On-fiber derivatization for direct immersion solid-phase microextraction. Part II.Acylation of amphetamine with pentafluorobenzoyl chloride for urine analysis

Abstract

On-fiber derivatization was used for solid-phase microextraction (SPME) in order to increase the detectability and extractability of drugs in biological samples. Amphetamine, which was used as a model compound, was derivatized with pentafluorobenzoyl chloride (PFBCl) and subjected to gas chromatography with electron capture or mass spectrometric detection. Extraction was performed by direct immersion of a 100 μm polydimethylsiloxane-coated fiber into buffered human urine. On-fiber derivatization was performed either after or simultaneously with extraction. The former procedure gave cleaner chromatograms but the latter turned out to be superior with respect to linearity and repeatability. For the on-fiber derivatization of amphetamine an excess of reagent is required. Because a considerable part of the PFBCl loaded on to the fiber is used up by reaction with matrix compounds and water, a reagent loading time of 5 min was needed to obtain a linear range (r = 0.9756) from 250 pg mL⁻¹ to 15 ng mL⁻¹. Due to an interfering matrix compound, the limit of detection was also found to be dependent on the reagent loading time, i.e., the limit of detection for a PFBCl loading time of 5 min is 250 pg mL⁻¹ whereas that for a 1 min loading time it is 100 pg mL⁻¹. The relative standard deviation (n = 7) of the method was about 11% at an amphetamine concentration of 1 ng mL⁻¹. The applicability of the method for the determination of drugs in biological samples is shown.