PHYTOCHEMICAL STANDARDIZATION, TLC FINGERPRINTING AND ANTIOXIDANT EVALUATION OF AN OPTIMIZED POLYHERBAL FORMULATION

Abhay Kumar Mourya*, Manju Makhija, Shanti Bhushan Mishra, Mahesh Gupta

In the present study, a polyherbal formulation with high antioxidant activity has been formulated and standardized with the help of medicinal plants used in traditional medicine. Twelve medicinal plants were used to prepare their hydroalcoholic extracts by cold percolation method in the ratio of 70% ethanol: 30% water. Three different polyherbal formulations (F1, F2 and F3) were prepared and analysed for phytochemical composition, TLC fingerprinting and in vitro antioxidant activity. The preliminary phytochemical screening of all the formulations showed presence of flavonoids, phenolics, tannins, glycosides, terpenoids and carbohydrates. TLC fingerprinting was done on silica gel 60 F254 plates using toluene:ethyl acetate:formic acid (5:4:1 v/v/v) as mobile phase, with well separated chromatographic profiles and typical Rf values that suggest the presence of several bioactive phytoconstituents. The antioxidant activity was conducted using DPPH radical scavenging assay, Ferric Reducing Antioxidant Power (FRAP) assay and Potassium Ferricyanide reducing power assay with the concentrations varying from 20 to 100 µg/ml. All formulations showed antioxidant activity in all experimental models with the highest activity observed for F3. The DPPH assay revealed that the formulation F3 exhibited 89.1 ± 1.5 % inhibition at 100 µg/mL while significant ferric reducing activity was obtained from formulation F3 in FRAP assay and significant reducing power activity was obtained from formulation F3 in potassium ferricyanide assay. The increased antioxidant activity of the formulation F3 can be explained by co-action of the phytoconstituents present in formulation and rich in these phenolics and flavonoids. Optimized formulation showed significant antioxidant activity and can be used as a potential natural source of antioxidant phytoconstituents.