US3979262A
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US
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Compositions and methods for the determination of oxidizing agents
Assignee: Hoffmann La Roche Inc
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Filing: 1975-02-11
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Granted: 1976-09-07
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Expires: 1995-02-11
Abstract
Abstract Compositions and methods are described for the determination of specific components in a biological fluid by a quantitative colorimetric determination of an oxidizing agent, e.g., hydrogen peroxide, produced in the biological fluid by an appropriate enzyme, said compositions comprising an aromatic amine component, a hydrazone component and a citric and/or maleic acid component.
Inventors
- 👤 Paul Hunziker
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 02:39
CPC Classifications
G; G01; G01N; G01N31/00; G01N31/22; G01N31/228; C; C12; C12Q; C12Q1/00; C12Q1/26; C12Q1/28; C12Q1/54; C12Q1/60; C12Q1/62; C12Q2326/00; C12Q2326/32
Claims Analysis
Total Claims: 27
Claim 1
Independent Claim
144 words
A composition for the colorimetric determination of oxidizing agents comprising: a. as the first component, an amine selected from the group consisting of an aromatic amine and an acid addition salt thereof with the proviso that substituents on the aromatic amine are not simultaneously in the ortho and para positions; b. as the second component, a compound, capable of oxidatively coupling with the amine of the first component to form a colored entity, selected from the group consisting of a hydrazone and an acid addition salt thereof; and c. as the third component, a buffer selected from the group consisting of citric acid, maleic acid and mixtures of citric and maleic acid wherein the amount of the buffer component in the composition is such as to provide for a pre-selected pH range of from about 3.2 to about 5.5 in a resulting colored medium.
Claim 2
Dependent Claim
144 words
A composition for the colorimetric determination of oxidizing agents comprising: a. as the first component, an amine selected from the group consisting of an aromatic amine and an acid addition salt thereof with the proviso that substituents on the aromatic amine are not simultaneously in the ortho and para positions; b. as the second component, a compound, capable of oxidatively coupling with the amine of the first component to form a colored entity, selected from the group consisting of a hydrazone and an acid addition salt thereof; and c. as the third component, a buffer selected from the group consisting of citric acid, maleic acid and mixtures of citric and maleic acid wherein the amount of the buffer component in the composition is such as to provide for a pre-selected pH range of from about 3.2 to about 5.5 in a resulting colored medium.
Claim 3
Dependent Claim
29 words
A composition as in claim 1 wherein the buffer component is citric acid and the weight ratio range of buffer component to amine component to hydrazone component is 25-240/0.5-2/0.5-2.
Claim 4
Dependent Claim
28 words
A composition as in claim 1 wherein the buffer component is maleic acid and the weight ratio range of buffer component to amine component to hydrazone is 20-240/0.5-6/0.5-6.
Claim 5
Dependent Claim
20 words
A composition as in claim 1 wherein the aromatic amine component is N,N-dimethylaniline and the hydrazone component is N-methyl-2-benzothiazolinone-hydrazone hydrochloride.
Claim 6
Dependent Claim
13 words
A composition as in claim 4 wherein the buffer component is citric acid.
Claim 7
Dependent Claim
13 words
A composition as in claim 4 wherein the buffer component is maleic acid.
Claim 8
Dependent Claim
20 words
A composition as in claim 1 wherein the aromatic amine component is m-phenylenediamine and the hydrazone component is N-methyl-2-benzothiazolinone-hydrazone hydrochloride.
Claim 9
Dependent Claim
13 words
A composition as in claim 7 wherein the buffer component is citric acid.
Claim 10
Dependent Claim
13 words
A composition as in claim 7 wherein the buffer component is maleic acid.
Claim 11
Dependent Claim
37 words
The composition as in claim 1 wherein the aromatic amine component is 30 mg. of m-phenylenediammonium dichloride, the hydrazone component is 30 mg. of 3-methyl-2-benzothiazolinone-hydrazone hydrochloride and the buffer component is 3.6 g. of citric acid powder.
Claim 12
Dependent Claim
37 words
The composition as in claim 1 wherein the aromatic amine component is 30 mg. of m-phenylenediammonium dichloride, the hydrazone component is 30 mg. of 3-methyl-2-benzothiazonehydrazone hydrochloride and the buffer component is 1.55 g. of maleic acid powder.
Claim 13
Dependent Claim
37 words
The composition as in claim 1 wherein the aromatic amine component is 30 mg. of N,N-dimethylaniline hydrochloride, the hydrazone component is 30 mg. of 3-methyl-2-benzothiazolinone-hydrazone hydrochloride and the buffer component is 3.6 g. of citric acid powder.
