An extended method for separating and quantitating molecular species of phospholipids

AN improved and extended method for separating and quantitating molecular species of four phospholipid classes is presented. Crude lipid extract is first separated into phospholipid classes on a silica column. Each phospholipid class is then separated into molecular species without derivatization using high-performance liquid chromatography on columns packed with octadecyl silica. Quantitation of individual species is achieved by measuring absorbance at 205 nm. Factors for converting absorbancies to mol fractions have been determined. Quantitation by absorbance at 205 nm agrees well with quantitation by gas chromatography which is preferred to quantitation by phosphate analysis. One hundred phospholipid species have been identified. A table of relative retention times of molecular species is provided. Examples of quantitative analyses of species composition are presented.