Rational selection of reverse phase columns for high throughput LC-MS lipidomics.
Human blood plasma was chosen as a representative complex natural lipidome with large variance of lipid classes, species and lipid concentrations. Lipids were separated by RPLC on five different reverse phase columns with different types of stationary phase particles, size and chemistry. High mass accuracy MS analysis and data-dependent MS/MS analysis were performed using a Q Exactive HF Hybrid Quadrupole-Orbitrap Mass Spectrometer to identify individual lipid molecular species. This workflow was applied to evaluate the separation capability of each column and to identify the lipidomics profile in highly complex biological samples. As a result, we report more than 600 lipid species covering 18 lipid classes in human blood plasma and provide suggestions to the selection of the appropriate reverse phase column for the analysis of specific lipidomes. In each folder are reported the 6 replicates (3 in positive, 3 in negative ion mode) for the 5 column used: - Accucore C18 (2.1 x 150 mm, 2.6 µm) - HypersilGOLD C18 (2.1 x 150 mm, 1.9 µm) - Accucore C30 (2.1 x 150 mm, 2.6 µm) - Acclaim C30 (2.1 x 150 mm, 1.9 µm) - Acclaim C30 (2.1 x 250 mm, 3.0 µm)