Strontium (Sr), hydrogen (H) and oxygen (O) in vent fluids are important for understanding the water–rock interaction and hydrothermal flux in hydrothermal systems. We have analyzed the Sr, H and O isotopic compositions of seawater, vent fluid and hydrothermal plume samples in the Kueishantao hydrothermal field, as well as their calcium (Ca), total sulfur (S), Sr, arsenic (As), stibium (Sb), chlorine (Cl) and manganese (Mn) concentrations for understanding the origin and processes of fluids. The results suggest that most As, Sb and Mn are leached from andesitic rocks into the fluids, and most Ca and Cl remained in the deep reaction zone during the fluid–andesitic rock interaction. The ranges of 87Sr/86Sr, δDV-SMOW and δ18OV-SMOW values in the yellow spring, white spring and plumes are small. The 87Sr/86Sr, δDV-SMOW and δ18OV-SMOW values of fluids and plumes are like those of ambient seawater, indicating that the Sr, H and O of vent fluids and hydrothermal plumes are derived primarily from seawater. This suggests that the interaction of andesite and subseafloor fluid is of short duration and results in the majority of As, Sb and Mn being released into fluids, while most Ca and Cl remained in the deep reaction zone. In addition, there was no significant variation of Sr, H and O isotopic compositions in the upwelling fluid, keeping the similar isotopic compositions of seawater. There are obvious correlations among the pH values, As and Sb concentrations, and H isotopic compositions of the vent fluids and hydrothermal plumes, implying that the As and Sb concentrations and H isotopic compositions can trace the dispersion of plumes in the ambient seawater. According to the Sr concentrations and 87Sr/86Sr values, the water/rock ratios are 3076~8124, which is consistent with the idea that the interaction between fluid and andesite at the subseafloor is of short duration. The hydrothermal flux of Sr discharged from the yellow spring into the seawater is between 2.06 × 104 and 2.26 × 104 mol/yr, and the white spring discharges 1.18 × 104~1.26 × 104 mol/yr Sr if just andesites appear in the reaction zone.