Extraction protocols and liquid chromatography coupled with mass spectrometry (LC-MS) and tandem mass spectrometry methods (LC-MS/MS) for the measurement of five lipid categories, namely; glycerophospholipids, glycerolipids, sphingolipids, sterol lipids and fatty acyls in human plasma are described here. Step-by-step instructions are provided for the liquid/liquid extraction methods, including solution preparation and the non-endogenous lipid internal standards used. All instrumental conditions, chromatography columns and solutions required for the LC-MS and LC-MS/MS methods are also detailed.