title: Development and validation of a UHPLC-MS/MS bioanalytical method to quantify in plasma the analgesic candidate PT-31 following a preliminary pharmacokinetic study in rats.
abstract: A selective and sensitive UHPLC-MS/MS bioanalytical method to determine PT-31, an analgesic drug candidate, in rat plasma was developed and validated. Analyses were performed using a UHPLC-MS/MS system equipped with an electrospray ionization interface operating in the positive ionization mode using a C18 reversed-phase column with a mobile phase of water:acetonitrile (68:31, v/v) containing 0.1% acetic acid eluting in a gradient mode with a flow rate of 0.3 mL/min. Plasma samples were deproteinized with cold acetonitrile containing 0.01% TFA (1:2, v/v) and 50 μL of the supernatant were injected into the system. PT-31 and phenytoin (internal standard) retention times were roughly 1.0 and 1.5 min, respectively. Linear standard curves were plotted for the 0.01-10 µg/mL concentration range, with a coefficient of determination > 0.99. The method's precision was over 88%. Maximum intra- and inter-day relative standard deviations were 14.6% and 11.6%, respectively. Interfering substances were not detected in the chromatogram, indicating that the method was specific. PT-31 stability was assessed under different temperature and storage settings. The method was used to characterize PT-31 plasma pharmacokinetics following administration of 5 mg/kg i.v. to Wistar rats. Therefore, the method described is sensitive, linear, precise and specific enough to determine PT-31 in preclinical pharmacokinetic investigations. Copyright © 2015 John Wiley & Sons, Ltd.
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
authors.forename: T C
authors.initials: TC
authors.lastname: Bessegato
authors.affiliation: Núcleo de Pesquisa de Produtos Naturais e Sintéticos, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: M
authors.initials: M
authors.lastname: Niehues
authors.affiliation: Núcleo de Pesquisa de Produtos Naturais e Sintéticos, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: G A
authors.initials: GA
authors.lastname: Buqui
authors.affiliation: Núcleo de Pesquisa de Produtos Naturais e Sintéticos, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: N P
authors.initials: NP
authors.lastname: Lopes
authors.affiliation: Núcleo de Pesquisa em Inovação Terapêutica, Universidade Federal de Pernambuco, Recife, PE, Brazil.
authors.forename: I R
authors.initials: IR
authors.lastname: Pitta
authors.affiliation: Núcleo de Pesquisa em Inovação Terapêutica, Universidade Federal de Pernambuco, Recife, PE, Brazil.
authors.forename: S L
authors.initials: SL
authors.lastname: Galdino
authors.affiliation: Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
authors.forename: T
authors.initials: T
authors.lastname: Dalla Costa
mesh_terms: D000700:Analgesics / Q000097:blood* / Q000493:pharmacokinetics; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D048289:Imidazolidines / Q000097:blood / Q000493:pharmacokinetics; D057230:Limit of Detection; D051381:Rats; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: C549017:3-(2-chloro-6-fluorobenzyl)-imidazolidine-2,4-dione; D000700:Analgesics; D048289:Imidazolidines
keywords: PT- 31; UHPLC-MS/MS PT-31; analgesic; bioanalytical method validation; preclinical pharmacokinetics