HPLC法测定盐酸吉西他滨原料及其注射用制剂的有关物质

目的采用梯度洗脱的HPLC法测定盐酸吉西他滨原料及其注射用制剂的有关物质，并利用LC-MS对碱破坏后得到的未知杂质进行进一步研究。方法使用CB色谱柱，以不同比例的0．14mol·L^-1磷酸盐缓冲液（pH2．5±0．1）和甲醇分别为流动相A和B，梯度洗脱，流速1．0mL·min^-1，二极管阵列检测器，检测波长275nm。结果吉西他滨主峰和各杂质峰均达到基线分离；胞嘧啶和吉西他滨分别在质量浓度0．4676—4．676mg·L^-1和0．4164—4．164mg·L^-1内呈良好线性，r分别为1．0000和0．9998；盐酸吉西他滨的最低检测限为0．1249ng。LC-MS结果显示碱破坏后得到的两个未知杂质为吉西他滨的同分异构体。结论本方法灵敏、准确，专属性强，为分析各个厂家的药品质量提供了一个科学合理的评价体系。

Determination of related substances in gemcitabine hydrochloride raw materials and gemcitabine hydrochloride for injection by HPLC

AIM To establish a method for the determination of related substances in gemcitabine hydrochloride raw materials and injection preparations by using gradient elution of HPLC and to make further studies on unknown impurities of base treated system solution by LC-MS. METHODS Ca column was adopted. Mobile phase A and B respectively consisted of different proportions of phosphate buffer solution （pH 2.5 ± 0.1） and methanol. The flow rate with gradient elution was 1.0 mL· min^-1. The diode army detector was used to detect, and the detection wavelength was 275 nm. RESULTS Gemeitabine hydroehloride and its related substance were obviously separated under the chromatographic system deseribed above. A good relationship was observed in the concentration range of 0.467 6 - 4.676 mg- L^- 1 （cytosine） and 0.416 4 - 4. 164 mg·L^- 1 （gemeitabine）, with the coefficient of 1.000 0 （cytosine） and 0. 999 8 （gemcitabine）. The limit of detection was 0.124 9 ng （gemeitabine）. CONCLUSION The method is sensitive, accurate and specific, and can provide a scientific evaluation for quality control of gemeitabine hydroehloride raw materials and injection preparations from different companies.