柱切换柱后衍生高效液相色谱法测定前列地尔注射液中主药的含量

目的：建立一种高效、快速、样品在线预处理的柱切换柱后衍生高效液相色谱法，用以测定脂质体微球类药物前列地尔注射液中主药的含量。方法：采用Merck LiChroCART^® 25-4色谱柱（25 mm×4 mm，5 μm）为预处理柱，以无水乙醇为洗脱液，流速为2.0 mL·min^-1，柱温60 ℃；以LiChrospher^® 100 RP-18e C₁₈色谱柱（25 mm×4 mm，5 μm）为压力替换柱，用以平衡系统压力，柱温60 ℃；采用Agilent Zorbax SB C₁₈色谱柱（250 mm×4.6 mm，5 μm）为分析柱，以0.0067 mol·L^-1磷酸盐缓冲液（取磷酸二氢钾9.07 g，加水1000 mL使溶解，用0.1 mol·L^-1氢氧化钠溶液调节pH至6.3）-乙腈（5：2）为流动相，流速1.0 mL·min^-1，检测波长278 nm，柱温60 ℃；以1 mol·L^-1氢氧化钾溶液为柱后反应液，流速0.5 mL·min^-1；进样量20 μL。结果：前列地尔浓度在0.25~12.0 μg·mL^-1范围内线性关系良好（r=0.9993）；平均回收率（n=9）为99.25%。结论：本方法经方法学验证操作简便、准确，可作为前列地尔注射液含量测定的分析方法。

Objective: To establish an efficient and rapid HPLC method with automated sample pre-treatment by post-column derivatization and column switching system for the determination of the content of alprostadil in alprostadil injection by HPLC. Methods: The on-line pretreatment was performed on a Merck LiChroCART^® 25-4(25 mm×4 mm,5 μm),the pretreatment column eluent was anhydrous ethanol at a flow rate of 2.0 mL·min^-1,the column temperature was set at 60 ℃. The column for pressure correction was achieved with a LiChrospher^® 100 RP-18e (25 mm×4 mm,5 μm),the column temperature was set at 60 ℃. The determination was carried out on an Agilent Zorbax SB C₁₈ column (250 mm×4.6 mm,5μm) with the mobile phase of a mixture of 0.0067 mol·L^-1 phosphate BS (9.07 g potassium dihydrogen phosphate added with 1000 mL water,the pH value adjusted to 6.3 with 0.1 mol·L^-1 sodium hydroxide solution)-acetonitrile (5:2) at a flow rate of 1.0 mL·min^-1,the detection wavelength was 278 nm,the column temperature was set at 60 ℃. The post-column reaction mixture consisted of 1 mol·L^-1 potassium hydroxide solution at a flow rate of 0.5 mL·min^-1. The injection volume was 20 μL. Results: Good linearity was obtained for alprostadil ranged 0.25-12.0 μg·mL^-1(r=0.9993),and the recovery (n=9) of this method was 99.25%. Conclusion: The method methodology validation shows that this can be used to determine the content of alprostadil in alprostadil injection.