Lipid profiling of FPLC-separated lipoprotein fractions by electrospray ionization tandem mass spectrometry.
Glycerophospholipid and sphingolipid species and their bioactive metabolites are important regulators of lipoprotein and cell function. The aim of the study was to develop a method for lipid species profiling of separated lipoprotein classes. Human serum lipoproteins VLDL, LDL, and HDL of 21 healthy fasting blood donors were separated by fast performance liquid chromatography (FPLC) from 50 microl serum. Subsequently, phosphatidylcholine (PC), lysophosphatidylcholine, sphingomyelin (SM), ceramide (CER), phosphatidylethanolamine (PE), PE-based plasmalogen (PE-pl), cholesterol, and cholesteryl ester (CE) content of the separated lipoproteins was quantified by electrospray ionization tandem mass spectrometry (ESI-MS/MS). Analysis of FPLC fractions with PAGE demonstrated that albumin partially coelutes with HDL fractions. However, analysis of an HDL deficient serum (Tangier disease) showed that only lysophosphatidylcholine, but none of the other lipids analyzed, exhibited a significant coelution with the albumin containing fractions. Approximately 60% of lipoprotein CER were found in LDL fractions and 60% of PC, PE, and plasmalogens in HDL fractions. VLDL, LDL, and HDL displayed characteristic lipid class and species pattern. The developed method provides a detailed lipid class and species composition of lipoprotein fractions and may serve as a valuable tool to identify alterations of lipoprotein lipid species profiles in disease with a reasonable experimental effort.
Wiesner
Philipp
P
Institute of Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany.
Leidl
Katharina
K
Institute of Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany.
Boettcher
Alfred
A
Institute of Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany.
Schmitz
Gerd
G
Institute of Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany.
Liebisch
Gerhard
G
Institute of Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany. Electronic address: gerhard.liebisch@klinik.uni-regensburg.de.
D000328:Adult; D001774:Blood Chemical Analysis; D002784:Cholesterol / Q000097:blood; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D020404:Glycerophospholipids / Q000097:blood; D006801:Humans; D008055:Lipids / Q000097:blood* / Q000302:isolation & purification; D008074:Lipoproteins / Q000097:blood* / Q000737:chemistry* / Q000145:classification; D008297:Male; D021241:Spectrometry, Mass, Electrospray Ionization; D013107:Sphingolipids / Q000097:blood; D014280:Triglycerides / Q000097:blood; D055815:Young Adult
D020404:Glycerophospholipids; D008055:Lipids; D008074:Lipoproteins; D013107:Sphingolipids; D014280:Triglycerides; D002784:Cholesterol