Soduim Hydroxide (NaOH) 4N 1 WU/mL = 1U/L x 2.

07
1 U/L = 1WU/mL x 0.483
SAMPLES
Serum, free of hemolysis. NORMAL VALUES

PROCEDURE
GOT GOT GOT/AST = 8-40 WU/ml (3.85-19.3 U/L)
Substrate GOT (R.1) 0.5 ml
Preincubate for 5 min at 37°C.
Colorimetric test. Reitman-Frankel Serum 100 µL REFERENCES
Mix. Return to bath for 60 min 1. Reitman S., Frankel S.,Am. Clin. Pathol., 28,56 (1957)
2. Tietz, NW.,Fund of Clinical Chem., 446 (1970)
For in vitro diagnostic use only DNPH R.2 0.5 ml
3. Schmidt, E., Enzymology Biol.Clin., 3,1 (1963)
Mix. Allow to stand for 20 min at room temperature.
Store at 2-8ºC NaOH 0.4 N 5.0 ml
MIX. Let stand for 15 min at room temperature.
INTENDED USE Read at 505 nm against a water blank.
For the determination of GOT concentration in human serum. The colour is stable at least 60 minutes.

PRINCIPLE
The glutamic transaminase enzymes, serum glutamic oxalacetic CALCULATIONS ATLAS Medical
(GOT) and serum glutamic pyruvic (GPT), catalyze the transfer of From absorbencies, read units of GOT from corresponding William James House,
the amino group of glutamic acid to oxalacetic acid and pyruvic curves. Cowley Road, Cambridge, CB4 0WX, UK
acid in reversible reactions. The transaminase activity is Tel: ++44 (0) 1223 858 910
proportional to the amount of oxaloacetate pyruvate formed CALIBRATION (mL) Fax: ++44 (0) 1223 858 524
over a definite period of time and is measured by a reaction with
2.4-dinitrophenylhydrazine (DNPH) in alkaline solution. 1 2 3 4 5 6 PPI769A01
Water 0.2 0.2 0.2 0.2 0.2 0.2 Rev A (28.10.2015)
REAGENTS GOT R.1 1.0 0.9 0.8 0.7 0.6 0.5 Catalogue Number Store at
Pyruvic stand. -- 0.1 0.2 0.3 0.4 0.5 For In-Vitro Diagnostic
Materials Provided Caution
DNPH R.2 1.0 1.0 1.0 1.0 1.0 1.0 use
Content Concentration Mix. Allow to stand for 20 min at room temperature. Number of tests in the Read product insert
NaOH 0.4 N 10 10 10 10 10 10 pack before use
Reagent -1 Substrate GOT (AST)
L-aspartate 100 mmol/L Lot (batch) number Manufacturer
MIX. Allow to stand for at least 15 min.
Ketoglutarate 2 mmol/L Read against water blank at 505 nm. Fragile,
Expiry date
Plot a calibrator curve of the absorbances found vs. the handle with care
corresponding units, on a graph paper, according to the following
Manufacturer fax Do not use if
concentrations: number package is damaged

Reagent -2 Colour reagent Manufacturer telephon

GOT WU/mL 0 22 55 95 150 215
2.4-dinitrophenylhydrazine 1 mmol/L e number
U/L 0 11 27 46 72 104

