.alpha.-amylase assay utilizing modified starch as the...

C - Chemistry – Metallurgy – 09 – K

Patent

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150/15.2, 150/3.

C09K 3/00 (2006.01) C12Q 1/40 (2006.01) G01N 33/48 (2006.01)

Patent

CA 1068586

Abstract of the Disclosure A specific and sensitive method for the determination of .alpha.-amylase activities in a sample is disclosed. The method utilizes a partially modified starch substrate which contains blockages to the action of exo- carbohydrases but not to .alpha.-amylase, an endo-carbohydrase. Cleavage of the modified substrate by .alpha.-amylase leads to free nonreducing ends which can then be attacked by a suitable exo-carbohydrase serving as the coupling enzyme. The reaction sequence is as follows: Blocked starch .alpha.-amylase unblocked starch phosphoglucomutase inorganic phosphate glucose-1-phosphate phosphoglucomutase glucose-6-phosphate glucose-6-phosphate dehydrogenase, NAD NADH. The rate of increase of absorbance due to NADH formation can be measured to determine the level of .alpha.-amylase activity in a sample, particularly one of serum or urine origin. The values obtained are useful in clinical diagnosis.

281158

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