Gloriosa superba L is an important and cultivated medicinal plant. The seed and tubers are containing alkaloids such as Colchicine and Colchicoside. In the present investigations, the phyto-compounds from the different accessions of tubers of Gloriosa superba L cultivars from Sirumalai (GA1) region, Mulanoor (GA2)region, Thuraiyur (GA3)region, Konganapuram (GA4) region and Vedaranyan (GA5) region were extracted by using ethanolic extract and the chemical composition and its concentration in the tubers of Gloriosa superba were determined by the method of HPLC analysis. The methanolic extract is used for to find out various phytochemical constituents. The five different accessions of different agro climatic zone cultivated Gloriosa superba L selected for present investigation. The result shows that Colchicine concentration in all five different accessions.