Despite advances in the last few years, the use of supercritical fluid chromatography (SFC) in combination with mass spectrometry (MS) for lipidomic analysis has not reached the popularity of LC-MS. However, SFC presents clear advantages that can be exploited, such as fast, reproducible and class-based separations including nonpolar lipid classes, such as cholesterol esters, triacylglycerols, diacylglycerols, monoacylglycerols and cholesterol. In this study we show how SFC can be used for comprehensive lipidomic analyses after optimization of parameters such as back-pressure regulator (BPR), column temperature or overfeed volume of injection. We also compare the performances of triple quadrupole (QqQ) and quadrupole time-of-flight (QTOF) mass spectrometers coupled to SFC in terms of qualitative and quantitative analyses.