US5094943A
Expired / Lapsed
US
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Process and reagent for the improved quantitative colorimetic determination of hydrogen peroxide
Assignee: Roche Diagnostics GmbH
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Filing: 1988-01-19
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Granted: 1992-03-10
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Expires: 2008-01-19
Abstract
Abstract The present invention provides a process for the improved colorimetric determination of hydrogen peroxide as formed by a hydrogen peroxide-producing oxidase, by addition of a chromogenic system and measurement of the colored material formed, wherein superoxide dismutase (E.C. 1.15.1.1) is added to the reagent solution. The present invention also provides a reagent for the improved colorimetric determination of hydrogen peroxide, comprising a hydrogen peroxide-producing oxidase, a chromogenic system, a buffer and optionally adjuvant enzymes, wherein it also contains superoxide dismutase.
Inventors
- 👤 Joachim Siedel
- 👤 Albert Roder
- 👤 Joachim Ziegenhorn
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 15:58
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/26; C12Q1/28
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
53 words
A process for colorimetric determination of hydrogen peroxide formed by a hydrogen peroxide producing oxidase which does not produce superoxide comprising adding to a non-superoxide containing sample containing said hydrogen peroxide producing oxidase a chromogenic system and superoxide dismutase in an amount sufficient to increase sensitivity of colorimetric determination of said chromatogenic system.
Claim 2
Dependent Claim
53 words
A process for colorimetric determination of hydrogen peroxide formed by a hydrogen peroxide producing oxidase which does not produce superoxide comprising adding to a non-superoxide containing sample containing said hydrogen peroxide producing oxidase a chromogenic system and superoxide dismutase in an amount sufficient to increase sensitivity of colorimetric determination of said chromatogenic system.
Claim 3
Dependent Claim
18 words
Process according to claim 1, wherein 10 to 150 μg. of superoxide dismutase/ml. of reagent solution are added.
Claim 4
Dependent Claim
18 words
Process according to claim 2, wherein 25 to 75 μg. of superoxide dismutase/ml. of reagent solution are added.
Claim 5
Dependent Claim
13 words
Process according to claim 1, wherein the superoxide dismutase is Cu, Zn-superoxide dismutase.
Claim 6
Dependent Claim
55 words
Process according to claim 1, wherein the chromogenic system is a Trinder reagent, said reagent containing 4-aminoantipyrine, peroxidase, and one member of the group consisting of phenol a phenol derivative or an aniline derivative, wherein said phenol, phenol derivative and aniline derivative oxidatively couple to 4-aminoantipyrine in the presence of H 2 O 2 .
Claim 7
Dependent Claim
19 words
Process according to claim 5, wherein, as phenol derivative, there is used 2,4,6-triiodo-3-hydroxybenzoic acid or the analogous tribromo compound.
Claim 8
Dependent Claim
15 words
Process according to claim 5, wherein, as aniline derivative, there is used N-ethyl-N-β-sulphoethyl-m-toluidine or N-ethyl-N-β-hydroxyethyl-m-toluidine.
Claim 9
Dependent Claim
35 words
Process according to claim 1, wherein the chromogenic system comprises peroxidase, an aniline derivative which oxidatively couples to 4-amino-antipyrine in the presence of H 2 O 2 and 3-methyl-2-benzo-thiazolinone hydrazone (MBTH) or 3-methyl-2-benzo(2'-sulfo)-thiazolinone hydrazone (MBTH-S).
Claim 10
Dependent Claim
18 words
Process according to claim 8, wherein, as aniline derivative, there is used N,N-dimethylaniline, N,N-dimethylaminobenzoic acid, N-ethyl-N-β-sulphoethyl-m-toluidine or N-ethyl-N-β-hydroxyethyl-m-toluidine.
Claim 11
Dependent Claim
17 words
Process according to claim 1, wherein the hydrogen peroxide-producing oxidase is sarcosine oxidase, uricase or oxalate oxidase.
Claim 12
Dependent Claim
40 words
A reagent for colorimetric determination of hydrogen peroxide comprising a hydrogen peroxide-producing oxidase, a chromogenic system, a buffer optionally adjuvant enzymes, and superoxide dimutase in an amount effective to improve the sensitivity of the colorimetric determination of the chromogenic system.
Claim 13
Dependent Claim
40 words
A reagent for colorimetric determination of hydrogen peroxide comprising a hydrogen peroxide-producing oxidase, a chromogenic system, a buffer optionally adjuvant enzymes, and superoxide dimutase in an amount effective to improve the sensitivity of the colorimetric determination of the chromogenic system.
Claim 14
Dependent Claim
13 words
Reagent according to claim 11, wherein the superoxide dismutase is Cu, Zn-superoxide dismutase.
Claim 15
Dependent Claim
57 words
Reagent according to claim 11 wherein the chromogenic system is a Trinder reagent said reagent containing peroxidase, 4-aminoantipyrine and at least one member of the group consisting of phenol a phenol derivative and an aniline derivative wherein said phenol, phenol derivative or aniline derivative oxidatively couple to 4-aminoantipyrine in the presence of H 2 O 2 .
Claim 16
Dependent Claim
18 words
Reagent according to claim 13, wherein, as phenol derivative, it contains 2,4,6-triiodo-3-hydroxybenzoic acid or the analogous tribromo compound.
Claim 17
Dependent Claim
14 words
Reagent according to claim 13, wherein, as aniline derivative, it contains N-ethyl-N-β-sulphoethyl-m-toluidine or N-ethyl-N-β-hydroxyethyl-m-toluidine.
Claim 18
Dependent Claim
34 words
Reagent according to claim 11, wherein the chromogenic system contains peroxidase, an aniline derivative which oxidatively couples to 4-amino-antipyrine in the presence of H 2 O 2 and 3-methyl-2-benzo-thiazolinone hydrazone (MBTH) or 3-methyl-2-benzo(2'-sulfo)-thiazolinone (MBTH-S).
Claim 19
Dependent Claim
16 words
Reagent according to claim 16, wherein the aniline derivative is N,N-dimethylaniline, N,Ndimethylaminobenzoic acid, N-ethyl-N-β-sulphoethyl-m-toluidine or N-ethyl-N-hydroxyethyl-m-toluidine.
Claim 20
Dependent Claim
17 words
Reagent according to claim 11, wherein the hydrogen peroxide-producing oxidase contains sarcosine oxidase, uricase or oxalate oxidase.
Claim 21
Dependent Claim
17 words
Reagent according to claim 12, wherein said reagent contains 10 to 150 ug. superoxide dismutase per ml.
Claim 22
Dependent Claim
19 words
Reagent according to claim 11 wherein said reagent is present on a solid, inert single or multi-layer carrier material.
References Cited (Backward Citations)
US4454094A
Cited
JPS5966899A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1995-03-10 | 1995-09-09 | 1996-03-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1999-03-10 | 1999-09-09 | 2000-03-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2003-03-10 | 2003-09-09 | 2004-03-10 | $7700.00 | Window Closed / Lapsed |