The measurement of zeta potential (ZP) has important applications in a wide range of industries including ceramics, pharmaceuticals, medicine, mineral processing, electronics, cement industry and water treatment. It provides new information on cement hydration, gel structure formation and the effects of chemical and mineral admixtures. In this study, ZP and pH measurements of Portland cement suspension prepared at wt% 1 cement/water ratio in the presence of four different types of waters, i.e., pure water, tap water, and salted (NaCl and CaCl2) water, were carried out depending on the time. Also, vicat tests of Portland cement pastes prepared at a water-to-cement ratio of 0.34 were performed. After the vicat tests, X-ray diffraction analysis and scanning electron microscopy investigations were performed on the samples. The most important result obtained from this study is that setting time and time-dependent ZP values of Portland cement paste have different values depending on the used mixing water type. Among the water types, water with CaCl2 gives the lowest ZP as absolute value, so this leads to minimum initial and final setting times for the cement paste prepared with this type of water.