US4233403A
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Amylase assay
Assignee: Dade International Inc
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Filing: 1978-02-27
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Granted: 1980-11-11
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Expires: 1998-02-27
Abstract
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.
Inventors
- 👤 Robert C. Menson
- 👤 Venkatachalam Narayanswamy
- 👤 Richard C. Burns
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 04:56
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/34; C12Q1/40; C12Q2334/00; C12Q2334/10; Y; Y10; Y10S; Y10S435/00; Y10S435/81
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
88 words
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.
Claim 2
Independent Claim
88 words
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.
Claim 3
Dependent Claim
74 words
The method of claim 1 wherein R is a substituted aromatic radical selected from the group consisting of ##STR11## in which X and Y are individually selected from the group consisting of H, NO 2 , halogens, alkyls of from 1 to 4 carbon atoms, OR' or CO 2 R', where R' is an alkyl of from 1 to 6 carbon atoms, and at least one of X and Y is NO 2 .
Claim 4
Dependent Claim
9 words
The method of claim 2 wherein R is ##STR12##
Claim 5
Dependent Claim
10 words
The method of claim 1 wherein the maltase is α-glucosidase.
Claim 6
Dependent Claim
9 words
The method of claim 1 wherein R is 4-nitrophenyl.
Claim 7
Dependent Claim
40 words
The method of claim 5 wherein the solution is maintained at a substantially constant pH in the basic range and a substantially constant temperature and wherein the spectral absorbance is within one hour of adding the sample to the substrate.
Claim 8
Dependent Claim
25 words
The method of claim 2 wherein the solution is maintained at a substantially constant pH in the basic range and at a substantially constant temperature.
Claim 9
Dependent Claim
55 words
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 α-(4-nitrophenyl) glucoside of maltotetraose, maltopentaose or maltohexaose.
Claim 10
Dependent Claim
55 words
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 α-(4-nitrophenyl) glucoside of maltotetraose, maltopentaose or maltohexaose.
Claim 11
Dependent Claim
62 words
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.
Claim 12
Dependent Claim
62 words
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.
Claim 13
Dependent Claim
75 words
The test kit of claim 9 wherein R is a substituted aromatic radical selected from the group consisting of ##STR14## in which X and Y are individually selected from the group consisting of H, NO 2 , halogens, alkyls of from 1 to 4 carbon atoms, OR' or CO 2 R', where R' is an alkyl of from 1 to 6 carbon atoms, and at least one of X and Y is NO 2 .
Claim 14
Dependent Claim
11 words
The test kit of claim 10 wherein the maltase is α-glucosidase.
Claim 15
Dependent Claim
10 words
The test kit of claim 10 wherein R is ##STR15##
Claim 16
Dependent Claim
10 words
The test kit of claim 10 wherein R is 4-nitrophenyl.
Claim 17
Dependent Claim
32 words
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 α-(4-nitrophenyl) glycoside of maltotetraose, maltopentaose, or maltohexaose.
Claim 18
Dependent Claim
32 words
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 α-(4-nitrophenyl) glycoside of maltotetraose, maltopentaose, or maltohexaose.
Claim 19
Dependent Claim
48 words
A process for determining the α-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 α-glucosidase and determining amylase activity by the release of p-nitrophenol.
Claim 20
Dependent Claim
48 words
A process for determining the α-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 α-glucosidase and determining amylase activity by the release of p-nitrophenol.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1983-11-11 | 1984-05-12 | 1984-11-11 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-11-11 | 1988-05-12 | 1988-11-11 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-11-11 | 1992-05-12 | 1992-11-11 | $7700.00 | Window Closed / Lapsed |