Validation of lipidomic analysis of human plasma and serum by supercritical fluid chromatography–mass spectrometry and hydrophilic interaction liquid chromatography–mass spectrometry

• Published in: Analytical and bioanalytical chemistry Vol. 412; no. 10; pp. 2375 - 2388
• Main Authors: Wolrab, Denise, Chocholoušková, Michaela, Jirásko, Robert, Peterka, Ondřej, Holčapek, Michal
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.04.2020; Springer; Springer Nature B.V
• Subjects: Analytical Chemistry; Biochemistry; Biological materials; Biological properties; Biological samples; Blood plasma; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Chromatography; Comparative analysis; Current Progress in Lipidomics; Food Science; Hydrophilicity; Laboratories; Laboratory Medicine; Lipids; Liquid chromatography; Mass spectrometry; Mass spectroscopy; Monitoring/Environmental Analysis; Optimization; Quantitation; Research Paper; Scientific imaging; Selectivity; Separation; Spectroscopy; Spectrum analysis; Supercritical fluids; Germany; Hydrophilic interaction liquid chromatography; Validation; Plasma; Lipidomics; Serum; Matrix effect; Quantitation; Mass spectrometry; Supercritical fluid chromatography
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-020-02473-3

Ultrahigh-performance supercritical fluid chromatography–mass spectrometry (UHPSFC/MS) has a great potential for the high-throughput lipidomic quantitation of biological samples; therefore, the full optimization and method validation of UHPSFC/MS is compared here with ultrahigh-performance liquid chromatography–mass spectrometry (UHPLC/MS) in hydrophilic interaction liquid chromatography (HILIC) mode as the second powerful technique for the lipid class separation. First, the performance of six common extraction protocols is investigated, where the Folch procedure yields the best results with regard to recovery rate, matrix effect, and precision. Then, the full optimization and analytical validation for eight lipid classes using UHPSFC/MS and HILIC-UHPLC/MS methods are performed for the same sample set and applied for the lipidomic characterization of pooled samples of human plasma, human serum, and NIST SRM 1950 human plasma. The choice of appropriate internal standards (IS) for individual lipid classes has a key importance for reliable quantitative workflows illustrated by the selectivity while validation and the calculation of the quantitation error using multiple internal standards per lipid class. Validation results confirm the applicability of both methods, but UHPSFC/MS provides some distinct advantages, such as the successful separation of both non-polar and polar lipid classes unlike to HILIC-UHPLC/MS, shorter total run times (8 vs. 10.5 min), and slightly higher robustness. Various types of correlations between methods (UHPSFC/MS and HILIC-UHPLC/MS), biological material (plasma and serum), IS (laboratory and commercially mixtures), and literature data on the standard reference material show the intra- and inter-laboratory comparison in the quantitation of lipid species from eight lipid classes, the concentration differences in serum and plasma as well as the applicability of non-commercially available internal standard mixtures for lipid quantitation.