually ensure that drug elimination is adequate. Different gentamycin therapeutic ranges have also ben reported by other investigators.
9
symptoms.

values.
Linearity
part Antibiotic TDM Multi-Calibrator 1 and reassayed. The value obtained on reassay should be derived as follows:
Actual Value = (2 x diluted value) – Concentration of Antibiotic TDM Multi-Calibrator 1.
Precision
20 days.

2.1 0.08 3.8 0.11 5.3
4.4 0.09 1.9 0.15 3.5
6.5 0.17 2.6 0.25 3.8
Method Comparison
Patient serum samples were used to compare this Gentamycin OSR6420 assay on the AU640 against another commercially available gentamycin assay.
y = 0.984x + 0.13 r= 0.999 n = 135 Sample range = 0.2– 17.3 µg/mL
Sensitivity
The lowest detectable level on an AU640 analyser was estimated at 0.13 µg/mL.
The lowest detectable level (LDL) represents the lowest measurable level of gentamycin that can be distinguished from zero. It is calculated as the
Interference
Bilirubin ≤ 1026 µmol/L IgM ≤ 26 g/L
Haemoglobin ≤ 10 g/L Total Protein ≤ 120 g/L
IgA ≤ 27 g/L Triglyceride ≤ 11.3 mmol/L
IgG ≤ 51 g/L Rheumatoid Factor ≤ 1270 IU/mL

2009-08
Specificity
The following compounds when tested with the Gentamycin OSR6420 assay, yielded the following percent cross-reactivity results:
5-Fluorocytosine 0.06 < 0.6
Amikacin 6.00 < 1.0
Amphotericin 0.24 < 0.1
Ampicillin 6.00 < 0.1
Carbenicillin 6.00 < 0.1
Cefamandole Nafate 6.00 < 0.1
Cephalexin 6.00 < 0.1
Cephaloglycin 6.00 < 0.1
Cephaloridine 6.00 < 0.1
Cephalosporin C 6.00 < 0.1
Cephalothin 6.00 < 0.1
Chloramphenicol 6.00 < 0.1
Clindamycin 6.00 < 0.1
Dideoxykanamycin 6.00 < 0.1
Erythromycin 0.36 < 0.1
Ethacrynic Acid 0.60 < 1.0
Furosemide 0.72 < 1.0
Fusidic Acid 0.60 < 1.0
Kanamycin A 6.00 < 0.1
Kanamycin B 6.00 < 0.1
Lincomycin 6.00 < 0.1
Methicillin 6.00 < 0.1
Methotrexate 0.60 < 1.0
Methylprednisolone 0.60 < 1.0
Neomycin 6.00 < 0.1
Netilmicin 0.52 1.15
Oxytetracycline 1.00 < 0.6
Penicillin G 6.00 < 0.1
Penicillin V 0.72 < 1.0
Prednisolone 0.72 < 1.0
Rifampicin 0.60 < 1.0
Sisomicin 0.01 62.1
Spectinomycin 6.00 < 0.1
Streptomycin 6.00 < 0.1
Sulfadiazine 0.60 < 1.0
Sulfamethoxazole 0.72 < 1.0
Sulfanilamide 6.00 < 0.1
Sulthiame 6.00 < 0.1
Tetracycline 6.00 < 0.1
Ticarcillin 6.00 < 0.1
Tobramycin 6.00 < 0.1
Trimethoprim 0.18 < 0.6
Vancomycin 0.60 < 1.0
Limitations
