Development, Phytochemical Characterization (Alkaloids, Flavonoids, Terpenoids), and Chromatographic Standardization of a Polyherbal Formulation of Azadirachta indica, Ocimum sanctum, and Tinospora cordifolia with Experimental Antidiabetic Evaluation

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Background: Diabetes mellitus is a rapidly growing metabolic disorder associated with chronic hyperglycemia and multiple systemic complications. Conventional therapies, although effective, are often limited by adverse effects, high cost, and reduced long-term efficacy. Polyherbal formulations offer a promising alternative due to their multitargeted action and synergistic therapeutic potential. Objective: To develop, phytochemically characterize, and chromatographically standardize a polyherbal formulation of Azadirachta indica, Ocimum sanctum, and Tinospora cordifolia, and to evaluate its antidiabetic activity through in vitro and in vivo experimental models. Methods: Plant materials were extracted using hydroalcoholic solvents and combined in optimized ratios to formulate a polyherbal blend. Preliminary phytochemical screening and quantitative estimation of alkaloids, flavonoids, and terpenoids were performed. Chromatographic standardization was carried out using HPTLC and HPLC techniques with marker compounds. In vitro antidiabetic activity was assessed through α-amylase and αglucosidase inhibition assays. In vivo efficacy was evaluated in streptozotocin-induced diabetic Wistar rats by monitoring blood glucose levels, lipid profile, liver enzymes, and histopathological changes in pancreatic tissue. Results: The polyherbal formulation showed a high yield of bioactive constituents, with significant levels of flavonoids and terpenoids. Chromatographic analysis confirmed the presence of key marker compounds with well-resolved peaks and consistent retention times. The formulation demonstrated potent enzyme inhibitory activity with IC₅₀ values comparable to standard drugs. In vivo studies revealed a significant reduction in blood glucose levels, improvement in lipid profile, and restoration of pancreatic β-cell architecture, particularly at higher doses. Conclusion: The developed polyherbal formulation exhibits significant antidiabetic potential, supported by its rich phytochemical profile and multi-mechanistic action. Chromatographic standardization ensures quality, safety, and reproducibility. The findings suggest that the formulation may serve as a promising alternative or adjunct to conventional antidiabetic therapy, warranting further clinical investigation.

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