Quantitative determination of trace phenazopyridine in human urine samples by hyphenation of dispersive solid-phase extraction and liquid-phase microextraction followed by gas chromatography/mass spectrometry analysis

Magnetic dispersive solid-phase extraction followed by dispersive liquid-liquid microextraction coupled with gas chromatography/mass spectrometry was applied for the quantitative analysis of phenazopyridine in urinary samples. Magnetic dispersive solid-phase extraction was carried out using magnetic graphene oxide nanoparticles modified by poly(thiophene-pyrrole) copolymer. The eluting solvent of this step was used as the disperser solvent for the dispersive liquid-liquid microextraction procedure. To reach the maximum efficiency of the method, effective parameters including sorbent amount, adsorption time, type and volume of disperser and extraction solvents, pH of the sample solution, and ionic strength as well as desorption time, and approach were optimized, separately. Characterization of the synthesized sorbent was studied by utilizing infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray analysis. Calibration curve was linear in the range of 0.5-250 ng/mL (R-2 = 0.9988) with limits of detection and quantification of 0.1 and 0.5 ng/mL, respectively. Intra- and interday precisions (RSD%, n = 3) of the method were in the range of 4.6-5.4% and 4.0-5.5%, respectively, at three different concentration levels. Under the optimal condition, this method was successfully applied for the determination of phenazopyridine in human urine samples. The relative recoveries were obtained in the range of 85.0-89.0%.