ACCUCARETM
ALPHA-AMYLASE
Kinetic colorimetric test
Catalog No: Product Description: Pack Size:
AMY 10 ALPHA-AMYLASE 10 X 1.1 ML
AMY 30 ALPHA-AMYLASE 10 X 3 ML


METHOD
Kinetic colorimetric test


PRINCIPLE
The alpha-amylase hydrolyses p-nitro-phenylmalto-heptaoside producing glucose polymers and p-nitrophenyl oligosaccharides with shorter chain. These are further hydrolysed by gluco amylase and alpha-glucosidase to release p-nitrophenol. The increase in absorbance at 405 nm is proportional to the alpha-amylase activity.


SAMPLE:
Serum, plasma urine diluted 1:2.


REFERENCE VALUES
  SERUM URINE
30°C : upto 55 U/l 170 U/24 h
37°C : upto 90 U/l 450 U/24 h


The reference values are to be considered as indicative only. Every laboratory should establish its own normal ranges.


QUALITY CONTROL
Accutestrol N - H


REAGENTS
Reagent I : Buffer Reagent
Reagent II : Substrate Reagent


REFERENCES
Wohlegemuth, J. Biol. Chem., 29, 1 (1908)
Somogyi, M., J. Biol. Chem., 125, 399 (1938).
Street H.V., Close J.R., Clin. Chem. Acta, 1,256 (1956) .


REAGENT PREPARATION
Dissolve contents of one vial of substrate reagent with the
volume of buffer reagent as specified on the enzyme vial.
STABILITY :
60 days at 2-8°C. 5 days at R.T.

AUTOMATED PARAMETERS
Wave length 405 nm
Cuvette 1 cm light path
Reaction Temperature 37°C
Measurement Against distilled water
Reaction Type Kinetic test
Reaction Direction Increasing
Sample/Reagent Ratio 1:40
Delay/Lag/ time 60 Secs
Interval time 30 Secs
No: of Readings 04
Blank absorbance limit <0.7
Factor 4824
Low Normal at 37°C 0 U/L
High Normal at 37°C 90 U/L
Linearity 1200 U/L

MANUAL PROCEDURE

PIPETTE INTO TEST TUBE
SAMPLE 25 µl
REAGENT 1000 µl

Mix well and incubate for 1 min. at 37°C & read initial absorbance and start timer simultaneously. Measure absorbance increase every minute for 3 minutes p A/ min.

CALCULATION AND LINEARITY
  SERUM URINE
U/l 30°C : p A/ min. x 5256 15768
U/l 37°C : p A/ min. x 4824 14472

This method is linear upto a concentration of 1200 U/L (serum / plasma) or upto 3600 U/l (urine)

NOTES
Lipemia, Bilirubin, glucose do not interfere with the assay.
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