GPT (ALAT) IFCC fluid (5+1)

Kinetic determination of Glutamate Pyruvate Transaminase - IFCC version

REAGENT PACKAGE INFORMATION TESTING PROCEDURE

Product FURUNO Configuration Material provided
Product No. - Working solutions as described above
Name Analyzer R1 R2
GF05917084-F3 3 x 67mL 3 x 17mL Additional materials required
GPT (ALAT) CA-270 - Calibrators and controls as indicated below
GF05917084-F 6 x 67mL 6 x 17mL
IFCC fluid CA-400 - “Saline Diluent” is used as diluent:
GF05917084-F4 4 x 20mL 4 x 7mL
(5+1) CA-800 Product No. Product name Configuration
GF05917084-F8 8 x 20mL 8 x 7mL
FSD917070 Saline Diluent 5 x 67mL
INTENDED USE
In vitro test for the quantitative determination of GPT (ALAT) Alanine MEASUREMENT RANGE
Aminotransferase in human serum and plasma. 3.4 – 800 U/L in CA-270
1.7 – 730 U/L in CA-400
SUMMARY 1.7 – 900 U/L in CA-800
Alanine aminotransferase (glutamate-pyruvate-transaminase) belongs to the
group of transaminases which catalyze the conversion of amino acids to the CONVERSION FACTOR
correspond-ding α-ketoacids via the transfer of amino groups; they also catalyze U/L μkat/L Factor: 0.0167
the reverse process. Although higher activities exist in the liver, minor activity μkat/L U/L Factor: 60
can also be detected in the kidneys, heart, skeletal muscle, pancreas, spleen,
and lungs. Elevated levels of transaminases are indicative of myocardial EXPECTED VALUES
infarction, hepatopathies, muscular dystrophy, and damage to internal organs. Reference: Tietz et.al.
Increased ALT activity in the serum, however, is a rather specific indicator of U/L µkat/L
damage to the liver parenchyma, while AST is not necessarily a liver-specific male: < 45 < 0.77
parameter. female: < 34 < 0.58
In 1956, Wroblewski and LaDue described the first kinetic determination of ALT Each laboratory should investigate the transferability of the expected values to
in serum. In 1977 and 1980, the International Federation of Clinical Chemistry its own patient population and if necessary determine its own reference range.
(IFCC) recommended standardized methods for the determination of ALT with For diagnostic purposes, the GPT results should always be assessed in
optimized substrate concentrations, use of TRIS buffer*, simultaneous pre- conjunction with the patient medical history, clinical examination and other
incubation of serum with buffer (to avoid competing reactions with NADH), findings.
substrate start, and pyridoxal phosphate activation.
The method described here is derived from the IFCC reference method. REPRODUCIBILITY
*TRIS = Tris(hydroxymethyl)-amino methane. Reproducibility in CA-800
Within run (n= 20) Day-to-day run
MEASUREMENT PRINCIPLE
GPT
(Duplicate, 20days)
α-Ketoglutarate + L-Alanine L-Glutamate + Pyruvate Mean SD Mean SD
The increase in pyruvate is determined in an indicator reaction catalysed by LDH. CV% CV%
(U/L) (U/L) (U/L) (U/L)
Pyruvate + NADH + H+ L-lactate + NAD++ H2
LDH Low 43.6 1.506 3.45 44.7 0.578 1.29
+
NADH is oxidized to NAD . The rate of the NADH decrease is directly High 125.7 1.372 1.09 109.7 1.291 1.18
proportional to the rate of formation of pyruvate and thus the GPT activity.
TEST COMPARISON
CONCENTRATION OF WORKING SOLUTION Comparison with a test of other manufacturer’s reagent of GPT(ALT) in CA-800
Reagent 1: Buffer Y = 1.036 x + 0.030, R = 0.999, N = 105
TRIS-Buffer pH 7.5 70 mmol/L
L-Alanine 410 mmol/L QUALITY CONTROL
LDH ≥1.7 U/mL Multi QC sample “Centronorm” and ”Centropath” are used.
Reagent 2: Starter Product No. Product Name Configuration
NADH 0.3 mmol/L
CNORM005 5 x 5 mL
α-Ketoglutarate 18 mmol/L
CNORM010 Centronorm 10 x 5 mL
CNORM020 20 x 5 mL
REAGENT PREPARATION, STORAGE AND STABILITY
CPATH005 5 x 5 mL
R1: Ready to use
CPATH010 Centropath 10 x 5 mL
R2: Ready to use
The sealed reagents are stable up to the indicated expiry date if stored at +2° - CPATH020 20 x 5 mL
+8°C. The control intervals and limits must be adapted to the individual laboratory and
After opening the bottle, it is stable for 61 days when kept onboard in the country-specific requirements. Values obtained should fall within established
analyzer, if contamination is avoided. limits. Each laboratory should establish corrective measures to be taken if
values fall outside the limits
SAMPLE MATERIAL
Serum, heparin-/ EDTA-plasma. CALIBRATION
Stability: 24 hours at +20°C to +25°C Multi-calibrator “Centrocal” is used.
3 days at +2°C to +8°C S1: Saline, S2: Centrocal
Separate serum/plasma from clot/cells within 8 hours at room temperature or 48 Recommended calibration interval: Weekly
hours at +2°C to +8°C. Product No. Product Name Configuration
Centrifuge samples containing precipitate before performing the assay. CCAL005 5 x 3mL
CCAL010 Centrocal 10 x 3mL
PRECAUTIONS AND WARNINGS CCAL020 20 x 3mL
For in vitro diagnostic use only. Attend to the normal precautions required for
handling all laboratory reagents. The reagents contain sodium azide as LITERATURE
preservative. Do not swallow. Avoid contact with skin and mucous membranes.  Z. klin. Chem. u. klin. Biochem. 8 (1970) 658 and 10 (1972), 182
 Klein G, Lehmann P, Michel E, Regenauer H. Vergleich der IFCC-Methoden
INTERFERENCES für ALAT, ASAT und GGT bei 37 °C mit den eingeführten Standardmethoden
Free type of Bilirubin: No interference less than 20mg/dL bei 25 °C und 37 °C. Lab Med 1994;18:403-404.
Conjugate type of Bilirubin: No interference less than 20mg/dL  Carl A. Burtis, David E. Bruns: Tietz fundamentals of clinical chemistry and
Hemolytic hemoglobin: No interference less than 500mg/dL molecular diagnostics, 7th ed.
Chyle: No interference less than 1800 FTU (*)
*: FTU: Formazine turbidity unit

