Dachang ore field, located in the southwest margin of Jiangnan ancient land and the northeast side of Youjiang Basin, is an important non-ferrous metal ore producing area in China. Tongkeng deposit is a super-large tin polymetallic deposit with the largest scale, the most complex metallogenic elements and typical metallogenic characteristics in Dachang ore field, which is composed of upper tin polymetallic ore body and deep zinc-copper ore body. Sphalerite is researched from different types of ore bodies (tin polymetallic ore bodies, zinc-copper ore bodies) from copper pit deposits. In this paper, trace and rare earth element and H-O isotope analyses were conducted with the intention of further exploring the source of ore-forming materials and fluids in the area and deepening the understanding of the genesis of the deposit. The results show that sphalerite in different types of ore bodies of Tongkeng deposit is characterized by enrichment of Cu, Pb, Fe, Sb, In and poor Ge. The rare earth distribution patterns of sphalerite are mostly light rare earth enriched "V-shaped" right-sloping curve, characterized by high LREE/HREE values and negative Eu and Ce anomalies. The H-O isotopic composition of sphalerite indicates that the ore-forming fluid is mainly magma, accompanied by the mixing of atmospheric precipitation. The above characteristics show that the ore-forming materials and ore-forming fluids of Tongkeng deposit mainly come from magma, and the genesis of the deposit belongs to magmatic hydrothermal filling metasomatism.