title: An improved analytical strategy combining microextraction by packed sorbent combined with ultra high pressure liquid chromatography for the determination of fluoxetine, clomipramine and their active metabolites in human urine.
abstract: A powerful and sensitive method, by microextraction packed sorbent (MEPS), and ultra-high performance liquid chromatography (UHPLC) with a photodiode array (PDA) detection, is described for the determination of fluoxetine, clomipramine and their active metabolites in human urine samples. The MEPS variables, such as sample volume, pH, number of extraction cycles (draw-eject), and desorption conditions (solvent and solvent volume of elution) were optimized. The analysis were carried out using small sample volumes (500μL) and in a short time period (5min for the entire sample preparation step). Good linearity was obtained for all antidepressants with the correlation coefficients (R(2)) above 0.9965. The limits of detection (LOD) ranged from 0.068 to 0.087μgmL(-1). The recoveries were from 93% to 98%, with relative standard deviations less than 6%. The inter-day precision, expressed as the relative standard deviation, varied between 3.8% and 8.5% while the intra-day precision between 3.0% and 7.1%. In order to evaluate the proposed method for clinical use, the MEPS/UHPLC-PDA method was applied to analysis of urine samples from depressed patients.
authors.affiliation: CQM-UMa, Centro de Química da Madeira, Centro de Ciências Exactas e da Engenharia da Universidade da Madeira, Campus Universitário da Penteada, 9000-390 Funchal, Portugal; Faculdade de Medicina da Universidade de Coimbra, Azinhaga de Santa Comba, Celas, 3000-548 Coimbra, Portugal.
authors.forename: Vera
authors.initials: V
authors.lastname: Alves
authors.affiliation: CQM-UMa, Centro de Química da Madeira, Centro de Ciências Exactas e da Engenharia da Universidade da Madeira, Campus Universitário da Penteada, 9000-390 Funchal, Portugal; Faculdade de Medicina da Universidade de Coimbra, Azinhaga de Santa Comba, Celas, 3000-548 Coimbra, Portugal.
authors.forename: João
authors.initials: J
authors.lastname: Gonçalves
authors.affiliation: CQM-UMa, Centro de Química da Madeira, Centro de Ciências Exactas e da Engenharia da Universidade da Madeira, Campus Universitário da Penteada, 9000-390 Funchal, Portugal.
authors.forename: Carlota
authors.initials: C
authors.lastname: Conceição
authors.affiliation: Faculdade de Medicina da Universidade de Coimbra, Azinhaga de Santa Comba, Celas, 3000-548 Coimbra, Portugal; Instituto Nacional de Medicina Legal e Ciências Forenses, I.P., Delegação Centro, Largo da Sé Nova, 3000-213 Coimbra, Portugal.
authors.forename: Helena M
authors.initials: HM
authors.lastname: Teixeira
authors.affiliation: CQM-UMa, Centro de Química da Madeira, Centro de Ciências Exactas e da Engenharia da Universidade da Madeira, Campus Universitário da Penteada, 9000-390 Funchal, Portugal. Electronic address: jsc@uma.pt.
authors.forename: José S
authors.initials: JS
authors.lastname: Câmara
mesh_terms: D000928:Antidepressive Agents / Q000652:urine*; D002851:Chromatography, High Pressure Liquid; D002997:Clomipramine / Q000652:urine*; D005473:Fluoxetine / Q000652:urine*; D006801:Humans; D057230:Limit of Detection; D052617:Solid Phase Microextraction
chemical_list: D000928:Antidepressive Agents; D005473:Fluoxetine; D002997:Clomipramine
keywords: Antidepressants; Human urine samples; Microextraction by packed sorbent (MEPS); Ultra high performance liquid chromatography (UHPLC)