梯度洗脱高效液相色谱法检测他克莫司有关物质

 目的建立他克莫司有关物质检测的方法,并观察样品在不同溶剂中可能存在的异构体的转化情况,同时液-质联用技术进一步确证有关物质。方法以辛烷基硅烷键合硅胶为填充剂,以0.02mol·L^-1磷酸二氢钾(pH3.5)与乙腈为流动相梯度洗脱;检测波长为210nm,柱温60℃;利用液-质联用技术确证可能存在的杂质。结果样品在质子性的溶剂中确实存在异构体的转化且该转化能达到平衡,但在乙腈溶剂中基本无转化,该溶剂可作为他克莫司有关物质的测定溶剂,样品中的主要杂质为发酵副产物。结论本法简便、准确,重复性好,能有效地检测样品中的有关物质。

Determination of Related Substances of Tacrolimus by HPLC with Gradient Elution

OBJECTIVE To establish a HPLC method for determining the related substances of tacrolimus.METHODS The determination was performed on a C₈ reversed phase column and the constitutes were gradiently eluted with a mixture consisting of acetonitrile-0.02 mol·L^-1 phosphates buffer(pH 3.5).UV Detection was set at 210 nm.The temperature of column was set at 60 ℃.The related substances of tacrolimus were confirmed by LC-MS.RESULTS Tacrolimus was transformed into its epimers in the solution but not in CH₃CN.CONCLUSION A sensitive and specific method for detecting the related substances from tacrolimus is developed and used to the quality control of tacrolimus.