The Central Hunan metallogenic province, located in the southeastern part of the Yangtze Block, South China, is one of the most important Sb–Au metallogenic belts in China. The Baiyunpu gold deposit is one of the important Carlin‐type gold deposits occurring in Devonian sedimentary strata in Central Hunan. Optical microscopy and in situ laser ablation (multi‐collector) inductively coupled plasma mass spectrometry (LA‐[MC]‐ICP‐MS) are used to determine the texture, trace element compositions and sulfur isotope for pyrites from the pre‐ore and syn‐ore stages to constrain the sources of reduced sulfur and metals and the mechanism of gold precipitation. Four generations of pyrite were identified by petrography and pyrite compositions. Pre‐ore Py1 is characterized by relatively low concentrations of As, Au, Sb and Cu and high concentrations of Co, Ni, Bi and Pb. Py1 has positive δ34S values of 6.5–10.1‰, which we ascribe to a bacterial reduction of marine sulfate in closed marine systems, or from weak Rayleigh fractionation by bacterial sulfate reduction (BSR) or thermochemical sulfate reduction (TSR) during sedimentary‐diagenetic processes. Pre‐ore Py2 presents as rims of Py1 and contains relatively high As (69,530 ppm), Sb (123.2 ppm) and low Au (4.89 ppm). Compared with Py2, Py3 has relatively high Au (194.4 ppm) and Cu (24.6 ppm) and low As (40,801 ppm). We interpret that Py2 and Py3 could have precipitated from an Au‐free but As‐rich early hydrothermal fluids or from low‐Au and As‐rich fluids at the beginning of the ore stage. The δ34S values of Py2 and Py3 in the Baiyunpu deposit are concentrated in a narrow range of −1.2 to 1.2‰, suggesting the involvement of magmatic sulfur. Syn‐ore pyrites (Opy) are enriched in Au (517.8 ppm), As (39,719 ppm), Cu (261.9 ppm) and Sb (171.4 ppm). Irregular boundaries between Py1 and Opy suggest that the replacement may have been important for the formation of pyrites at the syn‐ore stage. The variation of As concentration in Opy1, Opy2 and Opy3 appear to have recorded fluctuations in the chemical composition of ore fluids. The δ34S values of Opy (0.1–5.9‰) are slightly higher than Py2 and Py3 due to mixing with sedimentary components with elevated δ34S values in late‐stage hydrothermal fluids. Profile analyses for Baiyunpu auriferous pyrite grains suggest that Au was positively correlated with Cu, but not with As. The correlation between Au, Cu and As in Opy may be dominated by their concentrations of ambient evolving fluids.