SWIFS Controlled Substances Resources Documents v2.1 (02.2009) 76 Pages PDF

Controlled Substances Resource Documents Version 2.
Controlled Substances Resource Documents Version 2.1 Revisions and Corrections Effective Date 2/26/09 Authorized by GC/MS 5973 Quick Reference Operation and Calibration: ELT Manufacturers relative abundance target for mass 219 was updated, autotunes were replaced by standard spectra autotunes, and the maintenance schedule was updated. (version 2.1) GC/MS 5973 Calibration and Maintenance: Manufacturers relative ELT abundance target for mass 219 was updated, autotunes were replaced by standard spectra autotunes, and the maintenance schedule was updated. (version 2.1) IR Calibration Criteria was removed and replaced with FTIR ELT Operation, Version 2.0. (version 2.1) Description
2/26/09
2/26/09
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory
Revisions and Corrections Resource Documents, Version 2.1
Under development
Texas Controlled Substances and Related Statutes Click the following link and refer to Sections: 481 Texas Controlled Substances Act 482 Simulated Controlled Substances 483 Dangerous Drugs 485 Abusable Volatile Chemicals http://tlo2.tlc.state.tx.us/statutes/hs.toc.htm
GAS CHROMATOGRAPHY AND GAS CHROMATOGRAPHY/MASS SPECTROMETRY Gas Chromatography (GC) Chromatography is a physical method of separation that involves the differential distribution of a substance between two phases: one phase is a stationary bed of large surface area and the other phase is a mobile phase, which percolates through the stationary bed. In GC, the mobile phase is a gas, and in order for a substance to be amenable to GC analysis, it must be capable of being volatilized in the GC inlet. The chromatographic separation process occurs as a result of repeated sorption-desorption of a substance between the stationary and mobile phase as the substance travels along the stationary bed. Separation of two or more substances occurs when the distribution coefficients between the mobile and stationary phases are different for the substances. As a substance elutes from the end of a GC column, it is detected by a flame ionization detector, mass spectrometer, or other analytical detector. In a flame ionization detector, the gas exiting the column is mixed with hydrogen and air and burned. Ions and electrons form in the flame and decrease resistance in the electrode gap. This allows current to flow in the detector circuit. Current flow is monitored and charted as a chromatogram. The amount of current generated is dependent upon the type and number of ions formed in the flame. Under consistent instrument conditions, a particular substance will burn in the same manner; therefore, the amount of current generated is proportional to the amount of substance burned allowing quantitation of the material when compared to known standards. To improve accuracy and consistency of quantitation, an internal standard is often added to a sample and the relative response between sample and internal standard is used for quantitation. GC can also give a very good indication of the identity of a substance; although it does not usually provide conclusive identification of a substance. Under consistent instrument conditions, movement of a specific material through a GC column will take a constant period of time. Thus, the time it takes a substance to travel through a GC column (retention time) is consistent for a particular substance. Therefore, retention time on the GC column provides a very good indication of substance identity when compared to a standard material. Gas Chromatography/Mass Spectrometry (GC/MS) A mass spectrometer may be the detector for a GC; in this case, the instrumentation is called GC/MS. Mass spectrometry provides a unique identification of the materials tested; it can also be used for quantitation. As a substance elutes from the GC and enters the MS, it is bombarded with an electron beam. The molecule fragments into characteristic ions depending upon the relative strength of its chemical bonds and chemical structure. Positively charged fragments of the molecule are directed down the analyzer and impinge upon the detector of the MS.
GC/MS Overview Version 2.0
Plotting the relative abundance of each mass fragment identified by the analyzer then generates a mass spectrum. A mass spectrum provides a unique identification of the material being tested. Unknown spectra are identified using in-house library spectra files, commercial computer libraries, and reference materials containing published mass spectra. Under consistent instrumental conditions, the mass ions generated are consistent for a particular substance allowing qualitative identification, and the abundance of ions generated are proportional to concentration of the substance within the instruments linear range allowing quantitation.
GC/MS Overview Version 2.0
Directions for Operation of the HP 6890 GC with G2070AA Chemstation

