Women are susceptible to urinary tract infections (UTIs), especially during pregnancy and in the perimenopausal and postmenopausal stages. Significant disabilities and fatalities are linked to UTIs and they have a direct effect on the patients' quality of life. The aim of the current study was to prepare the aqueous extract of Dioscorea alata, Embelia ribes and their formulation extract and analyze the efficacy of antimicrobial activity against UTI pathogens and its cytotoxic effect. In the research work, the extract was prepared and the antimicrobial efficacy of D. alata, E. ribes and their formulation was analyzed using the agar well diffusion technique (50 µL) and the time-kill curve assay. The extracts and their formulation were used to determine toxicity and cytotoxic effects using brine shrimp lethality analysis. D. alata, E. ribes and their formulation was prepared and color of the extract was confirmed. In antimicrobial activity against UTI pathogens, especially Pseudomonas spp., a higher inhibitory zone was observed in D. alata, E. ribes and their formulation extract at the concentration (50 µL) and the cytotoxic effect showed lower toxicity in all extract at the higher concentration (80 µL). The formulation extract showed significant extract antimicrobial activity with lower toxicity, suggesting its potential use as an antimicrobial agent for the treatment of UTIs.