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US4116774A
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Method for the determination of enzyme activities
Assignee: Sankyo Co Ltd
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Filing: 1977-05-20
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Granted: 1978-09-26
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Expires: 1997-05-20
Abstract
Abstract Improvement in determination of an enzyme activity in a substance having an enzymatic activity which comprises employing an aromatic primary amine derivative as a substrate and an iron complex treated with a peroxide as a color reagent.
Inventors
- 👤 Sadamasa Minato
- 👤 Atsushi Hattori
- 👤 Yoshihiko Baba
- 👤 Yuichiro Yabe
- 👤 Seigo Ueda
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 01:03
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/34
Claims Analysis
Total Claims: 8
Claim 1
Independent Claim
134 words
In a method for the colorimetric determination of enzyme activity in a substance wherein the substance is treated with a mixture of a substrate and color reagent and is then colorimetrically analyzed, the improvement which comprises: the substrate being an aromatic primary amine derivative selected from the group consisting of L-glutamic acid γ-p-dimethylaminoanilide, L-glutamic acid γ-p-diethylaminoanilide, L-glutamic acid γ-p-methoxyanilide, L-glutamic acid γ-p-hydroxyanilide, S-benzyl-L-cysteine p-dimethylaminoanilide, S-benzyl-L-cysteine p-hydroxyanilide, S-benzyl-L-cysteine p-methoxyanilide, L-leucine p-dimethylaminoanilide, L-leucine p-hydroxyanilide, L-leucine p-methoxyanilide and N-benzyl-L-tyrosine p-dimethylaminoanilide and said color reagent is an iron complex selected from the group consisting of sodium pentacyanoammineferroate and potassium pentacyanoammineferroate which is treated with a peroxide selected from the group consisting of sodium periodate, potassium periodate, potassium permanganate and hydrogen peroxide, and wherein said iron complex is further blended with an alkali metal hydrogen-carbonate and low molecular dextran.
Claim 2
Dependent Claim
134 words
In a method for the colorimetric determination of enzyme activity in a substance wherein the substance is treated with a mixture of a substrate and color reagent and is then colorimetrically analyzed, the improvement which comprises: the substrate being an aromatic primary amine derivative selected from the group consisting of L-glutamic acid γ-p-dimethylaminoanilide, L-glutamic acid γ-p-diethylaminoanilide, L-glutamic acid γ-p-methoxyanilide, L-glutamic acid γ-p-hydroxyanilide, S-benzyl-L-cysteine p-dimethylaminoanilide, S-benzyl-L-cysteine p-hydroxyanilide, S-benzyl-L-cysteine p-methoxyanilide, L-leucine p-dimethylaminoanilide, L-leucine p-hydroxyanilide, L-leucine p-methoxyanilide and N-benzyl-L-tyrosine p-dimethylaminoanilide and said color reagent is an iron complex selected from the group consisting of sodium pentacyanoammineferroate and potassium pentacyanoammineferroate which is treated with a peroxide selected from the group consisting of sodium periodate, potassium periodate, potassium permanganate and hydrogen peroxide, and wherein said iron complex is further blended with an alkali metal hydrogen-carbonate and low molecular dextran.
Claim 3
Dependent Claim
23 words
A method according to claim 1 wherein said iron complex is a freeze-dried preparation further blended with sodium hydrogencarbonate and low molecular dextran.
Claim 4
Dependent Claim
25 words
A method according to claim 1 wherein said substance is human serum and a weakly acidic buffer with a pH of 3.0-7.0 is further present.
Claim 5
Dependent Claim
72 words
A method according to claim 1 wherein said aromatic primary amine derivative is a member selected from the group consisting of L-glutamic acid γ-p-dimethylaminoanilide, L-glutamic acid γ-p-diethylaminoanilide, L-glutamic acid γ-p-methoxyanilide, L-glutamic acid γ-p-hydroxyanilide, S-benzyl-L-cysteine p-dimethylaminoanilide, S-benzyl-L-cysteine p-hydroxyanilide, S-benzyl-L-cysteine p-methoxyanilide, L-leucine p-dimethylaminoanilide, L-leucine p-hydroxyanilide, L-leucine p-methoxyanilide and N-benzyl-L-tyrosine p-dimethylaminoanilide, and said iron complex is a freeze-dried preparation of a blend of sodium pentacyanoammineferroate treated with hydrogen peroxide, sodium hydrogencarbonate and low molecular dextran.
Claim 6
Dependent Claim
58 words
A diagnostic composition kit for determining an enzyme activity in a body fluid which comprises (a) an aromatic primary amine derivative as a substrate, (b) a freeze-dried preparation of a blend of an iron complex treated with a peroxide, an alkali metal hydrogencarbonate and low molecular dextran, and (c) a weakly acidic buffer with a pH of 3.0-7.0.
Claim 7
Dependent Claim
58 words
A diagnostic composition kit for determining an enzyme activity in a body fluid which comprises (a) an aromatic primary amine derivative as a substrate, (b) a freeze-dried preparation of a blend of an iron complex treated with a peroxide, an alkali metal hydrogencarbonate and low molecular dextran, and (c) a weakly acidic buffer with a pH of 3.0-7.0.
Claim 8
Dependent Claim
60 words
A method according to claim 1 wherein said aromatic primary amine derivative is a member selected from the group consisting of L-glutamic acid γ-p-dimethylaminoanilide, L-glutamic acid γ-p-diethylaminoanilide, L-glutamic acid γ-p-methoxyanilide, L-glutamic acid γ-p-hydroxyanilide, S-benzyl-L-cysteine p-dimethylaminoanilide, S-benzyl-L-cysteine p-hydroxyanilide, S-benzyl-L-cysteine p-methoxyanilide, L-leucine p-dimethylaminoanilide, L-leucine p-hydroxyanilide, L-leucine p-methoxyanalide and N-benzyl-L-tyrosine p-dimethylaminoanilide, said iron complex is sodium pentacyanoammineferroate and said peroxide is hydrogen peroxide.
References Cited (Backward Citations)
US3769173A
Cited
US3892631A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1981-09-25 | 1982-03-27 | 1982-09-26 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1985-09-25 | 1986-03-27 | 1986-09-26 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1989-09-25 | 1990-03-27 | 1990-09-26 | $7700.00 | Window Closed / Lapsed |