HPLC-ELSD法测定复合磷脂固醇脂质中磷脂酰胆碱的含量

HPLC-ELSD determination of phosphatidylcholine in phospholipid cholesterol complex lipids

目的:采用 HPLC-ELSD 法测定复合磷脂固醇脂质中磷脂酰胆碱(PC)的含量。方法:采用 Waters Nova-pak(?) silica60(?)径向柱(100 mm×8 mm,4 μm),流动相:己烷-异丙醇(3:4)为流动相 A,己烷-异丙醇-水(3:4:0.75)为流动相 B,梯度洗脱,流速1.5 mL·min^(-1);蒸发光散射检测,ELSD 漂移管温度:40℃,雾化气:氮气,载气压力:340 kPa。结果:在选定的色谱条件下,棕榈酸甘油三酯、磷脂酰乙醇胺、溶血磷脂酰乙醇胺、磷脂酰胆碱、鞘磷脂、溶血磷脂酰胆碱各成分之间可达到很好分离。磷脂酰胆碱进样量在21.2～127.2 μg范围内与峰面积的线性关系良好(r=0.9969),最低检测限为424 ng(S/N=3),3个浓度水平下磷脂酰胆碱的平均同收率(n=9)为100.6%。结论:方法灵敏,快速,准确,重复性好,可用于产品的质量控制。

Objective：To determinate phosphatidylcholine（PC） in phospholipid cholesterol complex lipids by HPLC - ELSD. Methods：The separation was performed on a Waters Nova - pak silica 60_＆ column（ 100 mm × 8 mm,4 μm） using gradient elution with solution A of hexane - isopropanol （3：4 ） and solution B of hexane - isopropanol - water（3：4： 0. 75 ）as mobile phase ;The flow rate was 1.5 mL·min^-1. An evaporative light scattering detector was used. The temperature of the drift tube was set at 40℃. The nebulizing gas was N2 and the pressure was 340 kPa. Results： Good separations of tripalmitin, phosphatidylethanolamine, lysophosphatidylethanolamine, phosphatidylcholine,sphingomyelin and lysophosphatidylcholine could be achieved under above system. The calibration curve was linear in the range of 21.2 - 127.2μg with correlation coefficient of 0. 9969. The detection limit was 424 ng（ S/N =3 ）. The average recovery（ n = 9 ） was 100. 6% for three different levels. Conclusion： This method is sensitive, rapid, accurate, good reproducibility, and can be used for the quality control of this product.