The bacterial resistance shown by the microorganisms in the presence of antimicrobial agents has prompted the search for new materials to overcome this public health problem. Gambir (Uncaria gambir, Roxb.) leaves extract has high catechin content thereby potentially antibacterial. This research aimed to examine the effect of purified gambir extract on inhibiting growth and killing bacteria. The antibacterial activity of Purified gambir extract (Uncaria gambir Roxb.) with concentrations of 32%, 16%, and 8% was tested against S. aureus and P. aeruginosa using the agar diffusion method. Purified gambir extract at concentrations of 8%, 16%, and 32% showed antibacterial activity against Staphylococcus aureus with an average inhibition zone diameter of 19.25 mm, 23.45 mm, and 26.25 mm, respectively. Purified gambir extract at concentrations of 8%, 16%, and 32% also showed antibacterial activity against Pseudomonas aeruginosa with an average inhibition zone diameter of 11.55 mm, 15,4 mm, and 18,5 mm, respectively. In conclusion, purified gambir extract (Uncaria gambir Roxb.) has very strong antibacterial activity against Staphylococcus aureus bacteria and strong antibacterial activity against Pseudomonas aeruginosa.