Determination of free fatty acids in plasma by gas chromatography

Ken'ichi Ichihara; Chihiro Kohsaka; Naohiro Tomari; Yoshihiro Yamamoto; Takehiro Masumura

doi:10.1016/j.ab.2020.113810

Determination of free fatty acids in plasma by gas chromatography

Anal Biochem. 2020 Aug 15:603:113810. doi: 10.1016/j.ab.2020.113810. Epub 2020 Jun 6.

Authors

Ken'ichi Ichihara¹, Chihiro Kohsaka², Naohiro Tomari², Yoshihiro Yamamoto², Takehiro Masumura³

Affiliations

¹ Kyoto Integrated Science & Technology Bio-Analysis Center, Kyoto Municipal Institute of Industrial Technology and Culture, 134 Cyudoji-Minamicho, Kyoto, 600-8813, Japan. Electronic address: ichihara@kpu.ac.jp.
² Kyoto Integrated Science & Technology Bio-Analysis Center, Kyoto Municipal Institute of Industrial Technology and Culture, 134 Cyudoji-Minamicho, Kyoto, 600-8813, Japan.
³ Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Shimogamo, Kyoto, 606-8522, Japan.

PMID: 32511966
DOI: 10.1016/j.ab.2020.113810

Abstract

A method was developed for determination of free fatty acids (FFAs) in plasma by gas chromatography. Plasma was extracted with 3 vol of methanol. Most cholesterol esters and triacylglycerols did not dissolve in the aqueous methanol. FFAs in the crude lipid solution were directly and selectively methylated with (trimethylsilyl)diazomethane at room temperature. Fatty acid methyl esters (FAMEs) formed were extracted with hexane, and nonreactive phospholipids were washed out with 95% methanol. The partially purified FAME preparation was analyzed by gas chromatography. The composition and amount of plasma FFAs closely approximated those obtained using two different methods.

Keywords: Blood lipids; Free fatty acid; Gas chromatography; Nonesterified fatty acid.

MeSH terms

Chromatography, Gas / methods*
Esters / chemistry
Fatty Acids, Nonesterified / blood*
Hexanes
Methanol / chemistry
Methylation
Temperature

Substances

Esters
Fatty Acids, Nonesterified
Hexanes
Methanol