Amylase assay
AssigneeE. I. Du Pont de Nemours and Company
InventorsRobert C. Menson, Venkatachalam Narayanswamy, Richard C. Burns
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Abstract
Disclosed herein are a method and a reagent test kit, both using an improved substrate to measure the amylase content of a sample. The substrate used is a glycoside consisting of a defined polysaccharide glycosyl residue and a substituted aromatic radical attached to the terminal unit of the glycoside. When detached from the polysaccharide, the aglycone exhibits a different spectral absorbance than the substrate.
Claims15
A method for the rapid determination of the amylase content of a sample comprising the amylase content of a sample comprising the steps of:
- (a) adding to a solution containing a measured amount of the sample a defined polysaccharide substrate having the following formula:
- ##STR10## where n is 2, 3, or 4, and R is a substituted aromatic radical which, as a detached aglycone, exhibits a different spectral absorbance than the substrate;
- (b) adding a maltase to the solution;
- and
- (c) monitoring the spectral absorbance of the solution.
In a method for rapidly determining the amylase content of a sample comprising the steps of adding a maltase and a substrate to a solution containing a measured amount of the sample and monitoring the change in spectral absorbance of the solution, the improvement wherein the substrate is .alpha.-(4-nitrophenyl) glucoside of maltotetraose, maltopentaose or maltohexaose.
A reagent test kit for rapidly determining the amylase content of a sample comprising:
- (a) a defined polysaccharide substrate having the following formula:
- ##STR13## wherein n is 2, 3 or 4, and R is a substituted aromatic radical which, when detached from the polysaccharide in the form of a phenolate anion, exhibits a different spectral absorbance than the substrate;
- and
- (b) maltase.
In a test kit for rapidly determining the amylase content of a sample comprising a maltase and a substrate, the improvement wherein the substrate is .alpha.-(4-nitrophenyl) glycoside of maltotetraose, maltopentaose, or maltohexaose.
A process for determining the .alpha.-amylase content of a sample comprising the steps of adding an oligosaccharide substrate, having 4-6 glucose units and a terminal 4-nitrophenyl group to a solution containing a measured amount of said sample and .alpha.-glucosidase and determining amylase activity by the release of p-nitrophenol.
Dependencies are inferred from the claim text.