HPLC法测定11种PC和PE类合成磷脂

Determination of Eleven Synthetic PC and PE Phospholipids by HPLC

建立了HPLC法测定11种磷脂酰胆碱(PC)和磷脂酰乙醇胺(PE)类合成磷脂。结合11种合成磷脂的结构特点和在色谱中的保留行为,首次将孔径为12和15 nm(120和150Å)的C8色谱柱串联,串联顺序为Welch-Topsil-C8柱(4.6 mm×250 mm,5μm,15 nm)在前,Ultimate XB-C8柱(4.6 mm×250 mm,5μm,12 nm)在后,以水-甲醇-乙酸-三乙胺为流动相A,以甲醇-异丙醇-乙酸-三乙胺为流动相B,梯度洗脱。结果显示,11种PC和PE类合成磷脂的最小分离度为1.0,7种PC类合成磷脂在质量浓度0.1~1.0 mg/m L内线性关系良好,4种PC类合成磷脂在质量浓度0.03~0.30mg/m L内线性关系良好,精密度RSD均小于2.5%。并且,此分离方法被成功应用于2种脂质体注射液和1种辅料中合成磷脂的测定。

An HPLC method was established to determine eleven synthetic phosphatidylcholine(PC)and phosphatidylethanolamine(PE)phospholipids.Consideration of the structural characteristics and retention behaviors in chromatography of the phospholipids,C8 chromatography columns with pore sizes of 12 and 15 nm(120 and 150Å)were connected in series for the first time.The serial sequence was Welch Topsil C8 column(4.6 mm×250 mm,5μm,15 nm)at front and Ultimate XB C8 column(4.6 mm×250 mm,5μm,12 nm)at back.The analysis was carried out in the gradient elution mode with water-methanol-acetic acid-triethylamine as mobile phase A,and methanol-isopropanol-acetic acid triethylamine as mobile phase B.The results showed that the minimum separation degree of eleven synthetic PC and PE was 1.0.It was linear for seven synthetic PC phospholipids in the concentration of 0.1-1.0 mg/mL,and it was linear for four synthetic PC phospholipids in the concentration of 0.03-0.30 mg/mL.RSDs of precision were all less than 2.5%.And this separation method was successfully applied to the determination of synthesized phospholipids in two types of liposome injections and one excipient.