In order to investigate the bioavailability of kaolin adsorbed copper on gills of carpio, and to explain its possible mechanism in terms of speciation, exposure experiments with constant concentration of water soluble copper and increased concentration of kaolin adsorbed copper was carried on, and the speciation analyses was developed using MINTEQA2. The results of the exposure experiment indicated that expose to kaolin adsorbed copper would increase the metal accumulation in the fish gills. The shift of the copper speciation was demonstrated via chemical equilibrium calculation. To be specific, the kaolin adsorbed copper would be partially desorbed in the fish gill microenvironment in the alkaline water environment.