Gamma Glutamyl Transferase Reagent (5X Concentrate)

Gamma Glutamyl Transferase Reagent
(5X Concentrate)
V
PRODUCT SUMMARY
Stability : 14 days at 2-8°C
Linear Range : Up to 600 U/L
Specimen Type : Serum
Method : Kinetic
Reagent Preparation : Add 7 mL of distilled or
deionised water.
INTENDED USE
This reagent is intended for the in vitro quantitative determination gammaglutamyltransferase
(GGT) [(γ-Glutamyl) - Peptide: Amino Acid γ-Glutamyltransferase,
EC2.3.2.2], in human serum.
SUMMARY
Although gamma-glutamyltransferase (GGT) is present in a variety of tissues, the
serum enzyme appears to be primarily from the hepato-biliary system. Consequently,
GGT is elevated in all forms of liver disease or damage. It is clinically useful in
detecting obstructive jaundice, cholangitis, and cholecystitis.
Elevated levels are also observed with drug use (alcohol, sedatives, anticonvulsants
and tranquilizers). 1
PRINCIPLE
The first commercially available kinetic methods for the determination of GGT were
based on the work of Szasz 2 , Rosalki and Tarlow 3 . These methods utilised γ-glutamylp-nitroanilide
(Glu-4-NA) as the substrate, however the poor solubility and stability of
Glu-4-NA was a major limitation. In order to improve the method Persijn 4 investigated
further with Glu-4-NA derivatives and found that γ-glutamyl-3-carboxy-4-nitroanilide
(Glucana) was superior to the Glu-4-NA with respect to both solubility and stability.
The Glucana substrate now forms the basis of the IFCC and ECCLS recommended
procedures. The GGT- Soluble Substrate method utilises Glucana in a one step
method in which the reaction is initiated with the addition of sample. GGT present
in the sample catalyzes the transfer of the glutamyl group from the substrate to
glycylglycine forming glutamylglycylglycine and 5-amino-2-nitrobenzoate.
L-γ-glutamyl-3-carboxy-4-nitroanilide + glycylglycine
SYMBOLS IN PRODUCT LABELLING
For in vitro diagnostic use
Batch code/Lot number
Catalogue number
Consult instructions for use
Xn - Harmful
STORAGE AND STABILITY
1. The unopened reagents are stable until the expiration date stated on the label
when stored at 2-8°C.
2. After reconstitution the reagent is stable for 14 days when stored on the Olympus
Demand or Reply at 2-8°C.
DETERIORATION
1. The dry powder reagent should be a white to off-white, free flowing powder.
2. The reconstituted reagent should be a clear, slightly yellow solution.
3. Failure to achieve assay values on freshly prepared control sera could indicate
deterioration.
SPECIMEN COLLECTION
Clear, non-hemolyzed serum is the recommended sample. 5
SAMPLE STORAGE
GGT is stable for 7 days when stored at 2-8°C. 6
INTERFERING SUBSTANCES
1. No interference from Lipemia, measured as triglycerides, up to 200 mg/dL.
2. No interference from Conjugated Bilirubin up to 60 mg/dL.
3. No interference from Free Bilirubin up to 48 mg/dL.
4. No interference from Hemoglobin up to 350 mg/dL.
5. Young has reviewed drug effects on serum GGT levels. 6
PROCEDURE
Test Parameters
Refer to the Thermo Reagent Applications.
Temperature Limitation
Use by/Expiration Date
CAUTION. CONSULT INSTRUCTIONS
FOR USE.
Manufactured by
GGT
MATERIALS PROVIDED
GGT Reagent
10 x 7 mL
L-γ-glutamylglycylglycine + 5-amino-2-nitrobenzoate
The rate of formation of 5-amino-2-nitrobenzoate is proportional to the activity of GGT
present in the sample and can be measured bichromatically at 410/660nm.
REAGENTS
Reactive Ingredients Initial Conc. Final Conc.
Tris buffer 550 mmol/L 110 mmol/L
Glycylglycine 550 mmol/L 110 mmol/L
L-γ-glutamyl-3-carboxy-4-nitroanilide 16 mmol/L 3.2 mmol/L
Also contains non-reactive fillers and stabilizers
pH 8.20 ± 0.1 at 20°C
PRECAUTIONS
Do not ingest. Avoid contact with skin and eyes. If spilt thoroughly wash affected
area with water. Reagent contains sodium azide which may react with copper or
lead plumbing. Flush with plenty of water when disposing. For further information
consult the Bicarbonate Reagent Material Safety Data Sheet. The Packaging of
This Product Contains Dry Natural Rubber.
