Nakagawa Y, Waku K
J Chromatogr. 1986 Sep 5;381(2):225-31. doi: 10.1016/s0378-4347(00)83588-4.
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.
我们描述了一种快速高效的高效液相色谱（HPLC）方法，用于分离源自磷脂酰胆碱的二甲基磷脂酸的分子种类，磷脂酰胆碱天然存在于哺乳动物组织中。通过磷脂酶D将磷脂酰胆碱衍生化为二甲基磷脂酸，随后进行重氮甲烷处理，然后在反相柱上完成分离。通过反相HPLC在40分钟内实现了二甲基磷脂酸主要分子种类的分离。大鼠肝脏磷脂酰胆碱中的二甲基磷脂酸被分离为12个独立的峰。通过气相色谱测定鉴定了大鼠肝脏磷脂酰胆碱中的13种不同分子种类。这种改进的方法适用于使用放射性前体研究各种组织中磷脂分子种类磷酸基团的代谢。
