Clinical laboratory methods lecture notes

for midwifery students

Section V clinical chemistriy

By Henok.B (MLT,HI)

January 2018
DDU

1
1. Understand Explain the basics of clinical
chemistry
2. Understand, interpret and explain carbohydrate
metabolism disorders along with the method used
in laboratory Dx methods
3. Understand, interpret and explain RFT
4. Understand, interpret and explain LFT
5. know the basics of body fluid analysis

2
Clinical Chemistry
 is defined as an area in laboratory medicine
that deals with chemical analysis of body
fluids such as blood, urine, spinal fluid as
well as feces, tissue, calculi and other
materials.
◦ Links the knowledge of general, organic,
Inorganic analytical & biochemistry with
an understanding of human physiology.
 Analyte
 Analysis
 Types of Analysis
◦ Qualitative
◦ Semi-quantitative
◦ Quantitave
 Analysis is defined as the procedural steps
performed to determine the kind or amount of
analyte in a specimen.
 Analyte is a substance or constituent in which the
lab conducts tests.
 Qualitative analysis is defined as a test that is
used to detect presence or absence of a particular
analyte from a given sample. Results are reported
as negative or positive.

 Semi-quantitative analysis is type of analysis that
is used to give a rough estimate of concentration
of a particular analyte from the given sample.
Results are graded as 0, 1+,2+ etc
 Quantitive analysis is the type of analysis that
involves accurate measurement of a particular
analyte from a given sample. The results are
expressed in mass units per given volume of
specimen.
 Analyzer
 Analyzer is defined as: an instrument used to
perform analysis
 Reagent
 Reagent is defined as: Chemicals (solution or
powder) that are use to convert the analyte from
the sample into a measurable form
 To assess the physiological function of our
body systems or organs
 To diagnose and monitor diseases
 To follow up response to treatment
 Provides biochemical testing of patient
sample
◦ Glucose
◦ Protein
◦ Bilirubin
◦ Creatinine
◦ Lipids
◦ Enzymes
◦ Electrolytes
◦ And Other biochemicals
Introduction
 Constructed from
o Carbon - carbo-
o Hydrogen & O2 – (hydrates, or
water)
 General formula Cn(H2O)n

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-Energy source- 1g= 4kcal/17kj
 by conversion of glucose CO2 +H2O + ATP
 Stored as glycogen in the liver or
triglyceride in adipose tissue
-Cell structure (plants-cellulose & animals-
chitin)
-Recognition markers
- eg. A,B,O blood types
-Structural component of nucleic acids
-Part of plasma membrane

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 Based on 6 different properties

1. Size of base carbon chain:

2. Number of sugar units:

3. Location of C=O group:

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4. Chemical reactivity:
reducing & non- reducing

5. Based on stereochemistry

13
 Structural polysaccharides
a. Cellulose

o Major component of plant cell

walls
o Polymers of glucose
o Very few organisms produce
cellulose, enzyme that hydrolyze
cellulose

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 Made of glucose with a nitrogen containing
group
 Major component of arthropod exosketeon
& fungal cell walls

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I. Hormone that decrease plasma glucose
- Insulin
II. Hormoes that increase blood glucose
- Glucagon
- Epinephrine/adrenalin
- Growth hormone & ACTH
(adrenocorticotropic hormone)
- Glucocortico steroids
-Thyroid hormone
- Somatostatin

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1.Hyperglycemia
Effects: immediate effects
Long term effect
2. Hypoglycemia-
3. A normal or decreased plasma glucose
concentration often with excretion of a
non glucose - reducing sugar in the urine
(inborn errors of CHO metabolism)

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Definition:
 is a group of metabolic disorders of CHO
metabolism in which glucose in under
utilized
 characterized by hyperglycemia resulting
from defects in insulin production,
secretion &/or action

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Classification of Diabetes mellitus

-Type I or IDDM (Juvenile diabetes)

-Type II- NIDDM

-Gestational diabetes mellitus (GDM)
- Malnutrition related (MRDM)

