Sestak T L, Subbaiah P V, Jaskowiak N T, Bagdade J D
Department of Biochemistry, Rush Medical College, Chicago, Illinois 60612.
Anal Biochem. 1990 Nov 15;191(1):156-9. doi: 10.1016/0003-2697(90)90402-u.
Plasma disaturated phosphatidylcholine (DSPC) concentration has been implicated as a risk factor for atherosclerosis. However, suitable methods for the estimation of these compounds in plasma are not available. In this paper, a method for the estimation of DSPC using argentation thin-layer chromatography and high-performance liquid chromatography is described. It is quantitative for the measurement of individual and total DSPC species and is not dependent on fatty acid chain length. The method employs hydrolysis of total plasma phosphatidyl choline by phospholipase C, followed by benzoylation of the diacylglycerols. The benzoates are then fractionated on silver nitrate-impregnated silica gel thin-layer chromatography plates, and the disaturated species separated and quantitated by high-performance liquid chromatography. The method is sensitive and reproducible and allows many samples to be done at once. With this method, the amounts of DSPC were found to be significantly higher in a group of normolipidemic diabetic subjects, compared to age-matched controls.
血浆不饱和磷脂酰胆碱（DSPC）浓度被认为是动脉粥样硬化的一个危险因素。然而，目前尚无适用于估计血浆中这些化合物的方法。本文描述了一种使用银化薄层色谱法和高效液相色谱法估计DSPC的方法。该方法可对单个和总DSPC种类进行定量测量，且不依赖于脂肪酸链长度。该方法采用磷脂酶C水解总血浆磷脂酰胆碱，然后对二酰基甘油进行苯甲酰化。然后将苯甲酸盐在硝酸银浸渍的硅胶薄层色谱板上进行分离，不饱和种类通过高效液相色谱法进行分离和定量。该方法灵敏且可重复，可同时处理多个样品。使用该方法发现，与年龄匹配的对照组相比，一组血脂正常的糖尿病患者的DSPC含量显著更高。