• Samples containing antibodies to E. coli β-galactosidase may result in artificially high results which will not fit the clinical profile. The incidence of
patients having such antibodies is extremely low.
• Sisomicin shows significant cross reaction (>50%) with the Gentamycin assay. Gentamycin results from patients receiving this drug should be
interpreted with care.
• Carryover from reagents containing gentamycin may cause spuriously high results. Please pay specific attention to the contamination parameters
provided in the setting sheet. In order to optimise workflow it is recommended to assay specimens for gentamycin in batches.
† Antibiotic TDM Multi-Calibrator Cat. No.: ODC6413
∝ Values set for working in µg/mL. To work in SI units (µmol/L) multiply by 2.09
‡ To prevent contamination by gentamycin containing reagents instigate the following corrective procedure.
Prepare a 20% aqueous solution of Wash Solution Cat No. OSR0001 and enter the following settings in Parameter – Special - Contamination
Parameters.
Bibliography
1. Barza M, Scheife T. Drug therapy reviews: Antimicrobial spectrum, pharmacology and therapeutic use of antibiotics-part 4: amimoglycosides. American Journal
of Hospital Pharmacy. 1977:34: 723-737.
2. Sande MA, Mandell GL. Antimicrobial Agents, The Aminoglycosides. In: Gilman AG, Goodman LS, Gilman A, eds. The Pharmacological Basis of Therapeutics.
New York: MacMillan Publishing Company, 1980:1162-1180.
3. Kahlmeter G. Gentamicin and Tobramycin, Clinical Pharmacokinetics and Nephrotoxicity Aspects on Assay Techniques. Scandinavian Journal of Infectious
Disease 1979:132 (suppl. 18).
4. Barza M, Lauermann M. Why Monitor Serum Levels of Gentamicin? Clinical Pharmacokinetics 1978;3:202-215.
5. Henderson DR, Friedman SF, Harris JD, Manning WB, Zoccoli MA. CEDIA™, A New Homogeneous Immunoassay System. Clin. Chem. 1986;32(9): 1637-
1641.
6. Mattie H. The Clinical Importance of Determination of Aminoglycodise Concentrations, Aminoglycoside Assay Methods and Clinical Relevence. Proceedings of
Amsterdam Symposium, 1978.
7. Baselt RC, Cravey RH. Disposition of Toxic Drugs and Chemicals in Man. 1990, 3rd edition, pp. 805-807.
8. Dipersio, J.R.: Gentamicin and Other Aminoglycosides, in Pesce, A.J. and Kaplan, L.A. (eds) Methods in Clinical Chemistry, C.V. Mosby Co., 1987.
9. Hallworth M, Capps N, eds. Therapeutic drug monitoring and clinical biochemistry, London:ACB Venture Publications, 1993.

