BXC0201B BXC0201C BXC0201D BXC0201E For In Vitro Diagnostics Use Only

Lot Number
10x20ml 10x10ml 5x50ml 20x2ml Catalogue Number
Storage Temperature
AST (GOT) STORE AT 2-8°C
Expiry Date (Year / Month)
PRODUCT CODE: BXC0201 Warning, Read Enclosed Documents
INSTRUCTIONS FOR USE
Instructions For Use

FOR IN-VITRO DIAGNOSTIC USE ONLY Manufactured By

serum to allow side reactions with NADH to occur, substrate start, and Manual Procedure:
AST (GOT) optional pyridoxal phosphate activation.
This method is derived from the IFCC reference method.
Wavelength
Hg340 nm,
Temperature Cuvette Measurement

IFCC UV – A25/A15 APPLICATION *TRIS = Tris(hydroxymethyl)-aminomethane 334 nm or Hg

+25oC/+30oC/
1 cm light path
Against Air or
37oC Distilled water
365 nm
Reagent Concentration:
Kit Contents: BXC0201B BXC0201C BXC0201D BXC0201E Tris Buffer pH 7.8 80 mmol/l Pipette into test tubes as follows:
R1
L-Aspartate 200 mmol/l Macro Semi Micro
R1 Buffer 2 x 105ml 1 x 105ml 5 x 50ml 1 x 45ml NADH 0.18 mmol/l Working Reagent 2000 µl 1000 µl
R2 Reagent 10 x 20ml 10 x 10ml 5 x 50ml 20 x 2ml LDH 800 U/l Sample 200 µl 100 µl
R2
MDH 600 U/l Mix, incubate for 1 min. at assay temperature and start stopwatch
Oxoglutarate 12 mmol/l simultaneously. Read again after exactly 1, 2 and 3 minutes. If the
Intended Use: A/min. is between 0.11 and 0.25 at Hg 334 nm/ 340 nm or 0.06 and
In vitro test for the quantitative determination of aspartate amino- Reagent Handling and Preparation:
R1 Buffer: Ready to use, stable up to the expiry date when stored at 0.013 at Hg 365 nm use only the values for the first 2 minutes for the
transferase (AST) in human serum and plasma. calculation.
2-8oC.
Test Principle: R2 Reagent: Reconstitute enzyme reagent R2 with the corresponding Calculation:
Method recommended by the IFCC: volume of buffer R1. Wait for at least 15 min before use. Macro Micro
Cat. No. BXC0201D, reconstitute one vial of Enzyme Reagent (R2) with 340 nm A/min x 1746 1746
GOT
a portion of Buffer (R1) and then transfer the entire contents to R1 Hg 334 nm A/min x 1780 1780
2-Oxoglutarate + L-Aspartate Glutamate +
bottle rinsing R2 bottle several times. Hg 365 nm A/min x 3235 3235
Oxaloacetate
This reagent is stable for 14 days at +2 to +8°C.
MDH
Linearity:
Oxaloacetate + NADH + H+ Malate + NAD+ Sample:
Up to 440 U/l
Serum, heparinised or EDTA plasma
If the change of absorbance per minute is higher than 0.250 at 340 nm
Clinical Significance : Specimen: or 0.130 at 365 nm the sample has to be diluted with 0.9% NaCI or
Aspartate aminotransferase (glutamate oxaloacetate transaminase) Collect serum using standard sampling tubes. distilled/deionised water (e.g. 1+9). Multiply the result by the
belongs to the transaminases, which catalyze the interconversion of Heparin or EDTA plasma. appropriate dilution factor (e.g. factor 10).
amino acids and -ketoacids by transfer of amino groups. Aspartate
aminotransferase is commonly found in human tissue. Although heart 24 hours at +20oC to +25 oC Sensitivity:
Stability:
muscle is found to have the most activity of the enzyme, significant 7 days at +2oC to +8 oC Detection limit: 4 U/l or 0.07 µkat/l
activity has also been seen in the brain, liver, gastric mucosa, adipose The lower detection limit represents the lowest measurable AST
Separate serum/plasma from clot/cells within 8 hours at room
tissue, skeletal muscle, and kidneys. concentration that can be distinguished from zero.
temperature or 48 hours at +2°C to +8°C.
AST is present in both the cytoplasm and mitochondria of cells. In cases Centrifuge samples containing precipitate before performing the
involving mild tissue injury, the predominant form of AST is that from the Imprecision:
assay.
cytoplasm, with a smaller amount coming from the mitochondria. Reproducibility was determined using quality controls. The following
Severe tissue damage results in more of the mitochondrial enzyme results were obtained.
Testing Procedure:
being released. Elevated levels of the transaminases can signal Materials Provided:
myocardial infarction, hepatic disease, muscular dystrophy, and organ Intra Assay – Within run
 Working Solutions as described above
damage. In 1955, Karmen et al described the first kinetic determination Additional Materials Required: MW SD CV
Sample
of AST activity in serum.  Controls as indicated below U/l U/l %
The International Federation of Clinical Chemistry (IFCC)  0.9% NaCI Level 1 33.4 0.76 2.29
recommended in 1977 and 1980 standardized procedures for AST Level 2 116 1.56 1.34
determination, including optimization of substrate concentrations,
employment of TRIS* buffers, preincubation of combined buffer and