Users are referred to the appropriate instrument manuals for additional information.
There are two configurations for each instrument. One is ONLINE and one is OFFLINE. The online must be selected to run a sequence or to operate the instrument. The offline mode is to look at data or reprint reports while the instrument is in operation. The offline mode looks exactly like the online mode; however, it will not run samples or control the instrument in any way. FID Detector: FID detectors are used on the instrument. The output should be constant and stable. This may change with instrument usage or with maintenance of the detector. If the flame is out, the instrument will automatically light the flame when a method is loaded. At this time, there is no need to manually light the flame. If the flame will not light, seek assistance from a supervisor. Pausing the sequence: If the sequence must be paused during an analytical run to change an air tank or for another purpose, simply choose RUN CONTROL from the menu bar and select PAUSE SEQUENCE. When finished, resume the sequence by choosing RUN CONTROL from the menu bar and RESUME SEQUENCE. It will begin at the next line from where it stopped. The sequence does not stop immediately. It will not stop until the current sample run is complete.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Operation of the HP 6890 GC Version 2.0
Partial Sequence: Partial sequence may be used to rerun a sequence or start a sequence in the middle after stopping. This is not done after pausing it. Partial sequence may also be used in the offline mode to reprocess certain parts of a sequence in which regeneration of a report or group of reports is needed. Unless absolutely necessary, do not use the partial sequence when running a sample. Once the partial sequence has been started, samples cannot be added to the sequence; this makes the instrument unusable by other users until the sequence is complete. Starting a Run: 1. Check the gases. If low, change before starting the analytical run. If the gases run out during a run or become low during a run see the instructions for pausing a sequence. At this point a sequence must either be created or added to: A. Adding to a sequence: 1. A single click with the left mouse button on the carousel picture will bring up the sequence table. Selecting SEQUENCE from the menu bar, and then selecting SEQUENCE TABLE may also access the sequence table. 2. Using the instructions for Sequence Table, add unknown samples. B. Creating a sequence: 1. Select SEQUENCE from the menu bar and select: A. SEQUENCE PARAMETERS. Follow instructions for sequence parameters. B. SEQUENCE TABLE. Add samples using the instructions for sequence table. C. SAVE SEQUENCE AS. Save the sequence in the following format: Month (3 letters), day (2 numbers), and letter (a, b, c); for example: SEP23A 2. Go back to sequence table and select Run Sequence button to start the sequence, or select START located above carousel picture. The order of these is not important. They all must be done to properly store the data in the proper locations. The sequence will run if not saved; however, if the instrument locks up or the computer needs to be rebooted for any reason, the sequence will be lost, and sample information will need to be re-entered. Therefore, save the sequence.
2.
Operation of the HP 6890 GC Version 2.0
Sequence Parameters Screen: This screen can be accessed through the menu bar under SEQUENCE. The subdirectory is the name of the data path in which the computer will store the data accumulated from the sequence. Enter the subdirectory information in the following format: month (3 letters), day (2 numbers), and letter (a, b, c); for example, SEP23A Currently initials are not entered in the operator box. Everything else in this screen is a default value and should not be change or updated at this time. When finished simply hit the OK button to exit.
Sequence Table Screen: This screen is where the user enters the sequence to be run and sample information. This screen may be accessed by a single click of the left mouse button on the carousel picture or by selecting it under the SEQUENCE item on the menu bar. The critical information that must be present is the vial location, sample name, method name, inj/vial, and sample type. All other cells should be left blank. Any numbers in these blank cells could cause the sample to be injected improperly, calculated wrong or any number of other errors. To select a line move the cursor to the line number cell on the screen. This will produce an arrow. Click the left mouse button on this arrow and it will select the entire line (darkening the line). Drag the mouse down any number of lines to select the entire group of lines. Once a line or group of lines has been selected, the user may cut, copy, insert, insert vial range, or append line. The Insert button will insert a blank line above or in front of the line that you have selected.
The Insert Vial Range button will prompt you to enter information. Type in the appropriate information and hit the OK button. DO NOT put any number in the injection volume box. This will override the method and inject that amount, in microliters, onto the column. If left blank, it will default to the method for that information. The information that is inputted will be inserted above or in front of the line that has been selected.
The Append Line will put a blank line after the line that has been selected. The Cut button will cut the selected line. It is stored until another action takes place, i.e.: another cut or a copy. It may be pasted into another place in the sequence once it has been cut or simply discarded. The Copy button will copy the selected line but leave the line in place. It may then be pasted into another place in the sequence. The Paste button will paste the selected cut or copied line above or in front of the newly selected line. At this point, if the sequence has been saved, select the Run Sequence button to start a run. If the sequence has not been saved, select the OK button to exit this screen and save the sequence before returning. If the sequence is already running and samples are added to the sequence then simply select the OK button after the samples have been added. Then save the sequence (no need for the save as function).
Reports are printed as a sample run is completed using macros written into the method parameters. If a report must be reprinted or reprocessed, use the following instructions: Reprinting a report: If the instrument is running a sequence, use the offline mode of the instrument. From the Data Analysis screen select: Select File from the menu bar and select Load Signal 1. In the Folders: box, selected the correct data path where the signal is stored. For example, C:\hpchem\1\data\aug05c. 2. In the File name: box, selected the correct data signal to be analyzed, by clicking on the name with the left mouse button, or retype the filename of the signal desired to print. Select OK. 3. If the Integrate and print report after load button is selected, the report will be automatically generated. If it is not selected, the chromatograph can be manually integrated utilizing the Chemstation software. This will print out a formatted report. Another way to reprint a report is go into the offline mode and load the sequence for the report that you want to reprint. Select the partial sequence. At this point simply mark the line or lines that you want to regenerate and it will print the report in the proper format. This is the preferred way to reprocess a batch or reprocess more than a single sample.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Operation of the HP 6890 GC Version 2.0
Gas Chromatography CALIBRATION AND MAINTENANCE Agilent/HP 6890 Gas Chromatograph Coupled with Flame Ionization Detectors
All chemists are responsible for ensuring the proper operation of gas chromatographs (GC), running QC samples, evaluating instrument calibration, and providing general housekeeping of instrumentation as needed. I. Instrument Calibration:
Calibration of the GC will be evaluated each day that the GC is used. This will be accomplished by running quality control samples (QC) for routine drugs (methamphetamine, cocaine, heroin, mdma, etc.) and evaluating QC results as described in the applicable analytical procedure. If the QC sample is not within the set limits of +/- 5% of the target concentration, additional action must be taken to obtain an acceptable QC result such as re-running QC, re-extracting and rerunning QC, making new stock with re-extraction and re-analysis, re-calibrating (run curve), and/or consulting a supervisor. Additional information may be found in the appropriate analytical procedure. Calibration of retention time (RT) will also be evaluated for these drugs and internal standard each day the GC is used. Retention time is properly calibrated when these drugs fall into the expected retention time window established by the appropriate GC method. If retention time falls outside the expected window, the chemist will attempt to determine the cause by reviewing recent preventive maintenance records or discussing the situation with a supervisor. Based upon the retention time of known standards, the Calibration Table for the drug and/or internal standard will be updated, the method saved, and the chromatogram reprocessed as necessary. A supervisor will be notified if appropriate or if problems persist. Documentation of instrument calibration is found in the Calibration Curve/Response Factor Log Book and the results of the daily QC are documented on the GC Calibration/QC Log located in the Drug Laboratory. II. Instrument Maintenance:
Routine preventive maintenance activities such as changing a gas cylinder, septum, and/or inlet liner is documented in the GC Maintenance Log located near each instrument. Activities other than scheduled preventive maintenance should be approved in advance by a supervisor.
GC Maintenance HP6890 Version 2.0
GC Maintenance Schedule Daily/with use Change wash vial solvents Check gases Change septum Change liner and o-ring Backup data Change gold seal Trim column Change gas tanks Replace column Clean split side arm Clean FID and jet
Weekly
Monthly As needed
Maintenance activities should be consolidated to minimize instrument down time. For example, if it is time to change the gold seal and cut the column, it may be more prudent to change the liner and septum at that time or hold off changing the gold seal until the liner and septum are changed. The maintenance interval is not a rigid schedule and should be based upon workflow. Unless there is an immediate problem, a chromatographic run should continue to completion, and the maintenance performed at the end of a run. Where duplicate instrumentation exists, only one instrument at a time should be scheduled for preventive maintenance to avoid multiple instruments being out of service at the same time. Ultraclean Technique: Liners, gold seals, and certain other parts must be handled using ultraclean techniques to avoid contamination of the part with oils from the skin, plasticizers from plastic laboratory benchcoats, etc. Do not handle an ultraclean part with your hands; wear gloves or use a fresh Kimwipe. Lay the ultraclean part on a clean Kimwipe. Changing the Wash Vial Solvent: Discard waste solvent into the hazardous waste. Replace spent solvent in wash vials with fresh solvent(s) and replace the vials in the appropriate location on the instrument autosampler. Changing the Liner and Septum: Tools needed: inlet wrench and tweezers.
NOTE: Turn off carrier gas; if the septum nut is removed without turning off the carrier gas, the glass wool plug may be blown out. Use ultraclean technique at all times when handling the new liner. 1. Cool the inlet to < 80 oC to minimize oxidation and prevent burns. 2. Turn off the carrier gas on the GC by using the keypad on the front panel of the GC: From the front panel on the GC, push the FRONT INLET button, scroll down using the button to the Total flow line. Turn off the flow by pressing the OFF button. Once maintenance is complete, turn the flow back on by pressing the ON button. 3. Once the inlet is cool and the gas is off, remove the septum nut. Remove and replace the septum. Reinstall and tighten the septum nut. These are not ultraclean parts but care should be taken to avoid any unnecessary contamination. 4. Remove the nut covering the liner. Remove the old liner and discard. 5. Using ultraclean technique, carefully slide an o-ring over the end of the new liner, approximately from the top of the liner, and slide into the inlet. The glasswool plug should be nearest the bottom of the liner. 6. Replace and tighten the nut and turn the inlet temperature and carrier gas flow on. 7. A blank should be run to bake out the new liner before analyzing samples. Changing the Gold Seal: Tools needed: appropriate screw driver (Phillips/Torx/etc.), 9/16 wrench, wrench Note: The gold seal is located at the bottom of the inlet. It is accessed through the nut in the oven where the column comes out of the inlet. Use ultraclean technique at all times when handling the gold seal. 1. Cool the inlet to < 80 oC to minimize oxidation and prevent burns. 2. Turn off the carrier gas flow (follow the procedure listed above for Changing the Liner and Septum). 3. Remove the column from the inlet. (Once the column is removed, the ferrule will need to be replaced and the column cut.) 4. Remove the insulator cup (if present) to reveal the nut housing the gold seal. 5. Remove the nut and discard the used gold seal and washer. 6. Insert the new washer and gold seal into the nut: a. Insert the washer into the nut; the washer goes between the gold seal and the nut. b. Insert the gold seal into the nut with the grooves visible on top. (These grooves are the exits for the split gas during split injection and for the purge flow after a splitless injection.) 7. Reinstall the nut containing the gold seal and washer, and tighten. 8. Replace the insulator cup over the nut (if present). 9. Replace the ferrule and trim the column as described below. 10. Reinstall the column and check for leaks with a leak detector. (If you turned off the carrier gas, turn it back on and let it flow for a few minutes before you check for leaks.) 11. Turn on the inlet temperature.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory GC Maintenance HP6890 Version 2.0
12. A blank should be run to bake out the system before analyzing samples. Trimming the Column and Installing a New Column (Inlet/Detector): Tools needed: column cutting wafer, inch wrench NOTE: Hydrogen and airflow to the detector must be turned off any time the detector end of the column is removed from the FID unit. Any time a column is removed from the inlet/detector, the ferrule should be replaced and the column should be trimmed. The inlet end of the column may be cut several inches to remove active sites and restore separation capacity. 1. 2. 3. 4. 5. 6. Cool the inlet and detector to < 80 oC. This will minimize oxidation and prevent burns. Turn off the carrier flow following the procedures in Changing the Liner and Septum. Remove the column from the inlet and detector by loosening the column nuts. Remove the column nut(s) and ferrule(s). The ferrule, which is made of graphite, may be stuck to the nut; remove all ferrule particles. Slide a septum, column nut, and ferrule onto the free ends of the column (tapered end of the ferrule should point away from the column nut, the flat side toward the nut). Cut two or three inches off the end of the column: a. The column should be cut by scoring one side with a wafer and then snapping the column at the score. b. Inspect this cut with a magnifying glass. This cut must be clean and contain no rough edges. If there are rough edges, repeat until a clean cut is achieved or seek guidance. c. Wipe the end of the column with a Kimwipe and methanol, or hexane. d. Place the column back into the inlet or detector. When installed, the column should protrude 5 mm (4 mm to 6 mm) beyond the ferrule into the inlet. Use the septum as a guide for this measurement. e. Tighten the column nut so that the column does not slide with a gentle tug. The detector end of the column should be prepared in the same manner. Insert the column into the detector until it comes to a stop, finger tighten the nut, pull the column back approximately 1 mm or as noted by the manufacturer. Use the septum as a guide for this measurement. Tighten the column nut. Turn on the carrier gas, and check for leaks with a leak detector.
7. 8.
9.
Conditioning the Column: If conditioning is not done properly the column may be ruined. 1. Allow the carrier gas to flow through the column for approximately one hour with the GC oven at room temperature. 2. Ramp the oven temperature at 10-15 degrees per minute to the final conditioning temperature. The final conditioning temperature should be 10 degrees higher than the maximum oven temperature to be used in the method but should not exceed 10 degrees below the maximum operating temperature of the column as recommended by the manufacturer.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory GC Maintenance HP6890 Version 2.0
3. Condition the column several hours or overnight. Cleaning FID Jets or Replacement: Refer to HP 6890 Series Gas Chromatograph Maintenance and Troubleshooting Manual. Cleaning the Split Side Arm: The split vent side arm is the exhaust for split gasses and compounds that are purged off during an injection. This can become very dirty and without maintenance can cause deleterious results. 1. 2. 3. 4. 5. Cool the injector temperature and turn off the gasses. Remove the liner from the inlet as specified in the Changing the Liner and Septum section. Remove the autosampler tray, tower, fan cover, and the top rear instrument cover. Remove the split side arm from the inlet to the filter located at the rear of the instrument. Inspect the ends of the side arm to make sure they are not clogged. If they are, use an old syringe to unclog the ends. 6. Using vacuum, pull a solvent such as chloroform through the side arm and into waste. 7. Repeat step 4 using a solvent such as methanol and then air to dry. 8. Using a small brush or Q-tip dipped in chloroform clean out the inlet arm where the side arm attaches. 9. Reconnect the side arm to the inlet and the filter. 10. Replace the covers and autosampler tray and tower. 11. Change the liner, o-ring and septum. 12. Turn on the temperatures and gases. 13. Check for leaks.
QUICK REFERENCE ROUTINE CALIBRATION AND OPERATION AGILENT AND HP 5973 MASS SELECTIVE DETECTOR Standard Spectra Autotune: For each day the MS is used a Standard Spectra Autotune shall be performed prior to routine instrument operation and after instrument maintenance: From the Instrument Control Screen select Instrument, select Perform MS Autotune, select Standard Spectra Tune from the Select Tune Type menu (see attached). Tune is automatically saved in the Stune.u directory. Review the Tune Report and file for future reference. Criteria for the Standard Spectra Tune are as follows: 1. 2. 3. 4. 5. 6. 7. Low background: less than 150 peaks Low water and air: less than 10% Correct mass assignments 0.2 amu (69, 219, 502) Symmetrical, smooth mass peak shapes Consistent mass peak widths (0.55 0.1) (Default value of Drug MS is 0.55) Isotope mass assignments should be 1 amu greater than parent peaks Appropriate EM voltage: 1000-2800 electron volts. If the voltage is not within these limits, review the history of the electron multiplier or consult a supervisor. The EM voltage will increase over time as the source becomes dirty with use. Cleaning the source should return the EM voltage to a normal operating level. 8. Mass 69 abundance should be 150,000-400,000 9. Typical relative abundance: - Mass 69 = 100 % - Mass 219 = 35-130% - Mass 502 = 2-5 % 10. Proper isotope ratios: - Mass 70/69 = 0.5-1.6 % - Mass 220/219 = 3.2-6.4 % - Mass 503/502 = 7.9-12.3 %
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 1
GC/MS Quick Reference Version 2.1
Target percent relative abundances are set for certain other PFTBA masses. The system will come as close as possible to the values shown: - Mass 50 1 % - Mass 131 55 % - Mass 414 3.5 % If the tune meets the criteria listed, the chemist who performed the tune will initial and circle the initials. All other chemists using the instrument that day will review the tune for acceptance of data and initial. Autotune: From the Instrument Control Screen select Instrument, select Perform MS Autotune, select Autotune from the Select Tune Type menu (see attached). Tune is automatically saved in the Atune.u directory. Review the Tune Report and file for future reference. Acceptable criteria for the Autotune are as follows: 1. 2. 3. 4. 5. 6. 7. Low background: less than 150 peaks Low water and air: less than 10 % Correct mass assignments 0.2 amu (69, 219, 502) Symmetrical, smooth mass peak shapes Consistent mass peak widths (0.55 0.1) (Default value of Drug MS is 0.55) Isotope mass assignments should be 1 amu greater than parent peaks Appropriate EM voltage: 1000-2800 electron volts. If the voltage is not within these limits, review the history of the electron multiplier or consult a supervisor. The EM voltage will increase over time as the source becomes dirty with use. Cleaning the source should return the EM voltage to a normal operating level. 8. Mass 69 abundance should be 150,000-400,000 9. It is normal at times to have a base peak of 219 instead of 69 10. Relative abundance: - Mass 69 = 100 % - Mass 219 = 35-130% - Mass 502 = 2 % 11. Isotope ratios: - Mass 70/69 = 0.5-1.6 % - Mass 220/219 = 3.2-6.4 % - Mass 503502 = 7.9-12.3 %
