Fractionation of the phytexponent polyherbal formulation by column chromatography and qualitative antioxidant screening

Abstract

Medicinal plants are a rich source of natural healthcare products for alleviating various diseases. They have been reported to be of great importance due to their endless therapeutic properties that are useful in curing many chronic diseases. Their usage in the management of many profound compared with synthetic drugs that have been associated with side effects. Many natural products including polyherbal formulations are derived from plants. Various synthetic products presently in the market are arguably costly and associated with adverse side effects, therefore warranting a need for viable alternatives. Medicinal plants have various bioactive compounds, which exhibit diverse pharmacological properties. However, many of the bioactive compounds present in these plants have not been separated making their isolation and characterization a challenge. Phytexponent, a polyherbal formulation, is composed of alcoholic extracts of Viola Tricolor, Echinacea purpurea, Allium sativum, Triticum repens, and Matricaria chamomilla. Phytexponent has been widely used in the complementary medicine to treat inflammatory disorders, mange stress, boost the body immunity, manage blood pressure and many more other conditions. However, the components present in the phytexponent have not been separated and the pure compounds isolated and characterized. Hence this study, evaluated the column chromatographic fractionation and qualitative antioxidant evaluation of the phytexponent. The phytexponent was fractionated by adopting the normal phase column chromatography. Column was packed with the silica gel slurry prepared in the petroleum ether, followed by loading of the phytexponent mixed in few grams of silica and then eluted with various organic solvents starting with those of lower polarity to high. The collected fraction that were pooled together based on their properties and then qualitatively evaluated for the antioxidant activity using 1,1-diphenyl-2- picryhydrazyl free radical assay. The silica gel pre-coated thin layer chromatography plates were spotted with the collected fractions and then developed in the mobile phase and then air dried followed by spraying with 1,1-diphenyl-2-picryhydrazyl methanolic solution. Chromatographic separation of the Phytexponent yielded 285 fractions which were further separated by thin layer chromatography. Based on their thin layer results, they were combined in 13 fractions. Visualization was achieved under Ultraviolet light at 254 nm and 365 nm, respectively, whereby various colours were observed. The minerals could be barite, calcites, corundum and halite that fluorescents red, pectolite and margarite that fluorescents blue and apatite that fluorescents purple. Under short ultraviolet albite, bentonite and calcite that fluorescents blue was present. The antioxidant activity was only present in the petroleum ether: Dichloromethane (50:50) and Dichloromethane (100 %) fractions in which yellow spots on the purple background was noted. Therefore, the phytexponent polyherbal formulation contain various compounds that are non-polar in nature as most of the isolated compounds were from the non-polar solvent. Similarly, the phytexponent has antioxidant compounds and can be used as a natural source of free radical scavengers