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Complications of Diabetes mellitus
- Ketoacidosis
- Neuropathy
- Retinopathy
- Angiopathy
- Nephropathy
- Infection
20

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 depends on the demonstration of
hyperglycemia

Diagnostic criteria (Type I)

1. history
2. Classic symptoms of diabetes
3. Demonstration of significant
hyperglycemia
Note: diagnosis of type II is more difficult
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 Glucose is among most common of clinical
laboratory analyses because of the high prevalence
of diabetes mellitus
 Hyperglycemia is the common characteristics of
group of disorders called diabetes
 Four types: I & II DM, gestational diabetes and
other causes of hyperglycemia
 Other carbohydrates present transiently in human
plasma, only glucose has reference ranges
Polyuria-
Polydipsia-
Polyphagia –
Unexplained weight loss-

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 Considerations:
Type of specimen
Normal values
Methods:
 There are different test methods for the
detection of both types of diabetes and
hypoglycemia

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 Serum and plasma are the most common
specimens for quantitative glucose analysis
 Glucose is also measured in CSF and urine
 Whole blood capillary glucose limited to
patient administered (self) or point-of-care
glucose monitoring. Not acceptable for
diagnosis of DM
 Serum (red top or
serum separator tube)
 Plasma (heparin,
citrate, EDTA)
 Fluoride-oxalate (grey-
top) will inhibit
glycolysis
 Serum/ plasma should
be separated from cells
after centrifugation
 Glucose is reduced 5-
7% per hour in
uncentrifuged/
unseparated sample
Specimen collection time options for
blood glucose measurements

-Random blood sugar (RBS)

-FBS (fasting blood sugar )
-Two- hours post prandial (2h.pp)

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4. Glucose tolerance test (GTT)
-GTT is a test used to diagnose mild or hidden
cases of diabetes
-Non-preferred method of diabetes diagnosis

i-Oral glucose tolerance

-Test procedure and interpretation
ii. Intravenous glucose tolerance test
Glucose tolerance test.docx

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1. Quantity of glucose administered
2. Rate of absorption
◦ low- in mal-absorption
◦ High-in Hyperthyroidism
3. Age
- Elderly people have decreased CHO
tolerance

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 Reference Ranges- from fasting
◦ Serum---------------------------74-106
mg/dl
◦ whole blood------------------- 65 -95 mg/dl
◦ CSF----------------------------- 40-70 mg/dl

 Compare the patient results with the

reference range to assess for hyper- or
hypoglycemia
 Non protein nitrogenous (NPN) substances: are end
products of metabolism that contains nitrogen
 Azotemia: An excess of urea or other nitrogenous
compounds in the blood
 Anti diuretic hormone (ADH): is a posterior
pituitary gland hormone, important for
reabsorption of water from the kidneys.
 Diabetic insipidus: A disorder associated with
secretion and metabolism of anti diuretic hormone
(ADH), manifested by excessive urine production.
 Nephron: functional units of kidney
 Gout: Group of disorders of purine
metabolism
 Renal failure: Acute or chronic decline in renal
function
 Filtration of small molecules
 Reabsorption of essential substances
 Secretion into urine from blood stream
 Excretion
 Hormonal regulation: erythropoietin, ADH,
aldosterone
 Homeostasis
 Non-selective filtration across semi-
permeable membrane
 Low molecular weight substances will pass
into the urine
 Filtrate lacks large molecular weight
substances such as protein, protein-bound
substances, cells
 GFR: ~120 mL/min
 Passive vs. active transport mechanisms
◦ Conserves water and essential substances
 Water: always passively reabsorbed
 Glucose, HCO3: active transport
 Eliminates waste products not filtered by
glomerulus
◦ Drugs that are protein-bound (protein remains in
bloodstream)
◦ Organic waste:
 Blood urea nitrogen = BUN
 Creatinine
 Uric acid
◦ Uromodulin protein secreted into tubules
◦ Acid-base regulation: H+, HCO3
 Defined as the plasma concentration of a
substance that when exceeded, the kidney
tubules will not reabsorb any more into the
bloodstream, resulting in the substance being
excreted into the urine
 Example: glucose ~160-180 mg/dl
 Glomerular disease
 Cystic renal disease
 Diabetes
 Renal calculi
 Toxic nephropathy
 Obstructive uropathy
 Miscellaneous renal diseases
 These are compounds that contain nitrogen,
but are not proteins
 Often called NPN compounds
 Include end products of metabolism
 Kidneys act to excrete metabolic waste into
urine
 Because the kidneys act to excrete these
compounds into urine, measurement of NPN
compounds in plasma is useful for
assessment of kidney function
 Include >15 compounds
◦ Amino acids
◦ Ammonia
◦ Blood urea nitrogen (BUN)

