The lipidomics method utilises an RPC approach that separates individual lipids and other hydrophobic metabolite species (as opposed to class-based lipid separations and direct infusion methods ). The method used herein was designed as an adaptation of a now-widely used approach (seemingly originating in the work of Castro-Perez et al ) which employs a C18 stationary phase and 90/10 mixture of 2-propanol and acetonitrile as the strong eluent. In an effort to enable higher solvent flow rate and improved peak capacity (benefitting isomeric structure separation), the strong eluent 2-propanol/acetonitrile ratio was modified from the original method (90/10) to 50/50, resulting in a less viscous eluent better suited to high flowrate delivery for higher peak capacity. The lower elution strength of the resulting mix was compensated by use of a shorter acyl chain bonded phase (C8 instead of C18), allowing the clean elution of triglycerides and longer chain lipids. Phosphoric acid was added to the mobile phase for the chromatographic peak shape enhancement of acidic phospholipids. The addition of phosphoric acid requires special care of and attention to the composition of the LC pumps and pump heads (in our case, replacing more common grade 316 stainless steel parts with MP35N alloy parts available from the original equipment manufacturer) to reduce pump surface corrosion. The method is used with both positive and negative mode ion detection.