The Chengchao iron deposit, the largest high-grade skarn iron deposit in southeastern Hubei Province, contains considerable amounts of magnetite and by-product anhydrite. To obtain better understanding of the ore-formation process, this study carried out He-Ar-S-Pb multi-isotopic analyses on the pyrites formed during two stages of mineralization. The results indicate that the δ34S values (ranging from 14.0‰ to 17.6‰) of pyrites formed from the two stages have no obvious differences, suggesting that they were not derived from a single magmatic sulfur source. The δ34S values of anhydrite mostly range from 21.9‰ to 28.4‰, similar to that of the Middle Triassic sedimentary anhydrite in the Middle-Lower Yangtze River metallogenic belt (MLYRB). The Pb isotopic compositions of the pyrites of both stages are homogeneous, with values of 208Pb/204Pb, 207Pb/204Pb, and206Pb/204Pb being 38.006–38.257, 15.523–15.556, and 17.806–18.052, respectively, indicating a mixed crust-mantle source. The He-Ar results exhibit different compositions of the two stages: the 3He/4He (R/Ra) and 40Ar/36Ar values for the early-stage pyrite are 0.46–0.63 and 311–322, respectively, whereas the values for late-stage pyrite are 0.23–0.34 and 305–361, respectively. Both stages of pyrites indicate the multiple sources of the ore-forming fluids, with decreasing amount of magmatic water and increasing amount of modified meteoric water(MASW) during fluid evolution. The Triassic evaporites played an important role in the mineralization process.