メチルシリカカラムを用いた高速液体クロマトグラフィーによるホスファチジルコリン及びホスファチジルエタノールアミンの定量法
1984 年 33 巻 1 号 p. 34-39
詳細
1984 年 33 巻 1 号 p. 34-39
Reversed phase high performance liquid chromatography (HPLC) on methyl silica column was used for the separation of phospholipids including phosphatidyl choline (PC), phosphatidyl ethanolamine (PE), phosphatidyl inositol (PI) and phosphatidyl serine (PS) in naturally occurring samples.
Hexane/2-propanol/water (1 : 60 : 39) was one of the best eluents for the separation of these phospholipids which were detected at 210nm.
The molar extinction coefficient of each phospholipid is not yet known and it is very difficult to obtain an authentic sample, so that purified soybean phospholipids, quantitated by Reinecke's salt method for PC and by 2, 4-dinitro-1-fluorobenzene (DNFB) method for PE, were used as standard samples for the determination of PC and PE.
The calibration curves between the amount and the peak height in the HPLC were obtained with good linearity ranging from 3.7 to 74μg for PC and from 0.7 to 6.6μg for PE.
PC and PE in some phospholipids such as soybean, egg yolk, pig liver and dry yeast (Saccharomyces cerevisiae sp.) phospholipids were determined with reliable accuracy under the conditions employed.
The present method can be used for the routine quantitation of PC and PE in various phospholipids prepared from plant and animal sources.
