This systematic overview of lipidomic analysis by the president and vice president of the Lipidomics Society covers international standards of lipid analysis, available technology, key lipid classes, as well as applications in basic research, medicine, pharma and food industry. Aspects of sample preparation, separation methods, different mass spectrometry modes, as well das identification and quantitation are covered, including the use of bioinformatics tools for data analysis. Analytical approaches for all major classes of biological lipids, from fatty acids to phospholipids to sterols are discussed, and a wide range of novel and established lipidomic applications from fundamental research to clinical diagnostics and food and crop science are introduced.