◦ Creatinine
Protein  amino acids  ammonia  urea
◦ Uric acid
Muscle breakdown product

Nucleic acid catabolism

 NPN compound present in highest
concentration in blood and urine
 U= Urea
 Blood Urea Nitrogen = BUN
 Urea contains 2 nitrogen atoms: 28 g
nitrogen/mole of urea
 BUN x 2.14 = urea
 Formation and excretion
◦ Synthesized in the liver: ammonia  urea

Protein  amino acids  ammonia [LIVER]  urea

◦ Conversion of ammonia to urea is last liver function

to fail in end stage liver disease
 Plasma ammonia levels rise
 Reference Range:
 For adults (Serum/plasma)……………….. 6-20 mg/dl

 New borne upto one week( Serum/plasma)……… 3-

25mg/dl
 Adult over 60 (Serum/plasma) ……………………..8-
23mg/dl
 Urine, 12-20 g/24hrs
Convert 22 mg/dL BUN to urea mg/dL

BUN 22 x 2.14 = Urea 47 mg/dL

 Clinical Significance:
 Plasma levels are dependent upon
◦ Diet
◦ Liver function
◦ Kidney function
◦ State of hydration
Clinical Significance:
Increased urea (BUN):
 Azotemia or uremia
◦ Increased protein intake (leads to increased urea
formation)
◦ Decreased kidney function (decreased excretion
into urine results in elevated plasma levels)
◦ Dehydration (lack of body water results in increased
levels)
 Clinical Significance:
Decreased urea (BUN):
◦ Decreased protein intake (leads to decreased urea
formation)
◦ Decreased liver function (decreased conversion of
ammonia to urea)
 Not a good test for GFR:
◦ Influenced by diet, liver function
 Enzymatic (indirect) method
 Chemical (direct) method
 Specimen collection/handling
◦ Serum or heparinized plasma
◦ Stability: up to 24 hours at room
temperature; 1 week at 2-4oC
 Specimen collection/handling
◦ Urine
 Timed collection preferred
 Must be diluted prior to measurement
 1:10, 1:20
 Stability: up to 1 week stored in refrig when pH<5
 Some bacteria are able to hydrolyze urea to ammonia
resulting in falsely decreased urine urea levels
 As ammonia increases in the urine, the pH becomes
more alkaline
 Formation and Excretion
◦ Spontaneously derived from creatine in muscle
 High energy ATP storage and use in muscle
◦ Produced at a constant rate day to day
◦ Excreted into urine through glomerular filtration;
not significantly reabsorbed or secreted by tubules
 Reference Range
 Serum Adult male: 0.6-1.1
mg/dl
Adult female: 0.5-0.8 mg/dl
Child: 0-0.6
mg/dl