If the tune meets the criteria listed, the chemist who performed the tune will initial and circle the initials. All other chemists using the instrument that day will review the tune for acceptance of data and initial. Failure of Tune To Meet Acceptable Criteria: If the daily Standard Spectra Autotune fails, the operating chemist will take note of any error messages generated by the Chemstation Software, check all sources of leaks for tightness, and inform a supervisor. The supervisor will evaluate instrument operation and attempt to correct any problem. If the problem is resolved, an Autotune and Standard Spectra Autotune shall be performed, and applicable technical, maintenance, and repair information documented in the GC/MS Maintenance Logbook. If the problem cannot be resolved, the instrument will be marked out of service and a supervisor will arrange for instrument repair. GC/MS Maintenance Schedule: GC/MS maintenance is documented in the GC/MS Maintenance Logbook located near the instruments. Daily/with use Tune (Standard Spectra Autotune) Change wash vial solvents Check gases Weekly Monthly Change liner and o-ring Change gold seal Trim column Back up data and sequences to CD or disc Check autocal vial Replace rough pump oil Replace septum (minimum-weekly) Trim column Change gas tanks Replace column
GC/MS Quick Reference Version 2.1 3
6 months Yearly As needed
Change filament Clean source Clean split side arm To Set Up and Run a Sequence on the GC/MS System: From the MS Top Screen select Sequence, select Edit Sample Log Table Click on blank line in the table. The box labeled as Type should read Blank for method blanks and Sample for unknown samples. Use the tab key or mouse to move to the Vial box and enter the corresponding vial number to the unknown sample in the autosample tray. Move to Method and enter the name of the method to be used for the current sample. (For a list of methods, hold the shift and ? keys in this field.) Supply the Sample Name and any Miscellaneous Information (if any), and the Expected Barcode. Use the Repeat, Cut, Copy, and Paste buttons as appropriate to add samples to the table. Repeat copies the highlighted line, increments the vial number, places the new line immediately after the highlighted one. Copy copies the highlighted line without change. Use Paste to position the copied line where you wish. When finished entering unknown samples into the sample log table, click OK. From the MS Top Screen select Sequence, and select Run. Enter correct data file name (i.e. C:\HPCHEM\1\DATA\ or C:\MSDCHEM\1\DATA\), insure that Full Method is checked under Method Sections to Run, that Inject Anyway is checked under On Barcode Mismatch, and click OK. Select Sequence and Save from the menu task bar. Enter correct sequence path name and click OK (ie. C:\HPCHEM\1\SEQUENCE\ or C:\MSDCHEM\1\SEQUENCE\). From the MS Top menu task bar select Sequence and Print Sequence. Print Full format. From the MS Top Screen select Sequence, and select Run. Start sequence table by clicking on Run Sequence.
To Analyze MS Data: To load a data file: From Data Analysis Screen menu task bar, select File, and select Load Data File. Select a data file name and click OK, or double click the left mouse button on the data file name. The total ion chromatogram (TIC) for the data file is loaded and displayed in window [2].
A data file must be loaded to perform any of the following tasks. To integrate a chromatogram: From Data Analysis menu task bar, select either Chromatogram/Integrate or Chromatogram/AutoIntegrate. To select a spectrum: Double-click the right mouse button on the time point of interest in the chromatogram. The mass spectrum appears in window [1].
To print a spectrum: From Data Analysis task bar menu, select File, and select Print. Choose either Select Window or TIC & Spectrum from one of the options and the correct window desired.
To zoom in: Position the pointer at one corner of the area you wish to expand in a chromatogram or spectrum. Press and hold the left mouse button while dragging the mouse to select the area you wish to expand. Release the mouse button. The selected area expands to fill the existing window. To zoom out: Position the pointer anywhere in the zoomed window. Double-click the left mouse button. To average spectra: Position the pointer in the chromatogram at the starting time for the range you want to average. Press the right mouse button while dragging the mouse to the end of the range you want to average. Release the mouse button. The spectra in the selected range are averaged and the averaged spectrum is displayed in window [1]. To subtract two spectra: Select a spectrum (double-click the right mouse button in the chromatogram). Select the spectrum to be subtracted (double-click the right mouse button in the chromatogram). Select Subtract from the Spectrum menu. The spectrum selected second is subtracted from the first spectrum and the resulting spectrum is displayed in window [1].
To Use Spectral Libraries: To search a single spectrum: In Data Analysis, load a data file. The TIC is displayed. Select a spectrum. The selected spectrum appears in a window below the chromatogram. Initiate the library search by double-clicking the right mouse button in the window containing the spectrum. When the search is complete, the search results appear on the screen. The spectrum for the unknown, the reference spectrum you select from the list of hits, and, if available, the chemical structure of the reference compound is displayed. To print library results: Click the Print button to print a copy of the displayed spectra. Click Done button to clear the library search results from the screen.
References: HP5973 MSD/HP6890 Series GC Quick Reference Calibration and Maintenance procedure in the GC/MS Operation Procedure notebook
GC/MS Quick Reference Version 2.1 5
GC/MSD Chemstation and Instrument Operation Student Manual Vol #1 Course #H4043A
Gas Chromatography/Mass Spectrometry CALIBRATION AND MAINTENANCE Agilent/HP 6890 GC with 5973 Series Mass Spectrometer Tuning the instrument is performed by the use of the autotune program which collects the mass and abundance for selected isotope fragments of the tuning compound, perfluorobutylamine (PFTBA). The autotune report can be used to monitor system performance over time and can indicate the need for maintenance. All chemists are responsible for ensuring the proper operation of the gas chromatograph/mass spectrometers (GC/MS) and general housekeeping of the instrumentation as needed. I. Calibration A. Standard Spectra Tune
The Standard Spectra Tune ensures standard response over the full mass range. This tune is recommended to optimize mass spectral library searches. Therefore, calibration of the GC/MS is performed each day of use by using the Standard Spectra Tune. Tuning should be performed after instrument maintenance and before operation. For the 5973 instruments tune as follows: 1. Select the instrument control screen. 2. Select the Instrument menu item. 3. Select Perform MS Autotune menu item. 4. Select tune to perform Standard Spectra Tune. 5. Review the tune using criteria listed below. Acceptable criteria for the Standard Spectra Tune are as follows: 1. 2. 3. 4. 5. 6. 7. Low background: less than 150 peaks Low water and air: less than 10% Correct mass assignments 0.2 amu (69, 219, 502) Symmetrical, smooth mass peak shapes Consistent mass peak widths (0.55 0.1) Isotope mass assignments should be 1 amu greater than parent peaks Appropriate EM voltage 1000-2800 electron volts. If the voltage is not within these limits, review the history of the electron multiplier or consult a supervisor. The EM voltage will increase over time as the source becomes dirty with use. Cleaning the source should return the EM voltage to a normal operating level. (If it does not, the EM may be going bad.) 8. Mass 69 abundance should be 150,000 - 400,000 9. Typical relative abundance: - Mass 69 = 100 %
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory GC/MS Maintenance HP6890 Version 2.1
- Mass 219 = 35-130% - Mass 502 = 2-5 % 10. Proper isotope ratios: - Mass 70/69 = 0.5-1.6 % - Mass 220/219 = 3.2-6.4 % - Mass 503/502 = 7.9-12.3 % In addition, the standard auto tune sets targets for the percent relative abundances for certain other PFTBA masses. The system will come as close as possible to the values shown: - Mass 50 1 % - Mass 131 55 % - Mass 414 3.5 % If the tune meets the criteria listed, the chemist who performed the tune will initial and circle the initials. All other chemists using the instrument that day will review the tune for acceptance of data and initial. B. Autotune Autotune maximizes instrument sensitivity over the mass range, using PFTBA masses 69, 219, and 502. This tune is used in troubleshooting instrument operation. The standard spectra tune should be run prior to routine instrument operation. Acceptable criteria for the Autotune is as follows: Low background: less than 150 peaks Low water and air: less than 10% Correct mass assignments 0.2 amu (69, 219, 502) Symmetrical, smooth mass peak shapes Consistent mass peak widths (0.55 0.1) Isotope mass assignments should be 1 amu greater than parent peaks Appropriate EM voltage: 1000-2800 electron volts. If the voltage is not within these limits, review the history of the electron multiplier or consult a supervisor. The EM voltage will increase over time as the source becomes dirty with use. Cleaning the source should return the EM voltage to a normal operating level. 8. Mass 69 abundance should be 150,000 - 400,000 9. It is normal at times to have a base peak of 219 instead of 69 10. Relative abundance: - Mass 69 = 100 % - Mass 219 = 35-130% - Mass 502 = 2 % 11. Isotope ratios: - Mass 70/69 = 0.5-1.6 % - Mass 220/219 = 3.2-6.4 % - Mass 503/502 = 7.9-12.3 %
1. 2. 3. 4. 5. 6. 7.
If the tune meets the criteria listed, the chemist who performed the tune will initial and circle the initials. All other chemists using the instrument that day will review the tune for acceptance of data and initial. C. Quick Tune
Quick Tune provides re-tuning for optimum response and resolution, and for accurate mass assignment. Only the mass axis, peak widths, and EM voltage are adjusted; the lenses are unaffected. This tune may be used to rapidly check tuning after maintenance. The Standard Spectra Tune should be run prior to routine instrument operation. D. Failure of a Tune to Meet Acceptable Criteria:
If the Autotune or Standard Spectra Autotune fails, the operating chemist will take note of any error messages generated by the Chemstation Software, check all sources of leaks for tightness, and inform a supervisor. The supervisor will evaluate instrument operation and try to correct the problem. If the problem is corrected, Standard Spectra Autotune shall be performed, and applicable technical, maintenance, and/or repair information documented in the GC/MS Maintenance Logbook. If the problem cannot be resolved, the instrument will be marked as out of service, and the supervisor will arrange for instrument repair. II. Maintenance GC/MS maintenance is documented in the GC/MS Maintenance Logbook located near the instruments. GC/MS Maintenance Schedule Daily/with use Tune: Standard Spectra Autotune Change wash vial solvents Check gases Change liner and o-ring Change gold seal Trim column Back up data and sequences to CD or disc Check autocal vial Replace rough pump oil Replace septum Trim column
GC/MS Maintenance HP6890 Version 2.1
Weekly Monthly
6 months Yearly As needed
Change gas tanks Replace column Change filament Clean source Clean split side arm Maintenance activities should be consolidated to minimize instrument down time. For example, if it is time to change the gold seal and cut the column, it may be more prudent to change the liner and septum at that time or hold off changing the gold seal until the liner and septum are changed. The maintenance interval is not a rigid schedule and should be based upon workflow. Unless there is an immediate problem, a chromatographic run should continue to completion, and the maintenance performed at the end of a run. Where duplicate instrumentation exists, only one instrument at a time should be scheduled for preventive maintenance to avoid multiple instruments being out of service at the same time. Ultraclean Technique: Liners, gold seals, and certain other parts must be handled using ultraclean techniques to avoid contamination of the part with oils from the skin, plasticizers from plastic laboratory benchcoats, etc. Do not handle an ultraclean part with your hands; wear cotton gloves or use a fresh Kimwipe. Lay the ultraclean part on a cloth wrapping or on a clean Kimwipe. Changing the Wash Vial Solvent: Discard waste solvent into the hazardous waste. Replace spent solvent in wash vials with fresh methanol, and replace the vials in the appropriate location on the instrument autosampler. Changing the Liner and Septum: Tools needed: inlet wrench and tweezers. NOTE: Turn off carrier gas; if the septum nut is removed without turning off the carrier gas, the glass wool plug may be blown out. Use ultraclean technique at all times when handling the new liner. 1. Cool the inlet to < 80 oC to minimize oxidation and prevent burns. 2. Turn off the carrier gas on the GC by using the keypad on the front panel of the GC: From the front panel on the GC, push the FRONT INLET button, scroll down using the button to the Total Flow line. Turn off the flow by pressing the OFF button. Once maintenance is complete, turn the flow back on by pressing the ON button. 3. Once the inlet is cool and the gas is off, remove the septum nut. Remove and replace the septum. Reinstall and tighten the septum nut. These are not ultraclean parts but care should be taken to avoid any unnecessary contamination.
4. Remove the nut covering the liner. Remove the old liner and discard. 5. Using ultraclean technique, carefully slide an o-ring over the end of the new liner, approximately from the top of the liner, and slide into the inlet. The glasswool plug should be nearest the bottom of the liner. 6. Replace and tighten the nut and turn the inlet temperature and carrier gas flow on. 7. A blank should be run to bake out the new liner before analyzing samples. Changing the Gold Seal: Tools needed: appropriate screw driver (Phillips/Torx/etc.), 9/16 wrench, wrench Note: The gold seal is located at the bottom of the inlet. It is accessed through the nut in the oven where the column comes out of the inlet. Use ultraclean technique at all times when handling the gold seal. 1. Cool the inlet to < 80 oC to minimize oxidation and prevent burns. 2. Turn off the carrier gas flow (follow the procedure listed above for Changing the Liner and Septum). 3. Remove the column from the inlet. (Once the column is removed, the ferrule will need to be replaced and the column cut.) 4. Remove the insulator cup (if present) to reveal the nut housing the gold seal. 5. Remove the nut and discard the used gold seal and washer. 6. Insert the new washer and gold seal into the nut: a. Insert the washer into the nut; the washer goes between the gold seal and the nut. b. Insert the gold seal into the nut with the grooves visible on top. (These grooves are the exits for the split gas during split injection and for the purge flow after a splitless injection.) 7. Reinstall the nut containing the gold seal and washer, and tighten. 8. Replace the insulator cup over the nut (if present). 9. Replace the ferrule and trim the column as described below. 10. Reinstall the column and check for leaks with a leak detector. (If you turned off the carrier gas, turn it back on and let it flow for a few minutes before you check for leaks.) 11. Turn on the inlet temperature. 12. A blank should be run to bake out the system before analyzing samples. Trimming the Column (Inlet): Tools needed: column cutting wafer, inch wrench Any time a column is removed from the inlet, the ferrule should be replaced and the column should be trimmed. The inlet end of the column may be cut several inches to remove active sites and restore separation capacity. 1. Cool the inlet to < 80 oC. This will minimize oxidation and prevent burns. 2. Turn off the carrier flow following the procedures in Changing the Liner and Septum. 3. Remove the column from the inlet.
4. Remove the column nut and ferrule. The ferrule, which is made of graphite or graphite/vespel, may be stuck to the nut; remove all ferrule particles. 5. Slide a septum, column nut, and ferrule onto the free end of the column (tapered end of the ferrule should point away from the column nut, the flat side toward the nut). 6. Cut two or three inches off the end of the column: a. The column should be cut by scoring one side with a wafer and then snapping the column at the score. b. Inspect this cut with a magnifying glass. This cut must be clean and contain no rough edges. If there are rough edges, repeat until a clean cut is achieved or seek guidance. c. Wipe the end of the column with a Kimwipe and methanol, or hexane. d. Place the column back into the inlet. When installed, the column should protrude 5 mm (4 mm to 6 mm) beyond the ferrule into the inlet. Use the septum as a guide for this measurement. e. Tighten the column so that the column does not slide with a gentle tug. 7. Turn on the carrier gas, and check for leaks with a leak detector. Installing a New Column: Tools needed: column cutting wafer, inch wrench, MSD installation tool 1. Cool the injector and vent the detector. After the MS has vented, open the vacuum manifold. 2. Turn off the carrier flow following the procedure in Changing the Liner and Septum. 3. Loosen the column nuts from the injector and the transfer line from the detector, and remove the column. 4. Place a nut and ferrule on each end of the column. (The flat side of the ferrule goes toward the inlet nut and tapered side out, and the tapered end slides into the MS transfer line nut, with the flat side out.) NOTE: These are different types of ferrules. See supervisor for instruction. 5. Cut about 2-3 inches from the inlet end of the column using techniques described in Cutting the Column. 6. Install the new column into the inlet. 7. An MS column, which has been developed for use in a mass selective detector has extremely low column bleed and can be conditioned while in the detector. MS columns can also be conditioned prior to installation into the GC/MSD interface. 8. The column can be installed in the mass spec detector either with an installation tool or without. a. Without the tool: Slide the column into the GC/MSD interface until it can be pulled through the vacuum manifold, trim 1-2 cm off the end of the column, clean the outside of the column with methanol or hexane, adjust the column so it protrudes 1-2 mm past the end of the interface, hand tighten the nut, readjust the column as necessary and tighten the nut to turn. (Check tightness after one or two vent cycles. b. With the tool: Slide the column into column installation tool, trim 1-2 cm off the end of the column, clean the outside of the column with methanol or hexane, adjust the column so it protrudes 1-2 mm past the end of the tool, hand tighten the nut, slide the septum to touch the end of the nut, use two wrenches to tighten the nut turn, remove the column and nut from the installation tool (total length from the septum to the end of the column is
176 mm), clean the outside of the column with methanol or hexane, insert the column into the interface, and tighten the nut to turn. (Check tightness after one or two vent cycles. 9. Close the vacuum chamber, and pump down the mass spectrometer. Conditioning the Column: Tools: Carrier gas, wrench If conditioning is not done properly the column may be ruined. Do not use hydrogen to condition a column. 1. Allow the carrier gas to flow through the column for approximately 5-15 minutes with the GC oven at room temperature 2. Ramp the oven temperature at 5-15 degrees per minute to the final conditioning temperature. The final conditioning temperature should be at least 10 degrees higher than the maximum oven temperature to be used in a method. Do not exceed the manufacturers recommended maximum operating temperature. 3. Hold this temperature and allow carrier gas to flow for several hours, or overnight. 4. Return the GC oven temperature to a low standby temperature. Cleaning the Split Side Arm: The split vent side arm is the exhaust for split gasses and compounds that are purged off during an injection. This can become very dirty and without maintenance can cause deleterious results. 1. 2. 3. 4. 5. Cool the injector temperature and turn off the gasses. Remove the liner from the inlet as specified in the Changing the Liner and Septum section. Remove the autosampler tray, tower, fan cover, and the top rear instrument cover. Remove the split side arm from the inlet and the filter located at the rear of the instrument. Inspect the ends of the side arm to make sure they are not clogged. If they are, use an old syringe to unclog the ends. 6. Using vacuum, pull a solvent such as chloroform through the side arm and into waste. 7. Repeat step 4 using a solvent such as methanol and then air to dry. 8. Using a small brush or Q-tip dipped in chloroform clean out the inlet arm where the side arm attaches. 9. Reconnect the side arm to the inlet and the filter. 10. Replace the covers and autosampler tray and tower. 11. Change the liner, o-ring and septum. 12. Turn on the temperatures and gasses. 13. Check for leaks. Cleaning the Source for Mass Spectrometers/Changing the Filament/Fill Autocal Vial/ Change Rough Pump Oil: Follow instructions provided in the Agilent/HP 5973 Hardware Manual.
Operation of the 5973 Series Mass Spectrometer

