Acnano is a polyherbal nano-emulsion indicated for the treatment of mild to moderate as well as chronic acne. It contains Tea tree oil, Rosemary oil & Mentha oil along with Citrus limon. This study was undertaken to determine the antimicrobial efficacy of Acnano against acne causing microbes such as Staphylococcus aureus & S. epidermidis by time kill curve studies. A rapid killing time was achieved by Acnano. Bacterial count was 2 Log10 cfu/mL after 6 h of study in both the organisms. No deviation in pattern of bacterial inhibition was found and there was no regrowth reported even after exposure for longer time under influence of Acnano. In conclusion, Acnano possesses good antimicrobial activity against S. aureus & S. epidermidis.