依帕司他原料药有关物质的HPLC测定方法改进研究

Improvement Research of Determination for the Related Substances in Epalrestat by HPLC

目的采用改进的高效液相色谱法测定依帕司他原料药有关物质。方法采用依利特HypersilBDSC18色谱柱(250 mm×4.6 mm,5μm),以乙腈-pH6.5磷酸盐缓冲液(0.025 mol·L-1 KH2PO4+0.025 mol·L-1 Na2HPO4,用H3PO4调节pH至6.5)为流动相,梯度洗脱,流速为1.0 mL·min-1,检测波长为396 nm和280 nm,柱温为30℃,进样量为20μL。结果依帕司他与相邻杂质能较好地分离。立体异构体杂质在浓度0.010~30μg·mL-1内线性关系良好,检测限和定量限分别为0.80,2.4 ng·mL-1。精密度、稳定性、准确度、耐用性均符合要求。结论该方法可用于依帕司他原料药有关物质的检查。

OBJECTIVE To establish an improved HPLC method for the determination of the related substances in epalrestat.METHODS The chromatographic seperation was achieved on an Elite Hypersil BDS C18 column(250 mm×4.6 mm,5μm)with the mobile phase consisted of acetonitrile-phosphate buffer salt(0.025 mol·L-1 KH2PO4+0.025 mol·L-1 Na2HPO4)at pH 6.5 adjusted with phosphoric acid,used gradient elution at the flow rate of 1.0 mL·min-1.The detection wavelengths were set at 396 nm and 280 nm,and the injection volume was 20μL.The column temperature was maintained at 30℃.RESULTS The stereoisomer had good linear relationship within the range of concentration of 0.010-30μg·mL-1,the LOD and LOQ of stereoisomer was 0.80 and 2.4 ng·mL-1,respectively.Other validation parameters,including precision,sample stability,accuracy and robustness were all within the acceptance range.CONCLUSION The developed method is successfully applied to the determination of related substances in epalrestat.