Use of a sparingly soluable salt of a heteropoly acid for the determination of an analyte, a corresponding method of determination as a suitable agent therefor

• US 5,521,060 A
• Filed: 10/12/1994
• Issued: 05/28/1996
• Est. Priority Date: 12/02/1989
• Status: Expired due to Term

• Current Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Original Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Inventors: Unkrig, Volker, Hoenes, Joachim, Wielinger, Hans
• Primary Examiner(s): Robinson, Douglas W.
• Assistant Examiner(s): LEARY, LOUISE N
• Application Number:
  US08/321,512
• Time in Patent Office: 594 Days
• Field of Search: 435/4, 435/14, 435/18, 435/25, 436/113, 436/111, 436/112, 436/63
• US Class Current: 435/4
• CPC Class Codes: C12Q 1/00 Measuring or testing processes involving enzymes, nucleic acids or microorganisms measuring or testing apparatus with condition measuring or sensing means, e.g. colony counters, C12M1/34; Compositions therefor; Processes of preparing such compositions
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: C12Q 1/34 involving hydrolase
• CPC Class Codes: C12Q 1/48 involving transferase
• CPC Class Codes: G01N 33/48 Biological material, e.g. blood, urine G01N33/02, G01N33/26, G01N33/44, G01N33/46 take precedence; Haemocytometers counting blood corpuscules distributed over a surface by scanning the surface G06M11/02
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements
• CPC Class Codes: Y10T 436/173845 Amine and quaternary ammonium
• CPC Class Codes: Y10T 436/174614 Tertiary amine
• CPC Class Codes: Y10T 436/175383 Ammonia

Abstract

A sparingly soluble salt of a heteropoly acid is used for the determination of an analyte which is an aromatic amine rich in electrons or together with another substance leads to such an amine. The analyte is determined by means of the heteropoly blue color formed.

Claims

1. A method for determining an enzymatically-oxidizable substrate, which comprises

a) oxidizing the substrate in a sample in the presence of an oxido reductase enzyme specific for the substrate and an electron acceptor which accepts electrons and forms an organic amine which is a stronger reducing agent than aniline;

b) contacting the resultant organic amine with a sparingly soluble salt of a heteropoly acid capable of forming heteropoly blue with said organic amine; and

c) determining the presence of the substrate by detecting heteropoly blue formation.

2. The method according to claim 1, wherein the substrate is glucose, ethanol, cholesterol, lactate or NAD(P)H.

3. The method according to claim 1, wherein the electron acceptor is an aromatic nitroso compound, oxime or hydroxylamine.