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 BXC0201 – AST (GOT) | Revision No.10 MAY/11 | Page 1 of 2
 Inter Assay – Between Run fall within established limits. Each laboratory should establish corrective BIOSYSTEMS A25/A15 APPLICATION:
Mean SD CV measures to be taken if values fall outside the limits
Sample
U/l U/l %
Level 1 33.2 0.67 2.02 Health & Safety: A 25 A 15
Level 2 82.9 1.18 1.43 This kit is designed for use by suitably qualified laboratory personnel
TEST NAME AST AST
only. Exercise the normal precautions required for the handling of
laboratory reagents. Do not ingest the material. Dispose of material ANALYSIS MODE MON.KINETIC MON.KINETIC
Method comparison:
A comparison of the Biorex AST (y) with a commercial obtainable according to local guidelines. SAMPLE TYPE SERUM SERUM
assay (x) gave the following result with 48 samples: UNITS U/L U/L
References: GENERAL
y = 0.992x - 0.022; r= 0.995 REACTION TYPE DECREASING DECREASING
DECIMALS 1 1
Limitations - interference: 1. International Federation of Clinical Chemistry, Scientific
NO.OF REPLICATES 1 1
Criterion: Recovery within  10% of initial value. committee. J Clin Chem clin Biochem 1980 18: 521-534.
2. Bablok W et al. A General Regression Procedure for Method TEST NAME IN PT REP
Icterus: No significant interference up to an index I of 70
(approximate conjugated and unconjugated bilirubin: 70 mg/dl) Transformation. J Clin Chem Clin Biochem 1988;26:783-790. PROCEDURE READING MONOCH MONOCH
Haemolysis: No significant interference up to an index H of 900 3. Bergmeyer HU, Herder M, Rej R. Approved recommendation VOLUMES SAMPLE 25 25
(approximate haemoglobin concentration: 900 mg/dl). (1985) on IFCC methods for the measurement of catalytic REAGENT 1 300 300
Lipaemia (Intralipid): No significant interference up to an index L of 450 concentration of enzymes. Part 2. IFCC Method for aspartate REAGENT 2
(approximate triglycerides concentration: 900 mg/dl) There is poor aminotransferase. J Clin Chem Clin Biochem 1986;24:49. WASHING 1.2 1.2
correlation between turbidity and triglycerides concentration. 4. Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of
PREDIL FACTOR
Lipaemia may cause absorbance flagging as a result of an Interferences in Clinical Chemistry Instrumentation. Clin Chem 1
986;32:470-474. POST DIL FACTOR 2 2
absorbance increase.
5. Greiling H, Gressner AM (Hrsg.). Lehrbuch der Klinischen Chemie FILTERS MAIN 340 340
Normal Values: und Pathobiochemie,3. Auflage. Stuttgart/New York: Schattauer REFERENCE
According to the IFCC method. Verlag, 1995 TIMES READING 1 90s 90s
6. Karmen A et al. J Clin Invest 1955;24:126. READING 2 255s 255s
7. Passing H, Bablok W. A New Biometrical Procedure for Testing the REAGENT 2
25oc 30oc 37oc Equality of Measurements from Two Different Analytical Methods. CALIBRATION TYPE MULTIPLE MULTIPLE
Women up to 16 U/l up to 22 U/l up to 31 U/l J Clin Chem Clin Biochem 1983;21:709-720.
CALIBRATOR REPLICATES 3 3
Men up to 19 U/l up to 26 U/l up to 38 U/l 8. Schmidt FW. Ref Med Ges, Marburg/Lahn, December 1959. CALIBRATION
Each laboratory should investigate the transferability of the 9. Thefeld W et al. Dtsch med Wschr 1974;99:343. BLANK REPLICATES 3 3
expected values to its own patient population and if necessary 10. Tietz NW (Hrsg.). Clinical Guide to Laboratory Tests, 3. CALIBRATION CURVE
determine its own reference range. For diagnostic purposes, the AST Auflage. Philadelphia, PA: WB Saunders, 1995:76-77. BLANK ABS LIMIT 1.100 1.100
results should always be assessed in conjunction with the patient’s OPTIONS KINETIC BLANK LIMIT
medical history, clinical examination and other findings.
LINEARITY LIMIT 400 400
Use on Automated Analysers:
This reagent is suitable for use on a range of automated analysers.
Specific instructions for these applications are available on request
from our technical department.
For automated use we recommend a serum based calibrator to
eliminate any matrix bias which may be observed with the aqueous
standard.
Biorex Calibration Serum cat. No BXC0321K/ L/ M

Quality Control:

Biorex Normal Bovine Assayed Control Cat No BXC0313A (10x5ml)

Biorex Elevated Bovine Assayed Control Cat No BXC0313B (10x5ml
Biorex Normal Human Assayed Control Cat No BXC0312A (10x5ml)
Biorex Elevated Human Assayed Control Cat No BXC0312B (10x5ml)

The control intervals and limits must be adapted to the individual

laboratory and country-specific requirements. Values obtained should
fall within established limits. Each laboratory should establish corrective
measures to be taken if values fall outside the limits.

The control intervals and limits must be adapted to the individual

laboratory and country-specific requirements. Values obtained should

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 BXC0201 – AST (GOT) | Revision No.10 MAY/11 | Page 2 of 2