Development and validation method for determination of fluoxetine and its main metabolite norfluoxetine by nonaqueous capillary electrophoresis in human urine.
A simple, rapid and sensitive procedure using nonaqueous capillary electrophoresis (NACE) to measure fluoxetine and its main metabolite norfluoxetine has been developed and validated. Optimum separation of fluoxetine and norfluoxetine, by measuring at 230nm, was obtained on a 60cm x 75mum capillary using a nonaqueous solution system of 7:3 methanol-acetonitrile containing 15mM ammonium acetate, capillary temperature and voltage 25 degrees C and 25kV, respectively and hydrodynamic injection. Paroxetine was used as internal standard. Good results were obtained for different aspects including stability of the solutions, linearity, and precision. Detection limits of 10mugL(-1) were obtained for fluoxetine and its metabolite. This method has been used to determine fluoxetine and it main metabolite at clinically relevant levels in human urine. Before NACE determination, the samples were purified and enriched by means of extraction-preconcentration step with a preconditioned C(18) cartridge and eluting the compounds with methanol.
Flores
J Rodríguez
JR
Department of Analytical Chemistry and Foods Technology, UCLM 13071, Ciudad Real, Spain.
Nevado
J J Berzas
JJ
Peñalvo
G Castañeda
GC
Diez
N Mora
NM