Polyaniline nanofiber films were fabricated on the surface of stainless steel wire via a controllable and simple electrophoretic deposition route from a nonaqueous colloidal suspension consisting of polyaniline nanofibers. The prepared coating material was then characterized by field emission scanning electron microscopy equipped with energy dispersive spectroscopy and elemental mapping analysis. The fabricated polyaniline film-coated stainless steel wire was then utilized as an effective and novel sorbent phase for solid-phase microextraction of tamoxifen for subsequent gas chromatography/flame ionization detection of this anticancer drug. Parameters consisting of the temperature, extraction time, salt concentration, agitation speed, pH, temperature and time of desorption were studied and optimized using a one-at-a-time strategy. Under the optimum conditions, detection limit (S/N = 3), the limit of quantification (10/3 limit of detection), linear dynamic range, repeatability and reproducibility values of 0.51 μg L -1 , 1.7 μg L -1 , 2-1,130 μg L -1 , 5.7% and 8.6% were attained, respectively. The prepared fiber can preserve 90% of its efficacy after 20 consecutive cycles, demonstrating the suitable thermal stability and cyclability of the proposed solid-phase microextraction coating material for the determination of tamoxifen by gas chromatography/flame ionization detection. The route was effectively utilized to determine tamoxifen in urine samples, with relative recoveries ranging from 89 to 106%., (© 2021 John Wiley & Sons, Ltd.)