Toxoplasma gondii is an obligate intracellular parasite, which can invade most nucleated cells and proliferate inside the cells. During the process of T. gondii invasion and proliferate inside host cells, the cellular signal transduction network of the infected cells undergoing extensive changes. Phosphorylation is one of the most important post-translational modifications of proteins, and plays an important role in the process of cell signal transmission. To clarify how T. gondii regulates the host cell signal transduction process, in this study, we used titanium dioxide (TiO2) affinity chromatography to enrich the phosphopeptides in feline small intestine at 10 days post infection of T. gondii Prugniuad (Pru) strain, and quantify the phosphopeptides by using iTRAQ technology.