Aim: Dasamula is an Ayurvedic formulation, which is used for the management of pain, arthritis, and inflammatory disorders. Even though this formulation has been widely used in the Ayurvedic system of medicine to cure inflammation-mediated diseases, no deeper research studies have been conducted to prove its antioxidant and anti-inflammatory activities. Materials and Methods: In the present study, Dasamula was prepared using the roots of available herbal ingredients of such as Bilva (Aegle marmelos L. Correa), Syonaka (Oroxylum indicum L. Benth. Ex Kurz), Gambhari/Kumil (Gmelina arborea Roxb.), Patala/Pathiri (Stereospermum suaveolens Roxb.), Agnimantha/Munjai (Premna serratifolia L.), Salaparni/Orilai (Desmodium gangeticum L. DC), Prisniparni/ Sithirapaladai (Uraria picta Jacq. Desv), Brhati/Sundai (Solanum indicum L.), Kantakari/Vaizhalanthai (Solanum xanthocarpum L.) and Goksura/Nerinjil (Tribulus terrestris L.) and screened for phytochemical profile, total phenol content, antioxidant activity (1,1-diphenyl-2-picryl-hydrazyl assay), and anti-inflammatory property (red blood cell membrane stabilization assay). Results and Discussion: From the results, it was seen that Dasamula had an impeccable anti-inflammatory activity. The Dasamula preparation contains various phytochemicals such as alkaloids, tannins, saponins, flavonoids and quinones, total phenolic content (567 mg gallic acid equivalent/100 g) and exhibits antioxidant (44.86%) and anti-inflammatory activities (32.56%), which are contributed by its herbal ingredients, especially A. marmelos and G. arborea. Conclusion: Among the roots of 10 different herbal ingredients are used in the preparation of Dasamula, only A. marmelos and G. arborea root extracts are found to be efficient as anti-inflammatory agents, and the remaining ingredients might have some medicinal roles in related therapeutic conditions/alleviating side effects.