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www.biomaghreb.com
IVD
GPT - ALAT
Kinetic Test. IFCC Without Pyridoxal phosphate
Quantitative determination of alanine amino transferase activity
(EC 2.6.1.2) in human serum
FT En 12 Date of updating: 10/ 2020 Version B
IN VITRO DIAGNOSTIC USE
Manufacturer Use by In Vitro Diagnostic Temperature
Limitation
Catalogue number See insert Store away from light Sufcient
for < n > essais
Batch number
CLINICAL SIGNIFICANCE
ALAT (alanine amino-transferase) formerly known as Glutamic Pyruvic Transaminase (GPT) is an
enzyme found mainly in liver cells, and to a lesser extent in kidney, heart, and muscle cells. The mea-
surement of ALAT activity allows the detection of liver damage. When the liver is damaged, ALAT is
released into the bloodstream in patients with cirrhosis, hepatitis, cancer and jaundice due to biliary
congestion). In general, ALAT activity values are compared with the activities of other enzymes such
as alkaline phosphatase (PAL), aspartate aminotransferase (ASAT) and bilirubin to accurately define
the origin of liver damage.
PRINCIPLE
The kinetic determination of ALAT activity is based on the method developed by Wrobleski and
Ladue, and optimized by Henry and Bergmeyer.
The reaction is initiated by adding the sample to the reagent according to the following reaction
scheme:
GOT
2 oxoglutarate + L-Aspartate Glutamate + oxaloacetate
MDH
Oxaloacetate + NADH + H
+
Malate + NAD
+
The rate of decrease in NADH concentration is directly proportional to the alanine amino transfer
activity in the sample.
LDH : Lactate Dehydrogenase
REAGENT COMPOSITION
Reagent 1
Buffer
Buffer Tris pH 7.5 à 30°C
Alanine
100 mmol/l
500 mmol/l
Reagent 2
Substrate
NADH
LDH
Oxoglutarate
0.18 mmo/l
1200 U/l
15 mmol/l
SAFETY CAUTIONS
Biomaghreb reagents are intended for use by qualified personnel for in vitro use (do not pipette by
mouth).
• Refer to the current MSDS available on request or on www.biomaghreb.com.
• Check the integrity of the reagents before use.
• Disposal of waste: comply with applicable legislation.
For safety reasons, treat any specimen or reagent of biological origin as potentially infectious. Ob-
serve the applicable legislation.
REAGENT PREPARATION
Working solution:
Mix the substrate with3 ml
REF
(11015) and
REF
(11039) or 10 ml
REF
(11022) of Buffer R1.
For
REF
(11046) reconstitute each R2 with one vial R1.
SAMPLE COLLECTION AND HANDLING
Heparinized serum or plasma without hemolysis.
PRESERVATION AND STABILITY
Stored in the original, tightly stoppered bottle at 2-8°C, the reagents are stable if used and stored
under the recommended conditions:
• Before opening: Until the expiry date indicated on the label of the box at +4°C;
• After opening : (Working Solution):
24 hours at 20-25°C;
7 days at 2-8°C.
ADDITIONAL EQUIPMENT
• Basic equipment of the medical analysis laboratory ;
• Spectrophotometer or Clinical Biochemistry Analyzer.
LIMITS
Hemolysis can interfere.
QUALITY CONTROL
External quality control program.
It is recommended to control in the following cases:
• At least one test per series.
• Change of reagent bottle.
• After maintenance work on the analyzer.
If a control value is outside the confidence limits, repeat the procedure using the same control.
Use normal and pathological control sera.
LINEARITY
If the ΔDO/min at 340 nm is greater than 0.15, repeat the test by diluting the sample 1:10 with a 9
g/l NaCl solution.
Multiply the result by 10.
PROCEDURE
Wavelength: 340 nm;
Temperature: 25 - 30 or 37 ° C;
Tank: 1 cm thick;
Adjust the spectrophotometer zero to air or distilled water.
Working solution
1 ml 3 ml
Pre-incubate at the selected temperature (25, 30 or 37°C).
Sample
100 µl 300 µl
Mix and incubate 1 minute.
Measure the decrease in optical density per minute for 1 to 3 minutes.
CALCULATION
At wavelength 340 nm,
Δ DO / min x 1750 = IU/l
REFERENCE VALUES
25°C 30°C 37°C
Women
Up to 16 Ul/l Up to 22 Ul/l Up to 31UI/l
Men
Up to 22UI/l Up to 29 Ul/l Up to 40UI/l
REFERENCES
Bergmeyer H. Schaibe and Walefeld. Clin. Chem. 24 58 - 73 (1978);
Bergmeyer and Horder Clin. Chem. Acta 105 147 F (1980) ;
Henry R, J, et al,, Am J clin Path (1960), 34, 381-398.
