HPLC同时测定羟喜树碱注射液的有关物质及含量

Simultaneous Determination for Related Substances and Content of Hydroxycamptothecin Injection by HPLC

关键词:  高效液相色谱法  羟喜树碱注射液  喜树碱  有关物质  含量测定

Key words:  HPLC  hydroxycamptothecin injection  camptothecin  related substances  content determination

摘要:

目的 建立以高效液相色谱法同时测定羟喜树碱注射液的有关物质和含量的方法。方法 采用Ultimate XB-C18柱(4.6 mm×250 mm，5 μm)，流动相为甲醇-水(60∶40)，检测波长为370 nm，流速为1.0 mL·min^-1，柱温35 ℃。结果 有关物质：羟喜树碱与杂质分离度良好，羟喜树碱和其主要杂质喜树碱的线性范围均在0.2~1.0 μg·mL^-1之间，相关系数均在0.999 7以上，最低检测限分别为0.2 ng和0.3 ng。含量测定：羟喜树碱在4.0~36.3 μg·mL^-1内线性相关系数为1.000 0，根据3个不同生产厂家的处方工艺分别进行低、中、高3种浓度的回收率试验，结果均在100.0%~102.0%之间，RSD均在0.02%~0.71%之间(n=3)。结论 本方法专属性强、准确性高，可全面系统地控制羟喜树碱注射液的质量。

Abstract:

OBJECTIVE To establish an HPLC method for simultaneous determination of related substances and content of hydroxycamptothecin injection. METHODS The chromatographic column was Ultimate XB-C18(4.6 mm×250 mm, 5 μm), and the mobile phase consisted of a mixture of methanol-water(60∶40). The flow rate was 1.0 mL·min^-1 and the detection wavelength was 370 nm. The column temperature was maintained at 35 ℃。RESULTS For the determination of related substances: the separations between hydroxycamptothecin and its related substances were very well. The calibration curves of hydroxycamptothecin and its main related substance, namely camptothecinthe, were both linear within the ranges of 0.2-1.0 μg·mL^-1(r≥0.999 7), and the LODs were 0.2 ng and 0.3 ng, respectively. For the determination of content: hydroxycamptothecin was linear within the range of 4.0-36.3 μg·mL^-1(r=1.000 0), the average recoveries of low, medium and high concentrations based on the formulations from manufactory A, B and C were between 100.0% and 102.0%, the RSDs were between 0.02% and 0.71%(n=3). CONCLUSION The method is specific and accurate, and can systematically be used for the quality control of hydroxycamptothecin injection.