 Urine Male: 800-2000 mg/24hr

Female: 600-1800 mg/24hr
 Amniotic fluid: 1-2 mg/dl
 Clinical Significance
◦ Endogenous substance
◦ Amount produced is constant day to day: levels vary
<10% per day
◦ Amount produced is proportional to muscle mass
◦ Filtered by glomerulus; not handled by tubules
 Good test for GFR
◦ Not affected by diet
 Clinical Significance
Increased serum creatinine:
◦ Renal disease = impaired renal function
◦ 50-60% renal function lost before serum
creatinine increased (renal reserve)
 Specimen collection/handling
◦ Serum or heparinized plasma
 Avoid hemolysis
 Avoid lipemia
 Stability: one week at refrigeration temps
 Urine
◦ Time collection preferred; random acceptable
◦ Stability: up to 4 days in refrigeration
◦ Longer when frozen
 Formation and excretion
◦ End product of purine metabolism (by liver)
◦ Purines are precursors to nucleic acids ATP and GTP
◦ Readily filtered by glomerulus, but then is handled
by tubules
 Not a good test for GFR
 Clinical Significance
◦ Increased uric acid
 Gout
 Increased breakdown of nucleic acids: chemotherapy
 Renal disease
◦ Decreased uric acid
 Severe liver disease
 Fanconi’s syndrome
 Over treatment with ‘allopurinol’
 Used by clinicians to differentiate causes of
azotemia:
◦ Pre-renal
◦ Post-renal
 Azotemia: condition of increased NPN in
blood
◦ Most often due to increased BUN and creatinine
◦ The major NPN used to evaluate kidney function
 Also uric acid
 Calculated: serum BUN (mg/dl)
serum creatinine (mg/dl)

 Normal ratio: 10-20 with majority around

12-16
 Differentiation of azotemia

 Increased ratio with

Tend to be caused by pre-
increased BUN,
renal conditions:
normal creatinine
Congestive Heart Failure
Shock, hemorrhage
Dehydration
Increased protein metabolism
Increased protein catabolism
 Differentiation of azotemia

 Increased ratio with

dysproportionate Tend to be caused by post-
renal conditions that obstruct
increased BUN,
urine flow:
slightly increased
creatinine Stone
Tumor
Sever infection
 Differentiation of azotemia

 Increased ratio with

Tend to be caused by - renal
increased BUN,
conditions that decrease
increased creatinine kidney function:
Acute renal failure
Chronic renal failure
Glomerulonephritis
Tubular necrosis
 Decreased ratio
with decreased Tend to be caused by
BUN conditions of decreased urea
production:
Low protein diet
Liver disease
 Renal clearance expresses volume of blood
cleared of a substance per unit of time
Example: mL of substance per minute
 Substance used to monitor GFR must meet
the following criteria:
◦ Filtered exclusively by glomerulus
◦ Not reabsorbed by kidney tubules
◦ Not secreted by kidney tubules
 Most often used = creatinine clearance
 Why is creatinine clearance most often used
to monitor GFR?
◦ Creatinine freely filtered by glomerulus
◦ Creatinine not ‘rehandled’ by tubules
◦ Creatinine is an endogenous substance
◦ Amount of creatinine produced per day is constant
◦ Amount of creatinine produced is proportional to
muscle mass
 Patient preparation
◦ Patient should be well hydrated
◦ Avoid coffee and tea (caffeine) on day of test
 Specimen collection/handling
◦ Timed urine collection: 24 hour preferred
 Measure total volume of urine collected
 Measure urine creatinine (mg/dl)
◦ Serum/heparinised plasma
 Collect blood specimen sometime during the urine
collection period
 Measure serum/plasma creatinine (mg/dl)
 Standard clearance formula:
UV U = urine creatinine (mg/dl)
P V = total volume of urine collected:
ml/min
P = plasma creatinine (mg/dl)
 Clearance corrected for body surface area:
UV x 1.73 A = body surface area (BSA)
P A 1.73 = average BSA
Consider the following data:
Serum creatinine: 1.8 mg/dl
Urine creatinine: 63 mg/dl
Total urine volume: 1680 ml/24hr = 1680
ml/1440 minutes
Patient height: 178 cm
Patient weight: 82 Kg
1. Calculate the urine creatinine (mg/24hr)
2. Calculate the CrCl
3. Calculate the corrected CrCl for body surface area
Consider the following data:
Serum creatinine: 1.8 mg/dl
Urine creatinine: 63 mg/dl
Total urine volume: 1680 ml/24hr = 1680
ml/1440 minutes

1. Calculate the urine creatinine (mg/24hr)

63 mg x 1680 ml x 1 dl = 1058.5 = 1058

mg
dl 24 hr 100 ml 24 hrs
Consider the following data:
Serum creatinine: 1.8 mg/dl
Urine creatinine: 63 mg/dl
Total urine volume: 1680 ml/24hr = 1680
ml/1440 minutes

