The chromium speciation procedure was optimized using magnetic polyaniline nanoparticles (Fe
3
O
4
@PANI NPs) solid-phase extraction coupled with microsampling injection-flame atomic absorption spectrometry (MIS-FAAS). Chromium speciation was successfully achieved by Fe
3
O
4
@PANI NPs at pH 8.0. The recoveries obtained for Cr(III) and Cr(VI) were above 95% and under 5%, respectively. Recoveries of over 95% for Cr(III) from 40.0 mL of the sample were obtained using 25 mg Fe
3
O
4
@PANI NPs and 500 μL of 0.2% (w/v) thiourea (TU) solution prepared in 2 mol L
−1
HCl as eluent. Total chromium as Cr(III) was extracted quantitatively after reducing the Cr(VI) to Cr(III). The linear range, detection limit, preconcentration factor, and precision of the optimized method for Cr(III) in aqueous solution were 2.5–94.0 μg L
−1
, 0.335 μg L
−1
, 80, and 3.07%, respectively. The validation of the method was controlled using SPS-WW2 Batch 114 wastewater and BCR-715 industrial wastewater as standard reference materials (SRMs) for environmental water, and the obtained results were in close agreement with the certified values.