Centronic GmbH
Am Kleinfeld 11, 85456 Wartenberg/Germany
Phone: 0049-8762724300, Fax: 0049-8762724312
Edition 10/2024, TV-045-CE-005FU
e-mail: [email protected]
web: www.centronic-gmbh.com
Revision history

Document Name, version Revised Revise content

20. ALT TV-045-CE-001FU 2020.09 Original
20. ALT TV-045-CE-002FU 2020.12 To add the phrase below the item title at the header
Kinetic determination of Glutamate Pyruvate Transaminase - IFCC version
20. ALT TV-045-CE-003FU 2021.06 Corrected 2nd reaction formula in MEASUREMENT PRINCIPLE
Corrected reproducibility SD value for Low of Day-to-day run from 0.878 to 0.578
20. ALT TV-045-CE-004FU 2023.11 ・Paragraph “MESUREMENT RANGE”
1.7 – 800 U/L in CA-270 → 3.4 – 800 U/L in CA-270
20. ALT TV-045-CE-005FU 2024.10 ・REAGENT PACKAGE INFORMATION
Corrected table widths to display product information correctly.

Centronic GmbH
Am Kleinfeld 11, 85456 Wartenberg/Germany
Phone: 0049-8762724300, Fax: 0049-8762724312
Edition 10/2024, TV-045-CE-005FU
e-mail: [email protected]
web: www.centronic-gmbh.com