Standard Pyruvic standard. 1.2 mmol/L LINEARITY

When GOT value exceeds 180 WU/Ml (87 U/L repeat test using a
Materials Required But Not Provided 1:10 dilution of serum with saline sol.(9g/L). Multiply the result
by 10.
Content Concentration UNITS
 The rate of decrease in concentration of NADH, measured PROCEDURE
photometrically, is proportional to the catalytic 1. Assay conditions:
concentration of AST Wavelength : ...........................................................340 nm
GOT ( A S T ) present in the sample'. Cuvette : ................................................... 1 cm. light path
NADH. Kinetic UV. IFCC .Liquid Constant temperature .............................25°C /30°C / 37°C
Quantitative determination of aspartate REAGENTS 2. Adjust the instrument to zero with distilled water or air.
aminotransferase TRIS pH 7.8 80 mmol/L 3. Pipette into a cuvette:
GOT (AST) R1
Buffer Lactate dehydrogenase (LDH) 800 U/L WR (mL) 1.0
For in -vitro diagnostic use only.
Malate dehydrogenase (MDH) 600 U/L Sample (μL) 100
Store at 2-8°C. 1. Mix, incubate for 1 minute.
L-Aspartate 200 mmol/L 2. Read initial absorbance (A) of the sample, start the
INTENDED USE R2 NADH 0.18 mmol/L stopwatch and read absorbances at 1 minute intervals
For the quantitative determination of aspartate Substrate thereafter for 3 minutes.
aminotransferase GOT (AST) in human serum or plasma. α-Ketoglutarate 12 mmol/L 3. Calculate the difference between absorbances and the
PREPARATION average absorbance differences per minute (ΔA/min).
INTRODUCTION • Working reagent (WR):
The AST is a cellular enzyme, is found in highest Mix:4 vol. (R1) Buffer + 1 vol. (R2) Substrate. CALCULATIONS
concentration in heart muscle, the cells of the liver, the • Stability: 21 days at 2-8°C or 72 hours at room ΔA/min x 1750 = U/L of AST
cells of the skeletal muscle and in smaller amounts in temperature (15-25°C). Units: One international unit (IU) is the amount of enzyme
other weaves. that transforms 1 μmol of substrate per minute, in
Although an elevated level of AST in the serum is not STORAGE AND STABILITY standard conditions. The concentration is expressed in
specific of the hepatic disease, is used mainly to • • All the components of the kit are stable until the units per litre of sample (U/L).
diagnostic and to verify the course of this disease with expiration Temperature conversion factors
other enzymes like ALT and ALP. Also it is used to control date on the label when stored tightly closed at 2-8°C, To correct results to other temperatures multiply by:
the patients after myocardial infarction, in skeletal muscle protected from light and contaminations prevented
disease and other''45. during Assay Conversion factor to
Clinical diagnosis should not be made on a single test their use. temperature 250C 300C 370C
result; it should integrate clinical and other laboratory • Do not use reagents over the expiration date. 250C 1.00 1.37 2.08
data. • Signs of reagent deterioration: 300C 0.73 1.00 1.54
• Presence of particles and turbidity. 370C 0.48 0.65 1.00
PRINCIPLE OF THE METHOD • Blank absorbance (A) at 340 nm < 1.00.
Aspartate aminotransferase (AST) formerly called
QUALITY CONTROL
glutamateoxaloacetate (GOT) catalyses the reversible EQUIPMENTS NEEDED BUT NOT PROVIDED
Control sera are recommended to monitor the
transfer of an amino group rom aspartate to • Spectrophotometer or colorimeter measuring at 340
performance of assay Procedures: ATLAS H Normal and
α-ketoglutarate forming glutamate and oxalacetate. nm.
Pathologic.
The oxalacetate produced is reduced to malate by malate • Thermostatic bath at 25°C, 30°C 37°C (± 0.1 °C)
If control values are found outside the defined range,
dehydrogenase (MDH) and NADH: • Matched cuvettes 1.0 cm light path.
check the instrument, reagents and technique for
• General laboratory equipment.
AST problems.
L-Aspartate + α-Ketoglutarate Glutamate + Oxalacetate
SAMPLES
MDH
Oxalacetate + NADH + H+ Malate + NAD+ Serum or plasma': Stability 7 days at 2-8°C.
Each laboratory should establish its own Quality Control A list of drugs and other interfering substances with AST
scheme and corrective actions if controls do not meet the determination has been reported by Young et. a12,3.
acceptable tolerances.
REFERENCES
REFERENCE VALUES' 1. Murray R. Aspartate aminotransferase. Kaplan A et al.
370C 250C 300C Clin Chem The
Men up to 19 U/L 26 U/L C.V. Mosby Co. St Louis. Toronto. Princeton 1984;
38 U/L 1112-116.
Women up to 16 U/L 22 U/L 2. Young DS. Effects of drugs on Clinical Lab. Tests, 4th ed
31 U/L AACC Press,
These values are for orientation purpose; each laboratory 1995.
should establish Its own reference range. 3. Young DS. Effects of disease on Clinical Lab. Tests, 4th
ed AACC 2001.
PERFORMANCE CHARACTERISTICS 4. Burtis A et al. Tietz Textbook of Clinical Chemistry, 3rd
Measuring range: ed AACC 1999.
From detection limit of 1 U/L to linearity limit of 260 U/L. 5. Tietz N W et al. Clinical Guide to Laboratory Tests, 3rd
If the results obtained were greater than linearity limit, ed AACC 1995.
dilute the sample1/10 with NaCl 9 g/L and multiply the
result by 10.
Precision: ATLAS MEDICAL
Intra – assay (n=20) Inter – assay (n=20) Unit 4, William James House
Cowley Rd, Cambridge, CB4 0WX
Mean 17.0 135 17.3 131 Tel: ++44 (0) 1223 858 910
(U/L) Fax: ++44 (0) 1223 858 524
SD 0.72 1.05 0.81 2.25 PPI445A01
CV (%) 4.27 0.77 4.68 1.72 Rev C ( 10.11.2015)
Sensitivity.
1 U/L = 0, 0048 ΔA/min. Catalogue Number Store at
Accuracy: For In-Vitro Diagnostic
Caution
Results obtained using ATLAS reagents (y) did not show use
systematic differences when compared with other Number of tests in the Read product insert
commercial reagents (X). pack before use
The results obtained using 100 samples were the Lot (batch) number Manufacturer
following:
Fragile, handle with
Correlation coefficient (r) : 0.9839 Expiry date
care
Regression equation: y= 0.9866 + 0.588.
Manufacturer fax Do not use if
The results of the performance characteristics depend on number
the analyzer Used. package is damaged
Manufacturer
telephone number
INTERFERENCES
Anticoagulants currently in use like heparin, EDTA, oxalate
and fluoride do not affect the results. Haemolysis
interferes with the assay'
 NADH 0.18 mmol/L the average absorbance differences per minute
R2 Lactate dehydrogenase (LDH) 800 U/L (ΔA/min).
Substrate Malate dehydrogenase (MDH) 600 U/L CALCULATIONS
-Ketoglutarate 12 mmol/L ΔA/min x 1750 = U/L of AST

