第 52 卷第 6 期 Vol. 52 No. 6 2022 年 12 月 Dec 2022

所属栏目：分析与测试 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

王 帆，杨 琪，李 帅，郭新艳，孙 清 （山东省药学科学院 山东省生物药物重点实验室，山东 济南 250101）

摘　要：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0.020~0.15 µg/mL，定量限0.061~0.45 µg/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。

关键词：阿伐那非；中间体；有关物质；高效液相色谱法

中图分类号：R977.1　　文献标识码：A　　文章编号：1009-9212（2022）06-0085-05

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

WANG Fan, YANG Qi, LI Shuai, GUO Xin-yan, SUN Qing (Shandong Academy of Pharmaceutical Sciences, Key Laboratory for Biological Drug Research of Shandong Province, Jinan 250101, China)

Abstract：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 0.020~0.15 μg/mL; the limit of quantification was 0.061~0.45 μg/mL. This method was accurate, specific, simple and easy to operate, which was suitable for the quality control of the M-1.

Key words：Avanafil; intermediates; related substances; HPLC

基金项目：山东省重点研发计划（重大科技创新工程项目）（2021ZDSYS07）。

作者简介：王 帆（1986—），女，辽宁朝阳人，主管药师，硕士，研究方向：新药研发（E-mail：sail-163@163.com）。

收稿日期：2022-07-07