A new colorimetric method for the determination of free fatty acids with acyl-CoA synthetase and acyl-CoA oxidase.
A rapid and sensitive spectrophotometric assay for free fatty acids using acyl-CoA synthetase and acyl-CoA oxidase is described. It is sensitive to as low as 5 nmol of free fatty acids, and the standard curve is linear up to 100 nmol. The assay consists of the measurement of H2O2 produced from free fatty acids by acyl-CoA synthetase and acyl-CoA oxidase. The quantity of H2O2 is determined by the absorbance at 550 nm in the presence of catalase and 4-amino-3-hydrazino-5-mercapto-1,2,4-triazole (AHMT). This method shows a broad specificity to long-chain fatty acids and the recoveries of added fatty acids (C12-C18) are more than 90%. The presence or absence of serum components or Escherichia coli cell-free extracts has no significant effect on the recovery of added palmitic acid.
D042962:Acyl-CoA Oxidase; D003066:Coenzyme A Ligases; D003124:Colorimetry; D005230:Fatty Acids, Nonesterified / Q000032:analysis* / Q000097:blood; D005260:Female; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis; D008297:Male; D010088:Oxidoreductases; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D000098859:Long-Chain-Fatty-Acid-CoA Ligase
D005230:Fatty Acids, Nonesterified; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D006861:Hydrogen Peroxide; D010088:Oxidoreductases; D042962:Acyl-CoA Oxidase; D003066:Coenzyme A Ligases; C090799:FAA2 protein, S cerevisiae; D000098859:Long-Chain-Fatty-Acid-CoA Ligase