2. Calculate the CrCl = UV/P

63 mg/dl x 1680 ml = 40.8 = 41 ml

1.8 mg/dl 1440 min min
Consider the following data:
Serum creatinine: 1.8 mg/dl
Urine creatinine: 63 mg/dl
Total urine volume: 1680 ml/24hr = 1680
ml/1440 minutes
Patient height: 178 cm
Patient weight: 82 Kg
Surface area = 2.00 m2
3. Calculate the corrected CrCl for body surface area

41 ml x 1.73 = 41 x 0.87 = 35.7 = 36 ml

min 2.00 min
 National Kidney Foundation recommends a
EGFR be calculated each time a serum
creatinine is reported
 Want to detect chronic renal disease earlier
 Predicts GFR based on patient age, sex, body
size, race, serum creatinine
 Do not need to collect timed urine: better for
patient
EGFR (ml/min) =

(140 - age) x (Weight in kg) x (0.85 if

female)
72 x Serum creatinine in mg/dl
 Reference Range

Adult male: 97-137 ml/min

Adult female: 88-128 ml/min

EGFR: >59 ml/min

 Clinical Significance
◦ Used to monitor GFR
◦ As renal function fails, CrCl decreases
◦ Dialysis indicated when CrCl critically low
(GFR ~ 10-20 mL/min)
 Correlates with Increased BUN/ Creatinine ratio
with increased BUN, increased plasma creatinine
and decreased ur. creatinine
 Right lobe Left lobe
 Carbohydrate
metabolism
 Protein metabolism
 Lipid metabolism
 Conjugation,
detoxification and
excretion
 Vitamin storage
 Digestion and formation
of bile
 Enzymes
 Bilirubin metabolism
Gall bladder Bile duct Pancreas
 Bile
 Bilirubin esters
 Bile Acid (salt or conjugates)
 Cholesterol
 Other
 Excreted waste products
 Bile emulsifies ingested fats for digestion
 Total and direct bilirubin
 Total protein
 Albumin,
 Cholesterol
 Triglycerides
 Urea
 Ammonia
 Liver Inflammatory Enzymes: AST, ALT,
ALP, GGT
 Cholesterol  Bile Acid
1. Hemoglobin released 5. Conjugated bilirubin
from red cells excreted in bile (via
2. Iron and globin are the gallbladder and
conserved common bile duct)
3. Insoluble albumin- 6. Enters sm. intestine as
bound unconjugated component of bile
bilirubin 7. Bacteria convert to
4. Albumin removed and urobilinogen (UBG)
conjugated to 8. ~15% UBG is
glucuronic acid by reabsorbed by
hepatocytes intestinal mucosa
1. Conjugated bili. (bili- 4.
rubin gluconoride)
enters the high pH of
small intestine. 5.
2. ß-glucoronidase
converts bilirubin
back to unconjugated 6.
form.
3. Bacteria reduce
biliribin to three
compounds known as
urobilinogen (UBG)
UBG is colorless
15% UBG is
reabsorbed and
returns to the liver
All re-excreted into
bile except 2-5% via
kidneys
Most (85%) UBG is
oxidized into urobilin,
the brown pigment of
feces
1. Jaundice: A condition characterized by a
brownish-yellow pigmentation of skin, sclera
& mucous membranes
2. Icterus: A condition characterized by an
increase of bilirubin in blood
3. Conjugated bilirubin (a.k.a.- direct): Bilirubin
coupled with glucuronic acid to enhance
solubility
1. Unconjugated bilirubin (a.k.a.- indirect):
Bilirubin that has not been coupled with
gluconic acid and is not water soluble until
bound to albumin
2. Total Bilirubin: Conjugated + unconjugated
bilirubin in the bloodstream
3. Delta Bilirubin: Conjugated bilirubin
covalently bound to albumin, usually a very
small fraction of total
 A symptom, not a disease
or disorder. The yellow
staining of connective
tissue from excess
bilirubin.
 Prominently - sclera of the Above photo is courtesy of the Centers for
Disease Control and Prevention
eyes and skin.
 Icterus describes the dark
yellow-brown color of
serum with increased
bilirubin.
 Normal serum
 Icteric serum
 Due to hemolytic anemia - acquired and inherited
 Caused by destruction of RBCs by either:
 Extravascular Hemolysis - RBCs sensitized w/ IgG
or complement, removed by phagocytes in tissues
(spleen and Kupffer cells of the liver).
 Intravascular hemolysis – less freq. than
extravascular.
 Laboratory Results: ↑ serum bilirubin (mostly
unconjugated), ↑ urine urobilinogen, ↓ Hgb ↓ Hct
MCV ↓ MCHC ↑ RDW & ↑ reticulocytes
Other Causes of Prehepatic Jaundice
 Neonatal physiologic jaundice
 Hemolytic disease of newborn

