The present work aims to formulate and evaluate the Eupatorium glandulosum Michx based topical gel for wound healing activity. The gel was formulated using the methanolic extract of the Eupatorium glandulosum michx and carbopol as base. The prepared gel was characterized for their physicochemical constants, preliminary phytochemical analysis, quantitative analysis, spreadability, pH, viscosity and extrudability. Antimicrobial and wound healing studies were carried out for the optimized formulation (F5). The physicochemical constants like total ash, acid insoluble ash, and water-soluble ash are specific identification. The methanolic extract was prepared by maceration method and the extract was formulated into topical gel formulation. The results of preliminary phytochemical investigation showed the presence of carbohydrates, cardiac glycosides, phenols and flavonoids. Quantitative analysis of polyphenolic compounds showed high content of flavonoids. Other parameters such as spreadability, pH, viscosity and extrudability were found to be well within the limits. The antimicrobial activity of optimized formulation was found to be effective in both gram positive and gram-negative organisms. The optimized formulation promotes better wound healing with comparison to that of the marketed product. From the present study, it was concluded that the prepared herbal gel formulation showed significant antimicrobial potential and found to posses better wound healing activity.