The aim of this work was to develop cellulose nanofibers to address some critical issues for the treatment of chromium in aqueous systems. Furthermore, the use of nanomaterials and how they relate to nanoscale operations at the biological level has generated considerable concerns in spite of their novel properties. This paper deals with the synthesis and characterizations of Cyanex-301coated cellulose nanofibers with the active compound being the thiol (SH) groups, that will suffice as a viable material for Cr(VI) determination from aqueous solutions. The proposed mechanism was the complexation between the thiol groups on Cyanex-301. The effect of different parameters on the determination including contact time, sorbent dosage, initial Cr(VI) ion concentration and solution pH was investigated by atomic absorbtion spectroscopy. In this work, water samples were collected from some industrial places in Babolsar city. The recovery of Cr(VI) at a spiked concentration of 100 ng kg−1 ranged from 88% - 93%. The repeatability and reproducability for the determination of Cr(VI) were 2.1% and 4.3%, respectively.