Simultaneous Identification and Quantitation of Fluoxetine and its Metabolite, Norfluoxetine, in Biological Samples by GC-MS

A sensitive method for the quantitation of fluoxetine and norfluoxetine in biological samples was developed. Blood, urine, and tissue samples were alkalinized and extracted with N-butyl chloride. The extracts were derivatized with pentafluoropropionic anhydride before gas chromatography-mass spectrometry (GC-MS). Selected ions were monitored at m/z 117 and 294 for fluoxetine; m/z 117, 176, and 280 for norfluoxetine; and m/z 122 and 299 for the internal standard fluoxetine-d5. The within-run and between-run precision as well as recovery were determined for both analytes. The empirical limit of detection was determined to be 12.5 micrograms/L for both fluoxetine and norfluoxetine, whereas the empirical limit of quantitation was 25 micrograms/L for both drugs. Calibration curves were linear in the range of 50-1000 micrograms/L for both analytes. Some drugs that were known or suspected of interfering with high-performance liquid chromatography and GC methods for fluoxetine and norfluoxetine were tested for interference. This is the only reported method that combines the use of a deuterated internal standard, selected ion monitoring by GC-MS, and derivatization for the identification and quantitation of fluoxetine and norfluoxetine.