HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

Papers In Press, published online ahead of print March 1, 2009
J. Lipid Res., doi:10.1194/jlr.D800028-JLR200

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: D800028-JLR200v1 D800028-JLR200v2 50/3/574    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Wiesner, P. Articles by Liebisch, G. Search for Related Content PubMed PubMed Citation Articles by Wiesner, P. Articles by Liebisch, G. Social Bookmarking                      What's this?

Journal of Lipid Research, Vol. 50, 574-585, March 2009
Copyright © 2009 by American Society for Biochemistry and Molecular Biology

Methods

Lipid profiling of FPLC-separated lipoprotein fractions by electrospray ionization tandem mass spectrometry

Philipp Wiesner, Katharina Leidl, Alfred Boettcher, Gerd Schmitz and Gerhard Liebisch¹

Institute of Clinical Chemistry and Laboratory Medicine, University of Regensburg, Germany

The online version of this article (available at http://www.jlr.org) contains supplementary data in the form of ten figures and four tables.

This work was supported by Deutsche Forschungsgemeinschaft (Li 923/2-1/2) and by the seventh framework program of the EU-funded "LipidomicNet" (proposal number 202272).

Published, JLR Papers in Press, October 9, 2008.

¹ To whom correspondence should be addressed. e-mail: gerhard.liebisch{at}klinik.uni-regensburg.de

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 µl 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.

Supplementary key words lipid species • lipoprotein fractionation • glycerophospholipids • sphingolipids • cholesterol • VLDL • LDL • HDL • fast performance liquid chromatography

Abbreviations: apoA-I, apolipoprotein A-I; CE, cholesteryl ester; CER, ceramide; ESI-MS/MS, electrospray ionization tandem mass spectrometry; FC, free cholesterol; FPLC, fast performance liquid chromatography; GC, gas chromatography; LPC, lysophosphatidylcholine; NBD, 7-nitrobenz-2-oxa-1,3-diazole; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PE-pl, PE-based plasmalogen; PL, total phospholipids; SM, sphingomyelin; TC, total cholesterol

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				S. Rutti, J. A. Ehses, R. A. Sibler, R. Prazak, L. Rohrer, S. Georgopoulos, D. T. Meier, N. Niclauss, T. Berney, M. Y. Donath, et al. Low- and High-Density Lipoproteins Modulate Function, Apoptosis, and Proliferation of Primary Human and Murine Pancreatic {beta}-Cells Endocrinology, October 1, 2009; 150(10): 4521 - 4530. [Abstract] [Full Text] [PDF]