title: Comparative evaluation of extraction methods for simultaneous mass-spectrometric analysis of complex lipids and primary metabolites from human blood plasma.
abstract: Metabolomic results on human blood plasma largely depend on the sample preparation protocols employed for protein precipitation and metabolite extraction. Five different extraction methods were examined, which can be grouped into two categories, liquid-liquid extraction and protein precipitation methods, including long-standing protocols such as the Folch extraction and Bligh-Dyer extraction in comparison to modern methods such as the Matyash protocol and two global metabolite extraction methods. Extracts were subjected to analysis of blood plasma lipids and primary metabolites by using chip-based direct infusion nanoelectrospray tandem mass spectrometry and gas chromatography coupled to time-of-flight mass spectrometry, respectively. Optimal extraction schemes were evaluated based on the number of identified metabolites, extraction efficiency, compound diversity, reproducibility, and convenience for high-throughput sample preparations. Results showed that Folch and Matyash methods were equally valid and robust for lipidomic assessments while primary metabolites were better assessed by the protein precipitation methods with organic solvent mixtures.
authors.affiliation: The Department of Bio and Fermentation Convergence Technology, Kookmin University, Seoul, 136-702, South Korea.
authors.forename: Do Yup
authors.initials: DY
authors.lastname: Lee
authors.forename: Tobias
authors.initials: T
authors.lastname: Kind
authors.forename: Young-Ran
authors.initials: YR
authors.lastname: Yoon
authors.forename: Oliver
authors.initials: O
authors.lastname: Fiehn
authors.forename: Kwang-Hyeon
authors.initials: KH
authors.lastname: Liu
mesh_terms: D000328:Adult; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D008055:Lipids / Q000097:blood*; D059625:Liquid-Liquid Extraction; D008297:Male; D055432:Metabolomics; D008875:Middle Aged; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D008055:Lipids