BXC0201B

BXC0201C

BXC0201D

BXC0201E

10x20ml

10x10ml

5x50ml

20x2ml

STORE AT 2-8C

AST (GOT)
IFCC UV

Kit Contents:

R1 AST Buffer
R2 AST Reagent
R1 AST Buffer
R2 AST Reagent

Reagent Concentration:
R1

BXC0201B

BXC0201C

2x105ml
10x20ml

1x105ml
10x10ml

BXC0201E

BXC0201D
5x50ml
5x50ml

1x45ml
20x2ml

Intended Use:
In vitro test for the quantitative determination of aspartate aminotransferase (AST) in human serum and plasma.
Test Principle:
Method recommended by the IFCC:
2-Oxoglutarate + L-Aspartate
Oxaloacetate + NADH + H+

FOR IN-VITRO DIAGNOSTIC USE ONLY

ISO 13485 accredited company

GOT

MDH

4 Glutamate + Oxaloacetate
4 Malate + NAD+

Summary:
Aspartate aminotransferase (glutamate oxaloacetate transaminase)
belongs to the transaminases, which catalyze the interconversion of
amino acids and -ketoacids by transfer of amino groups. Aspartate
aminotransferase is commonly found in human tissue. Although heart
muscle is found to have the most activity of the enzyme, significant
activity has also been seen in the brain, liver, gastric mucosa, adipose
tissue, skeletal muscle, and kidneys.
AST is present in both the cytoplasm and mitochondria of cells. In cases
involving mild tissue injury, the predominant form of AST is that from the
cytoplasm, with a smaller amount coming from the mitochondria.
Severe tissue damage results in more of the mitochondrial enzyme being
released. Elevated levels of the transaminases can signal myocardial
infarction, hepatic disease, muscular dystrophy and organ damage.
In 1955, Karmen et al described the first kinetic determination of AST
activity in serum. The International Federation of Clinical Chemistry
(IFCC) recommended in 1977 and 1980 standardized procedures for AST
determination, including optimization of substrate concentrations,
employment of TRIS* buffers, preincubation of combined buffer and
serum to allow side reactions with NADH to occur, substrate start and
optional pyridoxal phosphate activation.
This method is derived from the IFCC reference method.
*TRIS = Tris(hydroxymethyl)-aminomethane

R2

Tris Buffer pH 7.8

L-Aspartate
NADH
LDH
MDH
Oxoglutarate

BXC0201 AST (GOT) Page 1 of 2

80mmol/l
200mmol/l
0.18mmol/l
800U/l
600U/l
12mmol/l

Reagent Handling and Preparation:

R1 Buffer: Ready to use, stable up to the expiry date when stored at 28C.
R2 Reagent: Reconstitute enzyme reagent R2 with the corresponding
volume of buffer R1. Wait for at least 15 mins before use.
Cat No BXC0211D, reconstitute one vial of Enzyme Reagent (R2) with a
portion of Buffer (R1) and then transfer the entire contents to R1 bottle
rinsing R2 bottle several times.
This reagent is stable for 14 days at +2C to +8C.
Sample:
Serum, heparinised or EDTA plasma

Specimen:
Collect serum using standard sampling tubes.
Heparin or EDTA plasma
Stability:

24 hours at +20C to +25C

7 days at +2C to +8C

Separate serum/plasma from clot/cells within 8 hours at room

temperature or 48 hours at +2C to +8C.
Centrifuge samples containing precipitate before performing the assay.
Testing Procedure:
Materials Provided:
Working Solutions as described above
Additional Materials Required:
Controls as indicated
0.9% NaCI

Fortress Diagnostics Limited Unit 2C Antrim Technology Park, Antrim BT41 1QS (United Kingdom)
TEL: +44 (0) 2894 487676 FAX: +44 (0) 2894 469933 www.fortressdiagnostics.com

Manual Procedure:
Wavelength
Hg 340nm, 334nm
or Hg 365nm

Temperature
+25C/+30C/
+37C

Cuvette
1cm light path

Pipette into test tubes as follows:

Measurement
Against Air or
Distilled Water

Macro

Working Reagent

2500l

Sample

Micro
500l

250l
50l
Mix, incubate for 1 minute read initial absorbance at assay
temperature and start stopwatch simultaneously. Read again after
exactly 1, 2 and 3 minutes.
If the A/min is between 0.11 and 0.25 at Hg 334nm/340nm or 0.06
and 0.013 at Hg 365nm use only the values for the first 2 minutes for
the calculation.

