HPLC法测定马来酸氨氯地平原料药有关物质

Determination of related substances in amlodipine maleate raw material by HPLC

目的建立马来酸氨氯地平原料中有关物质的HPLC测定方法。方法采用Waters SunFire C18(250 mm×4.6 mm,5μm)色谱柱;以甲醇乙腈体积分数0.7%三乙胺溶液(取三乙胺7.0 mL,加水稀释至1000 mL,用磷酸调节pH值至3.0±0.1)(V∶V∶V=35∶15∶50)为流动相,流速1.0 mL·min^(-1),柱温30℃;检测波长237 nm。结果氨氯地平与各已知杂质及强降解产物均分离度良好,杂质Ⅰ、Ⅱ、Ⅲ、Ⅳ的质量浓度分别在0.29~9.51 mg·L^(-1)(r=1.000),0.29~9.51 mg·L^(-1)(r=0.9999),0.30~9.51 mg·L^(-1)(r=0.9995)和0.29~9.51 mg·L^(-1)(r=0.9999)范围内与峰面积呈良好的线性关系,校正因子分别为2.65、1.38、1.00、1.34;检测限分别为1.01、4.02、0.49、0.50 ng,定量限分别为4.03、10.01、1.46、1.98 ng。杂质I、杂质Ⅱ、杂质Ⅲ及杂质Ⅳ的回收率(n=9)依次为98.94%、101.63%、101.67%、101.79%,RSD依次为1.45%、1.87%、1.41%、1.43%。3批马来酸氨氯地平样品中已知杂质及未知杂质均小于0.1%,杂质总量均小于0.6%。结论该方法可用于马来酸氨氯地平原料药中有关物质的检测。

Objective To establish an HPLC method for the determination of related substances in amlodipine maleate raw material.Methods HPLC was adopted on a C18 column(250 mm×4.6 mm,5μm)with a mobile phase consisting of a mixture of methanol-acetonitrile-0.7%triethylamine solution(7.0 mL of triethylamine was dissolved in 1000 mL of water,and pH was adjusted to 3.0±0.1 with phosphoric acid)(V∶V∶V=35∶15∶50).The flow rate was 1.0 mL·min^(-1).The column temperature was maintained at 30℃and the detective wavelength was 237 nm.Results Amlodipine maleate was well separated from the specified impurities and the forced degradation products.There was a good linearity separately over the ranges 0.29-9.51 mg·L^(-1)(r=1.000)of impurityⅠ,0.29-9.51 mg·L^(-1)(r=0.9999)of impurityⅡ,0.30-9.51 mg·L^(-1)(r=0.9995)of impurityⅢ,and 0.29-9.51 mg·L^(-1)(r=0.9999)of impurityⅣ.The correction factors of 4 impurities were 2.65,1.38,1.00 and 1.34;the limits of detection were 1.01,4.02,0.49 and 0.50 ng,respectively;the limits of quantitation were 4.03,10.01,1.46,1.98 ng,respectively;the recovery rates were 98.94%,101.63%,101.67%and 101.79%,respectively;RSDs were 1.45%,1.87%,1.41%and 1.43%(n=9).The results of related substances in three batches of samples showed that the contents of specified impurities and unknown impurities were less than 0.1%,total impurities were less than 0.6%.Conclusion This method has good specificity and high sensitivity,and can be used for the determination of related substances in amlodipine maleate raw material.