SET OF RESPONSES FOR DETERMINATION OF ACTIVITY OF ASTARATAMINOTRANSPHERASE IN BLOOD MILK (METHOD OF RAYMAN - FRAENKEL)
APPOINTMENT
The kit is used to determine the activity of aspartate aminotransferase in human serum in clinical and diagnostic and biochemical laboratories and in scientific and research practice.
The set is designed for 60 macro, 125 semimicro or 250 micro values (including blank samples)
The linearity of the calibration should be in the range of 0.1
μmol / (hourly) to 2.5 μmol / (hrm) (from 0.028 μc / l to 0.7 μc / l).
The coefficient of variation of determination is no more than 6%.
Storage set - at a temperature from plus 2 ° С to plus 8 ° С.
The warranty period is 12 months from the date of manufacture. The kit is intended for use in vitro by a trained technician.
PRINCIPLE OF THE METHOD
As a result of the amination of 2-oxo-glutaric acid with L-aspartic acid, which occurs under the action of aspartate aminotransferase, L-glutamic and oxalic acid are formed, the latter is deliberately decarboxylated to the formation of pyruvic acid. The determination is based on the measurement of the optical density of 2,4-dinitrophenylhydrazones of 2-oxoglutaric and pyruvic acids in alkaline medium. Since the pyruvic acid hydrazone has a higher molar extinction coefficient, there is a direct proportional dependence of the optical density of the reaction solution on the activity of the enzyme.
COMPOSITION OF THE SET
1. Substrate-buffer solution of AsAT - 1 bottle with (50 ± 2) ml;
- phosphate buffer (0,100 ± 0,005) mol / l,
- L-aspartic acid (0.100 ± 0.005) mol / L,
- 2-oxo-glutaric acid (2.0 ± 0.1) mmol / liter
2. Stop-reagent - 1 bottle with (50 ± 2) ml;
2,4-dinitrophenylhydrazine (2.4 DNFH) (1.00 ± 0.05) mmol / L
3. Gauge solution - 1 vial with (5,0 ± 0,5) ml;
- pyruvic acid sodium (2,0 ± 0,1) mmol / liter,
(220 ± 11) μg / ml (corresponding to 176 μg / ml of pyruvic acid)
4. Sodium hydroxide solution (4,0 ± 0,2) mol / l or dry - 1 vial with (50 ± 2) ml
(WARNING, RADIO SUBJECT!) Or from (8,00 ± 0,32) g.
NORMAL GREATS
The activity of aspartate aminotransferase (0.1-0.45) μmol / (h) in addition to 37 ° C.
These values are approximate, it is recommended to determine their own normal values in each laboratory.
INTERFERENCE
Hemolysis, keto compounds, aspirin, barbiturates, opiates, penicillin overstate the result when determining AsAT.
Some medications and substances may influence the course of determination.
DIAGNOSTIC CHARACTERISTICS
Aminotransferases catalyze the formation of glutamic acid from 2-oxoglutarate by the transfer of amino groups.
As a rule, AsAT is present in many tissues, but the highest concentrations are determined in the liver, heart muscle, as well as in the kidneys and pancreas.
The level of serum ASAT increases with hepatitis and other liver diseases accompanied by necrosis of hepatocytes: infectious mononucleosis, cholestasis, cirrhosis, metastatic carcinoma of the liver, white fever, and the appointment of various drugs such as opiates, salicylates or ampicillin, after myocardial infarction, with severe coronary insufficiency, after attacks of paraxysmal tachycardia, with diseases of the skeletal muscles (for example, progressing muscular dystrophy), with acute pancreatitis and other diseases h2,3.
Clinical diagnosis should be based on the integration of clinical and laboratory data. Delivery term: 5 g.
Information updated: 02.03.2018