Utilizing G1701BA and G1701DA, Versions B.01.00 and D00.00 Mass Spectrometer Chemstation Software Overview The Chemstation software is a windows based application and usable on Windows 95, 98, NT and 2000. The following instructions are intended as a general guide and do not represent all of the ways that the software may be used. In depth literature is published by the manufacturer and should be used as the key reference and troubleshooting guide. Online help is also available at www.agilent.com. Users may begin using the Chemstation software by clicking on the Top icon located on the desktop. There are several ways to move around in the Chemstation software. The primary method of movement from one screen to another in the software is through the View menu item. If in the Top Level, Instrument Control or Data Analysis, the View menu item will allow the user to move to any of the other screens. Users are referred to the appropriate instrument manuals for additional information. Top Level Screen From the Top Level screen the user has the ability to edit, print, load, and save methods and sequences. This screen enables the user to begin instrument operation. Instrument Control Screen From the instrument control screen the user has the ability to modify instrument parameters such as injector, inlet, column and oven parameters, mass spec temperatures, and electron multiplier voltages. The user can also load, edit, save and print methods from this screen. This screen enables the user to monitor instrument parameters during a run. Instrument tuning is also done from this screen. Data Analysis Screen From the data analysis screen the user can edit the data analysis portion of a method, edit and select libraries, review total ion chromatograms, review individual spectra, compare unknown spectra to known libraries, and print reports. Data analysis is also the location of the quantitation databases used in quantitative methods. Storage of Data files, Methods and Sequences The hard drive is partitioned into two drives, drive C:\ and drive D:\. The C:\ drive is the location of the Chemstation and the operating system (Windows NT 4.0 or Windows 2000). The D:\ drive is designated for storage of other installed software. For the operation of the Mass Spectrometer Chemstation there are three key areas of storage.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Operation 5973 GC/MS Version 2.0
Data is stored in C:\hpchem\1\data\ or C:\msdchem\1\data\. Data consists of data.ms files containing total ion chromatograms and their spectra, pre_post.ini files containing information on the status of the instrument during the run of the indicated file, and log files containing information regarding the sequence that was run. Methods are stored in C:\hpchem\1\methods\ or C:\msdchem\1\methods\. Methods contain all of the information required to execute a run. Methods contain the required macros and instrument parameters such as oven ramping, pressure and flow parameters, injection temperatures and specifications, and instrument temperatures, as well as other information. Sequences are stored in C:\hpchem\1\sequence\ or C:\msdchem\1\sequence\. Sequences contain the information required to complete a series of injections utilizing the autosampler tower, tray and barcode reader. The information consists of vial number, sequence line number, sample name, method name, and operator information.
I. Sequences Loading a sequence or creating a new sequence From the top level: 1. Select Sequence menu item. 2. Select Load menu item. 3. Select the default.s or a previous sequence and OK. This sequence of events will load the selected sequence in the instruments memory. Proceed to Edit sequence section to edit the sequence or Running a sequence section to run the loaded sequence. Edit sequence Once the desired sequence has been loaded you may want to edit the sequence. From the top level: 1. Select Sequence menu item. 2. Select Edit Sample Log Table menu item. The edit screen will appear at this time with the sequence that the user had previously loaded. The user may edit, cut, copy, paste, or repeat lines in the sequence. Information that is required is Type, Vial, Method, and Expected Barcode. Line and Data File will be completed automatically by the Chemstation software. The user may wish to include a Sample Name and/or Miscellaneous Information.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 2 Operation 5973 GC/MS Version 2.0
Line the line number in the sequence completed by the Chemstation Type the type of sample: sample, blank, calibrator, etc. Data File unique number for a particular sequence generated by the Chemstation. This number is only unique for the specific sequence and may be repeated in other sequences. Method the method that the user wishes to employ. Sample Name the description of the sample. Repeat will increment the highlighted line by one number and will copy the highlighted line for all other information. Cut will delete the highlighted line (will hold it in memory for paste function until another cut or a copy is executed). Copy will copy the highlighted line to be used with the paste function. Paste will paste the highlighted line that has been cut or copied and insert it above the current highlighted line. Once the editing is complete select OK to exit. If you do not exit out of the edit screen the sequence will not continue.
Edit a sequence while it is running The user may also edit the sequence while a sequence is running to add specimen to the running sequence. If the sequence is running the menu items will not be available and the user must select the Edit Samp Log Tbl button from the center screen. At this time the edit sequence screen will appear.
Saving a sequence After the user has loaded and edited a sequence, the sequence must be saved. From the top level: 1. Select the Sequence menu item. 2. Select the Save menu item. The default location to save sequences is C:\hpchem\1\sequence\ or C:\msdchem\1\sequence\. If the location varies from the default location, locate the default and then save. The format in which to save the sequence is: month, day, and letter designation (ie: Jul29A).
Running a sequence There are several ways to begin a sequence. The user may select Run, Load and Run, or Position and Run. The Run menu item will begin the current sequence. The Load and Run sequence menu item will allow the user to load a new sequence
and then run it. The Position and Runmenu item will allow the user to begin the current sequence at a user specified location within the current sequence. To begin a sequence from the top level: 1. Select the Sequence menu item. 2. Select the Run, Load and Run, or Position and Run menu item. 3. The user will be prompted to enter the data path for the storage of the data collected during the sequence run. The following screen will appear:
4. Make sure that Full Method and Inject Anyway are selected. The user may or may not want to select the Overwrite Existing Data Files option. 5. Type the correct path for the storage of data. The default path is C:\hpchem\1\data\ or C:\msdchem\1\data\. The default location is where the Chemstation looks for data from the data analysis screen. If the user does not store the data in the default location be aware of the location that was selected. 6. Select Run Sequence button. If the sequence does not begin at this time, check the above screen and the selections to make sure that the proper selections have been made. If Load and Run or Position and Run are selected the user will be prompted to load a sequence or select a position within a sequence before the above screen appears. Using Keywords The Chemstation allows the use of keywords within a sequence. These keywords allow the user to perform certain functions within the sequence such as running a command or macro, tuning the instrument, or pausing the sequence. The most common keyword used is Pause. The Pause command will allow the user to pause the sequence after an injection or series of injections are made, or a method is complete. To use a keyword:
1. From the Sample Log Table: 2. Select the next line to be run, copy and paste so there are two identical lines. 3. Select the Keyword command in the Type box. 4. This will prompt the user to type the Keyword that is to be used. Select the keyword desired in the keyword box. 5. Select OK. When the current method is complete the keyword command will be executed.
II.
Methods
The method in the Chemstation software is the location of all of the instrument parameters and data analysis parameters that will take place during the execution of a method in a sequence. Methods should only be created or modified by those with knowledge of the required parameters or experience with instrument parameters. Creating a new method To create a new method the user may select menu items from either the Top Level screen or the instrument control screen. From either screen: 1. Select the Method menu item. 2. Select the Load menu item. 3. Load the default method or load an existing method and modify to create your new method. 4. Select the Save menu item to save the method with an appropriate name.
Operation 5973 GC/MS Version 2.0
Editing an existing method If the user needs to edit a method or check the contents of a method the method may be accessed from the Top Level screen or the instrument control screen. From either screen: 1. Select the Method menu item. 2. Select the Edit Entire Method menu item. 3. Edit or review the method. Save the method ONLY if aware of the changes that have been made. DO NOT save the method if unaware of the changes made.
III. Data Analysis The user may enter data analysis in several different ways. The user may open it from the icon on the desktop, from the Top Level screen during the execution of a sequence, or from the View menu item from any screen.
Loading a chromatogram (data file) The data file contains the total ion chromatogram and the spectra for the identified compounds in the chromatogram. To load a chromatogram: 1. Select the File menu item. 2. Select the Load Date File menu item. 3. Select the appropriate data file from the correct data path and select OK. (The data file will be in the location that was specified when the sequence was started. The default location is C:\hpchem\1\data\ or C:\msdchem\1\data\.)
Reviewing Data Once the chromatogram has been loaded it will appear in window [2]. It will appear as the total ion chromatogram (TIC) with peaks representing detector response to compounds and their retention times. There are many items that the user may want to use to review data. These include, but are not limited to, integrating, searching for specific ions, comparing unknown spectra to known libraries, generating reports, and printing spectra. The TIC will be normalized on the largest peak. This means that the largest peak will be completely visible from the top of the peak to the baseline. Large peaks in a TIC may make smaller peaks not visible when looking at the normalized chromatogram. If the user wishes to look at the smaller peaks the zoom function may be used. To do this left click and hold the mouse, drag a box around the area to be analyzed and release. This will zoom in on the area that the box was drawn around. Generating, comparing and printing spectra To generate a spectra at a specific TIC retention time double click the right mouse button at the point of interest. This will generate a mass spectrum of the area that was clicked on and it will appear below the TIC in window [1].
The method that is loaded will determine what library or libraries have been selected for use (see searching libraries for instructions on changing). To compare the generated spectrum with the library that is specified in the method: 1. Double click the right mouse button while the cursor is in window [1].
2. This will generate a comparative spectrum with the best match being shown first. This comparative spectrum is generated in window [24].
The spectrum for the unknown compound is located on top and the spectrum for the best match is located on the bottom. The library that the best match spectrum came from is listed in the PBM Search Results box. 3. To print the comparison, select the Print button located in the PBM Search Results box, or select the File menu item, Print and Selected Window. Type in the number 24 when prompted. The user may also want to print the TIC or the unknown spectrum without using the comparison method. To do this: 1. 2. 3. 4. Load chromatogram and generate spectrum. Select the File menu item. Select the Print menu item. The user will be prompted to enter the window to print. Enter the number of the window desired to print: 1 = spectrum, 2 = TIC.
Searching for specific ions When a specific ion needs to be isolated so the user may separate coeluting compounds or find a compound with a weak response, the Chemstation can search for specific ions within a TIC. To do this: 1. Select the Chromatogram menu item. 2. Select the Extract Ion Chromatograms menu item 3. This will prompt the user to enter the ion or ions that the Chemstation will look for and the retention time windows in which to look. 4. Enter the ions of interest. 5. Enter the retention time window in which to look. 6. Select the OK button. This will prompt a window displaying the extracted TIC and each ion will be displayed in a different color. Full spectrum may be generated by double clicking the right mouse button on the area of interest. It may be of value to the user to overlay these extracted ion chromatographs. To do this, select the Chromatogram menu item and Display Ion Chromatogram in Merged Format. Subtracting Spectra In the event that two compounds coelute or a compound with a weak response is hidden in the baseline it may be desirable to subtract interfering spectra. An example would be a drug that gives a weak response and is hidden in the baseline. In this instance the user would look for the ions in the compound of interest by a manual search or by searching for extracted ions. Once the compound is found the full spectrum may have ions from the baseline included in the ions for the compound. The Chemstation allows the user to subtract one spectrum from another to clean up or clarify a spectrum. To subtract a spectrum: 1. Obtain a mass spectrum of the desired compound (with the interfering ions). 2. Move the cursor to a location, usually just before or just after the compound of interest, and obtain a spectrum of that area. 3. Select Spectrum menu item. 4. Select Subtract menu item.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Operation 5973 GC/MS Version 2.0
10
The resulting spectrum will be the difference of the second obtained spectrum removed from the first obtained spectrum. Printing reports There are many ways to print reports and a number of reports are available in the Chemstation software. The most common way to print a report for a designated method is to: 1. Load the appropriate TIC. 2. Select the Method menu item. 3. Select the Run Method menu item. 4. This will run the data analysis portion of the method that has been selected in data analysis. It will print the report as if the sample had been run in a sequence. The user may also want to use the options available under the quantitate and tools menu items.
Using the Quantitate and Tools menu items the user will be able to use the Chemstation report format and to reprocess quantitative and qualitative reports linked to custom reports. Searching Libraries There are many different libraries that are available in the Chemstation ranging from inhouse libraries to commercially purchased libraries. The libraries are located on the C:\ drive of the computer in C:\database. To change the libraries that are automatically searched in a method:
11
1. Select the Spectrum menu item. 2. Select the Select Library menu item. 3. This will prompt the user to type in the name of the library to be used. This will affect the library that is used when the user is comparing an unknown spectrum to a library spectrum. Note: Mass Spectral libraries are used only as a tool in identifying an unknown. Chemists must consider a variety of factors before reaching a conclusion.
Addition of New Mass Spectra to Libraries If a new compound or standard is obtained, it is important to have this compounds mass spectra in a spectral library. The spectral library to place the new compound must be selected, usually it will be dcflab.l. (See instructions above to select the appropriate library.) To add a mass spectrum to a library: 1. Select the Spectrum menu item. 2. Select the Edit Library menu item. 3. Select the Add New Entry menu item.
