维兰特罗关键中间体含量分析方法及其有关物质研究

Study on the Content Analysis Method of Key Intermediates of Vilanterol and Its Related Substances

摘要: 目的：为建立一种新的HPLC法对维兰特罗中间体5及其有关物质分析检测的方法。方法：条件采用C18纳谱分析色谱柱(250 × 4.6 mm, 5 μm)，流动相乙腈(A)-水(B)，等度洗脱(80% A)，流速1.0 mL∙min⁻¹，检测波长220 nm，经制备或半制备色谱分离有关物质，通过HPLC以及LC-MS分析，实现对中间体5及其有关物质进行定性和定量分析。结果：研究表明，中间体5在一定浓度范围内呈现出良好的线性关系(Y = 38705X + 21.565)，平均加样回收率为100.61%~104.81%，RSD < 0.2%，重复性好(RSD < 0.5%)，12 h内供试品溶液稳定(RSD < 2.0%)。利用此方法对中间体5及其有关物质含量进行测定，中间体5含量为98.0%~99.5%，主要杂质总含量为0.3%~0.6%，经定性结构分析判定杂质I (RT 1.47 min)为对甲苯磺酸，杂质III (RT 2.82 min)为中间体4，为合成中未除净的原料。结论：本研究为工艺优化、有效降低或去除杂质及关键中间体的质量控制提供理论和方法支撑。

Abstract: Objective: In order to establish a new HPLC method for the determination of intermediate 5 of vilanterol and its related substances. Methods: The chromatographic analysis was performed on C18 nanoscopic column (250 × 4.6 mm, 5 μm) with mobile phase acetonitrile (A)-water (B) and isode-gree elution (80% A), and the flow rate was 1.0 min∙mL⁻¹. The detection wavelength was 220 nm, and the related substances were separated by preparative or semi-preparative chromatography. The qualitative and quantitative analysis of intermediate 5 and its related substances was realized by HPLC and LC-MS analysis. Results: The results showed that the intermediate 5 showed a good linear relationship in a certain concentration range (Y = 38705X + 21.565), the average recovery was 100.61%~104.81%, RSD < 0.2%, and the repeatability was good (RSD < 0.5%). The solution was stable within 12 h (RSD < 2.0%). The content of intermediate 5 and its related substances was determined by this method. The content of intermediate 5 was 98.0%~99.5%, and the total content of major impurities was 0.3%~0.6%. The impurity I (RT 1.47 min) was determined to be p-toluenesulfonic acid by qualitative structural analysis, and the impurity III (RT 2.82 min) was determined to be intermediate 4. It is the raw material not cleaned in synthesis. Conclusion: This study provides theoretical and methodological support for process optimization, effective reduction or removal of impurities and quality control of key intermediates.