Paeonol hydrazine was generated by the reaction of paeonol(2-hydroxy-4-methoxyphenylethanone) as raw material with hydrazine hydrate. Night novel N-benzylidene paeonol hydrazones were synthesized in the yields of 50.5%~91.9% by the reaction of paeonol hydrazine with various benzaldehydes respectively. The synthetic conditions were investigated preliminarily. The structures of the products were confirmed by FT-IR, 1H NMR, 13C NMR and LC-MS. The preliminary bioassay showed that some of the title compounds exhibited excellent antibacterial activity, in which the MIC of compound N-(4-fluorobenzylidene) paeonol hydrazones (3c) against S.aureus was 4μg/mL and compound N-(2-fluorobenzylidene) paeonol hydrazones (3b) similar effect against E.coli. than the standard reference drug (rifampin).