HPLC-DAD/MS法测定辛伐他汀胶囊中有关物质

Determination of related substances in simvastatin capsules by HPLC- DAD /MS

目的： 采用HPLC-DAD/MS法测定辛伐他汀胶囊中的有关物质，对已知杂质A、B、C、D、E、F、G进行方法学研究，并对本品中的有关物质进行归属分析。 方法： 采用C18柱（33 mm×4.6 mm，3 μm），以乙腈-0.1%磷酸溶液（50：50）为流动相A，0.1%磷酸的乙腈溶液为流动相B，梯度洗脱，流速为3.0 mL·min^-1；检测波长为238 nm。 结果： 辛伐他汀与各杂质及强制破坏产物能完全分离，主峰纯度因子均大于999.0；辛伐他汀及各杂质在低浓度范围内线性关系良好；各杂质加样回收率在98.8%-102.9%之间；进样精密度、重复性、中间精密度均符合要求；溶剂、空白辅料无干扰；辛伐他汀及杂质A、B、C、D、E、F、G的定量限分别为0.20、0.78、0.80、0.60、1.01、0.51、0.62、0.65 ng，相对校正因子分别为1.07、1.06、1.06、2.84、0.97、0.96、1.00。 结论： 本方法经方法学验证，可用于辛伐他汀及其制剂的有关物质检查。

Objective： To develop the HPLC-DAD/MS method for the determination of related substances in simvastatin capsules,including the validation of known impurities A,B,C,D,E,F and G and the attribution analysis of related substances in the product. Methods： The analysis was performed on a C18 column （33 mm×4.6 mm,3 μm） with mobile phases A （acetonitrile-0.1% phosphoric acid solution,50： 50） and B （0.1% phosphoric acid in acetonitrile） by gradient elution at the flow rate of 3.0 mL·min^-1,and the detection wavelength was set at 238 nm. Results： Simvastatin was separated completely from known impurities and degradation products under various conditions with the purity factor greater than 999.0.Simvastatin and impurities A,B,C,D,E,F,G revealed good linearities over the ranges of 0.7924-39.62 μg·mL^-1 （r=1.000）,0.3118-6.236 μg·mL^-1 （r=0.9999）,0.3204-6.408 μg·mL^-1 （r=0.9999）,0.3016-6.032 μg·mL^-1 （r=1.000）,0.3296-6.592 μg·mL^-1 （r=1.0000）,0.2032-4.064 μg·mL^-1 （r=0.9997）,0.2056-4.112 μg·mL^-1 （r=0.9999）,0.3232-6.464 μg·mL^-1 （r=0.9999）,respectively.The average recoveries of the above impurities were 101.9%,102.2%,99.8%,102.9%,98.8%,101.5%,99.2%,respectively.Sampling precision,repeatability and intermediate precision all met the requirements and the blank test showed no interference.The limits of quantification of simvastatin and impurities A,B,C,D,E,F,G were 0.20,0.78,0.80,0.60,1.01,0.51,0.62,0.65 ng and the relative correction factors were 1.07,1.06,1.06,2.84,0.97,0.96,1.00,respectively.Conclusion： This method can be used for the determination of simvastatin and its preparations by methodological validation.