This present study focuses on the in vitro biological activities of a polyherbal formulation composed of nine medicinal plants which were selected based on the traditional use in medicines. Hydroalcoholic extracts were prepared and evaluated for their in vitro efficacy by different models to shortlist for the final formulation. The extracts were tested for their antimicrobial, antioxidant and anti inflammatory activities. Among these, Andrographis paniculata, Coleus aromaticus, Tinospora cordifolia, Zingiber officianale and Piper nigrum extracts exhibited promising inhibitory activities against selected microorganisms. Extracts from Andrographis paniculata and Tinospora cordifolia exhibited significant scavenging activity against DPPH and NO free radicals. These two extracts exhibited strong anti-inflammatory response by inhibiting the LPS induced TNF in RAW cells. Among the tested extracts, five extracts from AP (Andrographis paniculata), CA (Coleus aromaticus), TC (Tinospora cordifolia), ZO (Zingiber officianale) and PN (Piper nigrum) were found to have promising antimicrobial, antioxidant and anti-inflammatory properties.