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Published on: June 1, 2017
Comprehensive Quantitative Profiling of Less Polar Lipids in Human Plasma Using Validated Reversed-Phase UHPSFC/MS/MS
Zuzana Lásko1, Veronika Šubrtová1, Ondřej Peterka1
1Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 53210 Pardubice, Czech Republic.
This study introduces a new method for analyzing less polar lipids in human plasma using reversed-phase ultrahigh-performance supercritical fluid chromatography-tandem mass spectrometry (RP-UHPSFC/MS/MS). The validated technique allows for high-throughput lipid profiling and accurate quantification of various lipid species.
Area of Science:
- Lipidomics
- Analytical Chemistry
- Biochemistry
Background:
- Lipid analysis is complex due to structural diversity and isomers.
- High-throughput methods are needed for lipidomic studies.
Purpose of the Study:
- To develop and validate a robust method for high-throughput profiling of less polar lipids in human plasma.
- To enable sensitive detection and accurate quantification of multiple lipid classes and species.
Main Methods:
- Reversed-phase ultrahigh-performance supercritical fluid chromatography-tandem mass spectrometry (RP-UHPSFC/MS/MS).
- Folch extraction, benzoyl chloride derivatization, and liquid-liquid extraction.
- Dual C18 column system for enhanced chromatographic resolution.
Main Results:
- Sensitive profiling of six major lipid classes (TG, DG, MG, SE, ST, FA) in under 18 minutes.
- Separation of isomeric species, including cis/trans and positional isomers.
- Accurate quantification of 147 lipid species in human plasma, with validated response factors for sterol esters.
Conclusions:
- The RP-UHPSFC/MS/MS method is robust, validated, and suitable for large-scale lipidomic studies.
- The method offers high sensitivity and resolution for analyzing less polar lipids.
- Accurate quantification necessitates the use of response factors, particularly for sterol esters.
