title: Highly sensitive assay for tiotropium, a quaternary ammonium, in human plasma by high-performance liquid chromatography/tandem mass spectrometry.
abstract: 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.
authors.affiliation: Research Center for Drug Metabolism, College of Life Science, Jilin University, Changchun 130023, China.
authors.forename: Jiang
authors.initials: J
authors.lastname: Wang
authors.forename: Yao
authors.initials: Y
authors.lastname: Jiang
authors.forename: Yingwu
authors.initials: Y
authors.lastname: Wang
authors.forename: Hao
authors.initials: H
authors.lastname: Li
authors.forename: J Paul
authors.initials: JP
authors.lastname: Fawcett
authors.forename: Jingkai
authors.initials: J
authors.lastname: Gu
mesh_terms: D000328:Adult; D001993:Bronchodilator Agents / Q000097:blood* / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid; D002976:Clenbuterol / Q000097:blood; D006801:Humans; D007202:Indicators and Reagents; D008297:Male; D012015:Reference Standards; D015203:Reproducibility of Results; D012602:Scopolamine Derivatives / Q000097:blood* / Q000493:pharmacokinetics; D053719:Tandem Mass Spectrometry; D000069447:Tiotropium Bromide
chemical_list: D001993:Bronchodilator Agents; D007202:Indicators and Reagents; D012602:Scopolamine Derivatives; D002976:Clenbuterol; D000069447:Tiotropium Bromide