High-Performance Thin-Layer Chromatography Coupled with Electrospray Ionization Tandem Mass Spectrometry for Identifying Neutral Lipids and Sphingolipids in Complex Samples

Abstract

Abstract High-performance thin-layer chromatography was directly combined with electrospray mass spectrometry (ESI-MS) for structural identification issues below the level of lipid classes in complex samples through a portable, automated, elution-based interface. For samples as diverse as biodiesel and human plasma, separation conditions using Automated Multiple Development were selected in each case to provide lipid classes as zones narrow enough to ensure a direct transfer of them to ESI-MS. The respective zone of interest can be selected at will. ESI+ spectra of neutral lipids and sphingolipids showed sodium adducts when recorded from the plate. By using the described technique and ion-trap technology, the respective sodium adducts were fragmented. Sodium remained as the charge of the fragment ions and, thus, was useful for their structural identification through MSn. In this way, composition profiles of each class by ESI+-MS, and further identification of individual lipids and the molecular species belonging to each of them, were obtained by MS/MS and/or high-resolution MS. Thus, mono and diacylglycerides in ESI+ and fatty acids (in ESI−) were identified as low-concentration impurities in a fatty acid methyl ester–based biodiesel sample. Likewise, molecular species of sphingomyelins and globotriaosylceramides were unequivocally identified in human plasma samples.