Reversed phase LC/MS/MS method for targeted quantification of glycerophospholipid molecular species in plasma

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• 作者：Olaf Uhl ; Claudia Glaser ; Hans Demmelmair ; Berthold Koletzko  ; ;   ; ^{office.koletzko@med.uni-muenchen.de}
• 关键词：Glycerophospholipids ; RP-HPLC ; MS/MS ; Plasma ; Fatty acids ; Quantification
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2011
• 出版时间：15 November 2011
• 年：2011
• 卷：879
• 期：30
• 页码：3556-3564
• 全文大小：318 K

文摘

The relationship between lipid status and metabolism, infant development and health has widely been studied, but the importance of individual glycerophospholipid species for biological functions in infants has hardly been considered. We developed a method for quantitative analyses of plasma glycerophospholipids from small sample volume. Proteins were precipitated with methanol, which eliminated further sample preparation. The supernatant was analysed by reversed-phase HPLC using a gradient of water, methanol and isopropanol as mobile phase. Electrospray ionisation in negative mode in combination with tandem mass spectrometry enabled detection of specific fatty acids as fragments of glycerophospholipid species. With this combination of chromatography and mass spectrometry, PC, lyso-PC, PE and lyso-PE species and their relevant isobaric compounds were quantified. Method validation showed a linear working range between 0.05 ¦Ìmol/L and 10 ¦Ìmol/L in diluted plasma samples. The intra-assay coefficients of variation (n = 6) ranged from 1.1 % to 13.9 % . Results were comparable with data of the human metabolome database and gas chromatographic fatty acid analyses. All quantitatively important PE and PC species are covered. The method can be applied for investigating dietary effects on plasma GP composition from small plasma volumes.