Quantitative determination of blood ammonia

• US 3,929,581 A
• Filed: 05/17/1974
• Issued: 12/30/1975
• Est. Priority Date: 06/07/1973
• Status: Expired due to Term

• Current Assignee: Friedrich Da Fonseca-Wollheim
• Original Assignee: Friedrich Da Fonseca-Wollheim
• Inventors: DA Fonseca-Wollheim, Friedrich
• Primary Examiner(s): Monacell, A. Louis
• Assistant Examiner(s): Fan, C. A.
• Application Number:
  US05/471,064
• Time in Patent Office: 592 Days
• Field of Search: 195/13.5R,99
• US Class Current: 435/26
• CPC Class Codes: C12Q 1/32 involving dehydrogenase
• CPC Class Codes: C12Q 1/58 involving urea or urease

Abstract

Blood ammonia is quantitatively determined by contacting a sample suspected of containing blood ammonia, with glutamate dehydrogenase, Alpha -ketoglutarate, and, as a coenzyme in the reduced state, nicotinamide-adenine-dinucleotide phosphate in reduced form.

Claims

1. PROCESS FOR THE QUANTITATIVE DETERMINATION OF BLOOD AMMONIA WHICH COMPRISES CONTACTING A SAMPLE SUSPECTED OF CONTAINING BLOOD AMMONIA WITH GLUTAMATE DEHYDROGENASE, A-KETOGLUTARATE, AND, AS A COENZYME IN THE REDUCED STATE, NICOTINAMIDEADENINE-DINUCLEOTIDE PHOSPHATE IN REDUCED FORM.

2. glutamate dehydrogenase;

and

3. 0.1 to 0.5 Mu Mol NADPH or a multiple thereof.

4. Process as claimed in claim 3, wherein the determination is carried out in a buffer of pH 8.0 to 9.0.

5. Process as claimed in claim 4, wherein the determination is carried out in a buffer of pH 8.5 to 8.7.

6. Process as claimed in claim 5, wherein the determination is carried out in a buffer of pH 8.6.

7. Process as claimed in claim 3, wherein the buffer used is one of tris buffer, triethanolamine buffer and phosphate buffer.

8. Reagent combination for carrying out the process as claimed in claim 1, comprising:

9. Reagent combination as claimed in claim 8, containing ADP dissolved in said buffer.

10. Reagent combination as claimed in claim 8, comprising:

11. Reagent combination according to claim 9, wherein the third component comprises:

0.1 to 0.5 Mu Mol NADPH 10 to 50 mg serum albumen 1 to 5 Mu Mol sodium bicarbonate 5 to 60 mg dextran or a multiple thereof.

12. Reagent combination as claimed in claim 10 also containing 2 to 30 mM alkali metal azide.