4. Enter information into appropriate cells: Name, Mol. Formula, Mol. Weight, etc. 5. Select the Include in search box.
6. After entering the information, select OK, and the Chemstation software will add the spectra into the library in the C:\database\ file path. 7. The Chemstation software will also assign the new spectra a number.
Parametric Retrieval is located under the View menu item in Data Analysis screen only. Parametric retrieval can be used to search a user designated library for compounds based upon name, molecular weight, CAS number, library entry number, etc. However, the information must be in the library before the search parameters will work. Occasionally all of the information may not be available with in-house libraries.
13
Tuning the 5973 The tuning instructions and parameters are located in the maintenance log and the reference collection found near each instrument.
14
OPERATION OF THE SPECTRUM ONE FTIR TABLE OF CONTENTS
1. How To Perform an Instrument Validation Check 2. How To Perform a System Suitability Check 3. How To Run a Background Scan 4. How To Run a Polystyrene Scan 5. How To Run a Negative Control 6. How To Analyze a Sample 7. How To Perform a Library Search 8. How To Perform Routine Maintenance 9. How To Change the Scale of a Spectrum 10. How To Print Reports using a Template 11. How To Add Spectra To a Library
FTIR Operation Version 2.0
OPERATION OF THE SPECTRUM ONE FTIR
Overview Users may begin using the Spectrum One software by clicking on the Spectrum icon located on the desktop. There are several ways to move around in the Spectrum One software. The primary method of movement from one screen to another in the software is through the various menus and the toolbar. The toolbar is the strip of icons that appears under the menus in the top of the Spectrum Window. The tools in the toolbar provide a quick way of executing some of the commands. Users are referred to the appropriate instrument manuals for additional information. Getting Started 1. Check to make sure the UATR with the KRS-5 crystal is in place. 2. Clean the top of the crystal and the probe using a Kimwipe and acetone. 3. Double click the Spectrum icon on the desktop. 4. Login: Login name = druglab. Click OK. 5. A pop-up window will appear, Accessory Ready Check, click on Cancel. 6. Click the Monitor icon on the toolbar, record the current energy in the FTIR QC Logbook.
1. Running the Instrument Validation Check: (Monthly) a. Carefully remove the ATR accessory from the instrument. (Raise the sample cover to the vertical position, press the release clip, slide the accessory out of the sample area. b. In the Instrument menu, go to Validate and click on Validate Instrument. c. Click OK. d. The report should print off automatically, if not, print it manually. Evaluate, initial, and place the report in the FTIR QC Logbook.
2. Running the System Suitability Check: (before use) a. Slide the ATR accessory back into the instrument. b. Clean the top of the crystal and the pressure arm using a Kimwipe and acetone. c. Check the energy of the system by clicking on the Monitor icon. The energy should be displayed in the window after a few seconds. d. Record this energy in the FTIR Maintenance Logbook. e. In the Instrument menu, go to Validate and click on System Suitability. f. Click OK. g. The report should print off automatically, if not, print it manually. Evaluate, initial, and place the report in the FTIR QC Logbook. 3. Running a Background Scan: (before use) a. Click on the Instrument icon. b. Under the Sample tab, name the scan. (Name = background 2-11-09). c. Under the Scan tab, the range start is 4000 cm-1, end is 515 cm-1, the scan type should be background, the units should be egy, the scan should not be minimized, and the scan number should be set to 32. d. Under the Instrument tab the resolution should be 4 cm-1. e. Click Apply. This button will then change to Scan. Click to begin background scan.
f. Print the background spectra, evaluate, initial, Logbook.
and place in the FTIR QC
4. Running a Polystyrene Scan: (Monthly) a. Click on the Instrument icon. b. Under the Sample tab, name the scan. (Name = polystyrene 2-11-09). c. Under the Scan tab, the range start is 4000 cm-1, end is 515 cm-1, the scan type should be sample, the units should be %T, the scan should not be minimized, and the scan number should be set to 32 scans. d. Under the Instrument tab, the resolution should be 4 cm-1. e. Under the Beam tab, click on the red disk with holes. Select polystyrene and click OK. f. Click Apply. This button will then change to Scan. Click to begin polystyrene scan. g. Integrate (click on Peaks icon) and print the polystyrene spectra. Evaluate it to determine if acceptable, initial it, and place in the FTIR QC Logbook.
5. Running a Negative and Positive Control a. Negative Control i. A negative control is run before each positive control and before each sample. ii. Follow procedure below for running a sample omitting the addition of sample (Step 4). b. Positive Control i. A positive control is run each day the instrument is used before samples are analyzed. ii. Follow procedure below for running a sample and use procaine HCl as the sample. 6. Running a Sample a. Click on the Instrument icon. b. Under the Sample tab, choose the desired scan type (sample); type in the necessary information about your sample. (Name = FL# + date analyzed). c. Click the Monitor icon within this window. d. Add sample to crystal. e. Move the pressure arm directly over the crystal. f. Monitor the force gauge as you tighten the pressure arm to about 75. g. Under the Scan tab, make sure the range start is 4000 cm-1, end is 515 cm-1, the scan type should be sample, the units should be %T, the scan should not be minimized, and the scan number should be 32. h. Under the Instrument tab, the resolution should be 4 cm-1.
i. Click on Apply. This button will then change to Scan. Click to begin sample scan.
7. Library Search a. IR Search helps you identify an unknown material. It will search through a library or libraries to find the spectra that best matches your unknown sample. It can also interpret your unknown spectrum directly by determining from the pattern of peaks (and absence of peaks) which functional groups are likely to be present. b. Even if your sample is not in the libraries, an interpretation will give you information on what functional groups are present in your sample and provide clues as to what it is. c. If you know that your sample is one of a range of materials or if you want to measure how similar your spectrum is to another spectrum, you should use Compare in the Process menu; this compares your sample spectrum with one other spectrum or several spectra and reports the degree of similarity between the spectra. d. There are three types of library search available: i. Interpretation An interpretation compares the peaks in your sample spectrum with the peaks that correspond to 896 Possible Structural Units (PSUs) or functional groups. A list of PSUs in your sample spectrum is displayed. You do not need to have a search library to perform an interpretation. ii. Euclidean Compares every point in your sample spectrum with the points in spectra in a library search. iii. Library search Compares the position of the peaks in your sample spectrum with the position of peaks in the spectra in a search library. e. Searching the Library i. Click on the IR search icon in the toolbar. ii. A match will automatically appear on the screen.
iii. To find a different match, click on the Euclidean Search Hit List window and double click on the desired match.
iv. To separate the spectra, click on View, then Overlay/Split, then Split.
8. Routine Maintenance a. FTIR maintenance is documented in the FTIR Maintenance Logbook located near the instrument. The maintenance interval is not a rigid schedule and should be based upon workflow. There may be long periods when the instrument is not used. If this is the case, an instrument validation check, system suitability check, and a polystyrene test should be performed every month at a minimum. b. FTIR Maintenance Schedule i. Daily/with use 1. System Suitability 2. Clean crystal and pressure arm 3. Check energy ii. Monthly 1. Polystyrene Standard 2. System Suitability 3. Instrument Validation iii. As needed 1. Replace dessicant c. Replacing the dessicant: i. Click on Intrument icon. ii. Click on the Tools icon, the Adjustments ToolBox will appear. iii. Click on the Maintenance button. iv. Remove the ATR accessory. v. Remove the two screws on the top panel using a Phillips screwdriver. vi. Note the position of the dessicant in its location before replacing it. vii. Reassemble the panel with the screws. viii. Re-install the ATR accessory. ix. Click on the Changed box under the Dessicant header. Click OK.
9. To change the scale of the spectrum: a. Make sure the spectrum view is set to overlay by going to View, Overlay/Split display, then click Overlay.
10
b. Then go to View and click on Format View. In the Ranges tab, change the scale of the abscissa and/or ordinate by typing in the desired ranges. Then click OK.
10. Printing Reports Using the Drug Lab Template a. Go to Setup and click on Report Template.
11
b. In the Report Builder Screen, go to File and click on Open Template.
c. Open Template called Drug Lab. d. Click on Top Graph Holder so the box turns purple.
12
e. Go back to Spectrum Screen with sample scan displayed and click RptBld. The sample scan should now be in the Top Graph Holder.
f. Go to library match screen and delete the sample scan from the screen by clicking on the scan name and hitting the delete button on the keyboard.
13
g. On the Report Builder Screen click on the Bottom Graph Holder so the box turns purple.
h. Go back to the Spectrum Screen with the library match displayed and click RptBld. The library match should now be in the Bottom Graph Holder.
14
i. On the Report Builder Screen go to File and click Print.
11. Adding Spectra To A Library a. The reference standard should be analyzed as described in the section on how to run a sample. b. When describing the sample, the name should be the name of the drug; the description should be the name of the drug, the manufacturer, and the lot number. c. Use the comment section to document the sampling technique (UATR) and the number of scans collected (32). d. Click on the drop down menu Setup, click IR Search (do not use the IR Search icon on the toolbar). e. Hi-light the appropriate library DCFLAB. f. Click Search. A new window opens. Click on Lib Utils from the drop down menu. g. Click on Add to Library, next click on top Browse button, find the spectrum name (cocaine HCl.sp or a specific case #) that was generated from your run. Hilight this file, click Open, verify the name, type in a New Spectrum ID (e.g. DL0001), and click OK. h. Seek supervisory assistance as needed.
15
PROCEDURE: ANALYTICAL BALANCE 1) Monthly Calibration a) At the beginning of each month, analytical balances should be calibrated and their linearity checked using certified weights that are located in the Drug Lab. b) Using the Balance Maintenance Log Calibration Check logsheet, record the date, actual weight, and the difference in the last decimal place measured. c) This difference must be within +/- 3 counts in the last place measured. d) If the difference is out of control, then the balance must be evaluated and recalibrated. i) Ensure that the balance is level and clean. ii) Refer to the balance manual for calibration procedure or seek supervisory assistance. e) Once the balance is recalibrated, the linearity should be checked using three certified weights (lower, middle and upper range), and the results recorded on the Balance Maintenance Log - Calibration Check. Also, include the chemists initials, the date, the certified weights used, actual weight measured, and the difference between the last decimal place measured and the target weight. 2) Daily Calibration Check a) Each day before the balance is used, the calibration must be checked. b) On the Balance Maintenance Log sheet record whether the balance is level and clean. c) Also record the difference (in counts) between the measured secondary weight and its target weight. d) The difference must be within +/- 3 counts in the last place measured. e) If the difference is out of control, then the balance must be recalibrated. i) Refer to the balance manual for the calibration procedure or seek supervisory assistance as needed. f) Once the balance is recalibrated, the linearity should be checked using three certified weights (lower, middle and upper range), and the results recorded on the Balance Maintenance Log Calibration Check logsheet. Also, include the chemists initials, the date, certified weights used, actual weights measured, and the difference in the last decimal place measured. 3) For balances with flashing recalibration messages: a) If the secondary weight is within +/- 3 counts after recalibration, no further action is necessary other than recording results in Balance Maintenance Log. 4) For balances with automatic recalibration: a) No additional action is needed other than daily calibration checks. 5) Seek supervisory assistance in any situation in which a balance cannot be calibrated properly or when balance operation appears unusual.
Analytical Balance Version 2.0
PIPET CALIBRATION Using Pipette Tracker Software Using the Pipette Tracker Software A summary of operation procedures is located at the end of this procedure. Pipette Tracker keeps information about pipettes in device files, which list details for each pipette. This program also uses separate methods, each defining how the calibration should be done. The method includes details on the number of expected volumes to be calibrated for the run, the number of samples to be tested for each expected volume, and the frequency of evaporation blanks. The method also specifies the testing criteria to be used in determining whether the calibration passes or fails. When a devices interval determines that it is due for a calibration, the device will appear automatically on the built-in WorkList. A time (reminder) has been specified for each interval, which will place the device on the WorkList before it is due to allow you to plan you your calibrations. When calibrating, the weights will be automatically collected from the balance and put into the computer. You simply follow the on-screen steps and the Pipette Tracker performs all the calculations, providing instant feedback if the device passes or fails the calibration and generates a report. This report will be kept by the laboratory Quality Manager. Logon/Logoff The program will ask for the users password each time the program is started. Each user must log on by entering his or her password in the [Logon] dialog box so that the program can record it. Once the log on has taken place, the program will automatically insert the users name into the operator field on the Calibration Run screen. The user can log off by pulling down the Password menu and select Logoff. Only certain components of the program will remain active. The next user may then log on by pulling down the Password menu and selecting Logon. Run Calibration There are three ways to get to the Run Calibration screen: From the WorkList, you can select a device and pull down Device menu and select Run Calibration. The currently highlighted device and method will be loaded. From the Inventory List, you can select a device and pull down Device menu and select Run Calibration. The currently highlighted device and the most due method (or the closest to being due) will be loaded. From the Device Setup screen, you can click on the Cal button to calibrate that particular device. The currently edited device and the most due method (or the closest to being due) will be loaded. Calibration Run Screen
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 1 Pipette Calibration Version 2.0
