title: Improved methods for the fatty acid analysis of blood lipid classes.
abstract: Two improved methods have been developed for preparation of fatty acid methyl esters (FAME) from major O-ester lipid classes in blood, i.e., cholesterol ester, triacylglycerol, and glycerophospholipids. The methods involve simple operations, and use neither harmful solvents such as chloroform or benzene nor highly reactive volatile reagents such as acetyl chloride. The FAME synthesis reaction proceeds under mild temperature conditions. The methods include (1) extraction of lipids from 0.2 ml of blood with 0.2 ml of tert-butyl methyl ether and 0.1 ml of methanol, (2) separation of the total lipids into lipid classes using a solid-phase extraction column or thin-layer chromatography, and (3) methanolysis of each lipid class at room temperature or at 45 °C. In all the operations, solvent concentration is performed only once prior to gas-liquid chromatography (GC). No noticeable differences in composition determined by GC have been found between FAME prepared by the present methods and those prepared by a conventional method involving lipid extraction with chloroform/methanol. The mild reaction and simplified procedures of the present methods enabled safe and reproducible analysis of the fatty acid compositions of the major ester-lipid classes in blood.
authors.affiliation: Department of Biomolecular Chemistry, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Shimogamo, Kyoto, 606-8522, Japan. ichihara@kpu.ac.jp
authors.forename: Ken'ichi
authors.initials: K
authors.lastname: Ichihara
authors.forename: Kumiko
authors.initials: K
authors.lastname: Yoneda
authors.forename: Ayuko
authors.initials: A
authors.lastname: Takahashi
authors.forename: Noriko
authors.initials: N
authors.lastname: Hoshino
authors.forename: Motoyoshi
authors.initials: M
authors.lastname: Matsuda
mesh_terms: D000465:Algorithms; D001774:Blood Chemical Analysis / Q000379:methods* / Q000592:standards; D005591:Chemical Fractionation / Q000379:methods; D002849:Chromatography, Gas; D002855:Chromatography, Thin Layer; D005227:Fatty Acids / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000145:classification; D006801:Humans; D008055:Lipids / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000145:classification*; D000432:Methanol / Q000737:chemistry / Q000494:pharmacology; D008738:Methyl Ethers / Q000737:chemistry / Q000494:pharmacology; D052616:Solid Phase Extraction / Q000379:methods; D012997:Solvents / Q000737:chemistry / Q000494:pharmacology
chemical_list: D005227:Fatty Acids; D008055:Lipids; D008738:Methyl Ethers; D012997:Solvents; C043243:methyl tert-butyl ether; D000432:Methanol