title: Chlorpromazine quantification in human plasma by UPLC-electrospray ionization tandem mass spectrometry. Application to a comparative pharmacokinetic study.
abstract: In the present study a method to quantify chlorpromazine in human plasma using cyclobenzaprine as the internal standard (IS) is described. The analyte and the IS were extracted from human plasma by a liquid-liquid extraction with diethyl ether/dichloromethane (70/30, v/v) and analyzed by an ultra performance liquid chromatography (UPLC) coupled to an electrospray tandem triple quadrupole mass spectrometer in positive mode (UPLC-ES(+)-MS/MS). Chromatography was performed isocratically on an Aquity UPLC BEH C18 1.7 μm (50 mm × 2.1 mm i.d.) operating at 40°C. The mobile phase was a mixture of 65% water+1% formic acid and 35% of acetonitrile at a flow-rate of 0.5 mL/min. The lowest concentration quantified was 0.5 ng/mL and a linear calibration curve over the range 0.5-200 ng/mL was obtained, showing intra-assay precisions from 2.4 to 5.8%, and inter-assay precisions from 3.6 to 9.9%. The intra-assay accuracies ranged from 96.9 to 102.5%, while the inter-assay accuracies ranged from 94.1 to 100.3%. This analytical method was applied in a relative bioavailability study in order to compare a test chlorpromazine 100 mg simple dose formulation versus a reference in 57 volunteers of both sexes. The study was conducted in an open randomized two-period crossover design and with a fourteen days washout period. Plasma samples were obtained over a 144-h interval. Since the 90% CI for both C(max), AUC(last) and AUC(0-inf) were within the 80-125% interval proposed by the Food and Drug Administration and ANVISA, it was concluded that chlorpromazine 100 mg/dose was bioequivalent to the reference formulation, according to both the rate and extent of absorption.
authors.affiliation: Synchrophar Assessoria e Desenvolvimento de Projetos Clínicos S/S Ltda, Campinas, SP, Brazil. medney@synchrophar.com
authors.forename: Ney Carter
authors.initials: NC
authors.lastname: Borges
authors.forename: Vinicius Marcondes
authors.initials: VM
authors.lastname: Rezende
authors.forename: Jose Marcos
authors.initials: JM
authors.lastname: Santana
authors.forename: Ricardo Pereira
authors.initials: RP
authors.lastname: Moreira
authors.forename: Roberto Fernandes
authors.initials: RF
authors.lastname: Moreira
authors.forename: Patrícia
authors.initials: P
authors.lastname: Moreno
authors.forename: Diego Carter
authors.initials: DC
authors.lastname: Borges
authors.forename: José Luiz
authors.initials: JL
authors.lastname: Donato
authors.forename: Ronilson Agnaldo
authors.initials: RA
authors.lastname: Moreno
mesh_terms: D000293:Adolescent; D000328:Adult; D000639:Amitriptyline / Q000031:analogs & derivatives; D019540:Area Under Curve; D001682:Biological Availability; D002746:Chlorpromazine / Q000097:blood* / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D018592:Cross-Over Studies; D004355:Drug Stability; D005260:Female; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D008875:Middle Aged; D011897:Random Allocation; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000639:Amitriptyline; C004704:cyclobenzaprine; D002746:Chlorpromazine