溴己新LC-MS/MS法测定及大鼠吸入给药后分布动力学研究  

Determination of bromhexine by LC-MS/MS and comparative study of distribution kinetics in rats after inhalation administration

作　　者：杨俊凤 刘全国 赵琳 李冬芝 辛飞洋 夏媛媛 胡金芳 YANG Junfeng;LIU Quanguo;ZHAO Lin;LI Dongzhi;XIN Feiyang;XIA Yuanyuan;HU Jinfang(Tianjin Tiancheng New Drug Evaluation Co.,Ltd.,Tianjin 300301,China;National Key Laboratory of Druggability Evaluation and Systematic Translational Medicine,Tianjin Pharmaceutical Research Institute Co.,Ltd.,Tianjin 300462,China;Research Unit for Drug Metabolism,Chinese Academy of Medical Sciences,Beijing 100050,China;Hainan Huluwa Pharmaceutical Group Co.,Ltd.,Haikou 570100,China)

机构地区：[1]天津天诚新药评价有限公司,天津300301 [2]天津药物研究院有限公司药物成药性评价与系统转化全国重点实验室,天津300462 [3]中国医学科学院药物代谢新技术创新单元,北京100050 [4]海南葫芦娃药业集团股份有限公司,海南海口570100

出　　处：《药物评价研究》2024年第9期2082-2089,共8页Drug Evaluation Research

基　　金：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)。

摘　　要：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.

关 键 词：LC-MS/MS 溴己新 吸入给药 分布动力学 

分 类 号：R969.1[医药卫生—药理学]