title: Hydrophilic interaction and reversed phase mixed-mode liquid chromatography coupled to high resolution tandem mass spectrometry for polar lipids analysis.
abstract: A hydrophilic interaction liquid chromatography (HILIC) fused-core column (150×2.1mm ID, 2.7μm particle size) and a short reversed-phase liquid chromatography (RPLC) column (20mm×2.1mm ID, 1.9μm) were serially coupled to perform mixed-mode chromatography (MMC) on complex mixtures of phospholipids (PL). Mobile phase composition and gradient elution program were, preliminarily, optimized using a mixture of phosphatidylcholines (PC), phosphatidylethanolamines (PE), their corresponding lyso-forms (LPC and LPE), and sphingomyelins (SM). Thus a mixture of PC extracted from soybean was characterized by MMC coupled to electrospray ionization (ESI) high-resolution Fourier-transform mass spectrometry (FTMS) using an orbital trap analyzer. Several previously undiscovered PC, including positional isomers (i.e. 16:0/19:1 and 19:1/16:0) of PC 35:1 and skeletal isomers (i.e. 18:1/18:2 and 18:0/18:3) of PC 36:3 were identified. Therefore, high-resolution MS/MS spectra unveiled the occurrence of isomers for several overall side chain compositions. The proposed MMC-ESI-FTMS/MS approach revealed an unprecedented capability in disclosing complexity of an actual lipid extract, thus representing a very promising approach to lipidomics.
authors.affiliation: Dipartimento di Chimica, Università degli Studi di Bari Aldo Moro, Campus Universitario, Via E. Orabona, 4-70126 Bari, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Granafei
authors.affiliation: Dipartimento di Chimica, Università degli Studi di Bari Aldo Moro, Campus Universitario, Via E. Orabona, 4-70126 Bari, Italy.
authors.forename: Pietro
authors.initials: P
authors.lastname: Azzone
authors.affiliation: Dipartimento di Chimica, Università degli Studi di Bari Aldo Moro, Campus Universitario, Via E. Orabona, 4-70126 Bari, Italy.
authors.forename: Vito Alessandro
authors.initials: VA
authors.lastname: Spinelli
authors.affiliation: Dipartimento di Chimica, Università degli Studi di Bari Aldo Moro, Campus Universitario, Via E. Orabona, 4-70126 Bari, Italy; Centro di Ricerca Interdipartimentale S.M.A.R.T., Università degli Studi di Bari Aldo Moro, Campus Universitario, Via E. Orabona, 4-70126 Bari, Italy.
authors.forename: Ilario
authors.initials: I
authors.lastname: Losito
authors.affiliation: Dipartimento di Chimica, Università degli Studi di Bari Aldo Moro, Campus Universitario, Via E. Orabona, 4-70126 Bari, Italy; Centro di Ricerca Interdipartimentale S.M.A.R.T., Università degli Studi di Bari Aldo Moro, Campus Universitario, Via E. Orabona, 4-70126 Bari, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Palmisano
authors.affiliation: Dipartimento di Chimica, Università degli Studi di Bari Aldo Moro, Campus Universitario, Via E. Orabona, 4-70126 Bari, Italy; Centro di Ricerca Interdipartimentale S.M.A.R.T., Università degli Studi di Bari Aldo Moro, Campus Universitario, Via E. Orabona, 4-70126 Bari, Italy. Electronic address: tommaso.cataldi@uniba.it.
authors.forename: Tommaso R I
authors.initials: TRI
authors.lastname: Cataldi
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D005583:Fourier Analysis; D057927:Hydrophobic and Hydrophilic Interactions; D010713:Phosphatidylcholines / Q000032:analysis; D010743:Phospholipids / Q000032:analysis*; D013025:Glycine max / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D010713:Phosphatidylcholines; D010743:Phospholipids
keywords: Hydrophilic; Hydrophobic; Mixed-mode chromatography; Phosholipids; Polar lipids; Serially coupled columns; Tandem mass spectrometry