Pyrite commonly incorporates a wide range of trace and minor elements, which in turn may modify some of the mineral’s physical and chemical properties. Published band position data for the Raman spectra of pyrite show a wide variation, and the relationship between band position and the trace/minor element incorporation in pyrite is poorly constrained until now. This prompted a case study on pyrite with varying As content from the Shizilishan Sr-(Pb-Zn) deposit, eastern China, combining electron probe microanalysis with Raman spectroscopy. Results show a significant correlation, with a major downshift (~10 cm–1) of the positions of all three Raman bands with increase of As content from below 0.05 to 4.89 wt % in pyrite. This phenomenon is attributed to increased bond lengths and local distortions within an expanded pyrite crystal structure. Results highlight the potential that Raman spectroscopy offers to estimate the contents of trace/minor elements—especially As—in pyrite. Given that substitution of As into pyrite often facilitates co-incorporation of both economically useful (Au) and environmentally significant elements (Hg and Tl), Raman methodology could provide valuable, rapid assessment of pyrite chemistry in both gold deposit exploration and environmental science, although the impact of laser heating and mechanical polishing needs to be avoided or effectively reduced. Raman spectroscopy may also find a valuable future role within semiautomated multispectral analytical platforms that can generate close- to-real-time geologic information on freshly drilled core directly at the drill site or in outcrop.