In recent years, the genus Malassezia has been expanded based on molecular data; in addition to M. furfur and M. pachydermatis, five new species (M. sympodialis, M. globosa, M. obtusa, M. restricta, M. slooffiae) have been described. Apart from their lipid dependence, little is known about the metabolism and nutritional requirements of these new species. Defined inocula of Malassezia reference strains were cultured on selective agar for pathogenic fungi which was overlaid with olive oil. Samples of the olive oil overlay were taken at regular intervals and the lipid fractions were analysed by high performance thin layer chromatography. Depending on the time of incubation and the number of cells, M. sympodialis and the other recently described species produced a significant increase in free fatty acids. In addition, a band of an apolar substance was identified as a mixture of fatty acid ethyl esters. While showing growth, strains of M. furfur produced only small amounts of ethyl esters and free fatty acids. The growth kinetics of M. furfur and M. sympodialis were also different: for M. sympodialis, a clear lag phase was observed, possibly indicating the necessity of extracellular hydrolysis of the triglycerides. The significance of the synthesis of ethyl esters could not be clarified. For routine differentiation, this metabolic difference is only of limited usefulness because slight contamination of M. furfur strains with other lipophilic Malassezia species may lead to misinterpretation due to the high metabolic activity. These metabolic differences might be important in the pathogenesis of Malassezia infections.