High-performance liquid chromatographic analysis of serum short-chain fatty acids by direct derivatization.
The application of direct derivatization in conjunction with high-performance liquid chromatography is described for the analysis of short-chain fatty acids in serum. The method is based on the reaction of these acids with acidic 2-nitrophenylhydrazine hydrochloride, without complicated isolation steps, which produces their non-volatile hydrazine derivatives. The hydrazides of fourteen saturated and unsaturated, straight and branched, short-chain fatty acids were separated from other acid hydrazides and interfering components by a simple solvent extraction, and were eluted isocratically on a reversed-phase C8 column within 24 min. UV detection demonstrated that the detection limits for the acids were 200-400 fmol per injection with linearity over the range from 400 fmol to 5 nmol per injection. Analytical recoveries ranged from 96.8% to 103.1% and coefficients of variation ranged from 0.9% to 3.8%. The present method is simple, accurate and adequate for the analysis of short-chain fatty acids in biological fluids and tissues of patients suffering from organic acidemias.
Miwa
H
H
Faculty of Pharmaceutical Sciences, Fukuoka University, Jonan-ku, Japan.
Yamamoto
M
M
D002851:Chromatography, High Pressure Liquid; D005227:Fatty Acids / Q000097:blood*; D006801:Humans; D007202:Indicators and Reagents; D013056:Spectrophotometry, Ultraviolet
D005227:Fatty Acids; D007202:Indicators and Reagents