HPLC法测定索非布韦片的有关物质

Determination of related substances in sofosbuvir tablets by HPLC

目的:建立索非布韦片有关物质的HPLC法。方法:采用Waters Symmetry C18(4.6 mm×150 mm,5μm)色谱柱,流动相为0.01 mol·L^(-1)磷酸二氢钠溶液(用磷酸调节pH至3.0)-乙腈(75∶25),流速为1.0 m L·min^(-1),检测波长为210 nm,柱温30℃,进样量20μL,对有关物质进行定量分析。结果:在该色谱条件下,索非布韦与各杂质均能有效分离,杂质A(中间体)、杂质B(起始原料)、杂质C(降解产物)的定量限分别为1.012、2.005、2.010 ng,且在各自的线性范围内线性关系良好(r>0.999,n=7),杂质加样回收率(n=9)在96.11%~102.20%之间,重复性、精密度、稳定性等符合规定。对3批索非布韦片进行有关物质测定,结果总杂质量在0.108%~0.129%范围内。结论:方法学验证结果表明,本法可作为索非布韦片有关物质的测定。

Objective： To establish an HPLC method for determination of the related substances in sofosbuvir tablets. Methods： The test was based on an Waters Symmetry C18（5μm,4.6×150mm） column. The mobile phase consisted of 0.01 mol.L-1 sodium dihydrogen phosphate（adjusted to pH 3.0 with phosphoric acid） -acetonitrile （75：25） at a flow rate of 1.0 mL.min-1 and the detection wavelength was 210 nm.The column temperature was 30 ℃,and the volume of injection was 20μL. The related substances were quantified by the established method. Results： Sofosbuvir was separated completely from the impurities at this chromatographic conditions. The limits of impurity A（intermediate）, impurity B（material） and impurity C（degradation） were 1.012,2.005,2.010ng. The calibration curves of these impurities were linear in the self-concentration range（r〉0.999 ,n=7）.The recoveries of these impurities （n=9） were between 96.11% and 102.20%. The repeatability ,intermediate precision and stability were completely met the requirements. The total impurities were 0.108%～0.129% in 3 batches of sofosbuvir tablets. Conclusion： Methodology validation results proved that the established determination method can be used for quality control of sofosbuvir tablets.