BXC0213A BXC0213D BXC0213F For In Vitro Diagnostics Use Only

Lot Number
1x100ml 2x100ml 5x100ml Catalogue Number
Storage Temperature
ALT (GPT) STORE AT 2-8°C
Expiry Date (Year / Month)
PRODUCT CODE: BXC0213 Warning, Read Enclosed Documents
INSTRUCTIONS FOR USE
Instructions For Use

FOR IN-VITRO DIAGNOSTIC USE ONLY Manufactured By

The IFCC (International Federation of Clinical Chemistry)  Controls as indicated below

ALT (GPT) Recommendation for standardized procedures for ALT determinations
 0.9% NaCI
Manual Procedure:
IFCC UV (LIQUID STABLE) – A25 / A15 including: Wavelength Temperature Cuvette Measurement
APPLICATION 1. Optimization of substrate concentrations Hg 334 nm,
2. Employment of Tris buffers (instead of phosphate which has been Against Air or
340 nm or +30 oC/+37oC 1 cm light path
shown to inhibit recombination of the apoenzyme with pyridoxal Distilled water
Kit Contents: BXC0213A BXC0213D BXC0213F Hg 365 nm
phosphate).
R1 Enzyme Reagent 1 x 100ml 2 x 100ml 5 x 100ml
3. Preincubation of combined buffer and serum to allow side reactions Pipette into test tubes as follows:
R2 Oxoglutarate / 1 x 20ml 2 x 20ml 1 x 100ml
with NADH to occur. Sample Calibrator
Coenzyme+ 4. Substrate start (α-oxoglutarate). Working Reagent 1000 µl 1000 µl
5. Optional pyridoxal phosphate activation. This is an optimized
Sample 100 µl ---
Intended Use: standard method according to the concentrations recommended by
In vitro test for the quantitative determination of alanine Calibrator --- 100 µl
the IFCC.
amino-transferase (ALT) in human serum and plasma. Mix gently, incubate for 1 min. at assay temperature, measure initial
absorbance and start stopwatch simultaneously. Read again after
Reagent Concentration:
Test Principle: exactly 1, 2 and 3 minutes.
Tris Buffer pH 7.8 100 mmol/l
Method recommended by the IFCC: R1 L-Alanine 500 mmol/l
ALT LDH 1200 U/l Calculation:
2-Oxoglutarate + L-Alanine L-glutamate + pyruvate R2 NADH 0.18 mmol/l 340 nm A/min x 1746
LDH
Oxoglutarate 15 mmol/l Hg 334 nm A/min x 1780
pyruvate + NADH + H+ L-lactate + NAD+
Hg 365 nm A/min x 3235
Reagent Handling and Preparation:
Clinical Significance:
Mix contents of each bottle by gentle swirling before use. Linearity:
Alanine aminotransferase (glutamate pyruvate transaminase) belongs
Working Reagent: Linear up to 418 U/l
to the group of transaminases which catalyze the conversion of amino
Mix 1 volume of R2 with 5 volumes of R1. Wait for 5 minutes before At higher activities dilute the sample with 0.9% NaCl (e.g. 1:10). Multiply
acids to the corresponding -keto acids via the transfer of amino
use. the result by the appropriate dilution factor (e.g. 10)
groups; they also catalyze the reverse process. Although higher
This solution is stable: 35 days at +2 to +8°C Sensitivity:
activities exist in the liver, minor activity can also be detected in the
Detection limit: 3 U/l or 0.06 µkat/l
kidneys, heart, skeletal muscle, pancreas, spleen, and lungs. Elevated
Specimen: The lower detection limit represents the lowest measurable ALT
levels of transaminases are indicative of myocardial infarction,
Collect serum using standard sampling tubes concentration that can be distinguished from zero
hepatopathies, muscular dystrophy, and damage to internal organs.
Heparinized and EDTA-Plasma Imprecision:
Increased ALT activity in the serum, however, is a rather specific
Reproducibility was determined using controls. The following results
indicator of damage to the liver parenchyma, while AST is not
24 hours at 20°C to 25°C were obtained:
necessarily a liver-specific parameter. Stability:
In 1956, Wroblewski and LaDue described the first kinetic determination 3 days at 2°C to 8°C
Intra Assay – Within run
of ALT in serum. In 1977 and 1980, the International Federation of
Separate serum/plasma from clot/cells within 8 hours at room Mean SD CV
Clinical Chemistry (IFCC) recommended standardized methods for the Sample
temperature or 48 hours at 2-8°C. U/l U/l %
determination of ALT with optimized substrate concentrations, use of
TRIS buffer*, simultaneous preincubation of serum with buffer (to avoid Centrifuge samples containing precipitate before performing the Sample 1 36 1.4 4.0
competing reactions with NADH), substrate start, and pyridoxal phos- assay. Sample 2 141 2.1 1.5
phate activation.
The method described here is derived from the IFCC Reference Testing Procedure: Inter Assay – Between Run
method. Materials Provided: Mean SD CV
 Working Solutions as described above Sample
*TRIS = Tris(hydroxymethyl)-aminomethane U/I U/l %
Additional Materials Required:

Biorex Diagnostics Limited, Unit 2C Antrim Technology Park, Muckamore, BT41 1QS (United Kingdom)
Tel: +44 (0) 2894 468786 | Fax: +44 (0) 2894 469933 | Website: www.biorexdiagnostics.com BXC0213 – ALT (GPT) | Revision No.10 MAY/11 | Page 1 of 2
 Sample 1 36 1.47 4.1 6. Passing H, Bablok W. A New Biometrical Procedure for Testing the
Sample 2 136 2.68 1.95 Equality of Measurements from Two Different Analytical Methods.
J Clin Chem Clin Biochem 1983;21 :709-720.
Normal Values: 7. Thefeld W et al. Dtsch med Wschr 1974;99:343.
The IFCC-method: 8. Tietz NW (Hrsg.). Clinical Guide to Laboratory Tests, 3 rd
Philadelphia, Pa: WB Saunders, 1995:20-21.
30oC 37oC
9. Wallnöfer H, Schmidt E, Schmidt FW (Hrsg.). Synopsis der Leber-
Men up to 29 up to 41 krankheiten. Stuttgart: Georg Thieme Verlag, 1974.
U/l
Women up to 22 up to 31 10. Wroblewski F LaDue JS. Ann Intern Med 1956;45:801.
Men up to 0.48 up to 0.68 Wroblewski F, LaDue JS. Proc Soc Exp Biol Med
µkat/l
Women up to 0.37 up to 0.52 1956;91.
Each laboratory should investigate the transferability of the expected 11. Wroblewski F, LaDue JS. Proc Soc Exp Biol Med 1956;91 :569.
values to its own patient population and if necessary determine its
own reference range. For diagnostic purposes, the ALT results should BIOSYSTEMS A25/A15 APPLICATION:
always be assessed in conjunction with the patients medical history,
clinical examination and other findings.
A 25 A 15
Use on Automated Analysers:
TEST NAME ALT ALT
This reagent is suitable for use on a range of automated analysers.
ANALYSIS MODE MON.KINETIC MON.KINETIC
Specific instructions for these applications are available on request
from our technical department. SAMPLE TYPE SERUM SERUM
For automated use we recommend a serum based calibrator to UNITS U/L U/L
GENERAL
eliminate any matrix bias which may be observed with the aqueous REACTION TYPE DECREASING DECREASING
standard. DECIMALS 1 1
NO.OF REPLICATES 1 1
Biorex Calibration Serum cat. No BXC0321K/ L/ M
TEST NAME IN PT REP
Quality Control: PROCEDURE READING MONOCH MONOCH
VOLUMES SAMPLE 25 25
Biorex Normal Bovine Assayed Control Cat No BXC0313A (10x5ml) REAGENT 1 300 300
Biorex Elevated Bovine Assayed Control Cat No BXC0313B (10x5ml REAGENT 2
Biorex Normal Human Assayed Control Cat No BXC0312A (10x5ml) WASHING 1.2 1.2
Biorex Elevated Human Assayed Control Cat No BXC0312B (10x5ml) PREDIL FACTOR
POST DIL FACTOR 2 2
The control intervals and limits must be adapted to the individual
laboratory and country-specific requirements. Values obtained should FILTERS MAIN 340 340
fall within established limits. Each laboratory should establish corrective REFERENCE
measures to be taken if values fall outside the limits. TIMES READING 1 90s 90s
READING 2 255s 255s
Health & Safety:
REAGENT 2
This kit is designed for use by suitably qualified laboratory personnel
CALIBRATION TYPE MULTIPLE MULTIPLE
only. Exercise the normal precautions required for the handling of
laboratory reagents. Do not ingest the material. Dispose of material CALIBRATOR REPLICATES 3 3
CALIBRATION
according to local guidelines. BLANK REPLICATES 3 3
CALIBRATION CURVE
References: BLANK ABS LIMIT 1.100 1.100
1. International Federation of Clinical Chemistry, Scientific
OPTIONS KINETIC BLANK LIMIT
committee. J Clin Chem clin Biochem 1980 18: 521-534.
2. Bablok W et al. A General Regression Procedure for Method LINEARITY LIMIT 400 400
Transformation. J Clin Chem Clin Biochem 1988;26:783-790.
3. Bergmeyer HU, Herder M, Rej R. Approved recommendation
(1985) on IFCC methods for the measurement of catalytic
concentration of enzymes. Part 3. (FCC Method for alanine
aminotransferase. J Clin Chem Clin Biochem 1986;24:481-489.
4. Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of
Interferences in Clinical Chemistry Instrumentation. Clin Chem
1986;32:470-474.
5. Greiling H, Gressner AM (Hrsg.). Lehrbuch der Klinischen Chemie
und Pathobiochemie, 3rd Stuttgart/New York: Schattauer Verlag,
1995.

Biorex Diagnostics Limited, Unit 2C Antrim Technology Park, Muckamore, BT41 1QS (United Kingdom)
Tel: +44 (0) 2894 468786 | Fax: +44 (0) 2894 469933 | Website: www.biorexdiagnostics.com BXC0213 – ALT (GPT) | Revision No.10 MAY/11 | Page 2 of 2