title: Performance of tiloronoxim and tilorone determination in human blood by HPLC-MS/MS: method validation, uncertainty assessment and its application to a pharmacokinetic study.
abstract: A highly sensitive and selective HPLC-MS/MS method is presented for the quantitative determination of tiloronoxim and its metabolite tilorone in human blood. An aliquot of 200 microl human blood was extracted with a mixture of chloroform/ethyl ether (1/2, v/v), using metoprolol as the internal standard (the IS). Separation was achieved on an Xterra MS C18 column (50 mm x 2.1 mm, 5 microm) with a gradient mobile phase of methanol/water containing 15 mM ammonium bicarbonate (pH 10.5). Detection was performed using positive MRM mode on a TurboIonSpray source. The mass transitions monitored were m/z 426.3-->100.0, m/z 411.3-->100.0 and m/z 268.3-->116.1 for tiloronoxim, tilorone and the IS, respectively. The method was fully validated using total error theory, which is based on beta-expectation tolerance intervals and include trueness and intermediate precision. The method was found to be accurate over a concentration range of 1-100 ng/ml for both compounds. The measurement uncertainty based on beta-expectation tolerance intervals was assessed at each concentration level of the validation standards. This method was successively applied to a pharmacokinetic study of tiloronoxim in healthy volunteers.
authors.affiliation: Department of Pharmacy, Peking University Third Hospital, 49 North Garden Rd., Beijing 100191, PR China.
authors.forename: Xianhua
authors.initials: X
authors.lastname: Zhang
authors.forename: Jingli
authors.initials: J
authors.lastname: Duan
authors.forename: Suodi
authors.initials: S
authors.lastname: Zhai
authors.forename: Yiheng
authors.initials: Y
authors.lastname: Yang
authors.forename: Li
authors.initials: L
authors.lastname: Yang
mesh_terms: D000998:Antiviral Agents / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D057230:Limit of Detection; D013058:Mass Spectrometry / Q000379:methods*; D010091:Oximes / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics*; D012044:Regression Analysis; D015203:Reproducibility of Results; D013994:Tilorone / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics; D013997:Time Factors; D035501:Uncertainty
chemical_list: D000998:Antiviral Agents; D010091:Oximes; C539624:tiloronoxim; D013994:Tilorone