Method and reagent for the determination of glutamate-oxalacetate transaminase and glutamate-pyruvate transaminase

US4271265A · kind A · utility

Assignee

• Boehringer Mannheim GmbH · DE

Inventors

• Ulfert Deneke · Fürth, DE
• Peter Stahl · New York, US
• Walter Schneider · Schriesheim, DE

Key dates

• Filing date: Jul 20, 1979
• Grant date: Jun 2, 1981
• Expiry date: Jul 20, 1999

Classification

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

Abstract

In a method for the determination of glutamate-oxalacetate transaminase or glutamate-pyruvate transaminase by the reaction of oxalacetate or pyruvate with glutamate, with formation of .alpha.-ketoglutarate in a buffered solution, the .alpha.-ketoglutarate formed is reacted with .gamma.-aminobutyrate in the presence of .gamma.-aminobutyrate transaminase with formation of succinate semialdehyde, NADP is reduced with the latter in the presence of succinate semialdehyde dehydrogenase to give NADPH, and the latter is measured either directly or after conversion with a tetrazolium salt and an electron carrier such as diaphorase, phenantroline methosulphate or phenazine methosulphate to a formazane dye. A reagent suited for use with this method contains .gamma.-aminobutyrate transaminase, succinate semialdehyde dehydrogenase, .gamma.-aminobutyrate, glutamate and buffer, as well as either oxalacetate or pyruvate, and optionally a tetrazolium salt, an electron carrier and a surface-active agent.