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  

Determination of the content of alprostadil injection by HPLC with post-column derivatization and column switching

作　　者：周长明[1] 丁锐[1] 

机构地区：[1]北京市药品检验所,北京100035

出　　处：《药物分析杂志》2014年第3期402-406,共5页Chinese Journal of Pharmaceutical Analysis

摘　　要：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MerckLiChroCART@25—4色谱柱（25mm×4mm，5μm）为预处理柱，以无水乙醇为洗脱液，流速为2．0mL．min^-1，柱温60℃；以LiChrospher100RP-18eC18色谱柱（25mm×4mm，5μm）为压力替换柱，用以平衡系统压力，柱温60℃；采用AgilentZorbaxSBC18色谱柱（250mm×4．6mm，5μm）为分析柱，以0．0067mol·L^-1磷酸盐缓冲液（取磷酸二氢钾9．07g，加水1000mL使溶解，用0．1mol·L^-1氢氧化钠溶液调节pH至6．3）-乙腈（5：2）为流动相，流速1．0mL·min^-1，检测波长278nm．柱温60℃；以1mol·L^-1氢氧化钾溶液为柱后反应液，流速0．5mL·min^-1；进样量20μL。结果：前列地尔浓度在0．25～12．0μg·mL。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 alpros- tadil injection by HPLC. Methods： The on - line pretreatment was performed on a Merck LiChroCART 25 - 4 （25 mm x 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 x4 mm,5 μm） ,the column temperature was set at 60 ℃. The determination was carried out on an Agilent Zorbax SB Cls column （250 mm x4. 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） - aeetonitrile （5： 2） at a flow rate of 1.0 mL ·min^-1 , the de- tection wavelength was 278 nm, the column temperature was set at 60 ℃. The post - column reaction mixture con- sisted of 1 mol ·L^-1potassium 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 （ rt = 9 ） of this method was 99.25 %. Conclusion： The method methodology validation shows that this can be used to determine the content of alprostadilin alprostadil injection.

关 键 词：柱切换 柱后衍生 高效液相色谱 脂质体微球 前列地尔 含量测定 方法验证 

分 类 号：R917[医药卫生—药物分析学]