2009-08
GENTAMYCIN, AU400/AU640 Serum/Plasma Application GENTAMYCIN, AU600 Serum/Plasma Application
Test No # Test name GENT‡ ∇ Sample type Ser ∇ Page 1/2
Test Name: GENT‡ ∇ < > Type: Serum ∇ Operation: Yes ∇
Method: RATE1 ∇ Last L -2.00 Last H 2.50 Reaction + ∇ L 0.24∝ H 12∝
Measuring Point 1: First 24 Last 27 L 0.24∝ H 12∝ Point 2 Fst Lst B 0
Test No # Test name GENT ∇ Sample type Ser ∇ Page 2/2
Test Name: GENT ∇ < > Type: Serum ∇ Level L Level H
Normal range
1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # #
2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # #
3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # #
4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # #
5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # #
6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # #
7 Non select # #
8 Out of range # #
L H
Panic Value: # # Unit: µg/mL∝ Decimal places: # Panic value # #
General ISE
Test No # Test name GENT ∇
Test Name: GENT ∇ < > Type Serum ∇
Cal type 9 AA Count #
∇
Calibration Type: AA ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Formula 1 Y=AX+B Process Conc
∇ ∇
Point 1: # † -999999 9999999 Point 1 # ∇ † -999999 9999999
Point 2: # † -999999 9999999 Point 2 # † -999999 9999999
∇
Point 3:
Point 3 ∇
Point 4:
Point 4 ∇
Point 5:
Point 5 ∇
Point 6:
Point 6 ∇
Point 7:
Point 7 ∇
1-point cal. point
‡ To prevent contamination by Gentamycin containing reagents instigate the following corrective procedure
Prepare a 20% aqueous solution of wash Solution Cat No OSR0001 and enter the following settings in Parameter –Special – Contamination Parameters Select the function using the Function key or the Mouse
AU640 Preceding Following RGT.probe Wash cnt Cancel Mixer CUV ‡ To prevent contamination by Gentamycin containing reagents instigate the following corrective procedure
(ALL) GENT CLN 5 No Yes Yes Prepare a 20% aqueous solution of Wash Solution Cat No OSR0001 and enter the following settings in
Parameter – Special – Contamination Parameters
AU400 Preceding Reag Type Following Reag Type Reag Probe Wash Cnt Cancel Mixer Cuvette AU600 Preceding Following RGT . probe Clean cnt CUV Mixer
(ALL) R1 GENT R1 CLN-1/CLN-2 5 No Yes Yes ALL GENT CLN 5 Y Y
(ALL) R1 GENT R2 CLN-1/CLN-2 5 No Yes Yes # User defined ¤ Analyser default value
(ALL) R2 GENT R1 CLN-1/CLN-2 5 No Yes Yes † System Antibiotic TDM Multi-Calibrator Cat. No.: ODC6413
(ALL) R2 GENT R2 CLN-1/CLN-2 5 No Yes Yes ∝
Values set for working in µg/mL. To work in SI units (µmol/L) multiply by 2.09
TDM BSOSR6420.01
2009-08
GENTAMYCIN, AU2700/AU5400 Serum/Plasma Application GENTAMYCIN, AU680/AU480 Serum/Plasma Application
Test Name: GENT‡ ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: GENT‡ ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Test Name: GENT‡ ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: GENT‡ ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: GENT‡ ∇ < > Type Serum ∇ Test Name: GENT‡ Type Serum Use Serum Cal.
∇ < > ∇ ο
Point 1: # † -999999 9999999 Point 1: # ∇ † -999999 999999
1-Point Cal. Point: ο with CONC-0 Slope Check: None Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
‡ To prevent contamination by Gentamycin containing reagents instigate the following corrective procedure
Prepare a 20% aqueous solution of wash Solution Cat No OSR0001 and enter the following settings in Parameter – Special – Contamination Parameters Master Curve> OD Range
Ф AU680 Calibrator OD Conc Low High Stability
AU2700/ Preceding Following RGT.probe Wash Cleaning Mixer CUV
AU680 cnt Point-1 ∇ Reagent Blank 999 Day 0 Hour
(ALL) GENT CLN 5 No Yes Yes Point-2 ∇ Calibration 999 Day 0 Hour
AU480 Preceding Reag Type Following Reag Type Reag Probe Wash Cnt Cleaning Mixer Cuvette MB Type Factor: 1-Point Calibration Point ∇ ο with Conc-0
(ALL) R1 GENT R1 CLN-1/CLN-2 5 No Yes Yes
TDM BSOSR6420.01
2009-08
PARACETAMOL
3x R1 Lyo
3 x 20 mL R2
3 x 3 mL Calibrator
Intended Use
Enzymatic test for the quantitative determination of Paracetamol (Acetaminophen) in human serum and plasma on Beckman Coulter analysers.
Summary1,2,3,4,5
Paracetamol is a drug commonly used for its analgesic and antipyretic properties. However, excessive long term use has been reported to lead to
hepatotoxicity and nephrotoxicity. Paracetamol overdose can lead to severe hepatic damage and can result in hepatic failure and death if left untreated.
Early diagnosis of paracetamol overdose is important, as initiation of therapy within 16 hours of ingestion reduces the potential for hepatic injury and
decreases the rate of mortality.
Test Principle6
Paracetamol in the sample is hydrolysed by aryl acylamidase to yield p-aminophenol and acetic acid. The p-aminophenol formed reacts specifically with
o-cresol and ammoniacal copper sulphate, to form indophenol. The change in absorbance is directly proportional to the concentration of paracetamol
present in the sample, and is measured at 600nm. The assay is specific for the parent compound and does not detect paracetamol metabolites.
Reaction Principle
Aryl Acylamidase
Paracetamol p-Aminophenol + Acetic Acid
p-Aminophenol + o-Cresol + Ammoniacal Copper Sulphate Indophenol (blue)