 Phototherapy
 Liver inflammation and/ or cellular damage due to
various causes - viral hepatitis, parasites,
malignancy, drug-induced hepatitis. Clinically HAV
& HBV most common.
 Also, metabolic liver diseases, alcoholic cirrhosis,
autoimmune hepatitis.
 Degree of bilirubin uptake varies (normal to
decreased).
 Laboratory results: ↑ serum bili (unconjugated &
conjugated), positive urine bilirubin, liver enzymes
elevated (esp. ALT and AST)
 Transport Failure
◦ Dubin-Johnson syndrome
 Conjugation Failure
◦ Crigler-Najjar syndrome
 Intrahepatic obstruction
◦ Drug induced [e.g., chlorpromazine]
 An obstruction of the bile ducts, which serve as the
conduit of bile from the liver to the duodenum
 The most common cause is gallstones, but tumors
in or near to the bile ducts can also impede bile
flow into the small intestine
 Since conjugated bilirubin is normally excreted as a
component of bile, bilirubin accumulates in
circulation, leading to jaundice
 Post-hepatic jaundice includes increased
conjugated bilirubin, detected in both serum and
urine
The absence of bilirubin in
bile negatively effects
digestion
1. The brown pigment
urobilin is not produced -
feces become pale and
clay-colored
2. Causes poor absorption of
fats and fat-soluble
vitamins (A, D, E, K).
Deficiencies of these
nutrients are possible
 Non-hemolyzed serum or
heparinized plasma
 Fresh urine
 Protect from light (e.g.,
wrap collection tube in
aluminum foil)
 Light exposure will reduce
bilirubin and UBG detected
 Compare patient result with reference range
 * EU = “Ehrlich unit”, is equivalent to 1 mg/dL
 Newborn range for full term,1-2 days old

Patient conjugated total bilirubin Urine Urine

(direct) unconj. + bilirubin, urobilino-gen
bilirubin conjugated conjugated
Newborn Not < 12.0 mg/dL
(1-2 d old) applicable
Adult 0-0.2 mg/dL 0.3-1.2 negative <0.8 EU*
mg/dL
 The phosphatases include:
◦ Alkaline phosphatase (ALP)
◦ Acid phosphatase (ACP)
◦ Red cell phosphatase

Phosphatases catalyze the following:

 Organic phosphate monoester + water 
Alcohol + Phosphate ion
 Hydrolase enzyme catalyzes
dephosphorylation reactions:
 Removal of phosphate groups from
nucleotides, proteins, and alkaloids
 Alkaline pH improves activity.
Hepato- Hepato- Osteo- Intest- Placenta
cellular biliary blasts inal
Mucosa
(Bone)
NA ALP ALP ALP ALP
Alkaline phosphatase has two main forms:

 Bone sources

 Intestinal and liver sources

 Name of Isoenzyme Hepatic Bone

Heat Stability Stable at 560 C for 30 Labile: disappears

minutes at 560 C within 10
minutes

Electrophoretic Order Most anodic Intermediate

Chemical Inhibition Moderate inhibition by Strong inhibition by
urea but low inhibition urea but low inhibition
by l-phenylalanine by l-phenylalanine
 Intestinal Placental Other

