The focus of this present research is based on the analytical methodologies, which include the isolation of active ingredients from Celosia Spicata plant familyAmaranthaceae. This study emphasises the isolation of active compounds using column chromatography and with the help of TLC, solvent system identified which help in the characterisation of isolated compound. Furthermore characterisation performed via UV-visible Spectroscopy, FTIR (Fourier Transform Infrared spectroscopy), NMR (Nuclear Magnetic Resonance), and Mass Spectrometry with respect to standard compound. AdditionallyIn vitroantibacterial activity of Celosia Spicata plant was assessed byWell Diffusion assay.Quantitative analysis of total phenolics and flavonoids were determined by the well-known test protocol available in the literature.At the end result of all the analysis proven that the compound which was isolated from Celosia Spicata extract waspureStigma sterol.