o-Cresol 0.48%
Aryl Acylamidase ≥ 0.48 U/mL
Buffer pH 8.6
Sodium carbonate < 4.8%
Ammonium chloride < 0.48%
Copper Sulphate < 0.048%
Paracetamol Calibrator
Paracetamol (4-acetamidophenol) 2.0 mmol/L
Buffer pH 10.6
Preservative
To avoid the possible build-up of azide compounds, flush waste-pipes with water after the disposal of calibrator material.
Reagent Preparation
R1: Dissolve the contents of one vial of R1 Lyo (white cap) completely by adding approximately 10 mL of R1 buffer. When the R1 Lyo has completely
dissolved, pour the contents of the reconstituted enzyme vial back into the same R1 buffer bottle containing the remaining 10 mL of reagent. Mix gently by
inversion and place on board the instrument.
The reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C. Once prepared, reagents stored on board the instrument are
stable for 14 days.
Specimen7
Stable in serum and plasma for 14 days when stored at 2...8°C and 45 days when stored at -20°C.
Haemolysed, lipemic and icteric samples should be avoided.
Test Procedure
Calibration
Paracetamol Calibrator (2000 µmol/L; 302 mg/L) is included in the kit.
The calibrator paracetamol value is traceable to a gravimetrically prepared paracetamol standard.
Major preventative maintenance has been performed on the analyser or a critical part has been replaced.
The calibrator is stable, unopened, up to the stated expiry date when stored at 2…8°C. Once open, the calibrator is stable for 30 days, provided it is free
from contamination, tightly capped immediately after each use, and stored at 2…8°C.
Quality Control
BioRad Lyphochek Immunoassay Plus Controls or other control materials with values determined by this Beckman Coulter system can be used.
EN.01 BLOSR6x202.01 TDM
2009-08
The results obtained by any individual laboratory can vary from the given mean value. It is therefore recommended that each laboratory generates analyte
Calculation
The Beckman Coulter analysers automatically compute the paracetamol concentration of each sample.
Therapeutic Range
8
The therapeutic range for paracetamol varies and is reported as 66 – 199 µmol/L (10 – 30 mg/L).
The concentration of paracetamol in serum or plasma is dependent on the time interval post ingestion. Concentrations in serum of >993 µmol/L
(>150 mg/L), >497 µmol/L (>75 mg/L) and >265 µmol/L (>40 mg/L) at 4 hours, 8 hours and 12 hours post ingestion respectively, have been reported as
9
toxic.
For diagnostic purposes, results should always be assessed in conjunction with the patient's medical history, clinical examinations and other findings.
Data contained within this section is representative of performance on Beckman Coulter systems. Data obtained in your laboratory can differ from these
values.
Linearity
The test is linear within a concentration range of 99 – 2500 µmol/L (15 – 378 mg/L) for serum and plasma.
Precision
68 0.5 0.7 2.4 3.5
220 1.1 0.5 3.5 1.6
676 2.6 0.4 6.2 0.9
Sensitivity
The lowest detectable level on an AU2700 analyser was estimated as 2.57 µmol/L (0.39 mg/L)
The lowest detectable level represents the lowest measurable level of paracetamol that can be distinguished from zero. It is calculated as the absolute
Method Comparison
Patient serum samples were used to compare this Paracetamol OSR61202 assay on the AU640 against another commercially available paracetamol
y = 1.016x + 29.182 r = 1.000 n = 106 Sample range = 0 – 1943 µmol/L
Icterus: Interference less than 10% up to 8 mg/dL (137 µmol/L) bilirubin
®
N-Acetyl Cysteine: Interference less than 10% up to 0.5 g/L N-Acetyl Cysteine
Salicylic acid: Interference less than 3% up to 5 mmol/L Salicylic Acid
Acetylsalicylic acid: Interference less than 3% up to 1 g/L Acetylsalicylic Acid
10
In very rare cases gammopathy, especially IgM (Waldenström’s macroglobulinemia), may cause unreliable results.
.
11
Limitations
Haemolysed, icteric and lipemic samples should be avoided. Icteric and lipemic samples may cause false depression of results. Haemolytic samples may
cause false elevation of results. Consequently, results from any samples exhibiting these characteristics should be interpreted with care.
# User defined
¤ Analyser default value
† Paracetamol Calibrator included in the kit
* Values set for working in SI units (µmol/L). To work in mg/L, divide by 6.62.
BIBLIOGRAPHY
1. Porter WH. Clinical Toxicology. In: Burtis CA, Ashwood ER eds. Tietz Textbook of Clinical Chemistry Philadelphia: WB Saunders Company, 1999: 927-929.
2. Barker JD, de Carle DJ, Anuras S. Chronic excessive acetaminophen use and liver damage. Ann Intern Med 1997: 87; 299-301.
3. Prescott LF. The nephrotoxicity and hepatotoxicity of antipyretic analgesics. Br J Clin Pharmacol 1979: 7; 443-452.
4. Black M. Acetaminophen hepatotoxicity. Gastroenterol. 1980: 78; 382-392.
5. Ambre J, Alexander M. Liver toxicity after acetaminophen ingestion. J.AMA 1977: 238; 500-501.
6. Price CP, Hammond PM, Scawen MD. Evaluation of an enzymatic procedure for the measurement of acetaminophen. Clin Chem 1983: 29; 358-361.
7. Guder WG, Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of anticoagulants in diagnostic laboratory investigations and stability of blood,
plasma and serum samples. WHO/DIL/LAB/99.1 Rev.2, 2002.
8. Tietz NW In:Clinical Guide to Laboratory Tests 3rd. ed. Philadelphia: WB Saunders Company, 1999: 788pp.
9. Lewis RK, Poulocek FP. Assessment and treatment of acetaminophen overdose. Clin Pharm 1991: 10; 765-774.
10. Hullin DA. An IgM Paraprotein causing a falsely low result in an enzymatic assay for acetaminophen. Clin Chem. 1999 : 45; 155-157
TDM BLOSR6x202.01 EN.01

