Department of Analytical Chemistry, University of Pardubice, Faculty of Chemical Technology, Pardubice, Czech Republic.
Lipidica, a.s, Pardubice, Czech Republic.
Methods Mol Biol. 2025;2855:305-314. doi: 10.1007/978-1-0716-4116-3_18.
Ultrahigh-performance supercritical fluid chromatography-mass spectrometry (UHPSFC/MS) method is optimized for the high-throughput quantitation of lipids in human serum and plasma with an emphasis on robustness and accurate quantitation. Bridged ethylene hybrid (BEH) silica column (100 × 3 mm; 1.7 μm) is used for the separation of 17 nonpolar and polar lipid classes in 4.4 min using the positive ion electrospray ionization mode. The lipid class separation approach in UHPSFC/MS results in the coelution of all lipid species within one lipid class in one chromatographic peak, including two exogenous internal standards (IS) per lipid class, which provides the optimal conditions for robust quantitation. The method was validated according to European Medicines Agency and Food and Drug Administration recommendations. UHPSFC/MS combined with LipidQuant software allows a semiautomated process to determine lipid concentrations with a total run time of only 8 min including column equilibration, which enables the analysis of 160 samples per day.
超高效超临界流体色谱-质谱法（UHPSFC/MS）经过优化，可高通量定量分析人血清和血浆中的脂质，重点是确保稳健性和准确定量。采用桥联乙烯杂化（BEH）硅胶柱（100×3mm，1.7μm），在正离子电喷雾电离模式下，4.4 分钟内可分离 17 种非极性和极性脂质类。UHPSFC/MS 中的脂质类分离方法可使同一脂质类中的所有脂质物质在一个色谱峰中同时洗脱，包括每个脂质类中的两种外源性内标（IS），这为稳健定量提供了最佳条件。该方法按照欧洲药品管理局和美国食品和药物管理局的建议进行了验证。UHPSFC/MS 结合 LipidQuant 软件可实现半自动处理，确定脂质浓度的总运行时间仅为 8 分钟，包括柱平衡，每天可分析 160 个样本。
