High-performance liquid chromatographic analysis of serum short-chain fatty acids by direct derivatization

• Published in: Journal of chromatography Vol. 421; no. 1; p. 33
• Main Authors: Miwa, H, Yamamoto, M
• Format: Journal Article
• Language: English
• Published: Netherlands 1987
• Subjects: Chromatography, High Pressure Liquid; Fatty Acids - blood; Humans; Indicators and Reagents; Spectrophotometry, Ultraviolet
• DOI: 10.1016/0378-4347(87)80376-6

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.