Introduction/Objectives:Amaranthus spinosus ethanolicextract exhibits phenoliccompound presence and flavonoid presence, as well as a notable reduction in microbial growth suppression using the agar plate method. The existence of antimicrobial medicines with strong anti-microbial action, such as benzoic acid, O-amino benzohydroxamic acid, 2-aminobenzoyl hydrazide, and 2-hydroxy acetyl acetate, is supported by GC-mass studies. The primary goal of this study is to evaluate Amaranthus spinosus's ability to identify the chemical compounds needed to measure the antibacterial activity using various models. Methodology: The chemical molecule for the antibacterial activity was found using the GC-mass spectrometry equipment, phytochemical compounds, and the Agar Plate Method. Several models are utilized fordetermine the antimicrobial propertyof the Amaranthus Spinosus. Results: In this study data show ethanolic extract of the Amaranthus Spinosus show significant reduction in microbial growth inhibition using agar plate method, phenolic compound presence and flavonoid present. GC-mass study support the presence of antimicrobial drugs like Benzoic acid, O- Amino benzohydroxamic acid, 2-Aminobenzoyl hydrazide, Benzoic acid, 2-hydroxy acetyl acetate which have significant anti-microbial activity. Conclusion: The studies data support significant anti-microbial activity using ethanolic extract of Amaranthus Spinosus leaves for anti-microbial activity.The formation of suitable dosage form by isolate phytochemical compound which responsible for anti-microbial activity and use on the animalmodels will be increase the chance of better results. Keywords: Amaranthus Spinosus, GC-mass,Isolate Phytochemical Compound,Anti-microbial activity,