Claim 14
Independent Claim
159 words
A method for quantitatively determining the glucose content of a biological fluid which comprises: a. mixing the biological fluid with a glucoseoxidase solution having a pH of from about 7 to about 8; b. incubating the mixture from step (a) at room temperature; c. adding to the product from step (b) a composition comprising as the first component, an amine selected from the group consisting of an aromatic amine and acid addition salt thereof with the proviso that substituents on the aromatic amine are not simultaneously in the ortho and para positions, as the second component, a hydrazone and as the third component, a citric acid buffer; d. adding to the mixture from step (c) a buffered peroxidase solution having a pH of from about 7 to about 8 so that the pH of the final mixture ranges from 4.7 to 5.5; and e. measuring the glucose content of the fluid by colorimetrically determining the quantity of hydrogen peroxide.
Claim 15
Independent Claim
159 words
A method for quantitatively determining the glucose content of a biological fluid which comprises: a. mixing the biological fluid with a glucoseoxidase solution having a pH of from about 7 to about 8; b. incubating the mixture from step (a) at room temperature; c. adding to the product from step (b) a composition comprising as the first component, an amine selected from the group consisting of an aromatic amine and acid addition salt thereof with the proviso that substituents on the aromatic amine are not simultaneously in the ortho and para positions, as the second component, a hydrazone and as the third component, a citric acid buffer; d. adding to the mixture from step (c) a buffered peroxidase solution having a pH of from about 7 to about 8 so that the pH of the final mixture ranges from 4.7 to 5.5; and e. measuring the glucose content of the fluid by colorimetrically determining the quantity of hydrogen peroxide.
Claim 16
Dependent Claim
28 words
The method in accordance with claim 13 wherein the composition comprises 30 mg. of m-phenylenediammonium dichloride, 30 mg. of 3-methyl-2-benzothiazolinonehydrazone hydrochloride and 1.44 g. of citric acid powder.
Claim 17
Independent Claim
169 words
A method for quantitatively determining the uric acid content of a biological fluid which comprises: a. mixing the biological fluid with a uricase solution having a pH of from about 9 to about 10; b. incubating the mixture from step (a) at a temperature of from about 35°C. to about 40°C. c. adding to the product from step (b), a composition comprising as the first component, an amine selected from the group consisting of an aromatic amine and acid addition salt thereof with the proviso that substituents on the aromatic amine are not simultaneously in the ortho and para positions, as the second component, a hydrazone and as the third component, a citric acid buffer; d. adding to the mixture from step (c) a buffered peroxidase solution having a pH of from about 7 to about 8 so that the pH of the final mixture ranges from 3.2 to 4.7; and e. measuring the uric acid content of the fluid by colorimetrically determining the quantity of hydrogen peroxide present.
Claim 18
Independent Claim
169 words
A method for quantitatively determining the uric acid content of a biological fluid which comprises: a. mixing the biological fluid with a uricase solution having a pH of from about 9 to about 10; b. incubating the mixture from step (a) at a temperature of from about 35°C. to about 40°C. c. adding to the product from step (b), a composition comprising as the first component, an amine selected from the group consisting of an aromatic amine and acid addition salt thereof with the proviso that substituents on the aromatic amine are not simultaneously in the ortho and para positions, as the second component, a hydrazone and as the third component, a citric acid buffer; d. adding to the mixture from step (c) a buffered peroxidase solution having a pH of from about 7 to about 8 so that the pH of the final mixture ranges from 3.2 to 4.7; and e. measuring the uric acid content of the fluid by colorimetrically determining the quantity of hydrogen peroxide present.
Claim 19
Dependent Claim
28 words
The method in accordance with claim 15, wherein the composition comprises 30 mg. of m-phenylenediammonium dichloride, 30 mg. of 3-methyl-2-benzothiazolinonehydrazone hydrochloride and 3.6 grams of citric acid powder.