ATLAS GOT (AST) TEST EQUIPMENTS NEEDED BUT NOT PROVIDED Units: One international unit (IU) is the amount of enzyme that
(Kinetic)  Spectrophotometer or transforms 1 µmol of substrate per minute, in standard conditions.
For In-Vitro diagnostic and professional use only colorimeter measuring at 340 nm. The concentration is expressed in units per litre of sample (U/L).
Thermostatic bath at 25°C, 30°C o Temperature conversion factors
Store at 2-8C 37°C (± 0.1°C) – Matched. To correct results to other temperatures multiply by:
 cuvettes 1.0 cm light path.
 General laboratory equipment. Assay Conversion factor to
INTENDED USE
temperature 25°C 30°C 37°C
For the determination of GOT (AST) concentration in human PREPARATION
25°C 1.00 1.37 2.08
serum or plasma.  Working reagent (WR): 30°C 0.73 1.00 1.54
PRINCIPLE OF THE METHOD Dissolve ( ) one tablet of R 2 Substrate in 2 37°C 0.48 0.65 1.00
Aspartate aminotransferase (AST) formerly called glutamate mL of R1 Cap and mix gently to dissolve
oxaloacetate (GOT) catalyses the reversible transfer of an contents.
QUALITY CONTROL
amino group from aspartate to -ketoglutarate forming  Stability: 21 days at 2-8°C or 72 hours at room
Control sera are recommended to monitor the performance of
glutamate and oxalacetate. The oxalacetate produced is temperature (15-25°C).
assay procedures. If control values are found outside the
reduced to malate by malate dehydrogenase (MDH) and STORAGE AND STABILITY defined range, check the instrument, reagents and technique
NADH:  All the components of the kit are stable until the for problems.
Aspartate + -Ketoglutarate AST Glutamate + expiration date on the label when stored tightly Each laboratory should establish its own Quality Control
closed at 2-8°C, protected from light and scheme and corrective actions if controls do not meet the
Oxalacetate
contaminations prevented during their use. acceptable tolerances.
Oxalacetate + NADH + H .MDH Malate + NAD`  Do not use the tablets if appears broken.
REFERENCE VALUES'
 Do not use reagents over the expiration date.
25°C 30°C 37°C
The rate of decrease in concentration of NADH, measured  Sign of reagent deterioration:
photometrically, is proportional to the catalytic  Presence of particles and turbidity. Men up to 19 U/L 26 38 U/L
concentration of AST present in the sample'.  Blank absorbance (A) at 340 nm < 1.00. U/L
Women up to 16 U/L 22 31 U/L
INTRODUCTION SAMPLES U/L
The AST is a cellular enzyme, is found in highest Serum or plasma': Stability 7 days at 2-8°C.
concentration in heart muscle, the cells of the liver, the These values are for orientation purpose; each laboratory
cells of the skeletal muscle and in smaller amounts in other PROCEDURE
1. Assay conditions: should establish its own reference range.
weaves.
Although an elevated level of AST in the serum is not Wavelength: ……………………………340 nm
Cuvette :………………………………..1 cm. light path PERFORMANCE CHARACTERISTICS
specific of the hepatic disease, is used mainly to diagnostic Measuring range:
and to verify the course of this disease with other enzymes Constant temperature ………………25°C /30°C /37°C
2. Adjust the instrument to zero with distilled From detection limit of 5,44 U/L to linearity limit of 260 U/L. If
like ALT and ALP. the results obtained were greater than linearity limit, dilute
Also it is used to control the patients after myocardial water or air.
3. Pipette into a cuvette: the sample 1/10 with NaCl 9 g/L and multiply the result by
infarction, in skeletal muscle disease and other''4,s 10.
Clinical diagnosis should not be made on a single test result; WR (mL) 1.0
Sample (ML) 100 Precision:
it should integrate clinical and other laboratory data.

