GPT/ALAT IFCC

Cat.No Package Size REFERENCE VALUES (IFCC / 37°C)

142 016 7 x 10 ml 2 x 7 ml Women < 31 U/l
142 000 5 x 20 ml 2 x 10 ml Men < 41 U/l
142 007 (AS) 4 x 70 ml 3 x 20 ml Note: It is recommended for each laboratory to establish and
maintain its own reference values. The given data are only an
indication.
METHOD
Optimized, UV, Kinetic. PROCEDURES
Reagents can be used manually (see method below) and
PRINCIPLE on most analyzers. Applications are available on request.
Kinetic determination of Alaninaminotransferase
(ALAT) based upon IFCC recommendations: Wavelength : 340 nm, Hg 334 nm, Hg 365 nm
Temperature : 37°C,
L-Alanin+ 2-Oxoglutarat ALAT > L-Glutamat + Pyruvat Cuvette : 1 cm light path
Pyruvat + NADH + H+ < LDH > L-Lactat + NAD+ Read against air.

ALAT = Alaninaminotransferase  Sample Start Procedure / Monoreagent

LDH = Lactate dehydrogenase

Working reagent : 1 ml
REAGENTS (concentrations in the reagents) Sample : 100 µl

R1: TRIS-Buffer (pH 7,4) 100 mmol/l Mix and after a 1 minute incubation, measure the
L-Alanine 600 mmol/l
LDH  1500 U/l change of absorbance per minute (A/min) during
CAPs pH 9,6 85 mmol/l
3 minutes.
R2:
NADH 1,4 mmol/l  Start Reagent Procedure/R1-R2
2-Oxoglutarate 130 mmol/l

R1 : 1 ml
- PRECAUTIONS Sample : 100 µl
- For in vitro diagnostic use only. Mix and wait 1 minute
- The reagents contain < 0,95 g/l sodium azide.
Avoid contact with skin and/or mucous R2 : 200 µl
membranes ! Mix and after a 1 minute incubation, measure the
- Use clean or single use glass/plastic material change of absorbance per minute (A/min) during
only to avoid contaminations. 3 minutes.
STABILITY OF REAGENTS
When stored at 2 – 8 °C and protected from light,
the reagents are stable until the expiry date printed CALCULATION FACTORS
on the labels. Monoreagent Start reagent
340 nm : Activity (U/l) = A/min. x 1 745 2064
334 nm : Activity (U/l) = A/min. x 1 780 2103
PREPARATION OF WORKING REAGENT
365 nm : Activity (U/l) = A/min. x 3 235 3824
- For “Start Reagent-procedure”
reagents R1 and R2 are ready for use
- For “Sample Start-procedure” (monoreagent) CALIBRATORS & CONTROLS
- mix R1 + R2 = 5 + 1 For the calibration of automated analyzers Greiner
Stability : 3 weeks at 2 – 8 °C Multicalibrator is recommended, for quality control
5 days at 18 – 25 °C use Greiner normal and abnormal control, Unitrol I
and Unitrol II
SAMPLES
Serum free of hemolysis , plasma (EDTA/heparin)

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PERFORMANCE DATA (37°C) BIBLIOGRAPHY
- Analytical range 1. Thomas L. Alanine aminotransferase (ALT),
The reagent is linear up to ALAT activities Aspartate aminotransferase (AST). In: Thomas L,
corresponding to a A/min of 0,16 at 340 nm and editor. Clinical Laboratory Diagnostics. 1st ed.
334 nm,which corresponds to 1300 U/l. If A/min Frankfurt: TH-Books Verlagsgesellschaft; 1998. p.
55-65.
exceeds 0.16 at 340 nm, repeat the test using
2. Moss DW, Henderson AR. Clinical enzymology. In:
serum diluted 1/10 with sodium chloride solution (9 Burtis CA, Ashwood ER, editors. Tietz Textbook of
g/L).Multiply result by 10. Clinical Chemistry. 3rd ed. Philadelphia: W.B
Saunders Company; 1999. p. 617-721.
- Sensitivity 3. Bergmeyer HU, Horder M, Rej R. International
The sensitivity / lower detection limit is < 1U/l Federation of Clinical Chemistry (IFCC) Scientific
Committee, Analytical section: approved
- Precision recommendation (1985) on IFCC methods for the
measurement of catalytic concentration of enzymes.
Within-run reproducibility Part 3. IFCC method for alanine aminotransferase
Within series Mean Standard- CV (L-alanine:2-oxoglutarate aminotransferase, EC
n = 11 Value deviation [%] 2.6.1.2). J Clin Chem Clin Biochem 1986;24:481-95.
[U/l] [U/l]
4. Lorentz K, Röhle G, Siekmann L. Einführung der
Sample 1 38,4 0,51 1,32
neuen Standardmethoden 1994 zur Bestimmung der
Sample 2 143 1,95 1,36
Sample 3 111 1,27 1,14 katalytischen Enzymkonzentrationen bei 37 °C. DG
Klinische Chemie Mitteilungen 1995;26:190-2.
Between-run reproducibility
Day to day Mean Standard- CV
n = 11 Value deviation [%]
[U/l] [U/l]
Sample 1 36,0 0,78 2,15
Sample 2 146 0,94 0,65 SYMBOLS USED
Sample 3 109 1,40 1,28

- Correlation For in vitro diagnostic medical use

A comparative study has been performed between
the Greiner method and another commercial
reagent on 30 human serum samples. The Batch Code
parameters of linear regression are as follows:

Correlation coefficient r = 0.998 Use by

Linear regression: y = 0,967x + 3,365 U/l

Temperature limitation
INTERFERENCES
- Ascorbic Acid: no interference until 100 mg/dl
- Triglycerides: no interference until 1000 mg/dl
- Bilirubin: no interference up to 40 mg/dl
- Hemoglobin: no interference up to 400 mg/dl

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Greiner Diagnostic GmbH - Unter Gereuth 10 – D-79353 Bahlingen – Germany

www.greiner-diagnostic.com