Improved Analytical Approach Based on QuECHERS/UHPLC-PDA for Quantification of Fluoxetine, Clomipramine and their Active Metabolites in Human Urine Samples.
This paper reports, for the first time, the development of a modified Quick, Easy, Cheap Effective, Rugged and Safe (QuEChERS) combined with a dispersive SPE (d-SPE) based clean-up procedure as a new and powerful strategy for the simultaneous and efficient isolation of two different antidepressants, fluoxetine and clomipramine, and their active metabolites in human urine samples. A univariate experimental design with four independent variables such as sample volume, extraction solvent, buffered salts and clean-up step, was performed and used to investigate the effect of process variables on the extraction efficiency. Good linearity was achieved at the studied concentration range (0.1-5.0 µg mL-1), with correlation coefficients (R2) higher than 0.9961. Low detection limits, ranging between 0.060 and 0.092 μg mL-1were obtained for all analytes, whereas the lowest quantification limit was 0.1 μg mL-1, corresponding to the lowest concentration of the standard curve. The method also showed good results for accuracy, with values ranging from 91% to 105%. Intra- and inter-day precision, expressed as the relative standard deviation (RSD), were also satisfactory (<10%). Consistent recoveries of antidepressants ranging from 86% to 109% were observed when urine samples were fortified at three concentrations, namely 0.1, 2.5 and 5.0 μg mL-1In order to evaluate the proposed method for clinical use, the QuEChERS/UHPLC-PDA method was applied to analysis of 12 urine samples from depressed patients.
CQM- Centro de Química da Madeira, 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.
Vera
V
Alves
CQM- Centro de Química da Madeira, Universidade da Madeira, Campus Universitário da Penteada , 9000-390 Funchal, Portugal.
Carlota
C
Conceição
CQM- Centro de Química da Madeira, 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.
João
J
Gonçalves
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.
Helena M
HM
Teixeira
CQM- Centro de Química da Madeira, Universidade da Madeira, Campus Universitário da Penteada , 9000-390 Funchal, Portugal jsc@uma.pt.|Faculdade de Ciências Exactas e Engenharia, Universidade da Madeira, Campus Universitário da Penteada , 9000-390 Funchal, Portugal.
José S
JS
Câmara
D000928:Antidepressive Agents / Q000652:urine*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D002997:Clomipramine / Q000652:urine*; D004305:Dose-Response Relationship, Drug; D005473:Fluoxetine / Q000652:urine*; D006801:Humans; D057230:Limit of Detection; D013058:Mass Spectrometry; D015203:Reproducibility of Results
D000928:Antidepressive Agents; D005473:Fluoxetine; D002997:Clomipramine