2009-08
PARACETAMOL, AU400/AU640 Serum/Plasma Application PARACETAMOL, AU600 Serum/Plasma Application
System Reagent: OSR6x202 Reagent ID: 202 System Reagent: OSR6x202 Reagent ID: 202

Test No # Test name PARA ∇ Sample type Ser ∇ Page 1/2
Test Name: PARA ∇ < > Type: Serum ∇ Operation: Yes ∇
Test No # Test name PARA ∇ Sample type Ser ∇ Page 2/2
Level L Level H
Test Name: PARA ∇ < > Type: Serum ∇ Value/flag # ∇ # #
Normal range
Sex Age L Age H L H
Normal Ranges: Age L Age H 1 # # Y # M # Y # # #
∇ M→
2 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # #
3 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # #
4 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # #
5 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # #
6 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # #
7 Non select # #
ο 6. # ∇ # # # # # #
8 Out of range # #
L H
Panic value # #
Test No # Test name PARA ∇
General ISE
Test Name: PARA Type Serum Cal type 8 AB ∇ Count #
∇ < > ∇
Point 1: # † 1815* 3025* Point 2 ∇
1-Point Cal. Point: ο with CONC-0 Slope Check: ∇ Advanced Calibration: # ∇ MB type factor

† Paracetamol Calibrator included in the kit † Paracetamol Calibrator included in the kit
* Values set for working in SI Units (µmol/L). To work in mg/L divide by 6.62 * Values set for working in SI Units (µmol/L). To work in mg/L divide by 6.62
TDM BSOSR6x202.01
2009-08
PARACETAMOL, AU2700/AU5400 Serum/Plasma Application PARACETAMOL, AU680/AU480 Serum/Plasma Application
System Reagent: OSR6x202 Reagent ID: 202 System Reagent: OSR6x202 Reagent ID: 202
Test Name: PARA ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: PARA ∇ < > Type: Serum ∇ Operation: Yes ∇

μL μL
Reagent OD limit:
No Lag Time: ∇ On-board stability period: 14 Measuring Point1 First 10 Last 27 LIH Influence Check YES ∇
Test Name: PARA ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: PARA ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: PARA ∇ < > Type Serum ∇
Test Name: PARA ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † 1815* 3025* Point 1: # ∇ † 1815* 3025*
1-Point Cal. Point: ο with CONC-0 Slope Check: ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
† Paracetamol Calibrator included in the kit Calibrator OD Conc Low High Stability
* Values set for working in SI Units (µmol/L). To work in mg/L divide by 6.62 Point-1 ∇ Reagent Blank 7 Day 0 Hour
TDM BSOSR6x202.01
2009-08
Phenobarbital
2x R1 Lyo
2 x 16 mL R2 Buffer
2x R2 Lyo
Intended Use
Homogeneous enzyme immunoassay for the quantitative determination of phenobarbital in human serum and plasma on Beckman Coulter analysers. For
Summary
1,2
Phenobarbital has been widely prescribed for the treatment of epilepsy, particularly for controlling focal motor or sensory and grand mal seizures. After
3
oral doses of 2 to 3 mg/kg, phenobarbital is almost completely absorbed with peak levels achieved by 12 to 18 hours. Phenobarbital in circulation is
4
approximately 40% to 50% bound to plasma proteins with relatively low association constants. The major metabolic pathway of phenobarbital is
hydroxylation of the phenyl ring to p-hydroxyphenobarbital, an agent devoid of hypnotic activity, which is then excreted in the urine in equal amounts of the
4
free form and the form conjugated with glucuronic acid. Phenobarbital concentrations of 15-40 µg/mL in serum are normally considered to be within the
5,6
therapeutic range for maximum seizure control.
The need for monitoring phenobarbital concentrations is due to the narrow therapeutic index and the wide variability in individual rates of drug absorption,
7
metabolism, and clearance. Toxicity of phenobarbital therapy includes sedation, nystagmus, ataxia, paradoxical excitement, blood dyscrasia (including
coagulation defects in neonates of mothers given the drug during pregnancy), non-specific hepatic changes, rash (including severe exfoliative forms),
3,8-10
osteomalacia, the shoulder-hand syndrome and coma. In combination with other clinical information, monitoring serum or plasma phenobarbital levels
will provide physicians with an essential tool to aid in adjusting dosage and achieving optimal therapeutic effect, while avoiding both sub-therapeutic and
harmful toxic drug levels.
Test Principle11
be formed.
Reagent Composition
R1 Buffer MOPS (3-[N-morpholino]propanesulfonic acid buffer), buffer salts and preservative.
R1 Lyo Enzyme donor conjugated to phenobarbital, mouse monoclonal anti-phenobarbital antibody, chlorophenol red-β-D-galactopyranoside, buffer
salts and preservative.
R2 Buffer MOPS (3-[N-morpholino]propanesulfonic acid buffer), buffer salts, goat anti-mouse antibodies, stabilisers and preservative.
R2 Lyo Enzyme acceptor, releasing agent, buffer salts and preservative.
Safety Phrases:
Reagent Preparation

