Development and validation method for determination of Paroxetine and its metabolites by nonaqueous capillary electrophoresis in human urine. Experimental design for evaluating the ruggedness of the method.
A simple, rapid, and sensitive procedure using nonaqueous capillary electrophoresis (NACE) to measure Paroxetine (one of the mostly used antidepressants for mental diseases treatment) and three metabolites has been developed and validated. Optimum separation of paroxetine and metabolites was obtained on a 57 cm x 75 microm capillary using a nonaqueous buffer system of 9:1 methanol-acetonitrile containing 25 mM ammonium acetate and 1% acetic acid, with temperature and voltage of 25 degrees C and 15 kV, respectively, and hydrodynamic injection. Fluoxetine was used as an internal standard. Good results were obtained for different aspects including stability of the solutions, linearity, accuracy, and precision. Detection limits between 9.3 and 23.1 microg.L(-1) were obtained for paroxetine and its metabolites. A ruggedness test of the method was carried out using the Plackett-Burman fractional factorial model with a matrix of 15 experiments. This method has been used to determine paroxetine and its main metabolite B at clinically relevant levels in human urine. Prior to NACE determination, the samples were purified and enriched by means of an extraction-preconcentration step with a preconditioned C18 cartridge and eluting the compounds with methanol.
Rodríguez Flores
Juana
J
Department of Analytical Chemistry and Foods Technology, Ciudad Real, Spain. juana.rflores@uclm.es
Barzas Nevado
Juan J
JJ
Contento Salcedo
Ana M
AM
Cabello Díaz
Maria P
MP
D000085:Acetates; D000097:Acetonitriles; D001711:Biotransformation; D016903:Drug Monitoring / Q000379:methods* / Q000592:standards; D019075:Electrophoresis, Capillary / Q000379:methods* / Q000592:standards; D006801:Humans; D017374:Paroxetine / Q000378:metabolism* / Q000493:pharmacokinetics / Q000652:urine*; D015203:Reproducibility of Results; D012996:Solutions
D000085:Acetates; D000097:Acetonitriles; D012996:Solutions; D017374:Paroxetine; C018824:ammonium acetate; C032159:acetonitrile