Process and analytical agent for the determination of the activity of glutamate-oxalacetate-transaminase

US4591553A · kind A · utility

Assignee

• Boehringer Mannheim GmbH · DE

Inventors

• Ulfert Deneke · Fürth, DE
• Wolfgang Gruber · Bischofswerda, DE
• Hans-Georg Batz · Tutzing, DE

Key dates

• Filing date: Jun 1, 1983
• Grant date: May 27, 1986
• Expiry date: Jun 1, 2003

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y10S435/805
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention provides a process for the determination of the activity of glutamateoxalacetate-transaminase (GOT) in aqueous solutions in which PA0 (a) .alpha.-ketoglutarate is reacted with alanine-sulphinic acid, catalyzed by GOT, to give pyruvate, sulphur dioxide and glutamate, PA0 (b) the resultant pyruvate is reacted with phosphate and oxygen, catalyzed by pyruvate oxidase, to give acetyl phosphate, carbon dioxide and hydrogen peroxide, PA0 (c) the hydrogen peroxide and a chromogen is reacted, catalyzed by a peroxidase, to give a dyestuff and water and PA0 (d) the amount of dyestuff formed is determined, wherein the chromogen used is N-(3'-sulphonyl-4'-hydroxy-5'-chlorophenyl)-4-aminoantipyrine (SHYC-AA), 4,5-bis-[4-dimethylaminophenyl]-2-[3,5-dimethoxy-4-hydroxypheny l]-imidazole (B-DDH-imidazole) or a related substance. The present invention also provides an analytical agent for the determination of the activity of GOT, comprising .alpha.-ketoglutarate, alanine-sulphinic acid, a chromogen, peroxidase, pyruvate oxidase, soluble phosphate, buffer and water, wherein the chromogen is N-(3'-sulphonyl-4'-hydroxy-5'-chlorophenyl)-4-aminoantipyrine (SHYC-AA) or 4,5-bis-[4-dim…