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Quantitative and Qualitative Method for Sphingomyelin by LC-MS Using Two Stable Isotopically Labeled Sphingomyelin Species
Published on: May 7, 2018
LC-MS-based method for the qualitative and quantitative analysis of complex lipid mixtures
Ulf Sommer1, Haya Herscovitz, Francine K Welty
1Mass Spectrometry Resource, Department of Biochemistry, Boston University School of Medicine, Boston, MA, USA.
Journal of Lipid Research
|January 31, 2006
Summary
A new liquid chromatography-mass spectrometry (LC-MS) method analyzes human lipoprotein lipids. This robust technique differentiates lipid profiles in normal versus familial hypobetalipoproteinemia patients.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Lipidomics
Background:
- Lipidomic analysis is crucial for understanding lipoprotein function.
- Human lipoprotein-associated lipids play key roles in various physiological and pathological processes.
- Accurate characterization of lipid profiles is essential for disease diagnosis and management.
Purpose of the Study:
- To develop and demonstrate a simple, robust LC-MS methodology for comprehensive lipid mixture analysis.
- To apply this methodology to characterize lipids associated with human lipoproteins, specifically apolipoprotein B.
- To investigate lipid profile differences in individuals with normal lipid metabolism versus familial hypobetalipoproteinemia.
Main Methods:
- Utilized a multi-stage LC-MS approach involving normal-phase HPLC-MS for class separation and reversed-phase LC-MS/MS for individual lipid characterization.
- Employed an optional initial fractionation step on Silica 60.
- Applied nanospray MS for detailed structural elucidation of lipids.
- Used YMC PVA-Sil and Atlantis dC18 capillary columns for chromatographic separations.
Main Results:
- Successfully applied the LC-MS methodology to analyze human apolipoprotein B-associated lipids.
- Quantified low percentages of individual lipid molecular species with high accuracy.
- Demonstrated significant differences in lipid profiles between normal low-density lipoprotein (LDL) and familial hypobetalipoproteinemia-associated lipoproteins.
- Identified distinct lipid compositions linked to apolipoprotein B-100 versus apolipoprotein B-67.
Conclusions:
- The developed LC-MS methodology provides a robust and adaptable platform for comprehensive lipidomic analysis of lipoproteins.
- This approach enables sensitive detection and differentiation of lipid species, aiding in the study of lipid metabolism disorders.
- The findings highlight the utility of LC-MS in identifying lipid biomarkers for conditions like familial hypobetalipoproteinemia.