2009-08
Specimen
Stable in serum and plasma for 24 hours when stored at 2…8°C and 2 weeks when stored at -20°C.
12
Some therapeutic drug concentrations are reduced when a sample is stored in a separator tube for a prolonged period of time (>2 hrs).
Test Procedure
Calibration
Core TDM Multi-calibrator, Cat. No. ODC6411
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the phenobarbital concentration of each sample.
Expected Values
5
In most patients, a therapeutic response is achieved with phenobarbital concentrations in the 15-40 µg/mL range (65-172 µmol/L).
available information such as patient’s medical history, mode of drug administration, timing of sample collection, other drug treatments and clinical
symptoms.
values.
Linearity
Precision
20 days.
7.1 0.20 2.8 0.39 5.5
14.2 0.25 1.8 0.50 3.5
45.5 0.56 1.2 1.32 2.9
Method Comparison
Patient serum samples were used to compare this Phenobarbital OSR6413 assay on the AU640 against another commercially available phenobarbital
y = 0.964x + 0.73 r = 0.999 n = 139 Sample range = 1.2 – 71.4 µg/mL
Sensitivity
The lowest detectable level on an AU640 analyser was calculated as 0.6 µg/mL.
The lowest detectable level (LDL) represents the lowest measurable level of phenobarbital that can be distinguished from zero. It is calculated as the
interference
Bilirubin ≤ 1129 µmol/L Total Protein ≤ 130 g/L
Haemoglobin ≤ 10 g/L Triglyceride ≤ 11.3 mmol/L
Rheumatoid Factor ≤ 180 IU/mL
Specificity
The following compounds when tested with the Phenobarbital assay yielded the following percent cross-reactivity results:

2009-08
1,3-dimethylbarbituric acid 1000 < 0.12
2-Phenyl-2-ethylmalonamide 1000 < 0.12
5-(ρ-Hydroxyphenyl)-5-phenylhydantoin 1000 < 0.12
Amitriptyline 1000 < 0.12
Aprobarbital 1000 ≤ 5.3
Barbital 2000 ≤ 1.6
Butabarbital 1000 ≤ 2.3
Carbamazepine-10,11-epoxide 1000 < 0.12
Carbamazepine 1000 < 0.12
Chlorazepate 2000 < 0.12
Chlorpromazine 1000 < 0.12
Diazepam 1000 < 0.12
Ethotoin 1000 < 0.12
Ethosuximide 1000 < 0.12
Glutethimide 1000 < 0.12
Imipramine 2000 < 0.12
Mephenytoin 1000 < 0.12
Methsuximide 1000 < 0.12
Pentobarbital 1000 < 0.12
Phenytoin 400 ≤ 0.9
ρ-Hydroxyphenobarbital 2000 < 0.12
Primidone 1000 ≤ 0.5
Promethazine 1000 < 0.12
Secobarbital 2000 2.2
Sulthiame 1000 < 0.12
Valproic acid 2000 ≤ 0.7
Amobarbital (>20%) and Mephobarbital (>100%) show significant interference with the Phenobarbital assay.
Limitations
• Samples containing antibodies to E. coli β-galactosidase may result in artificially high results which will not fit the clinical profile. The incident of patients
having such antibodies is extremely low.
† Core TDM Multi-calibrator Cat. No.: ODC6411
* Values set for working in µg/mL. To work in SI units (µmol/L) multiply by 4.31.
Bibliography
1. Buchthal, F. Lennox-Buchthal, M.A.: Relation of Serum Concentration to Control Seizures, in Woodbury, D.M., Pentry, J.K., and Schmidt, K.P.
(eds):Antiepileptic Drugs, NY: Raven Press, 1972;335-343.
2. Hauptmann, A.: Luminal bei Epilepsy, Munch. Med. Wschr., 1912;59:1907-1909
3. Penry, J.K., Hewmark, M.E.: The use of anti-epileptic drugs, Annals of Internal Medicine, 1979;90:207-218.
4. Glazko, A.J.: Antiepileptic Drugs: Biotransformation, Metabolism, and Serum Half life, Epilepsia, 1975;16:367-391.
5. Buchthal, F., Svensmark,O.: Serum Concentrations of Diphenylhydantion (Phenytoin) and Phenobarbital and Their Relation to Therapeutic and Toxic
Effects, Psychiat. Neurol. Neurochir., 1971;74:117-135.
6. Kutt, He., Penry, K.: Usefulness of Blood Levels of Anti-Epileptic Drugs, Arch. Neurol.,1974;3:283-288.
7. Buchthal, F., Svensmark, O.: Aspects of the Pharmacology of Phenytoin (Dilantin) and Phenobarbital Relevent to Their Dosage in theTreatment of
Epilepsy, Epilepsia,1960;1:373-384.
8. Eadie, M.J., Tyrer, J.H.: Anticonvulsant Therapy: Pharmacological Basis and Practice, London: Churchill Livingstone, 1974.
9. Deut, C.E., Richens, A., Rowe, D.J.E., Stamp, T.C.B.: Osteomalacia with long-term anticonvulsant therapy in epilepsy, Brit. Med. J., 1970;4:69-72.
10. Farwell, J.R., Lee, Y.J., Hirtz, D.G., Sulzbacher, S.I., Ellenberg, J.H., Nelson, K.B.: Phenobarbital for febril seizures -effects on intelligence and on
seizure recurrence, N. Engl. J. Med., 1990;322:363-369.
11. Henderson, D.R., Friedman, S.F., Harris, J.D., Manning, W.B. and Zoccoli, M.A.: CEDIA, a New Homogeneous Immunoassay System, Clin. Chem.
1986;32(9):1637-1641.
12. Dasgupta, A., Dean, R., Saldand, S., Kinnaman G., McLawhon, R.: Absorption of Therapeutic Drugs by Barrier Gels in Serum Separator Blood
Collection Tubes. American Journal of Clinical Pathology. 1993:456-461.

