HPLC法测定利伐沙班片的含量和有关物质

目的: 建立利伐沙班片的含量和有关物质的HPLC法。方法: 采用RP-HPLC法测定利伐沙班片中利伐沙班的含量,并对其有关物质进行检测。色谱柱为Ultimate LP-C₁₈(250 mm×4.6 mm,5 μm),乙腈-0.01 mol·L^-1磷酸溶液为流动相梯度洗脱,流速1.0 mL·min^-1,柱温30 ℃,检测波长250 nm;同时采用NP-HPLC法测定利伐沙班片中R-利伐沙班的含量,色谱柱为CHIRALPAK AD-H(250 mm×4.6 mm,5 μm),以甲基叔丁基醚-异丙醇-二乙胺(70: 30: 0.1)为流动相,流速0.8 mL·min^-1,柱温30 ℃,检测波长250 nm。结果: RP-HPLC法中主峰和相邻杂质峰能完全分离,利伐沙班的线性范围为0.009 6~0.167 mg·mL^-1,平均回收率(n=9)为99.7%;NP-HPLC法中R-利伐沙班与利伐沙班能完全分离,R-利伐沙班的线性范围为1.78~5.35 μg·mL^-1,平均回收率(n=9)为100.7%。3批样品含量测定结果分别为99.1%、99.9%、100.2%,单个最大杂质分别为0.03%、0.06%、0.14%,总杂质分别为0.10%、0.13%、0.19%;3批样品中R-利伐沙班的含量分别为0.023%、0.021%、0.020%。结论: 经方法学验证,本方法可用于利伐沙班片的质量控制。

Objective: To establish an HPLC method of the content and related substances of rivaroxaban tablets. Methods: RP-HPLC was adopted to determine rivaroxaban,related substances,content uniformity and dissolution using an Ultimate LP-C₁₈ column(250 mm×4.6 mm,5 μm)with a gradient elution system of acetonitrile-0.01 mol·L^-1 phosphoric acid,while the column temperature was maintained at 30 ℃,the flow rate was 1.0 mL·min^-1 and the detection wavelength was 250 nm. NP-HPLC was adopted to determine R-rivaroxaban in rivaroxaban tablets using an CHIRALPAK AD-H column(250 mm×4.6 mm,5 μm)with a mobile phase consisting of tert-butyl methyl ether-isopropanol-diethylamine(70: 30: 0.1).The column temperature was maintained at 30 ℃,the flow rate was 0.8 mL·min^-1 and the detection wavelength was 250 nm. Results: Related substances were completely separated from the main constituents in RP-HPLC. The standard curve of rivaroxaban was linear over the range of 0.009 6-0.167 mg·mL^-1,with the average recovery of 99.7%(n=9). R-rivaroxaban was completely separated from rivaroxaban in NP-HPLC. The standard curve of R-rivaroxaban was linear over the range of 1.78-5.35 μg·mL^-1,with the average recovery of 100.7%(n=9). The content results of 3 batches were 99.1%,99.9% and 100.2%,respectively;the maximum single impurity was 0.03%,0.06% and 0.14%,respectively;the total impurities were 0.10%,0.13% and 0.19%,respectively. The content results of R-rivaroxaban in the 3 batches were 0.023%,0.021% and 0.020% respectively. Conclusion: The method is proved by the methodology validation that it can be used for the quality control of rivaroxaban tablets.