Improved procedure for the separation of the molecular species of dimethylphosphatidate by high-performance liquid chromatography.
We describe a rapid and efficient high-performance liquid chromatographic (HPLC) procedure for the separation of the molecular species of dimethylphosphatidic acid derived from phosphatidylcholine, which naturally occurs in mammalian tissues. The separation was accomplished on a reversed-phase column after derivatization of phosphatidylcholine to dimethylphosphatidate by means of phospholipase D and subsequent diazomethane treatments. Separation of the major molecular species of dimethylphosphatidate by reversed-phase HPLC was achieved within 40 min. Dimethylphosphatidate from rat liver phosphatidylcholine was resolved into twelve separate peaks. Thirteen different molecular species in rat liver phosphatidylcholine were identified by gas chromatographic determination. This improved method is applicable to studies on the metabolism of the phosphate group of the molecular species of phospholipid of a variety of tissues using a radioactive precursor.
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D008099:Liver / Q000032:analysis*; D010712:Phosphatidic Acids / Q000378:metabolism; D010713:Phosphatidylcholines / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D010712:Phosphatidic Acids; D010713:Phosphatidylcholines