Pharmacognostical, Phytochemical and Anticancer Activity of the Leaves of Asparagus Racemosus Willd., (Liliaceae)

AIM:The main aim of the present work is to study the anticancer activity of the leaves of Asparagus racemosus Willd.,OBJECTIVES:To standardize the leaves of Asparagus racemosus Willd., by carrying out the pharmacognostical and phytochemical studies. To evaluate the invitro anticancer activity of various extracts To evaluate the invivo anticancer activity Isolation and identification of Rutin SUMMARY AND CONCLUSION:Cancer is one of the leading causes of death worldwide. Currently available allopathic drugs for treating cancer causes number of side effects hence, people are now looking towards the herbal medicine. This paved a vital necessity for finding natural anticancer drug from herbal source. Asparagus racemosus Willd., is one such plant which is traditionally used in the treatment of cancer was selected for the present study. The literature survey showed only scrappy information on the leaves of this plant. With this scanty of information on the leaves, consistent expectation of unexplored phytochemical profile and pharmacological efficacy forms the rationale for the study.Pharmacognostical studies A perusal of literature showed paucity of pharmacognostical information on the leaves of this plant. Hence, it was carried out and reported for the first time. In pharmacognostical studies macroscopy, microscopy, determination of physicochemical constants, analysis of inorganic elements, heavy metals and pesticide residue of the leaves were carried out. Macroscopical study showed that the leaves are pine needle (phylloclades) shaped, sweet and bitter taste and dark green in colour. The various distinguishing features of leaves observed through anatomical studies were, Phyllodades 3-4 angled with lateral projections Radially oblong epidermal cells with prominent cuticle Circular chlorophyllous palisade cells Vascular strands with angular xylem and thick mass of phloem elementsThe microscopical analysis of powder showed the presence of lignified fibres, epidermis with palisade cells and stomata. Various physicochemical constants such as ash values, extractive values, loss on drying, crude fibre content, swelling index and foaming index were carried out. These values will help in confirming the identity and purity of the plant. Any significant deviation in the percentage of any parameters reported in this work may indicate adulteration or substitution in the drug. Qualitative estimation of inorganic elements, quantitative estimation of heavy metals and pesticide residue were carried out and it showed only trace amount of heavy metals (within the limits) and absence of pesticide residue. These pharmacognostical details observed from the present study might offer reliable clues for the correct identification of the leaves of this plant in crude as well as fragmentary form and also ensures its differentiation from its substitutes and adulterants. This is first report on the pharmacognostical standardization on the leaves of Asparagus racemosus Willd.,Phytochemical studies Phytochemical evaluation deals with chemical analysis of the extracts used for pharmacological screening. The successive solvent extraction was carried out with solvents like n-Hexane, chloroform, ethyl acetate and ethanol. Chloroform extract showed more percentage yield when compared to other extracts. Qualitative preliminary phytochemical analysis was aided in identifying the phytoconstituents present in different extracts. Quantitative estimation of flavanoid, tannin and phenolic content were carried out, for the extracts. Fluorescence analysis was carried out, to detect the fluorescent chromophores present in the powdered drug as well as in the extracts. No fluorescence was observed in powder and the extracts. Thin layer chromatography for the extracts was performed to identify the single or mixture of constituents in the extract. High Performance Thin Layer Chromatography- Finger print profile was carried out to detect the number of constituents present in the ethyl acetate extract. Invitro studies:The therapeutically active extract to carry out the invivo anticancer activity was selected based on invitro anticancer activity. The chloroform, ethyl acetate and ethanol extracts were subjected to invitro anticancer studies using HeLa cells line by MTT assay. Chloroform and ethyl acetate extracts showed significant anticancer activity by MTT assay and these two extracts were selected for invivo anticancer studies.Acute toxicity studies:Acute toxicity study was carried out for chloroform and ethyl acetate extracts according to OECD guidelines 423. The extracts are non-toxic up to 2000mg/kg body weight indicating the safety of the extracts.Invivo anticancer studies:Invivo anticancer activity of chloroform and ethyl acetate extracts were carried out on Ehrlich’s Ascites Carcinoma (EAC) bearing mice. The ethyl acetate extract showed significant anticancer activity at the dose of 400mg/kg body weight when compared to chloroform extract and the effects were comparable with the standard drug. Since, rutin was found to possess anticancer activity, an attempt was made to isolate rutin by partition separation method. The isolated compound was subjected to melting point, chemical test, TLC and IR spectroscopic studies. From the above mentioned studies, it can be concluded that the pharmacognostical standards generated for the leaves will be useful for the proper identification of the plant. With the support of phytochemical and invitro cytotoxic activity the chloroform and ethyl acetate extracts were selected and subjected to invivo anticancer activity. The ethyl acetate extract at the dose level of 400mg/kg showed significant anticancer activity. Further studies are focused on detailed spectral analysis of isolated rutin and isolation of other phytoconstituents which is responsible for anticancer activity of the leaves