The current study reports a novel fluorescent chemosensor based on the tagging agent 4,5,6,7-tetrachloro-2',4',5',7'-tetraiodofluorescein (Rose Bengal, RB) for detection of trace levels of Hg2+in environmental water. The established probe has been based upon liquid-liquid extraction (LLE) of the developed ternary complex ion associate {[Hg (bpy)2]2+.[RB]2-} of Hg2+ -2,2- bipyridyl complex [Hg (bpy)2]2+ and RB at pH 9.0 onto chloroform and measuring the resulting fluorescence enhancement signal intensity at λex/em = 570/ (580-600) nm. The limits of detection (LOD) and quantification (LOQ) of for Hg2+ was calculated to be 6.06 and 20 nM with a linear dynamic range (LDR) of 0.02-20µM, respectively. The stability constant, stoichiometry, chemical equilibria, and the thermodynamic parameters (ΔH, ΔS, and ΔG) of the developed ion associate were evaluated and assigned. Student's t and F tests at 95% confidence were fruitfully used for validation of the proposed methodology for Hg2+ detection in water samples with the aid of inductively coupled plasma-optical emission spectrometry (ICP-OES). The established strategy was successfully applied for detection of trace levels of Hg2+ in water samples with acceptable results. The proposed probe was satisfactorily applied for total determination and speciation of Hg in various water samples. Integrating the functional chelating agent onto associate formation to improve the selectivity and sensing properties of the LLE combining sensing probe towards target analyte in water represent the main interest.