An improved assay with 4-(2-thiazolylazo)-resorcinol for non-esterified fatty acids in biological fluids.
A simple and rapid colorimetric assay for non-esterified fatty acids (NEFA) was developed employing extraction of samples with 0.1 mol/l glycine (pH 2.7) and methoxyethanol : butyl ether, and formation of a copper-fatty acid soap detected with 4-(2-thiazolylazo)-resorcinol. The method developed is sensitive enough to detect as little as 11 nmol/0.25 ml sample. Results are not affected by marked molar excesses of phospholipids, cholesterol, cholesterol esters, triglycerides, bilirubin, or lactate. Recovery of palmitate in the extract averages 94.2 +/- 3.2% (S.D.) and other long chain fatty acids are recovered equally. The copper-NEFA-indicator complex formed is stable. Extracted samples can be stored for as long as 2 weeks without appreciable changes in measured NEFA content. The method developed is suitable for both manual and semi-automated procedures for determining NEFA content in biological fluids.
D000818:Animals; D001391:Azo Compounds; D003124:Colorimetry / Q000379:methods; D003300:Copper; D005230:Fatty Acids, Nonesterified / Q000032:analysis*; D012118:Resorcinols; D012756:Sheep
D001391:Azo Compounds; D005230:Fatty Acids, Nonesterified; D012118:Resorcinols; C026054:4-(2-thiazolylazo)resorcinol; D003300:Copper