The aim of this study is to investigate the anti-biofilm activity of 1,10-PHEN against E.coli and to analyze its anti-membrane mechanism.The bacteriostatic effect of 1,10-PHEN on E.coli was evaluated by measuring the time bactericidal curve,and the minimum biofilm inhibitory concentration(MBIC)of 1,10-PHEN on E.coli was measured.The inhibitory effect of 1,10-PHEN on E.coli biofilm under fluorescence microscope was observed.The effects of 1,10-PHEN on the development stage and EPS of E.coli biofilm were detected.The inhibitory effect of 1,10-PHEN on E.coli was concentration-dependent.The MBIC values of E.coli E-10 were 12.5 μmol/L,and 50.0 μmol/L could significantly destroy the mature biofilm of E.coli.1,10-PHEN could in-hibit the formation of biofilm by inhibiting the adhesion ability of E.coli at the early stage of bio-film formation.1,10-PHEN could inhibit the synthesis and secretion of EPS from E.coli biofilm.1,10-PHEN could inhibit the synthesis and secretion of EPS from E.coli biofilm,and the inhibition rates of EPS at 1/4 MIC were 32％and 78％,respectively.The inhibitory rates of 1,10-PHEN on eDNA of ATCC 29522 and E-10 at 1/4 MIC concentration were 72％and 61％,respectively,and the inhibitory rates of MIC concentration on extracellular protein were 69％and 56％.In conclusion,1,10-PHEN has anti-biofilm activity against E.coli,which provides a new idea for find-ing alternative antibiotics.