Intermediate labile: Stable at 560 C for 30 Regan isoenzyme:

disappears at 560 C minutes most stable
within 15 minutes

Cathodic -bone Migrates with hepatic Renal isoenzyme: rare

fraction or bone forms but most cathodic
Strong inhibition by l- Resistance to urea but Regan isoenzyme:
phenylalanine. Strong inhibition by l- Strong inhibition by l-
phenylalanine. phenylalanine.
 High concentration of ALP in hepatobiliary
cells
 Biliary inflammation or ductal obstruction
 Cellular inflammation and necrosis
 Increased with bone diseases of
osteoblastic activity
 Release of ALP into the circulation.
 Cholestasis may cause ALP increased 3-10 X
the normal levels.
 Serum total and direct bilirubin are increased.
 Analysis by the Bessey Lowry and Brock ALP
method
 p-nitrophenyl phosphate + H2O –(ALP, glycine
buffer, Mg2+, pH 10.5) p-nitrophenol + PO4 3+
-> yellow quininoid chromagen
measure increase in Abs. at 400 nm at 37 0 C
 Non-hemolyzed serum
 heparinized plasma
◦ Fresh or refrigerated
Reference ranges vary with method used:
 53 -128 U/L
 2x or more increases in serum or plasma:
 Bone cancer, bone disease (such as Paget’s)
 Hepatobiliary disease such as cholestasis,
cholelithiasis
 Catalyze interconversion of functional groups.
◦ Transaminase- Aminoacids to -oxoacids
◦ - insert for GGT

o Names:
- AST; formerly Glutamate Oxaloacetate
transaminase, GOT
-ALT; formerly Glutamate Pyruvate Transaminase,
GPT
Transaminase - transfers amino groups
 Other descriptive names:
◦ serum glutamic pyruvate transaminase (SGPT)
◦ alanine aminotransferase ( ALAT)
ALT catalyzes the reaction:
L-Alanine + -Oxoglutarate  Pyruvate +
L-Glutamate
Damage to tissue can release different types of
enzymes based on their location.
 Mild inflammation of the liver
 Reversibly increases the permeability of the cell
membrane
 Releases cytoplasmic enzymes: AST
 Necrosis releases mitochondrial ALT, AST
 Hepatocellular necrosis releases
mitochondrial ALT
◦ Associated with liver inflammation (hepatitis)
 Drugs overdose or toxicity
 Infections from Viruses or bacteria
 Alcohol
 Nonhemolyzed serum or plasma.
 Heparinized plasma
 < 2 day old samples
 Fasting specimen is preferred
Serum or plasma reference ranges vary with
method
 Reference ranges reflect the normal amount
in serum or plasma:
 Adult male <45 U/L
 Adult female <34 U/L
Serum glutamic oxaloacetic transaminase
(SGOT) or aspartate aminotransferase
(ASAT/AAT)
 Transfers amino groups to form oxaloacetate
 Found in serum from various tissues
 Associated with hepatocytes
 Hepatocellular inflammation releases
cytoplasmic AST
 Hepatocellular necrosis releases
mitochondrial AST.
◦ Associated with liver inflammation (hepatitis)
 Drugs overdose or toxicity
 Infections from Viruses or bacteria
 Alcohol
 Other organ diseases
◦ Myocardia infarction
HIV treatment, especially the reverse
transcriptase inhibitors are associated with
metabolic complications:
 Pancreatitis, hypertriglyceridemia, and lactic
acidosis
 Hepatomegaly, and hepatic inflammation
Patients taking these medications will have
liver enzyme levels monitored every few
months to monitor for these complications
 Nonhemolyzed serum or plasma.
 Heparinized plasma
 < 2 day old samples
 Nonfasting may falsely increase
 Involved in the transfer of glutamyl group
 Glutathione metabolism
 Cysteine product preserves intracellular
homeostasis of oxidative stress
 Found in biliary ducts of liver, kidney
tubules, and prostate
 Associated with disease in the liver such as
hepatobiliary obstruction or inflammation
 Elevated because of alcoholism
 Nonhemolyzed serum
 EDTA plasma