The top entries and the Environmental factors (Temp., Barom. P., Rel. Hum.) can be edited before Starting a Run. Once started, you are locked out of these areas. After the last weight is received, the Environmental factors can be revised again, if necessary. If these are changed after the last weight is received, the program will also recalculate the calibration results and statistics. After confirming the correct Input Mode (Balance), shown in the Balance ID field, ensure that the device and method are the ones you want before running the calibration. The following describes some of the components (others found in the manual) of the Run Calibration screen: Temp. -- This field contains the liquids temperature in degrees Centigrade. This information can be read from the meter found on the wall behind the balance. As noted earlier, this can only be edited at the start of a Calibration Run. Barom. P. -- This field contains the barometric pressure in mm of mercury ( inner most scale times 10). This information can be read from the barometer found on the wall behind the balance. As noted earlier, this can only be edited at the start of a Calibration Run. Rel. Hum. -- This field contains the value for the relative humidity, with the units as %. This information can be read from meter found on the wall behind the balance. As noted earlier, this can only be edited at the start of a Calibration Run. Abort Run -- Deletes all current weight, time and volume information for this run. Environmental factors are not changed. Device selection, operator name and method are not changed. Redo Sample -- This button becomes active once you have entered the Tare weight and the first calibration sample. Clicking on it cancels the results entered for the most recent sample only, and resets the Run Line to the previous sample so you can re-send the weight from the balance. Please note the Redo Sample button does not allow you to re-dispense the sample, only to correct an error in entry. If you have a problem with dispensing the sample, consider using the Void button at the end of the Run (see details below), or abort the calibration and start again. Void Sample -- This button becomes active only at the end of the Calibration Run, and before you click on Accept Run. If you experience problems dispensing one or more samples, you may choose to exclude these from the calculations for the Run, rather than abort the Run and start over. In effect, the calculations are performed with fewer sample(s). When the Run is finished, you will be able to move the cursor up and down through the cumulative weights shown on the Calibration Run display grid. Position the cursor over the sample to be voided, then click on the Void button. The calculated Volume for this sample will be replaced with the word VOID. You are able to Unvoid the sample by positioning your cursor over the voided cumulative weight and clicking on the Void button again. This reverses all effects of clicking Void. Accept Run -- This button only becomes active when the run is complete. Clicking on it accepts the Calibration Run, stores the results, then inactivates the button and the report will be automatically printed. Done -- Leaves the Calibration Run screen and goes back to the WorkList or Inventory List.
Pipette Calibration Version 2.0
Run Calibration Automatically (Through the Balance) First, ensure that the balance is calibrated properly by following the QA Procedure for the Analytical Balance, then proceed as follows from the Run Calibration screen: Making sure that the correct device (pipette) and method are listed, edit the Environment factors (Temp., Barom. P., Rel. Hum.) Start the calibration by clicking on Start Run button. Place your container with liquid on the balance. Tare the balance and click on Tare Wgt, which sends the weight over to the Calibration Run Table. Important: Tab over to Sample Wgt. Pipette a volume into the container. Once a stable weight has been obtained, press Enter, which sends the weight over to the Calibration Run Table. Continue until all volumes have been recorded. Using Abort, Void Sample and Redo as necessary. At the completion of the calibration , either click on Accept Run or Abort Run. Print a hard copy of the report, sign and file next to the balance or with the Quality Manager. Another device may be chosen by pulling down the device list. Once the calibration(s) are completed, click on Done. Log off by clicking on Password and choosing LogOff.
Calibration of Pipetters
Procedure Summary
There may be three reasons for checking calibration of a pipetter: Receiving a new pipetter into the laboratory. See the Quality Manager prior to calibration. Regularly scheduled calibration. Suspected error in calibration. A condensed summary follows: The computer, printer and balance should be on. If not, allow the balance to warm up for thirty minutes. With the 20 gram weight, check the balance calibration and record weight on the Balance Calibration Log. See Quality Manager if balance is not in calibration. Log on to the computer by pulling down the Password menu and selecting Logon. If you do not have a password, see Quality Manager. From the WorkList, select the pipetter that is to be calibrated. Pull down the Device menu and select Run Calibration. From the meters on the wall, enter the current temperature, barometric pressure and relative humidity. Begin the calibration by clicking on Start Run button. Place container with enough liquid to cover the bottom on to the balance. Tare the balance and click on the Tare Wgt button, which sends weight over to the computer Calibration Run Table. Important: Tab over to Sample Wgt button. Pipette a volume into the container. Once a stable weight has been obtained, press Enter, which sends the weight over to the computer Calibration Run Table. Continue until all volumes have been recorded. Using AbortRun, VoidSample and Redo as necessary. At the completion of the calibration, either click on Accept Run or Abort Run. Print a hard copy of the report, sign and file next to the balance or with the Quality Manager. Another pipetter may be chosen by pulling down the device list. Once the calibration(s) are completed, Click on the Done button. Log off the computer by pulling do down the Password menu and selecting Logoff. If pipetter performance is acceptable, the printed report is filed next to the balance or given to Quality Manager. If pipetter performance is not acceptable, immediately remove from service and notify Quality Manager.
DISPENSETTE CHECK
1) At the beginning of each week, solution dispensettes will be checked for volume accuracy. Materials needed for this check include volumetric flasks and the dispensette containing the solution to be dispensed. Dispense the solution into the flask and note the volume in the flask. The bottom of the meniscus should be at the line of the volumetric flask. Solution MeCl2 w/IS BuCl w/IS EtOH w/IS Dispensette Vol. 10 ml 5 ml 2 ml Volumetric Flask 10 ml 5 ml 2 ml
2) Using the Weekly Dispensette Volume Check log sheet, record the date, the initials of the chemist performing the check, the solution and volume dispensed, and the size volumetric flask used. Document whether or not the volume dispensed equales the volume of the flask. a. If the volume dispensed meets the volume of the flask, the dispensette may be used. b. If the volume dispensed does not meet the volume of the flask, advise a supervisor and remove the dispensette from service or inspect, clean, or adjust it as needed. i. The procedure for readjusting the dispensette can be found in the Calibration section of the Dispensette Operating Manual. ii. If adjustments are required, document these in the Comments section of the Weekly Dispensette Volume Check log. 3) If dispensing the solution from the dispensette becomes difficult, the dispensette may require cleaning. a. Follow the cleaning procedure found in the Dispensette Operating Manual. 4) If an air bubble forms in the dispensette filling valve, unscrew the dispensette from the bottle, remove the filling tube and insert a glass pipette to remove any liquid around the valve. If this does not remove the air bubble, the dispensette may need a more thorough cleaning. 5) Do not interchange dispensettes between different solutions. a. If it is necessary to change the type of solution in the dispensette, clean the dispensette thoroughly and document any changes on the logsheet.
Dispensette Check Version 2.0
THERMOMETER CALIBRATION CHECK 1. Annual Calibration Check 1.1. Reference Thermometers 1.1.1. NIST traceable reference thermometers are maintained under the direction of the Quality Manager. 1.1.2. Calibration of appropriate reference thermometers is verified annually by an outside vendor. 1.1.3. Records are maintained by the Quality Manager. 1.2. General Use Thermometers 1.2.1. Calibration of laboratory mercury and alcohol thermometers will be verified annually against a NIST traceable thermometer. 1.2.2. General use thermometers should read within +/- 2 divisions of the NIST thermometer. 1.2.2.1. If the general use thermometer does not meet the designated range, it will be replaced or repaired. 1.2.2.2. In the event that replacement or repair is not an option, the amount of deviation from the NIST thermometer is recorded on applicable log sheets and laboratory thermometer readings are adjusted accordingly. 1.2.3. Records of annual thermometer calibration checks will be maintained in the laboratory.
Thermometer Calibration Check Version 2.0
DRUG CASE REVIEW GUIDANCE 1. Administrative and technical review will be performed by the analyst and the reviewer prior to release of a case report. 1.1. The primary responsibility for ensuring the technical and administrative correctness and completeness of the case file lies with the analyst of record. 1.2. Case review by someone other than the analyst of record does not shift the responsibility for the scientific findings from the analyst to the reviewer. 2. The purpose of case review is 2.1. To ensure that the conclusions of the analyst are reasonable and within the constraints of scientific knowledge 2.2. To ensure that the conclusions detailed in the final report accurately and fairly represent the content in the case file 2.3. To ensure that a the case file contains adequate information so that another competent analyst can independently evaluate and interpret the data 2.4. To ensure that proper laboratory procedures were followed 2.5. To ensure that the report is editorially correct and free from typographical errors 3. Types of review 3.1. Administrative review 3.1.1. A procedure to check for consistency with laboratory policy and for editorial correctness. 3.2. Technical review 3.2.1. Review of notes, data, and other documents which form the basis for a scientific conclusion. 4. Technical and administrative review require an extensive and careful review of the case file and final report including but not limited to the following 4.1. Does each page of the case file have a case number and analyst signature or initials? 4.2. Are all entries in the case file clear and legible? 4.3. Are corrections handled properly (single strike through with initial)? There should be no write-overs. 4.4. Transmittal sheet and chemist report 4.4.1. Are the following items consistent on the final report 4.4.1.1.Case name 4.4.1.2.IFS case number 4.4.1.3.Agency number: service number, tag number, etc. 4.4.1.4.Agency name 4.4.1.5.Date received 4.4.1.6.Received by 4.4.1.7.Delivered by 4.4.1.8.Item received (transmittal sheet)/evidence submitted (final report) 4.4.2. Is all suspected drug evidence which is listed as an exhibit on the transmittal sheet accounted for on the final report? 4.4.3. Are heavy non-drug items which are listed on the transmittal sheet accounted for on the final report? 4.4.4. Is the internal evidence transfer complete from Evidence Registration to the analyst?
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 1 Drug Case Review Guidance Version 2.0
4.4.5. Are the submission weight and exit weight noted, are they in reasonable agreement, is the balance used listed? 4.4.6. Are the analysts initials and date listed to acknowledge the date the evidence was opened and that the evidence was received sealed? 4.4.7. Are the analysts intials and date listed acknowledging when the evidence was closed and that it was sealed? 4.4.8. If the evidence was reopened, is documentation provided for subsequent entry and exit weights, notation about the status of the evidence seal, individual opening the evidence, and a comment as to why the evidence was reopened. 4.5. Drug Identification Protocol Sheet 4.5.1. Are case number, exhibit number, and specimen description consistent between the protocol sheet, transmittal sheet, and final report. 4.5.2. Is it clear which evidence was tested? 4.5.3. Is the daily QC acceptable? 4.5.4. Is the retention time of the standard similar to the retention time of the sample tested. 4.5.5. Is the tentative identification consistent with preliminary testing and the final result? 4.5.6. Are all calculations accurate? 4.5.7. Was the proper analytical procedure used? 4.5.8. Are the instruments used in analysis noted? 4.5.9. Is the result consistent with testing in the case file? 4.5.10. Are the results on the protocol sheet (including drug, units, numbers, percentages, etc.) consistent with the final report and with lab reporting procedures? 4.5.11. Is the starting date and completion date noted. 4.6. Analytical output 4.6.1. Where applicable, did the barcode read properly? If not, is the case number verified? 4.6.2. Is the drug identified consistent with the final report and other documentation in the case file? 4.6.3. Are values used in calculations accurately transcribed to the protocol sheet? 4.6.4. Are all standard data acceptable? 4.6.5. Is it clear which instrument and what parameters were used? 4.6.6. Is data of sufficient quality to identify and/or quantitate a controlled substance? 4.7. Final report 4.7.1. Is all information on the final report consistent with the case record? 4.7.2. Does the final report accurately and fairly reflect the work performed? 4.7.3. Was the work performed consistent with lab procedures? 4.7.4. Is the case file complete? 4.7.5. Is the report signed? 4.7.6. Is the report free of typographical errors and is good grammar used?
Drug Case Review Guidance Version 2.0
METHOD VALIDATION GUIDANCE Validation and verification of new testing methods is summarized in the Quality Manual. The Supervisor and/or Section Chief will design a specific testing protocol to validate the accuracy of a new method or to verify the accuracy of a method transferred from one instrument to another. I. Transfer of a Method to New Instrumentation First, proper calibration and operation of new instrumentation is verified and documented using the applicable instrument calibration process. Secondly, analytical results are compared between the new and old instrumentation. Where possible, side by side operation of old and new instrumentation is compared using the same samples or extracts. Standards, internal standards, and quality control samples should meet their respective criteria described in the analytical procedure. Test specimens should also be run and should meet acceptability criteria established for quality control samples or similar proficiency test results. Replicate analytical runs should be made on a single day and also compared between multiple days; criteria should meet those established for standards, internal standards, and quality controls; test specimens should meet acceptability criteria established for quality control samples or similar proficiency test results. As applicable, analysts are trained in operation of new instrumentation. II. Development of a New Analytical Method A supervisor outlines proposed steps for a new analytical method based upon literature, other established procedures, or knowledge and experience of the materials to be analyzed. The target sensitivity and linear range is established based upon expected sample conditions and instrument performance. Standard curves are run to assess the applicability of the proposed analytical technique in comparison to the targeted analytical needs. Once it is determined that sensitivity and range are appropriate, the method is evaluated for interference from expected matrices or co-occurring substances. The assay is evaluated for stability by repeating analysis of single prepared samples and duplicate preparation of samples both within day and between day. Variability of standards, internal standards, quality control samples, and specimens are evaluated with respect to literature values and/or similar established procedures. Criteria for acceptable standard curves, internal standards, and/or quality control samples are established. A formal procedure is written, applicable analysts are trained, and results are reviewed for stability within analyst. Method results may also be evaluated against external standards or specimen results from reference laboratories; criteria for acceptability will be similar to that established for quality control samples.
Method Validation Guidance Version 2.0
WORKSHEET OF POSSIBLE VALIDATION STEPS

