J. Lipid Res. Neurobiology of Lipids (ISSN1683-5506)
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Originally published In Press as doi:10.1194/jlr.D300040-JLR200 on April 21, 2004

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Journal of Lipid Research, Vol. 45, 1355-1363, July 2004
Copyright © 2004 by American Society for Biochemistry and Molecular Biology

Methods

Bacterial phospholipid molecular species analysis by ion-pair reversed-phase HPLC/ESI/MS

Nicolas Mazzella*, Josiane Molinet*, Agung Dhamar Syakti*, Alain Dodi{dagger}, Pierre Doumenq1,*, Jacques Artaud* and Jean-Claude Bertrand§

* Laboratoire de Chimie Analytique de l'Environnement, Unité Mixte de Recherche 6171, Institut Fédératif de Recherche Pôle Méditerannéen des Sciences de l'Environement 112, Europôle de l'Arbois, BP 80, 13545 Aix-en-Provence Cedex 4, France
{dagger} Laboratoire d'Analyses Radiochimiques et Chimiques, Commissariat à l'Énergie Atomique, Centre d'études de Cadarache, 13108 St Paul lez Durance, France
§ Laboratoire de Microbiologie, Géochimie et Ecologie Marine, UMR 6117, case 901, Université de la Méditerranée, 163 avenue de Luminy, 13228 Marseille Cedex 09, France

1 To whom correspondence should be addressed. e-mail: pierre.doumenq{at}univ.u-3mrs.fr

This work set out to optimize the detection and separation of several phospholipid molecular species on a reversed-phase column with the use of an electrospray ionization/mass spectrometry-compatible counter-ion. An application of this technique concerned a qualitative and quantitative analysis of bacterial membrane phospholipids extracted from Corynebacterium species strain 8. The phospholipid classes of strain 8 were identified as phosphatidylglycerol, phosphatidylinositol, diphosphatidylglycerol, and a peculiar lipid compound, acyl phosphatidylglycerol.

Most of the molecular species structures were elucidated, and regarding phosphatidylglycerol, the fatty acid positions were clearly determined with the calculation of the sn-2/sn-1 intensity ratio of the fatty acyl chain fragments.

Abbreviations: APG, acyl phosphatidylglycerol; DMDS, dimethyl disulfide; DMPG, 1,2-dimyristoyl-sn-glycero-3-phospho-rac-glycerol; DPG, diphosphatidylglycerol; DPPE, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine; ELSD, evaporative light-scattering detector; ESI, electrospray ionization; FAME, fatty acid methyl ester; IPMS, intact phospholipid molecular species; MSM, mineral salt medium; PC, phosphatidylcholine, PE, phosphatidylethanolamine; PG, phosphatidylglycerol; PI, phosphatidylinositol; PLFA, phospholipid ester-linked fatty acid; POPE, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine; POPG, 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-rac-glycerol; PS, phosphatidylserine; TBAAc, tetrabutyl ammonium acetate; TBABr, tetrabutyl ammonium bromide; UV, ultraviolet light

Supplementary key words reversed-phase liquid chromatography/electrospray ionization/mass spectrometry • negative ionization • counter-ion • sn-1 and sn-2 fatty acid positions • Corynebacterium species


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Copyright © 2004 by the American Society for Biochemistry and Molecular Biology.