Wang Jiang, Jiang Yao, Wang Yingwu, Li Hao, Fawcett J Paul, Gu Jingkai
Research Center for Drug Metabolism, College of Life Science, Jilin University, Changchun 130023, China.
Rapid Commun Mass Spectrom. 2007;21(11):1755-8. doi: 10.1002/rcm.3021.
Tiotropium bromide, a long-acting inhaled bronchodilator analogous to ipratropium bromide, is currently undergoing development for the treatment of chronic obstructive pulmonary disease. To evaluate its systemic absorption in humans, we have developed a rapid and sensitive method for its determination in human plasma based on high-performance liquid chromatography with tandem mass spectrometric detection (HPLC/MS/MS). Reversed-phase chromatography of tiotropium and the internal standard clenbuterol was carried out using acetonitrile/10 mM ammonium acetate (1% formic acid) 40:60 as mobile phase in a run time of 3.0 min. The sample preparation involved deproteination with acetonitrile, extraction into dichloromethane and back-extraction into hydrochloric acid. The assay was linear over the concentration range 0.500-50.0 pg/mL with intra- and inter-day precision (as relative standard deviation) both <or=7.34%. The method was successfully applied to a pharmacokinetic study of systemic absorption in healthy male volunteers given a single 18 microg inhaled dose.
噻托溴铵是一种类似于异丙托溴铵的长效吸入性支气管扩张剂，目前正处于治疗慢性阻塞性肺疾病的研发阶段。为评估其在人体中的全身吸收情况，我们基于高效液相色谱-串联质谱检测（HPLC/MS/MS）开发了一种快速灵敏的方法来测定人血浆中的噻托溴铵。使用乙腈/10 mM醋酸铵（1%甲酸）40:60作为流动相，在3.0分钟的运行时间内对噻托溴铵和内标克伦特罗进行反相色谱分析。样品制备包括用乙腈进行蛋白沉淀，萃取到二氯甲烷中，然后反萃取到盐酸中。该测定法在0.500 - 50.0 pg/mL的浓度范围内呈线性，日内和日间精密度（以相对标准偏差计）均≤7.34%。该方法已成功应用于给予单次18微克吸入剂量的健康男性志愿者的全身吸收药代动力学研究。
