HPLC法测定盐酸乌拉地尔原料药中有关物质

Determination of related substances in urapidil hydrochloride by HPLC

目的建立盐酸乌拉地尔原料药中有关物质的HPLC检测方法。方法采用AgilentXDB C18色谱柱(250mm×4.6 mm,5μm),以0.05 mol·L^(−1)磷酸二氢铵溶液(含0.02%三乙胺,磷酸调pH值至6.0)为流动相A,乙腈为流动相B,梯度洗脱;体积流量为1.0 mL·min^(−1),柱温为30℃,检测波长为270、236 nm,进样量为10μL。进行方法专属性、稳定性、检测限与定量限、线性关系、精密度、耐用性考察。应用所建立的方法进行3批盐酸乌拉地尔原料药样品检测。结果盐酸乌拉地尔与相邻杂质之间、各杂质之间的色谱峰均分离良好;各个组分的检测限和定量限分别为0.0597~0.0900μg·mL^(−1)、0.1989~0.2999μg·mL^(−1)。各组分在相应浓度范围内均有良好的线性关系,r均在0.9998~1.0000。加样回收率均在92.2%~103.2%,RSD均≤5.37%。3批盐酸乌拉地尔原料药样品检测结果显示,已知杂质质量分数均小于0.15%、最大未知单杂均小于0.10%,杂质总质量分数小于0.50%。结论经方法学验证,所建立方法灵敏、高效、专属性强、准确度高,可用于盐酸乌拉地尔原料药有关物质的测定。

Objective To establish an HPLC method for the determination of related substances in urapidil hydrochloride.Method Agilent XDB C18(250 mm×4.6 mm,5μm)was chosen for analysis.Gradient elution was used to determine the related substances of urapidil hydrochloride with 0.05 mol·L^(−1)ammonium dihydrogen phosphate(containing 0.02%triethylamine,and pH adjusted to 6.0 with phosphoric acid),and acetonitrile as mobile phase B.The flow rate was 1.0 mL·min^(−1)and the column temperature was 30℃.The detection wavelengths were 270,236 nm,and the sample injection volume was 10μL.The specificity,stability,limit of detection and limit of quantification,linear relationship,precision and durability of the method were investigated.Three batches of urapidil hydrochloride API samples were detected by the established method.Results The resolution between urapidil and all other impurities were good.The limits of detection of urapidil hydrochloride and impurities were 0.0597—0.0900μg·mL^(−1)and the limits of quantitation of them were 0.1989—0.2999μg·mL^(−1).All known impurities had good linearity in appropriate concentration ranges and the correlation coefficients were all in the range of 0.9998—1.0000.The average recoveries of all known impurities were in the range of 92.2%—103.2%,with RSD of less 5.37%.The determination results of three batches of urapidil hydrochloride samples showed that the known impurities and the maximum unknown impurity were less than 0.10%and the total impurities were less than 0.5%.Conclusion The method was simple,rapid and accurate,which can be used to control the quality of urapidil hydrochloride.