ABSTRACT:
The term chromatography refers to a number of highly efficient techniques for the separation of a wide variety of substances ranging from simple ions to biopolymers. The separation is carried out, by distributing the component mixtures between a fixed stationary and a moving mobile phase. At the end of 50’s thin layer chromatography (TLC) was introduced to the laboratory as a rapid technique for simple qualitative analysis. A major advancement of TLC is the high performance thin layer chromatography (HPTLC) requiring shorter time and better resolution. Now a day it is regarded as a most useful technique for qualitative and quantitative analysis. In the present study, extracts of root sand leaves were prepared by taking 1 gram of each powder of root and leaf accurately weighed was taken and macerated in 20 ml of methanol for 3 hours with occasional shaking. n-Butano, :Acetic acid : Water (4: 1: 5) was used as mobile phase. Vanillin in sulphuric Acid, Ferric chloride solution and Iodine vapours were used as detecting agents. HPTLC Plates (E.Merck No. 5548) of silica gel 60F was used for analysis. Separation at Rf values 0.09, 0.14, 0.18, 0.36, 0.41, 0.49, 0.57, 0.64 were observed. Common spots of roots, leaves and whole plant were observed at Rf- 0.36, 0.57 and 0.64. This further confirmed the absence of alkaloids. The Rf- value 0.58 in the leaf sample and 0.56 in the root sample showed the presence of beta-sitosterol on comparing with the Rf-value of the standard beta-sitosterol which was found to be 0.58. Presence of Rf- 0.0, 00.19, 0.55, and 0.7 in the leaf sample coincided with the Rf-value 0.01, 0.18, 0.53, and 0.70 of gallic acid so, the presence of tannins in reference with the gallic acid (standard) was confirmed. Ethyl acetate: Formic acid: Glacial acetic acid: Water (10:1.1:1.1:2.6) was used as the solvent system for estimation of flavanoids. The samples used in the present study showed quenching at UV 254 nm nd 366nm.