Submitting Unit: Validation Title: Prepared by/Date: 1. Type of validation (check applicable box): Equipment Modification of an existing protocol New protocol Other: 2. Is this an original or revised validation plan? Original Revised 3. Describe the purpose or goal of the validation including the model and serial number of any new equipment: 4. Attach a copy of the written procedure: current (if applicable) and new. 5. The method is Quantitative Semi-quantitative Qualitative 6. Attach a validation plan with proposed timetable and list staff participating in the validation. The plan should lay out a course of action to assess the following, as applicable, which will be included in the final validation report: a. Method validation is the process of testing an analytical method, technique, or instrument to determine its suitability for meeting its intended purpose and to document its reliability under expected conditions of use. Generally the validation process is expected to i. Evaluate whether a new testing method meets identified analytical needs and current scientific practices; ii. Compare the new test methods performance with existing laboratory methodology; iii. Describe the conditions under which a testing method will produce valid results; iv. Predict possible sources of error; v. Determine limitations of a testing method; and vi. Establish baseline characteristics of the testing method (linearity, accuracy, etc.) which serve as benchmarks to evaluate future method
Method Validation Variables:

1. Evaluate whether a new testing method meets identified analytical needs and current scientific practices. 2. Compare the new test methods performance with existing laboratory methodology. 3. Describe the conditions under which a testing method will produce valid results. 4. Predict possible sources of error. 5. Determine limitations of a testing method. 6. Establish baseline characteristics of the testing method which will serve as benchmarks to evaluate future method performance. These characteristics may include: a. linearity/linear range: b. accuracy/precision: Accuracy is the ability to quantitate the true value of a control, standard, or sample. Precision is the closeness of agreement between many quantitations. Both accuracy and precision will be calculated using known controls and/or standards at several concentrations. The relative standard deviation and/or coefficient of variation will be used to determine precision and accuracy. c. specificity/selectivity: The selectivity of a method is the extent to which the method can determine a particular analyte of interest accurately in the presence of other components that may be expected to be present in the sample matrix. This parameter will be evaluated using actual samples and comparing the results to known methodologies. d. ruggedness/robustness: Ruggedness is the degree of reproducibility of the results obtained under a variety of conditions. These would include different analysts, reagents, extraction days, etc. These parameters are similar to precision and accuracy and will be evaluated using the results obtained from repeat analysis from several analysts. Robustness is the capacity of a method to remain unaffected by small deliberate variations in the method parameters. These changes/variations will be selected based on possible deviations from the analytical procedure such as extraction time, dry down time, solvent volume, changes in LC buffers, etc. e. repeatability/reproducibility: Repeatability is the ability of the method to accurately repeat a quantitation or identification under the same conditions (same analyst, same day, same instrument, same reagents, etc.). Reproducibility is the ability of the method to accurately repeat a quantitation or identification under different conditions (different analyst, different day, different instrument, different reagents, etc.). Each will be evaluated and reported as a relative standard deviation and/or coefficient of variation.
f. recovery efficiency: The recovery efficiency of a method is calculated by comparing the detector response to a known amount of analyte added to the sample matrix to that of a pure standard of the same quantity (concentration). This parameter will be evaluated for both compounds and expressed as a percentage recovery. g. limit of detection/quantitation: The limit of detection is the lowest concentration of an analyte in a sample that can be detected, though not necessarily quantitated. The limit of quantitation is the lowest concentration of an analyte that can be determined with acceptable precision and accuracy or is the amount equal to the lowest point on the calibration curve that can be determined with acceptable precision and accuracy. h. carryover: Carryover will be evaluated using high control samples followed by blank analysis. The high control samples will begin at the high calibration curve point and move upward through concentrations that would far exceed those expected in casework.
Texas Controlled Substances Weight Ranges Texas Drug Law Penalty Group 1 (delivery, manufacture, and/or possession) < 1g > 1g but < 4g > 4g but < 200g > 200g but < 400g > 400g Penalty Group 2 (delivery, manufacture, and/or possession) < 1g > 1g but < 4g > 4g but < 400g > 400g Penalty Group 3 and 4 (delivery, manufacture, and/or possession) < 28g > 28g but < 200g > 200g but < 400g > 400g Marihuana (delivery) < oz > oz but < 5 lbs > 5 lbs but < 50 lbs > 50 lbs but < 2000 lbs > 2000 lbs Marihuana (possession) < 2 oz > 2 oz but < 4 oz > 4 ox but < 5 lbs > 5 lbs but < 50 lbs > 50 lbs but < 2000 lbs > 2000 lbs
State Weight Ranges Version 2.0
Federal Controlled Substances Weight Ranges Marihuana < 250 g > 250 g but <1 kg > 1 kg but <2.5 kg > 2.5 kg but <5 kg > 5 kg but <10 kg > 10 kg but <20 kg > 20 kg but <40 kg > 40 kg but <60 kg > 60 kg but <80 kg > 80 kg but <100 kg > 100 kg but <400 kg > 400 kg but <700 kg > 700 kg but <1000 kg > 1000 kg but <3000 kg > 3000 kg but <10,000 kg > 10,000 kg but <30,000 kg > 30,000 kg Heroin <5g >5 g but < 10 g >10 g but < 20 g >20 g but < 40 g >40 g but < 60 g >60 g but < 80 g >80 g but < 100 g >100 g but < 400 g >400 g but < 700 g >700 g but < 1 kg >1 kg but < 3 kg >3 kg but < 10 kg >10 kg but < 30 kg > 30 kg
Cocaine < 25 g > 25 g but <50 g > 50 g but <100 g > 100 g but <200 g > 200 g but <300 g > 300 g but <400 g > 400 g but <500 g > 500 g but <2 kg > 2 kg but <3.5 kg > 3.5 kg but < 5 kg > 5 kg but <15 kg > 15 kg but <50 kg > 50 kg but <150 kg > 150 kg
Cocaine base < 250 mg > 250 mg but <500 mg > 500 mg but <1 g > 1 g but <2 g > 2 g but <3 g > 3 g but <4 g > 4 g but <5 g > 5 g but <20 g > 20 g but <35 g > 35 g but <50 g > 50 g but <150 g > 150 g but <500 g > 500 g but <1.5 kg > 1.5 kg
Methamphetamine < 2.5 g > 2.5 g but <5 g > 5 g but <10 g > 10 g but <20 g > 20 g but <30 g > 30 g but <40 g > 40 g but <50g > 50 g but <200 g > 200 g but <350 g > 350 g but <500 g > 500 g but <1.5 kg > 1.5 kg but <5 kg > 5 kg but <15 kg > 15 kg
Ice <250 mg >250 mg but < 500 mg >500 mg but < 1 g >1 g but < 2 g >2 g but < 3 g >3 g but < 4 g >4 g but < 5 g >5 g but < 20 g >20 g but < 35 g >35 g but < 50 g >50 g but < 150 g >150 g but < 500 g >500 g but < 1.5 kg > 1.5 kg
PCP <5g > 5 g but <10 g > 10 g but <20 g > 20 g but <40 g > 40 g but <60 g > 60 g but <80 g > 80 g but <100 g > 100 g but <400 g > 400 g but <700 g > 700 g but < 1 kg > 1 kg but <3 kg > 3 kg but <10 kg > 10 kg but <30 kg > 30 kg
Drug Evidence Disposal Facility Information
Sharps Environmental Services, Inc. P.O. Box 245 900 Lasalle Parkway Carthage, TX 75633 903-693-2525 903-693-3709 fax Contact: Judy Lawhorn Call two weeks prior to arrange disposal date and time. Certified peace officer or DEA licenses must accompany IFS staff.

