Determination of Fluoxetine in Blood Samples by High-Performance Liquid Chromatography Using Derivatization Reagent

A rapid and selective chromatographic method for determination of fluoxetine in blood samples has been developed. The method is based on a selective reaction between 7,7,8.8-tetracyanoquinodimethane (TCNQ) and molecules with a secondary amine moiety, resulting in a purple solution. Blood samples were spiked with fluoxetine and nortriptyline as an internal standard. then. both compounds were extracted applying a microwave-assisted extraction - a novel technique for isolation of an analyte from biological matrix After extraction, dried residues were dissolved in acetonitrilic solution of TCNQ and heated for 30 minutes at 80 degrees C. The obtained intense purple-blue colored solutions were then analyzed by high performance liquid chromatography with diode-array detection Chromatograms were recorded at 567 nm. Limits of detection and quantification of fluoxetine in blood were 0,03 and 0,10 mu g mL(-1). respectively It was concluded therefore that fluoxetine can be determined in blood in the therapeutic concentration range. The developed method has been applied to the analysis of whole blood samples collected from a female patient treated with Seronil (R) 20 (20 mg of fluoxetine)