Tāḷicāti vaṭakam (TSV), a polyherbal siddha drug was chosen, it was screened for antimicrobial study and also subjected to standardization parameters. The ingredients were procured, authenticated and prepared the drug as per standard operating procedure. The ethanolic extract of TSV was screened for nine bacteria and two fungi. Then the drug was investigated for the phytochemical profile, physicochemical parameters, thin layer chromatographic  photo documentation (TLC), high-performance thin-layer chromatography (HPTLC) finger print profile. Antimicrobial assay revealed  inhibitory activity against all test pathogens. TLC under UV  showed 8 bands at short wavelength, 13 bands at long wavelength; 8 bands showed post derivatization with vanillin Sulphuric acid reagent. The present investigation concluded that the siddha herbal preparation of TSV have great potential on antimicrobial against pathogens. This siddha formulation can be used to prevent the bacterial and fungal infections and the standards could be used for quality control of the drug.