Phytochemical Investigations on Dillenia Indica Linn, and Dillenia pentagyna Roxb. for Assement of their Anticancer Potential

Abstract

Quality as well as standardization of herbal medicines especially of ethno medicinal and folklore medicines are extremely important for evaluation of therapeutic potential of plants. Hereby considering lack of information and scientific evidences on indigenous medicines, it was endeavored to develop quality control parameters for herbal plants. A comprehensive study on bark and leaves of indigenous plants D. indica Linn and D.pentagyna Roxb. (known as ‘Karmal’ and ‘Mota karmal’ in Dang forest of Gujarat) has been carried out in accordance with WHO guidelines. Pharmacognostical work mentioned can successfully be applied in order to identify bark and leaves of both plants.Morphologically, Leaves and bark of both plant species are differentiated by its size and shape. Microscopically, bark and leaves of both plant showed raphides of calcium oxalates. Size of stone cells and presence of pigments in bark as well as size of trichomes in leaf were major differentiating characters of both species. The physicochemical evaluation ensures the quality of plants. In terms of distribution of active ingredients, these plant species include flavonoids, sterols and triterpenoid, tannins, glycosides etc. A potent anticancer compound, betulinic acid isolated using column chromatography as well as simple fractionation method. In addition, two recent technologies super critical fluid and microwave assisted extraction were designed and optimized for the extraction of betulinic acid which has been compared with conventional method i.e., soxhlet extraction. The percentage yields of betulinic acid obtained by soxhlet extraction, SFE, MAE were 0.28-36 %w/w, 0.69 %w/w and 0.91 %w/w respectively. The developed HPTLC and RP-HPLC is simple, rapid and specific techniques to quantify the betulinic acid from the bark and leaves of both plant species without compromising the accuracy. MTT assay has been performed on three different cancer cell lines HCT-15, DU145 and A-375. The content of betulinic acid in plant extracts and IC50 values on cell lines are positively related to each other as IC50 value is decreasing with increase in amount of betulinic acid in both species which was also checked against standard anticancer drugs Doxorubicin and Colchicine.Developed standardized parameters, various described methods for extraction, method development serve the purpose of quality assessment. Moreover, these two species are proved to possess good anticancer potential as well as contain appreciable quantity of betulinic acid which can be utilized further for exploration of its use.