High-performance thin-layer chromatography (HPTLC) is a widely used, fast and relatively inexpensive
method of separating complex mixtures. It is particularly useful for smaller, apolar compounds and offers
some advantages over HPLC. This review gives an overview about the special features as well as the
problems that have to be considered upon the HPTLC analysis of lipids. The term “lipids” is used here in a
broad sense and comprises fatty acids and their derivatives as well as substances related biosynthetically
or functionally to these compounds. After a short introduction regarding the stationary phases and the
methods how lipids can be visualized on an HPTLC plate, the individual lipid classes will be discussed and
the most suitable solvent systems for their separation indicated. The focus will be on lipids that are most
abundant in biological systems, i.e. cholesterol and its derivates, glycerides, sphingo- and glycolipids as
well as phospholipids. Finally, a nowadays very important topic, the combination between HPTLC and
mass spectrometric (MS) detection methods will be discussed. It will be shown that this is a very powerful
method to investigate the identities of the HPTLC spots in more detail than by the use of common staining
methods. Future aspects of HPTLC in the lipid field will be also discussed.