目的: 建立雷芬那辛雾化吸入溶液的有关物质控制方法。方法: 采用2D LC-HRMS对降解杂质进行结构确证,结合可能的降解途径,拟定控制的特定杂质。采用InfinityLab Poroshell 120 C18（250 mm×4.6 mm,4 μm）色谱柱,以0.02 mol · L-1磷酸二氢钾溶液（用2 mol · L-1氢氧化钾溶液调pH至7.5）-甲醇（93 ∶ 7）为流动相A,乙腈为流动相B,梯度洗脱,流速1.0 mL · min-1,检测波长230 nm,柱温35 ℃,采用加校正因子的主成分自身对照法进行杂质的定量分析。结果: 本品主要降解途径为氧化反应和水解反应,主要降解产物为杂质A、杂质B和杂质C。在拟定的色谱条件下,雷芬那辛主成分色谱峰与各杂质峰完全分离,各辅料成分不干扰主成分及有关物质的测定;杂质A、杂质B和杂质C的相对校正因子分别为1.27、1.08、1.06。多批次样品0 d未检出大于报告限度的杂质,在高温放置30 d,杂质B含量为0.31%,最大未知杂质含量为0.15%,总杂含量为1.06%。结论: 该方法专属性强,重复性好,可用于雷芬那辛雾化吸入溶液中有关物质的检查。
Objective: To establish a methodology for the control of related substances in revefenacin nebulizer solution. Methods: 2D LC-HRMS was employed to confirm the structure of the degraded impurities, and specific impurities were selected based on the possible degradation pathways. An InfinityLab Poroshell 120 C18(250 mm× 4.6 mm, 4 μm) column was utilized, with 0.02 mol · L-1 potassium dihydrogen phosphate solution (pH was adjusted to 7.5 by 2 mol · L-1 potassium hydroxide solution)-methanol (93 ∶ 7) serving as mobile phase A. Acetonitrile was employed as mobile phase B, with gradient elution at a flow rate of 1.0 mL · min-1. The detection wavelength was set at 230 nm and the column temperature was maintained at 35 ℃. The impurities were quantitatively analyzed by the principal component self-control method with correction factor. Results: The main degradation pathways were oxidation reaction and hydrolysis reaction, and the main degradation products were impurity A, impurity B and impurity C. Under the proposed chromatographic conditions, the principal component peak of revefenacin was completely separated from each impurity peak, and each excipient component did not interfere with the determination of the principal component and related substances. The relative correction factors of impurity A, impurity B and impurity C were 1.27, 1.08 and 1.06, respectively. No impurities exceeding the reported limit were detected in multiple batches of samples on 0 d, and the content of impurity B was 0.31%, the maximum unknown impurity content was 0.15%, and the total impurity was 1.06% after 30 d in high temperature. Conclusion: The method exhibits strong specificity and good repeatability, and can be applied on the detection of related substances in revefenacin nebulizer solution.
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