2009-08
PHENOBARBITAL, AU400/AU640 Serum/Plasma Application PHENOBARBITAL, AU600 Serum/Plasma Application

Test No # Test name PHB ∇ Sample type Ser ∇ Page 1/2
Test Name: PHB ∇ < > Type: Serum ∇ Operation: Yes ∇
Method: RATE1 ∇ Last L -2.00 Last H 2.50 Reaction + ∇ L 1.2* H 80*
Reaction slope: + ∇ Dynamic Range: Point 1 Fst 24 Lst 27 Correlation factor A 1.05
Linearity : % A 1.05 B 0
General LIH ISE Range Test No # Test name PHB ∇ Sample type Ser ∇ Page 2/2
Test Name: PHB ∇ < > Type: Serum ∇ Level L Level H
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
Panic Value: # # Unit: µg/mL* Decimal places: # Panic value # #
General ISE Test No # Test name PHB ∇
Test Name: PHB ∇ < > Type Serum ∇ Cal type 9 AA Count #
∇
Calibration Type: AA ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇
Point 1 # ∇ † -999999 9999999
Point 1: # † -999999 9999999
Point 2 # ∇ † -999999 9999999
Point 2: # † -999999 9999999
MB type factor
# User defined
† Core TDM Multicalibrator Cat. No.: ODC6411
† Core TDM Multicalibrator Cat. No.: ODC6411
TDM BSOSR6413.01
2009-08
PHENOBARBITAL, AU2700/AU5400 Serum/Plasma Application PHENOBARBITAL, AU680/AU480 Serum/Plasma Application
Test Name: PHB ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: PHB ∇ < > Type: Serum ∇ Operation: Yes ∇
μL μL
Reagent OD limit:
Measuring Point 1: First 24 Last 27 L 1.2* H 80* Wavelength Pri 570 ∇nm Sec. 660 ∇nm Factor for Maker A 1.05 B 0
Linearity : % A 1.05 B 0 Reaction Slope + ∇ Onboard Stability Period 60 Day 0 Hour
Test Name: PHB ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: PHB ∇ < > Type: Serum ∇
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
General ISE
General ISE
Test Name: PHB ∇ < > Type Serum ∇
Test Name: PHB ∇ < > Type Serum ∇ ο Use Serum Cal.
Point 1: # † -999999 9999999 Point 1: # ∇ † -999999 999999
Point 9 ∇
† Core TDM Multicalibrator Cat. No.: ODC6411 Calibrator OD Conc Low High Stability
* Values set for working in µg/mL. To work in SI units (µmol/L) multiply by 4.31 Point-1 ∇ Reagent Blank 999 Day 0 Hour
TDM BSOSR6413.01
2009-08
Phenytoin
2x R1 Lyo
2 x 16 mL R2 Buffer
2x R2 Lyo
Intended Use
Homogeneous enzyme immunoassay for the quantitative determination of phenytoin in human serum and plasma on Beckman Coulter analysers. For
Summary
Phenytoin (diphenylhydantoin) is one of the most widely prescribed anti-convulsant drugs for the treatment of epilepsy, particularly grand mal epilepsy
1
(major motor), cortical focal seizures and temporal lobe epilepsy.
2-7 2
Phenytoin concentrations of 10 to 20 µg/mL for adults and 6 to 14 µg/mL for children in serum are considered to be the therapeutic range for maximum
5,6
seizure control. The toxicity of phenytoin is often dose-related and affects mainly the central nervous system. Monitoring phenytoin concentrations in
serum is essential during therapy by providing physicians with an index for adjusting dosage, to achieve optimal therapeutic effect and avoid both
1,7
subtherapeutic and harmful toxic drug levels.
Test Principle8
be formed.
Reagent Composition
R1 Buffer MOPS (3-(N-morpholino)propanesulfonic acid buffer), mouse monoclonal anti-phenytoin antibody, buffer salts, stabiliser and preservative.
R1 Lyo Enzyme acceptor, releasing agent, detergent, buffer salts and preservative.
R2 Buffer MOPS (3-(N-morpholino)propanesulfonic acid buffer), buffer salts and preservative.
R2 Lyo Enzyme donor conjugated to phenytoin, chlorophenol red-β-D-galactopyranoside, buffer salts, stabiliser and preservative.

