In the present study, simple phytochemical screening procedures were carried out to find the various constituents present in the ethanolic extract of Dioscorea villosa tubers. Further,estimation of Diosgenin in Dioscorea villosa tubers, was carried out by HPTLC technique. The preliminary screening showed presence of proteins, flavonoids tannins, alkaloids, phenolic compounds saponins and glycosides. The extract was chromatographed on silica gel GF254 plates with Toluene: Ethyl Acetate: Acetic Acid: Formic Acid (4: 3: 1:1) as mobile phase. Detection and quantification were performed by densitrometric scanning, at 366 nm. The average recovery of diosgenin was found to be 0.48 %. The HPTLC technique has provided a good resolution of diosgenin from other constituents present in the ethanolic extract.