Aim of the present study was to develop a high performance thin layer chromatography (HPTLC) method to determine the diosgenin content variation in geographically different fenugreek seeds from Sudan and India. Chloroform extracts of fenugreek seeds were used to quantify diosgenin and compare with standard diosgenin (1 mg/ml). Different compositions of the organic solvents were tried to develop new solvent system, chloroform: methanol: toluene (5:1:1). Standard calibration curve was prepared by increasing the concentration of standard compound with the standard equation as y = 1427.x + 1372, R2 = 0.961. The standard equation was used to calculate the diosgenin content. HPTLC results revealed Rf value of diosgeninas 0.29, 0.30 in various tracks. The scanning of the TLC plates was carried out at 550 nm. The Sudanese sample consists of 0.12% (w/w) diosgenin which is approximately double of Indian sample 0.069% (w/w). HPTLC technique was found to be the best analytical technique to quantify diosgenin with excellent precision at low cost. The analysis is effective to determine the content variation in different geographical regions which could be the best approach to target the best source with high content of secondary metabolites.

Keywords: HPTLC, Diosgenin, Trigonellafoenum-graecum L.