R22 Harmful if swallowed.
R36/38 Irritating to eyes and skin.
S26 In case of contact with eyes, rinse immediately with plenty of water
and seek medical advice.
REAGENT PREPARATION
Add 7 mL of freshly distilled or deionized water to each vial. Swirl gently to dissolve
the contents. Allow to stand 10 minutes at room temperature before use.
MATERIALS REQUIRED BUT NOT PROVIDED
1. Clinical chemistry analyser capable of maintaining constant temperature
(37°C), using 5x concentrated reagents and measuring absorbance at
405 nm (405 - 420 nm)
2. Thermo Reagent Applications for the Olympus Demand or Reply.
3. Thermo Data-Trol N and Data-Trol A (Cat. No. 1902-050 or TR40001 and
1901-050 or TR41001) or equivalent.
STABILITY OF FINAL REACTION MIXTURE
The instrument automatically computes every determination at the same time
interval.
CALIBRATION
A calibrator is not required for the GGT Procedure. One U/L of GGT activity is that
amount of enzyme which produces one mmol/L of p-nitroaniline per minute.
LINEARITY
Linearity extends to 600 U/L. Samples exceeding linearity should be diluted with
normal saline and repeated. Multiply the result by the dilution factor when calculating
the unknown.
QUALITY CONTROL
Normal and abnormal control sera of known concentrations of GGT should be
analyzed routinely with each group of unknown samples. Data-Trol N and Data-Trol
A (Cat. No. 1902-050 or TR40001 and 1901-050 or TR41001) are recommended
for this purpose.
CALCULATION OF RESULTS
Results, expressed as U/L at 37°C, are automatically calculated.

LIMITATIONS
See Storage and Stability, Deterioration, Specimen Collection, Interfering Substances,
Sample Storage, and Linearity sections for limitations to this procedure.
EXPECTED VALUES 3
At 37°C : Males: < 50 U/L
Females: < 30 U/L
The quoted values are representative of the expected range for this method and
should serve as a guide only. It is recommended that each laboratory verify this
range or derives a reference interval for the population that it serves. 7
PERFORMANCE CHARACTERISTICS
Precision
WITHIN-RUN Level 1 Level 2
No. of data points 20 20
Mean U/L 20 67
SD 0.41 1.00
CV% 2.05 1.49
BETWEEN DAY Level 1 Level 2
No. of data points 20 20
Mean U/L 22 67
SD 1.80 2.05
CV% 8.18 3.06
COMPARISON STUDIES
A comparison of the Thermo GGT reagent (y) with a commercial reagent of the same
methodology (x) was performed on 147 human samples in a range of 2 - 831 U/L.
A correlation coefficient of 0.999 was obtained; the linear regression equation was
y = 0.99 x + 2.7.
ANALYTICAL SENSITIVITY
Based on an instrument resolution of A= 0.001, this Thermo GGT Procedure has
a sensitivity of 2.6 U/L.
REFERENCES
1. Kachmar JF, Moss DV. “Enzymes” in Fundamentals of Clinical Chemistry. Tietz
NW (Ed) WB Saunders Co. Philadelphia 1976; page 621-3.
2. Szasz G. Clin Chem 1969; 15: 124-36.
3. Rosalki SB, Tarlow D. Clin Chem 1974; 20: 1121-4.
4. Persijn JP and van der Slik W. J.Clin.Chem.Clin Biochem. 1976; 14: 421-7.
5. Tietz, N.W., Clinical Guide to Laboratory Tests, W.B. Saunders, Philadelphia,
1983, p. 244.
6. Young DS. Effects of Drugs on Clinical Laboratory Tests. Third Edition. 1990;
3: 183-5.
7. Wachtel M et al, Creation and Verification of Reference Intervals. Laboratory
Medicine 1995; 26:593-7.
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Reorder Information and Technical Support
Catalogue No.
Configuration
7200-016A 10 x 7 mL
Australia International U.S.A.
Phone 1800 333 110 61 3 9790 4100 (800) 558 9115
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P/N: PI191041.01 Rev 12/2006