Optimizing a ultrahigh pressure liquid chromatography-time of flight-mass spectrometry approach using a novel sub-2 μm core–shell particle for in depth lipidomic profiling of Caenorhabditis elegans
Journal of Chromatography A
September 12, 2014
Lipid profiling or lipidomics is currently applied in many different research fields. It refers to the global analysis of a samples lipid content using different analytical chemistry methods, with mass spectrometry as the mostly employed technology. We developed a comprehensive in-depth analysis method for the lipidome of the soil-dwelling nematode Caenorhabitis elegans, a widely used model organism. Four different columns were compared with a generic gradient and a novel sub-2-μm core–shell column, Waters Cortecs C18, showed superior performance in case of chromatographic peak characteristics, e.g. plate numbers and number of detected lipid features. Retention time deviation was generally less than 1% within one column and below 5% for columns from different batches. Intensity variation was lower than 30% for most detected features. Improved chromatographic separation showed enhanced resolution for isomeric lipids and allowed collection of highly detailed MS/MS spectra for lipid identification. In total 1304 lipid features were detected in positive ionization mode and 265 in negative mode. Lipids from different classes were annotated and MS/MS spectra obtained by data dependent fragmentation were used for identification purposes.