REAGENTS Intra-assay (n=20)

4. Mix, incubate for 1 minute.
R1 TRIS pH 7.8 80 mmol/L 5. Read initial absorbance (A) of the sample, start the Mean U/L 17.4 128
Buffer L-Aspartate 200 mmol/L stopwatch and SD 0.68 1.35
6. read absorbances at 1 minute intervals thereafter CV(%) 3.91 1.05
for 3 minutes.
7. Calculate the difference between absorbances and Inter-assay (n=20)
17.1 128
Atlas Medical
0.72 1.28
William James House,
4.20 0.99
Cowley Road, Cambridge, CB4 0WX, UK
Tel: ++44 (0) 1223 858 910
Sensitivity:
Fax: ++44 (0) 1223 858 524
1 U/L = 0,0017 ΔA/min.
Accuracy:
PPI192A01
Results obtained using ATLAS reagents (y) did not show
Rev C (10.11.2015)
systematic differences when compared with other
commercial reagents (x). The results obtained using 50
Catalogue Number Store at
samples were the following:
Correlation coefficient (r):0,99. For In-Vitro Diagnostic
Caution
Regression equation: y= 0,96x + 1,33. use
The results of the performance characteristics depend on Number of tests in the Read product insert
the analyzer used. pack before use

Lot (batch) number Manufacturer

INTERFERENCES
Anticoagulants currently in use like heparin, EDTA, oxalate Fragile, handle with
Expiry date
and fluoride do not affect the results. Haemolysis interferes care
with the assay' Manufacturer fax Do not use if
A list of drugs and other interfering substances with AST number package is damaged
determination has been reported by Young et. al2,3. Manufacturer
telephone number
REFERENCES
1. Murray R. Aspartate aminotransferase. Kaplan
A et al. Clin Chem The C.V. Mosby Co. St Louis.
Toronto. Princeton 1984; 1112-116.
2. Young DS. Effects of drugs on Clinical Lab. Tests,
4th ed AACC Press, 1995.
3. Young DS. Effects of disease on Clinical Lab.
Tests, 4th ed AACC 2001.
4. Burtis A et at. Tietz Textbook of Clinical
Chemistry, 3rd ed AACC 1999.
5. Tietz N W et al. Clinical Guide to Laboratory
Tests, 3rd ed AACC 1995