HPLC法测定阿伐那非中间体4-(3-氯-4-甲氧基苄胺基)-2-甲硫基-5-乙氧羰基嘧啶的有关物质

Determination of Related Substances in 4-(3-Chloro-4-methoxybenzylamino)-2-methylthio-5-ethoxycarbonyl pyrimidine as Intermediate of Avanafil by HPLC

建立高效液相色谱法测定阿伐那非中间体4-(3-氯-4-甲氧基苄胺基)-2-甲硫基-5-乙氧羰基嘧啶(M-1)的有关物质。以C18为填充物的不锈钢柱及紫外检测器,在254 nm波长下,以0.02 mol/L磷酸二氢钾水溶液(用1 mol/L氢氧化钾溶液调节pH=5.5)-乙腈进行梯度洗脱,以主成分自身对照法计算。M-1与4个已知杂质均可达到基线分离,在各自浓度范围内线性关系良好,精密度试验的RSD均不大于2%,平均回收率99.16%~100.44%,RSD 0.69%~1.03%,检测限0.020~0.15μg/mL,定量限0.061~0.45μg/mL。该方法操作简便,准确度、专属性良好,可满足中间体M-1的质量控制要求。

To establish a HPLC method for the determination of related substances in 4-(3-chloro-4-methoxybenzylamino)-2-methylthio-5-ethoxycarbonyl pyrimidine(M-1)as an intermediate of Avanafil.The determination was performed on a C18 column and UV absorbance detector at a wavelength of 254 nm,0.02 mol/L potassium dihydrogen phosphate solution(pH was adjusted to 5.5 with 1 mol/L potassium hydroxide solution)and acetonitrile were used as the gradient elution reagents.The principal component self-control comparison method was used to calculate the content of the impurities.M-1 and four known impurities could achieve a baseline separation,and the linear relationship was good within their respective concentration ranges;RSD of precision test was no more than 2%,the average recovery rate was 99.16%~100.44%;and the RSD was 0.69%~1.03%;the limit of detection was 0.020~0.15μg/mL;the limit of quantification was 0.061~0.45μg/mL.This method was accurate,specific,simple and easy to operate,which was suitable for the quality control of the M-1.