The methanolysis of small amounts of purified phosphoglycerides and sphingomyelin was studied and a quantitative comparison of five methods for the methanolysis of standard phosphoglycerides was made. These methods were based on methanolysis with boron trifluoride-methanol, methanolic sodium methoxide (at ambient temperature and with heating) and methanolic sulphuric acid. A further method was based on saponification with methanolic sodium hydroxide and subsequent esterification with boron trifluoride-methanol. Under the experimental conditions, only the sodium methoxide-catalysed method at ambient temperature gave complete methanolysis of phosphoglycerides. For methanolysis of sphingomyelin, boron trifluoride-methanol, methanolic sulphuric acid and methanolic hydrochloric acid were used. It was found that complete methanolysis of sphingomyelin takes 15 h at 90 degrees C. Based on these results, procedures for the methanolysis of phosphoglycerides and sphingomyelin separated by high-performance liquid chromatography are presented.