NP-HPLC法测定二氯二茂钛原料药的含量及有关物质

NP-HPLC determination of titanocene dichloride and its related substances

目的:建立正相高效液相色谱法测定有机金属抗癌原料药二氯二茂钛的含量及有关物质。方法:采用 Shimadzu LC-10A 液相色谱系统,色谱条件:以氰基 Lichrosorb—CN(4 mm×250 mm,5 μm)为色谱柱,以正己烷-二氯甲烷(50:50)为流动相,流速:1.0 mL·min^(-1),紫外检测波长:254 nm,柱温:25℃。以ω-溴代苯乙酮为内标测定二氯二茂钛原料药的含量;以自身对照法测定有关物质。结果:样品中待测组分与内标物分离良好,线性范围为2.5～50μg·mL^(-1),回归方程为 Y=0.0710X+0.0562,r=0.9999(n=7);日内和日间精密度 RSD 均小于1.0%,最低检测限为2 ng;方法的平均回收率(n=9)为99.9%。结论:方法简便、快速,结果准确,专属性强,灵敏度高,可同时测定有机金属类抗癌原料药二氯二茂钛的含量及有关物质。

Objective：To establish a normal -phase high -performance liquid chromatography（ NP- HPLC）method to determine the content of organometallic antitumor crude drug titanocene dichloride and its impurities. Methods：A Shimadzu LC - 10A HPLC system was adopted with Lichrosorb - CN column （4 mm ×250 mm,5 μm） as the stationary phase. The mobile phase consisted of hexane- dichloromethane（50： 50） with flow rate 1.0 mL · min^ -1 and column temperature was at 25 ℃. The UV detection wavelength was set at 254 nm. ω- Hypnone bromide was selected as an internal standard to determine the content of titanocene dichloride, and control method was used to determine the related substances. Results：Under the selected chromatographic conditions, there was a good separation efficiency between titanocene dichloride and degradation products of the sample. The linear range was 2. 5 - 50 μg · mL^-1 ,and regression equation was Y =0. 0710X ＋0. 0562 ,r =0. 9999（n =7）. The RSD of inter - day and intra- day was less than 1.0%. The lowest limit of determination was 2 ng. The average recovery （n = 9） was 99.9 %. Conclusion： The analytical method is simple, quick, accurate, sensitive and selective for the analysis of titanocene dichloride.