HPLC-ELSD测定猪肺表面活性物质及其冻干粉中7种磷脂组分含量

Determination of Seven Phospholipid Components in Porcine Pulmonary Surfactant and Its Lyophilized Powder by HPLC-ELSD Method

目的建立猪肺表面活性物质及其冻干粉中7种磷脂组分的含量测定方法。方法采用硅胶柱,梯度洗脱,蒸发光散射检测器检测。结果磷脂酰甘油(PG)、磷脂酰乙醇胺(PE)、磷脂酰肌醇(PI)、磷脂酰丝氨酸(PS)、二棕榈酰磷脂酰胆碱(DP-PC)、鞘磷脂(SPH)、浴血磷脂酰胆碱(LPC)7种磷脂的线性回归方程的相关系数分别为0.999 0,0.998 4,0.999 0,0.999 3,0.999 8,0.998 8,0.998 1;日内精密度分别为2.1%,2.3%,2.8%,1.7%,1.5%,2.3%,2.3%;回收率分别为99.57%,101.03%,100.02%,100.17%,98.51%,100.65%,97.76%。结论本方法可同时测定猪肺表面活性物质及其冻干粉中7种磷脂组分的含量,专属性强、准确度高。

OBJECTIVE To develop a HPLC-ELSD method for the determination of seven phospholipid components in porcine pulmonary surfactant and its lyophilized powder. METHODS The samples were separated on a silica column with gradient elution and detected by evaporative light scattering detector. RESULTS The correlation coefficients of linear regression equation for phosphatidylglycerol （ PG）, phosphatidylethanolamine （ PE ）, phosphatidylinositol （ PI ）, phosphatidylserine （ PS ） , dipalmitoyl-phosphatidylcholine （ OPPC ）, sphingomyelin （ SPH ） and lysophophatidylcholine （ LPC ）, were 0. 999 0,0. 998 4,0. 999 0,0. 999 3,0. 999 8,0. 998 8 and 0. 998 1 respectively. The intraday precisions were 2. 1% ,2.3% ,2. 8% ,1.7% ,1.5% ,2. 3% and 2. 3% ,respectively. The recoveries were 99. 57%, 101.03% , 100. 02% , 100. 17% ,98. 51%, 100. 65% and 97.76% , respectively. CONCLUSION The method is accurate and specific, it can be used for the determination of seven phospholipid components in porcine pulmonary surfactant and its lyophilized powder simultaneously.