Title: Harnessing Indigenous Flora: Development and Pharmacological Validation of a Novel Anti-inflammatory Polyherbal Extract

Background: Chronic inflammatory diseases are a global health concern, and the side effects of conventional treatments have spurred interest in natural alternatives. Polyherbal formulations, which leverage the synergistic effects of multiple plant compounds, offer a promising and potent approach to developing new anti-inflammatory remedies.

Methods: A novel polyherbal extract was formulated using the leaves of four indigenous plants traditionally used for their anti-inflammatory properties: Tamarindus indica, Ricinus communis, Calotropis gigantea, and Vitex negundo. The combined extract was subjected to preliminary phytochemical screening to identify major constituent classes. Quantitative analyses were performed to determine the total phenolic and total flavonoid content using spectrophotometric methods. The presence and concentration of the key anti-inflammatory triterpenoid, lupeol, were quantified using a validated High-Performance Thin Layer Chromatography (HPTLC) method. The anti-inflammatory activity was assessed in vitro by evaluating the extract’s ability to inhibit protein denaturation, while antioxidant activity was determined using the DPPH radical scavenging assay.

Results: Qualitative screening confirmed the presence of diverse phytochemicals, including flavonoids and phenolics. The HPTLC analysis successfully identified and quantified lupeol in the polyherbal extract, with a concentration of 0.52 mug/mg. The extract exhibited significant dose-dependent anti-inflammatory activity, effectively inhibiting protein denaturation. Furthermore, it demonstrated potent antioxidant properties, with an IC50 value of 45.18 mug/mL.

Conclusion: This study successfully developed and pharmacologically validated a novel anti-inflammatory polyherbal extract. The strong antioxidant and anti-inflammatory activities observed are attributed to its rich phytochemical profile and the synergistic action of its components, particularly the high concentration of lupeol. This research provides

scientific evidence supporting the traditional use of these indigenous plants and highlights the importance of polyherbal formulations as a viable and effective strategy for developing new therapeutic agents. Future research should focus on in vivo studies and the standardization of the formulation to ensure consistent efficacy.