In this study, cellulose acetate (CA)/iron oxide nanocomposites (NC1 and NC2) were synthesized and utilized as extractors for the selective separation of fluorene from aqueous samples. The selectivity of nanocomposites to different organic compounds was assessed. Based on the results, fluorene was the most quantitatively adsorbed on NC2. Batch method was performed by varying the initial fluorene concentrations and contact time. Langmuir and Freundlich isotherms were used for modeling the experimental data and were best correlated by the Langmuir model. Adsorption data were also followed the pseudo-second-order kinetic model. Finally, validation of the developed method was achieved for fluorene determination in real water samples.