J. Lipid Res. Neurobiology of Lipids (ISSN1683-5506)
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Originally published In Press as doi:10.1194/jlr.M500506-JLR200 on January 28, 2006

Papers In Press, published online ahead of print April 1, 2006
J. Lipid Res., doi:10.1194/jlr.M500506-JLR200
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Journal of Lipid Research, Vol. 47, 804-814, April 2006
Copyright © 2006 by American Society for Biochemistry and Molecular Biology

LC-MS-based method for the qualitative and quantitative analysis of complex lipid mixturesboxs

Ulf Sommer*, Haya Herscovitz{dagger}, Francine K. Welty§ and Catherine E. Costello*,1

* Mass Spectrometry Resource, Department of Biochemistry, Boston University School of Medicine, Boston, MA
{dagger} Department of Physiology and Biophysics, Boston University School of Medicine, Boston, MA
§ Cardiovascular Division, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA

boxs The online version of this article (available at http://www.jlr.org) contains an additional figure.

Published, JLR Papers in Press, January 28, 2006.

1 To whom correspondence should be addressed. e-mail: cecmsms{at}bu.edu

A simple and robust LC-MS-based methodology for the investigation of lipid mixtures is described, and its application to the analysis of human lipoprotein-associated lipids is demonstrated. After an optional initial fractionation on Silica 60, normal-phase HPLC-MS on a YMC PVA-Sil column is used first for class separation, followed by reversed-phase LC-MS or LC-tandem mass spectrometry using an Atlantis dC18 capillary column, and/or nanospray MS, to fully characterize the individual lipids. The methodology is applied here for the analysis of human apolipoprotein B-associated lipids. This approach allows for the determination of even low percentages of lipids of each molecular species and showed clear differences between lipids associated with apolipoprotein B-100-LDL isolated from a normal individual and those associated with a truncated version, apolipoprotein B-67-containing lipoproteins, isolated from a homozygote patient with familial hypobetalipoproteinemia. The methods described should be easily adaptable to most modern MS instrumentation.

Supplementary key words low density lipoprotein • intermediate density lipoprotein • normal-phase high-performance liquid chromatography-mass spectrometry • reversed-phase liquid chromatography-tandem mass spectrometry • familial hypobetalipoproteinemia

Abbreviations: B67, apolipoprotein B-67 (N-terminal 67% of apolipoprotein B); B100, apolipoprotein B-100 (full-length apolipoprotein B); ESI, electrospray ionization; IDL, intermediate density lipoprotein; IPA, iso-propyl alcohol, 2-propanol; MALDI-TOF, matrix-assisted laser desorption/ionization time-of-flight; MeOH, methanol; MS/MS, tandem mass spectrometry; MTBE, methyl t-butyl ether; PC, glycerophosphocholine; PE, glycerophosphoethanolamine; PI, glycerophosphoinositol; QoTOF, quadrupole orthogonal time-of-flight; QQQ, triple quadrupole


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