For mercury speciation analysis in environmental and biological samples, novel Hg 2+ ion-imprinted polymers (IIPs) were synthesized by a sol-gel process using the chelating agent dithizone, and then dithizone-Hg 2+ chelate as a template and 3-aminopropyltriethoxysilane as a functional monomer, followed by solid-phase extraction (SPE) and atomic fluorescence spectroscopy (AFS) detection. The resultant Hg-IIPs offered high binding capacity, fast kinetics, and their adsorption processes followed aLangmuir isotherm and pseudo-second-order kinetic models. The IIPs displayed excellent selectivity . Furthermore, the analytical results for the spiked mercury species in real biological samples, such as human hair and fish meat, confirmed that the methods are practically applicable to speciation analysis. The IIPs-SPE-AFS demonstrated significant application perspectives for rapid and high-effective cleanup, enrichment and determination of trace mercury species in complicated matrices.