Method for assaying fatty acids (4349625)

Inventors:
Tsutomu Okada, Tadashi Hishida, Minoru Muramatsu
Company:
Mitsubishi Chemical Industries Limited
Issued Date:
1982-09-14
Tags:
fatty acid, carbon atom

Abstract

There is disclosed a method for assaying fatty acids in a system containing albumin together with the fatty acids using acyl-CoA synthetase, characterized by performing the assay in the presence of a water-soluble salt of a dibasic fatty acid having 10 to 18 carbon atoms or a benzenesulfonate optionally having one or more C.sub.1 -C.sub.5 side chains. [USPTO]

Claims

What is claimed as new and intended to be covered by Letters Patent is:

1. In a method for assaying fatty acids in an aqueous system containing albumin free fatty acid in a sample by reacting, the free fatty acid with adenosine triphosphate in the presence of coenzyme A catalyzed by acyl-CoA synthetase thereby forming AMP and acyl-CoA and measuring the acyl-CoA or AMP formed, the improvement comprising: adding from 0.01 to 5 mole percent, based on said fatty acid, of a water soluble salt of a dibasic fatty acid having 10 to 18 carbon atoms or a salt selected from the group consisting of the sodium, potassium and ammonium salts of benzene sulfonic acid, p-toluene-sulfonic acid, p-ethylbenzenesulfonic acid, p-n-propylbenzenesulfonic acid, p-n-amylbenzenesulfonic acid, 2,4-dimethylbenzenesulfonic acid or 2,5-dimethylbenzenesulfonic acid to the aqueous assay medium.

2. The method of claim 1, wherein said measurement is by optical measuring means.

3. The method of claim 2, wherein said optical measuring means utilizes visible light.

4. The method of claim 2 wherein said optical measuring means utilizes ultraviolet light.

5. The method of claim 3, wherein said salt is that of a dibasic fatty acid having 10 to 18 carbon atoms.

6. The method of claim 5, wherein the concentration of said salt in said assay solution ranges from 1.times.10.sup.-4 to 1.times.10.sup.-1 mole/l.

7. The method of claim 5, wherein said salt is selected from the group consisting of the sodium, potassium and ammonium salts of sebacic acid, 1,9-nonamethylenedicarboxylic acid, 1,10-decamethylenedicarboxylic acid, 1,11-undecamethylenedicarboxylic acid, 1,12-dodecamethylenedicarboxylic acid, 1,13-tridecamethylenedicarboxylic acid, 1,14-tetradecamethylenedicarboxylic acid, 1,15-pentadecamethylenedicarboxylic acid and 1,16-hexamethylenedicarboxylic acid.

8. In a method for assaying fatty acids in an aqueous system containing albumin by reacting free fatty acid in a sample with adenosine triphosphate in the presence of coenzyme A catalyzed by acyl-CoA synthetase thereby forming adenosine monophosphate and acyl-CoA in the presence of acyl-CoA oxidase as a catalyst thereby producing hydrogen peroxide as a product which in turn oxidizes a mixture of 4-aminoantipyrine and 3-acetamino-N,N-diethylaniline to quinone dye, and determining the amount of free fatty acid in said sample by measuring the amount of quinoid dye produced, the improvement comprising:

adding from 0.01 to 5 mole %, based on said fatty acid, of a water soluble salt of a dibasic fatty acid having 10 to 18 carbon atoms or a salt selected from the group consisting of the sodium, potassium and ammonium salts of benzenesulfonic acid, p-toluenesulfonic acid, p-ethylbenzenesulfonic acid, p-n-propylbenzenesulfonic acid, p-n-amylbenzenesulfonic acid, 2,4-dimethylbenzenesulfonic acid and 2,5-dimethylbenzenesulfonic acid to the aqueous assay medium.

9. In a method for assaying fatty acids in an aqueous system containing albumin by reacting free fatty acid in a sample with adenosine triphosphate in the presence of coenzyme A catalyzed by acyl-CoA synthetase thereby forming adenosine monophosphate and acyl-CoA, forming pyruvic acid from a series of reactions in a quantity dependent upon the amount of said adenosine monophosphate produced and determining the amount of free fatty acid in said sample by measuring the amount of NADH consumed in the reaction with pyruvic acid, the improvement comprising:

adding from 0.01 to 5 mole %, based on said fatty acid, of a water soluble salt of a dibasic fatty acid having 10 to 18 carbon atoms or a salt selected from the group consisting of the sodium, potassium and ammonium salts of benzenesulfonic acid, p-toluenesulfonic acid, p-ethylbenzenesulfonic acid, p-n-propylbenzenesulfonic acid, p-n-amylbenzenesulfonic acid, 2,4-dimethylbenzenesulfonic acid and 2,5-dimethylbenzenesulfonic acid to the aqueous assay medium.

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