The research was a quantitative descriptive study using laboratory experimental methods. We conducted this research to analyze the combination formula of Tambora leaves, Sembalit Angin leaves, and turmeric rhizomes using 3: 2: 1 ratio and determine its effectiveness in-inhibiting the-growth of Staphylococcus aureus bacteria in vitro. The width of the inhibition zone measured the growth of Staphylococcus aureus from the outer side of the paper disc, which has been immersed in methanol extract of Tambora leaves, Sembalit Angin leaves, and turmeric rhizomes in nutrient agar medium. Calipers did measurement, which were made during the incubation period of 24 hours, 48 hours, and 72 hours. The data obtained from the effect of a combination formula of 3:2:1 Tambora leaves, Sembalit Angin leaves, and turmeric rhizomes on the growth of Staphylococcus aureus were analyzed by Anava statistical test, and  Duncan 1% test. The results found that the combination formula of 3:2:1 bio herbal methanol extract of Tambora leaves, Sembalit Angin leaves, and Turmeric rhizomes had a significant effect on the growth of Staphylococcus aureus at 1% significance, proven by the sig value. 0.00 <0.01, so the 3: 2: we can recommend 1 combination formula as the most effective combination extract formulation in inhibiting the growth of Staphylococcus aureus.