Objectives: The aim of the current study was to evaluate the pharmacological activity of the ethanolic extract of phyllanthus amarus by using leaves for its antimicrobial activity against the clinically isolated human pathogens. The pathogens such as the Staphylococcus ( gram positive bacteria ) and E. coli ( gram negative bacteria ) were evaluated in the formulation of herbal ointment. Materials and methods: The leaves are collected and then powdered, the bioactive compound was obtained by using the ethanol in the maceration process. Simple ointment base ( B.P.) was prepared, then extract ( Phyllanthus amarus ) was incorporated to form a different concentration of semi-solid herbal ointment. Leaves extract was carried out , by the detection of phytochemical analysis and evaluation were done. In the antimicrobial study was done by the cup plate method by using the sensitivity of the organisms in ointment contains variety of concentrations of extract in the zone of inhibition. Keywords: Herbal Ointment, phyllanthus amarus , phytochemical analysis, antimicrobial activity.