Comparison of simple monophasic versus classical biphasic extraction protocols for comprehensive UHPLC-MS/MS lipidomic analysis of Hela cells

摘要: Abstract In this study, two monophasic isopropanol-water mixtures (IPA:H2O 75:25 v/v and IPA:H2O 90:10 v/v) were compared with traditionally employed biphasic methods of Bligh & Dyer Matyash et al. as extraction systems for lipidomics analysis in Hela cells. Samples analyzed by UHPLC-ESI-QTOF-MS/MS positive negative mode using sequential window acquisition all theoretical fragment ion spectra (SWATH) a relatively new software (MS-DIAL) was the processing data which includes detection peaks, MS/MS deconvolution, identification detected lipids alignment peaks through samples. The studied performance parameters such precision, recoveries isotopically labeled internal standards endogenous lipids, number extracted complexity procedure showed that performs similar to protocol better than well v/v. However, less complex is simpler implement can be executed plastic rather glass, make an excellent alternative classical protocols reversed phase LC-MS studies.