Calculation:
340nm
Hg 334nm
Hg 365nm

A/min x
A/min x
A/min x

Macro
1746
1780
3235

Micro
1746
1780
3235

Linearity:
Up to 440U/l
If the change of absorbance per minute is higher than 0.250 at 340nm or
0.130 at 365nm the sample has to be diluted with 0.9% NaCI or
distilled/deionised water (e.g. 1+9). Multiply the result by the appropriate
dilution factor (e.g. factor 10).
Sensitivity:
Detection limit: 4U/l or 0.07kat/l
The lower detection limit represents the lowest measurable AST
concentration that can be distinguished from zero.

Imprecision:
Reproducibility was determined using controls. The following results were
obtained:
Sample 1
Sample 2

Intra Assay - Within Run

MW U/l
SD U/l
33.4
116

REVISED OCT/09

0.76
1.56

CV%
2.29
1.34

BXC0201B

BXC0201C

BXC0201D

BXC0201E

10x20ml

10x10ml

5x50ml

20x2ml

STORE AT 2-8C
FOR IN-VITRO DIAGNOSTIC USE ONLY

ISO 13485 accredited company

Sample 1

Intra Assay - Between Run

MW U/l
SD U/l

Sample 2

33.2

0.67

148

2.06

82.9

Sample 2

1.18

CV%
2.02
1.43
1.40

Method comparison:
A comparison of the Fortress AST (y) with a commercial obtainable assay
(x) gave the following result with 48 samples:
y = 0.992x - 0.022; r= 0.995
Limitations - interference:
Criterion: Recovery within 10% of initial value.
Icterus: No significant interference up to an index I of 70
(approximate conjugated and unconjugated bilirubin: 70 mg/dl)
Haemolysis: No significant interference up to an index H of 900
(approximate haemoglobin concentration: 900 mg/dl).
Lipaemia (Intralipid): No significant interference up to an index L of 450
(approximate triglycerides concentration: 900 mg/dl)There is poor
correlation between turbidity and triglycerides concentration.
Lipaemia may cause absorbance flagging as a result of an absorbance
increase.
Normal Values:
According to the IFCC method.
Women
Men

25C

up to 16U/l
up to 19U/l

30C

up to 22U/l
up to 26U/l

37C

up to 31U/l
up to 38U/l

Each laboratory should investigate the transferability of the expected

values to its own patient population and if necessary determine its own
reference range. For diagnostic purposes, the ALT results should always
be assessed in conjunction with the patients medical history, clinical
examination and other findings.

BXC0201 AST (GOT) Page 2 of 2

fall within established limits. Each laboratory should establish corrective

measures to be taken if values fall outside the limits.

Health & Safety:

This kit is designed for use by suitably qualified laboratory personnel only.
Exercise the normal precautions required for the handling of laboratory
reagents. Do not ingest the material. Dispose of material according to
local guidelines.

References:
1. International Federation of Clinical Chemistry, Scientific committee.
J Clin Chem clin Biochem 1980 18: 521-534.
2. Bablok W et al. A General Regression Procedure for Method
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 2. IFCC Method for aspartate aminotransferase. J
Clin Chem Clin Biochem 1986;24:49.
4. Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of
Interferences in Clinical Chemistry Instrumentation. Clin Chem 1
986;32:470-474.
5. Greiling H, Gressner AM (Hrsg.). Lehrbuch der Klinischen Chemie
und Pathobiochemie,3. Auflage. Stuttgart/New York: Schattauer
Verlag, 1995
6. Karmen A et al. J Clin Invest 1955;24:126.
7. Passing H, Bablok W. A New Biometrical Procedure for Testing the
Equality of Measurements from Two Different Analytical Methods. J
Clin Chem Clin Biochem 1983;21:709-720.
8. Schmidt FW. Ref Med Ges, Marburg/Lahn, December 1959.
9. Thefeld W et al. Dtsch med Wschr 1974;99:343.
10. Tietz NW (Hrsg.). Clinical Guide to Laboratory Tests, 3.
Auflage. Philadelphia, PA: WB Saunders, 1995:76-77.

Quality Control:
Fortress Normal Bovine Assayed Control Cat No BCX0313A (10x5ml)
Fortress Elevated Bovine Assayed Control Cat No BCX0313B (10x5ml)

Fortress Normal Human Assayed Control Cat No BXC0312A (10x5ml)

Fortress 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

Fortress Diagnostics Limited Unit 2C Antrim Technology Park, Antrim BT41 1QS (United Kingdom)
TEL: +44 (0) 2894 487676 FAX: +44 (0) 2894 469933 www.fortressdiagnostics.com

REVISED OCT/09