目的 建立大鼠血浆及肺部组织中溴己新浓度测定的LC-MS/MS方法，并对大鼠给予盐酸溴己新雾化吸入溶液受试制剂与参比制剂后药物在血浆及肺部组织分布进行比较。方法 样品处理采用简便的蛋白沉淀法，内标为苯海拉明，采用ZORBAX Eclipse XDB-C₁₈色谱柱，有机相为甲醇，水相为0.5%甲酸-5%甲醇水溶液，采用梯度程序进行洗脱，在电喷雾离子化源（ESI）的正离子模式下以多重反应监测（MRM）方式进行检测。对血浆和肺组织中溴己新的分析方法分别进行了完整验证和部分验证。健康SD大鼠随机分为两组，分别雾化吸入给予盐酸溴己新雾化吸入溶液受试及参比制剂，定时采集血浆、肺、气管支气管并测定溴己新的浓度。结果 血浆及肺组织中溴己新浓度测定的分析方法验证各项均符合指导原则规定。受试制剂与参比制剂在大鼠血浆、肺、气管支气管中的分布速度、分布程度基本一致，在主要靶部位气管支气管中的相对生物利用度为97.4%。结论 建立的LC-MS/MS分析方法快速、灵敏、简便，两种制剂在大鼠血浆及肺部组织分布具有一致性，可为该新剂型的国产化提供数据支持。

Objective To establish an LC-MS/MS method for the determination of bromhexine in rat plasma and lung, and the distribution in plasma and lung tissue was compared between the test and the reference formulations of bromhexine hydrochloride inhalation solution in rats. Methods Samples were pretreated by a simple protein precipitation method with diphenhydramine as the internal standard. Chromatographic separation was performed on a ZORBAX Eclipse XDB-C₁₈ column using a gradient elution program with solvents consisting of methanol and 0.5% formic acid-5% methanol aqueous solution. The detection was operated in the positive ion mode using multiple reaction monitoring (MRM) with an electrospray ionization source (ESI). The analytical methods for plasma and lung tissue were fully and partially validated, respectively. Healthy Sprague-Dawley rats were divided into two groups randomly and administered by inhalation with test and reference formulations of bromhexine hydrochloride inhalation solution, respectively. The plasma, lung and tracheobronchus were collected at predetermined time points and the concentration of bromhexine was measured. Results Bioanalytical method validation results all met the criteria of the guidelines. The distribution rate and extent of bromhexine in plasma, lung and tracheobronchus between the two formulations were consistent. The relative bioavailability in the main target organ tracheobronchus was about 97.4%. Conclusion The LC-MS/MS method established in this study is rapid, sensitive and simple. The distribution of bromhexine between the two formulations in rat plasma and lung tissue is consistent, which can provide data support for the localization of the new formulation.

R969.1

中国医学科学院医学与健康科技创新工程项目资助（2019-I2M-5-020）