Safety Phrases:
Reagent Preparation

Specimen
Stable in serum and plasma for 7 days when stored at 2…8°C and 4 weeks when stored at -20°C.
2009-08
Test Procedure
Calibration
Core TDM Multi-calibrator, Cat. No. ODC6411
Quality Control
Calculation
The Beckman Coulter analysers automatically compute the phenytoin concentration of each sample.
Expected Values
Therapeutic Range (µg/mL)
Investigator Adult Children
1
Buchthal and Lennox-Buchthal 15 – 25
2
Finn and Olanow 10 – 20 6 – 14
3
Jusko 10 – 20
6
Buchthal and Svensmark 10 – 20
4
Sohn and Ferrendelli 10 – 25
7
Penry and Newmark 10 – 20
9
symptoms.

values.
Linearity
Precision
20 days.
4.3 0.24 5.6

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CLINICAL CHEMISTRY

Beckman Coulter, Inc., 250 S. Kraemer Blvd.,Brea, CA 92821, USA.

4. GENERAL PRECAUTIONS AND WARNINGS

APPENDIX 1 PRODUCT GROUPINGS

APPENDIX 2 TABLE OF CALIBRATORS AND CONTROLS

2.1 Guide Format:

Example: REF: OSR6287

BLOSR6x87.01 Î IFU reference and revision code.

2.2 Guide Updates:

Manufacturer Biological Risk

In Vitro Diagnostic Medical Device Contents

Consult Instructions for Use Protect from Light

Material safety data sheet available to

Catalogue Number Serial Number

Keep Upright Caution, consult accompanying documents

Batch code Product conforms to the IVD directive

Authorized Representative in the

4. GENERAL PRECAUTIONS AND WARNINGS

Therapeutic Drug Monitoring (TDM)

Drugs of Abuse in Urine (DAU)

OSR6x43 Apo B ODR3005 (3 pt) / ODR3022 (5 pt) ODC0003, ODC0004

Qualitative: ODC6316 (300 µg/L).

1-Naphthol + FRTR salt Azo dye

Contents, Reagent Composition in the Test

Precautions and Warnings

Storage and Stability

EN.01 BLOSR6x01.01 Enzyme

Specific performance characteristics

Enzyme BLOSR6x01.01 EN.01

Setting Sheet Footnotes

EN.01 BLOSR6x01.01 Enzyme

Test Name: ACP‡ < > Type: Serum ∇ Operation: Yes ∇

Sample: Volume 8 μL Dilution 0 μL Pre-Dilution Rate: 1

Specific Test Parameters

Test Name: ACP‡ ∇ < > Type: Serum ∇

Value/Flag: # ∇ Level L: # Level H: #

Test Name: ACP‡ ∇ < > Type Serum ∇

Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇

Cal. No. OD CONC Factor/OD-L Factor/OD-H

Test Name: ACPж ∇ < > Type: Serum ∇ Operation Yes ∇

Sample Volume 8 μL Dilution 0 μL OD Limit

Parameters Specific Test Parameters

Test Name: ACPж ∇ < > Type: Serum ∇

Value/Flag: # ∇ Low High

Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇

EN.01 BLOSR6x03.01 Enzyme

Enzyme BLOSR6x03.01 EN.01

Specific Test Parameters Specific test parameters

Specific Test Parameters Specific test parameters

Children Male (U/L) Female (U/L)

Enzyme BLOSR6x04.01 EN.01

Specific Test Parameters Specific test parameters

Test Name: ALP4P < > Type: Serum ∇ Operation: Yes ∇

Contents, Reagent Composition in the Test

Precautions and Warnings

Without pyridoxal phosphate activation

EN.01 BLOSR6x07.01 Enzyme

Specific performance characteristics

Method Comparison with pyridoxal phosphate activation

Setting Sheet Footnotes

EN.01 BLOSR6x07.01 Enzyme

Sample: Volume 5 Dilution 0 Pre-Dilution Rate: 1 Sample Volume 5 µL Dilution 0 µL OD Limit

Specific Test Parameters Specific test parameters