Phytochemical Evaluation and Antioxidant Potential of Polyherbal Extract Mixture-an In Vitro and In Silico Study.

Standardisation of polyherbal formulations plays a key role in estimating the quality and safety of drugs. In this study, a novel polyherbal formulation was prepared for the treatment of diabetic nephropathy and it was attempted to evaluate for its safety and efficacy against free radicals. To ascertain the safety of the formulation, individual herbs and polyherbal formulation have been screened for microbial load, heavy metals, pesticide residues and aflatoxins according to Ayush guidelines. Phytochemical analysis was carried out using standard procedures and GCMS analysis to identify the bioactive compounds. Antioxidant methods such as DPPH, nitric oxide, hydrogen peroxide, hydroxyl and superoxide radical scavenging assays were undertaken. To predict the mechanism behind the antioxidant efficacy, a molecular docking analysis was performed against Keap1/Nrf2 (PDB ID: 2FLU) protein using AutoDock Vina software. Phytochemical analysis showed the presence of alkaloids, tannins, flavones, glycosides, polyphenols, phytosterols and saponins. The results of in vitro antioxidant studies showed increase in percentage inhibition with increase in concentration and found no significant difference compare with standards. Among the 15 bioactive compounds identified from GCMS analysis, guanosine showed high binding affinity with Keap1/Nrf2 and interacting with similar residues as that of standard ascorbic acid which is analysed through molecular docking. Based on the results, it has been concluded that the safety parameters of polyherbal formulation were found within specified limits of Ayush guidelines and its efficacy against oxidative stress plays an effective role to treat diabetic nephropathy due to its synergistic antioxidant effect.