EP0123902A2
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EP
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Procedure for the determination of peroxidase activity by end dilution titration and means for its realization
Assignee: BYK-Sangtec Diagnostica GmbH and Co KG
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Filing: 1984-03-27
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Granted: —
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Expires: 2004-03-27
Abstract
Abstract An improved determination of peroxidase activity by end dilution titration is carried out with a benzidine derivative of the general formula I where R1 stands for a hydrogen atom, a halogen atom, an amino group, a (C 1 -C 4 ) alkyl group, a (C 1 -C 4 ) alkoxy group, a carboxy group, a sulfo group or a carboxypropyloxy group and where R2 stands for a hydrogen atom, a halogen atom, a (C 1 -C 4 ) alkyl group, or a salt of this benzidine compound, in the presence of a non-chromogenic or low-chromogenic phenylene derivative of the general formula II where R3 and R4 are equal or different and stand for a hydroxy group or an amino group, a (C 1 -C 4 ) alkyl amino group or a benzylamino group.
Inventors
- 👤 Dierck Dr. Lentfer
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-30 08:23
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/26; C12Q1/28; C12Q2326/00; C12Q2326/10; C12Q2326/12
Claims Analysis
Total Claims: 9
Claim 1
Independent Claim
192 words
Claim 1. In a procedure for the determination of peroxidase activity by end dilution titration under peroxidase end point dilution titration determing conditions wherein a peroxidase is reacted with a sufficient amount of a benzidine derivative, the improvement comprising conducting said procedure utilizing as the benzidine derivative a benzidine derivative of the aeneral formula (I) where Rl stands for a hydrogen atom, a halogen atom, an amino group, a (C l -C 4 ) alkyl group, a (C l -C 4 ) alkoxy group, a carboxy group, a sulfo group or a carboxypropyloxy group and where R2 stands for a hydrogen atom, a halogen atom, a (C 1 -C 4 ) alkyl group, or a salt of this benzidine derivative, in the presence of a color retarding amount of a non-chromogenic or low-chromogenic phenylene derivative of the general formula (II) where R3 and R4 are equal or different and stand for a hydroxy group,an amino group, a (C l -C 4 ) alkyl amino group or a benzyl amino group, to retard color development for a sufficient period of time to permit a sharp dilution end point, and thereafter determining peroxidase activity.
Claim 2
Dependent Claim
28 words
Claim 2. Procedure according to Claim 1, where Rl of the benzidine derivative I, stands for hydrogen, methyl, or methoxy and where R2 stands for hydrogen or methyl.
Claim 3
Dependent Claim
27 words
Claim 3. Procedure according to Claim 1, where Rl and R2 of the benzidine derivative I, stand for methyl in ortho position to the respective amino groups.
Claim 4
Dependent Claim
18 words
Claim 4. Procedure according to Claim 1, where the phenylene derivative II is hydroquinone, o-phenylene diamine, or 4-aminophenol.
Claim 5
Dependent Claim
14 words
Claim 5. Procedure according to Claim I, where the phenylene derivative II is hydroquinone.
Claim 6
Dependent Claim
20 words
Claim 6. Procedure according to Claim 1, where the color is retarded for a period of 5 to 60 minutes.
Claim 7
Dependent Claim
20 words
Claim 7. Procedure according to Claim 6, where the color is retarded for a period of 15 to 45 minutes.
Claim 8
Dependent Claim
163 words
Claim 8. A reagent for determining peroxidase activity by end dilution titration comprising a benzidine derivative of the general formula (I) where Rl stands for a hydrogen atom, a halogen atom, an amino group, a (C 1 -C 4 ) alkyl group, a (C 1 -C 4 ) alkoxy 0 group, a carboxy group, a sulfo group or a carboxypropyloxy group and where R2 stands for a hydrogen atom, a halogen atom, a (C l -C 4 ) alkyl group, or a salt of this benzidine derivative in a sufficient amount to permit the determination of peroxidase activity by color formantion and a phenylene derivative of the general formula (II) where R3 and R4 are equal or different and stand for a hydroxy group an amino group, a (C l -C 4 ) alkyl amino group or a benzyl amino group, in a color retarding amount to prevent color formation for a sufficient period of time to permit a sharp end point determination.
Claim 9
Dependent Claim
165 words
Claim 9. A kit for determining peroxidase activity by end dilution titration comprising (a) a benzidine derivative of the general formula (I) where Rl stands for a hydrogen atom, a halogen atom, an amino group, a (C 1 -C 4 ) alkyl group, a (C l -C 4 ) alkoxy group, a carboxy group, a sulfo group or a carboxypropyloxy group, where R2 stands for a hydrogen atom, a halogen atom, a (C 1 -C 4 ) alkyl group, or a salt of this benzidine derivative, (b) a phenylene derivative of the general formula (II) where R3 and R4 are equal or different and stand for a hydroxy group an amino group, a (C l -C 4 ) alkyl amino group or a benzyl amino group, (c) hydrogen peroxide or an adduct thereof, (d) a conjugate of horse radish peroxidase and an antigen or antibody, and (e) a solid support coated with an antigen or antibody which will react with the analyte to be determined.
References Cited (Backward Citations)
No citation records indexed.
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