Phytochemical evaluation and Biomarker isolation from a Poly-herbal formulation

Plant derived medicines are popularly used nowadays considering their natural origin and low side effects. Phytochemical compounds are responsible for their various biological activities. Qualitative evaluation of such formulations is a challenging task due to high variability of their chemical components. This work aimed at preparing and evaluating a polyherbal anti-stress medicine, formulated using four potent Ayurvedic drugs namely Withania somnifera, Valeriana wallichii, Bacopa monnieri and Piper longum and also isolating a bioactive marker compound from the same.The formulation was prepared as per classical methods of Ayurveda and analysed for physicochemical characters like loss on drying, total ash content, acid insoluble ash, water soluble extractive value and alcohol soluble extractive value by standard methods. Phytochemical analysis was performed for detecting presence of carbohydrates, phenols, flavonoids, tannins, alkaloids, steroids, Terpenoids, glycosides and saponins and for estimating active groups like total phenolic content, flavonoids, tannins and saponins. High Performance Thin Layer Chromatography fingerprint profiling was performed to confirm the presence of various phytocompounds. The biomarker compound was isolated by column chromatography method and characterized by UV-Visible, H1-NMR, C13-NMR and Mass spectroscopy methods. Physicochemical analysis showed presence of about 18% water soluble components and about 14% alcohol soluble components. Preliminary phytochemical analysis confirmed presence of carbohydrate, phenols, flavonoids, tannins, alkaloids, steroids, glycosides, phytosterols and saponins. Quantitative analysis showed presence of about 7.6% phenols, 1.3% flavonoids, 1.1% tannins and 3.5% saponins. Spectroscopic study indicated isolated compound is a phenolic compound 3, 4, 5-tri hydroxybenzoic acid.