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A polymetallic layer is usually developed at the bottom of the early Cambrian black shale in Guizhou Province. The mineral that makes up the polymetallic layer is related to the sedimentary facies. To analyze the differentiation mechanism between polymetallic deposits (Ni‐Mo and V), the Zhijin Gezhongwu profile located in the outer shelf and the Sansui Haishan V deposit located in the lower slope are selected to study the in situ sulfur isotopes and trace elements of pyrite. The results show that δ34S values of pyrite vary widely from −7.8 ‰ to 28 ‰ in the Gezhongwu profile, while the δ34S values are relatively uniform (from 27.8 ‰ to 38.4 ‰) in the Haishan profile. The isotopic S composition is consistent with the transition that occurs in the sedimentary phase from the shelf to the deep sea on the transgressive Yangtze platform; this indicates that the δ34SO42– values in seawater must be differently distributed in depositional environments. The sulfur in the Ni‐Mo layer is produced after the mixing of seawater and hydrothermal fluid, while the V layer mainly originates from seawater. Overall, the Ni‐Mo and V deposits have been differentiated primarily on the basis of the combined effect of continental weathering and hydrothermal fluid. [ABSTRACT FROM AUTHOR]