The research was aimed to investigate the antibacterial active ingredient of Chinese herbal formulation extract. Different concentration of ethanol was used to process the water extract of the Chinese herbal formulation, in order to remove the impurity in the water extract. Four polarity extracts were obtained by using different polarities of solvents such as n-hexane, ethyl acetate, n-butanol and water. The antibacterial ability of each polar parts was judged according to the size of the antibacterial circle. 70% concentration of ethano1can effectively remove the impurity in the water extract which can reduce the interference in the separation of active constituents. Different polarity fractions of extracts from the Chinese herbal formulation showed different bacteriostasis. N-butanol fraction and water fraction had better bacteriostasis than ethyl acetate fraction and n-hexane fraction. This study showed that the antibacterial active constituents are mainly high polarity fractions and the other polarity fractions may have synergy.