Selective extraction of Polycyclic Aromatic Hydrocarbons (PAH) without structural modification of the Natural Organic Matter (NOM) from a heavily contaminated sediment was investigated using various solvents mixtures. Structural modification of the NOM was monitored after extraction using 3D-fluorescence spectroscopy. Better results were obtained with a 15-h PAH extraction under reflux with a dichloromethane/cyclohexane 20/80 mixture. The experimental procedure was validated with NOM standard materials before to be applied on natural sediments. It could be applied to any environmental solid sample such as sediments and soils.