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Comparison of SFC/MS and UHPLC/MS techniques in the lipidomic characterization of biological samples
Authors: Holčapek Michal | Lísa Miroslav | Cífková Eva
Year: 2014
Type of publication: ostatní do riv
Page from-to: nestránkováno
Titles:
Language Name Abstract Keywords
cze Comparison of SFC/MS and UHPLC/MS techniques in the lipidomic characterization of biological samples Nowadays, supercritical fluid chromatography (SFC) has a renaissance in analytical chemistry due to the commercial availability of reliable SFC systems with potential applications for fast separations of compounds with low to medium polarity, such as lipids. SFC / mass spectrometry (SFC/MS) coupling brings some advantages over conventional (U)HPLC/MS systems, especially in terms of reduced analysis time and therefore higher throughput in the lipidomic analysis. The basic approach for the lipidomic quantitation is based on the direct infusion (shotgun) without any chromatography, but this technique could suffer from suppression effects and the lack of structural details, such as the differentiation of isobaric species. (U)HPLC/MS can be used either for lipid class or lipid species separation. Lipid class separation is achieved by hydrophilic-liquid chromatography (HILIC) for polar lipids or normal-phase (NP) HPLC for nonpolar lipids. Reversed-phase (RP) HPLC enables the resolution of lipid species differing in alkyl chain lengths and the number and positions of double bonds. The combination of HILIC and RP is two-dimensional separation in offline or online modes provides the most detailed characterization due to a good orthogonality of both separation modes. SFC can be successfully applied for the class separation of both polar and nonpolar lipid classes in one run. At least 16 lipid classes together with 2 internal standards can be separated and quantified within 6 minutes. The coupling with high-resolution Q-TOF mass analyzer equipped with ion mobility separation brings another dimension of separation and structural confirmation for complex lipidomic studies, which is applied for the analysis of tumor samples of patients with selected types of cancer. This work was supported by ERC CZ project No. LL1302 (MSMT, Czech Republic).
eng Comparison of SFC/MS and UHPLC/MS techniques in the lipidomic characterization of biological samples Nowadays, supercritical fluid chromatography (SFC) has a renaissance in analytical chemistry due to the commercial availability of reliable SFC systems with potential applications for fast separations of compounds with low to medium polarity, such as lipids. SFC / mass spectrometry (SFC/MS) coupling brings some advantages over conventional (U)HPLC/MS systems, especially in terms of reduced analysis time and therefore higher throughput in the lipidomic analysis. The basic approach for the lipidomic quantitation is based on the direct infusion (shotgun) without any chromatography, but this technique could suffer from suppression effects and the lack of structural details, such as the differentiation of isobaric species. (U)HPLC/MS can be used either for lipid class or lipid species separation. Lipid class separation is achieved by hydrophilic-liquid chromatography (HILIC) for polar lipids or normal-phase (NP) HPLC for nonpolar lipids. Reversed-phase (RP) HPLC enables the resolution of lipid species differing in alkyl chain lengths and the number and positions of double bonds. The combination of HILIC and RP is two-dimensional separation in offline or online modes provides the most detailed characterization due to a good orthogonality of both separation modes. SFC can be successfully applied for the class separation of both polar and nonpolar lipid classes in one run. At least 16 lipid classes together with 2 internal standards can be separated and quantified within 6 minutes. The coupling with high-resolution Q-TOF mass analyzer equipped with ion mobility separation brings another dimension of separation and structural confirmation for complex lipidomic studies, which is applied for the analysis of tumor samples of patients with selected types of cancer. This work was supported by ERC CZ project No. LL1302 (MSMT, Czech Republic). SFC/MS; UHPLC/MS; lipidomic characterization

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