Claim 20
Independent Claim
235 words
A method for quantitatively determining the cholesterol content of a biological fluid which comprises a. mixing the biological fluid with an aqueous suspension of cholesterolesterase at a pH of from about 7 to about 7.5, to form a control mixture. b. incubating the control mixture from step (a) at a temperature of from about 35°C. to about 40°C; c. treating an aliquot of the control mixture from step (b) with an aqueous suspension of cholesteroloxidase at a pH of from about 7 to about 7.5, to form a test mixture. d. incubating the test mixture of step (c) at a temperature of from about 35°C. to about 40°C.; e. adding to the test mixture of step (d) and the control mixture of step (b) a composition comprising as the first component, an amine selected from the group consisting of an aromatic amine and acid addition salt thereof with the proviso that substituents on the aromatic amine are not simultaneously in the ortho and para positions, as the second component, a hydrazone and as the third component, a citric acid buffer; f. adding to each of the mixtures from step (e) a peroxidase solution having a pH of from about 7 to about 7.5; g. maintaining the reaction for 10 minutes to allow the color to develop; and h. measuring the cholesterol content of the fluid by colorimetrically comparing the test mixture with the control mixture.
Claim 21
Independent Claim
235 words
A method for quantitatively determining the cholesterol content of a biological fluid which comprises a. mixing the biological fluid with an aqueous suspension of cholesterolesterase at a pH of from about 7 to about 7.5, to form a control mixture. b. incubating the control mixture from step (a) at a temperature of from about 35°C. to about 40°C; c. treating an aliquot of the control mixture from step (b) with an aqueous suspension of cholesteroloxidase at a pH of from about 7 to about 7.5, to form a test mixture. d. incubating the test mixture of step (c) at a temperature of from about 35°C. to about 40°C.; e. adding to the test mixture of step (d) and the control mixture of step (b) a composition comprising as the first component, an amine selected from the group consisting of an aromatic amine and acid addition salt thereof with the proviso that substituents on the aromatic amine are not simultaneously in the ortho and para positions, as the second component, a hydrazone and as the third component, a citric acid buffer; f. adding to each of the mixtures from step (e) a peroxidase solution having a pH of from about 7 to about 7.5; g. maintaining the reaction for 10 minutes to allow the color to develop; and h. measuring the cholesterol content of the fluid by colorimetrically comparing the test mixture with the control mixture.
Claim 22
Dependent Claim
28 words
The method in accordance with claim 15 wherein the composition comprises 30 mg. of m-phenylenediammonium dichloride, 30 mg. of 3-methyl-2-benzothiazolinonehydrazone hydrochloride and 1.44 mg. of citric acid powder.
Claim 23
Dependent Claim
128 words
A diagnostic test kit for the determination of hydrogen peroxide which comprises: a. a container having a composition comprising: i. as the first component, an amine selected from the group consisting of an aromatic amine and an acid addition salt thereof with the proviso that substituents in the aromatic amine are not simultaneously in the ortho and para positions; ii. as the second component, a compound, capable of oxidatively coupling with the amine of the first component to form a colored entity, selected from the group consisting of a hydrazone and an acid addition salt thereof; and iii. as the third component, a buffer selected from the group consisting of citric acid, maleic acid and mixtures of citric and maleic acid; and b. A container having buffer-peroxidase mixture.
Claim 24
Dependent Claim
128 words
A diagnostic test kit for the determination of hydrogen peroxide which comprises: a. a container having a composition comprising: i. as the first component, an amine selected from the group consisting of an aromatic amine and an acid addition salt thereof with the proviso that substituents in the aromatic amine are not simultaneously in the ortho and para positions; ii. as the second component, a compound, capable of oxidatively coupling with the amine of the first component to form a colored entity, selected from the group consisting of a hydrazone and an acid addition salt thereof; and iii. as the third component, a buffer selected from the group consisting of citric acid, maleic acid and mixtures of citric and maleic acid; and b. A container having buffer-peroxidase mixture.
Claim 25
Dependent Claim
29 words
A diagnostic test kit for the determination of uric acid in biological fluids which comprises, in addition to the containers of claim 19, a container having a buffer-uricase mixture.
Claim 26
Dependent Claim
27 words
A diagnostic test kit for the determination of glucose in biological fluids, which comprises in addition to the containers of claim 19, a container having buffer-glucoseoxidase mixture.
Claim 27
Dependent Claim
37 words
A diagnostic test kit for the determination of cholesterol in biological fluids which comprises, in addition to the containers of claim 19 a. a container having a buffer-cholesterolesterase mixture; and b. a container having a buffer-cholesteroloxidase mixture.
References Cited (Backward Citations)
US2970945A
Cited
US3862885A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1979-09-07 | 1980-03-08 | 1980-09-07 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1983-09-07 | 1984-03-08 | 1984-09-07 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1987-09-07 | 1988-03-08 | 1988-09-07 | $7700.00 | Window Closed / Lapsed |