SWIFS Controlled Substances Resources Documents v2.1 (02.2009) 76 Pages PDF

Hochgeladen von

Dokumentinformationen

Originaltitel

Copyright

Verfügbare Formate

Dieses Dokument teilen

Dokument teilen oder einbetten

Freigabeoptionen

Stufen Sie dieses Dokument als nützlich ein?

Sind diese Inhalte unangemessen?

Copyright:

Verfügbare Formate

SWIFS Controlled Substances Resources Documents v2.1 (02.2009) 76 Pages PDF

Hochgeladen von

Copyright:

Verfügbare Formate

Controlled Substances Resource Documents Version 2.

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Revisions and Corrections Resource Documents, Version 2.1

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

GC/MS Overview Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

GC/MS Overview Version 2.0

Directions for Operation of the HP 6890 GC with G2070AA Chemstation

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Operation of the HP 6890 GC Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Operation of the HP 6890 GC Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Operation of the HP 6890 GC Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Operation of the HP 6890 GC Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

GC Maintenance HP6890 Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

GC Maintenance HP6890 Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

GC Maintenance HP6890 Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 1

GC/MS Quick Reference Version 2.1

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 2

GC/MS Quick Reference Version 2.1

6 months Yearly As needed

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 4

GC/MS Quick Reference Version 2.1

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 6

GC/MS Quick Reference Version 2.1

6 months Yearly As needed

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Operation of the 5973 Series Mass Spectrometer

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Operation 5973 GC/MS Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Operation 5973 GC/MS Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Operation 5973 GC/MS Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Operation 5973 GC/MS Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Operation 5973 GC/MS Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Operation 5973 GC/MS Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

Operation 5973 GC/MS Version 2.0

OPERATION OF THE SPECTRUM ONE FTIR TABLE OF CONTENTS

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

FTIR Operation Version 2.0

OPERATION OF THE SPECTRUM ONE FTIR

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

FTIR Operation Version 2.0

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

FTIR Operation Version 2.0

f. Print the background spectra, evaluate, initial, Logbook.

and place in the FTIR QC

Dallas County Institute of Forensic Sciences Controlled Substances Laboratory

FTIR Operation Version 2.0

FTIR Operation Version 2.0