Abstract:The goal of this study was to investigate the constituents and antimicrobial activity of industrial hemp（Cannabis sativa L.）leaves against Listeria monocytogenes as a potential food preservative.The leaves were first extracted with ethanol，further partitioned with ethyl acetate，and then subjected to silica gel gradient fractionation with petroleum ether-ethyl acetate.The main active ingredients were initially separated based on the minimum inhibitory concentration as measured by broth dilution and then identified by gas chromatographymass spectrometry.Following extraction and identification，antimicrobial activity was investigated in detail by comparing changes in cell wall integrity，cell membrane permeability，energy metabolism and oxidative damage of the eluted fractions.The antimicrobial components were mainly distributed in the 1∶1 petroleum ether：ethyl acetate fraction from the gradient extraction（Fr.1：1），with a minimum inhibitory concentration of 62.5 μg/mL.A total of 24 volatile components were identified，among which methyl linolenate（40.79%）and methyl hexadecanoate（13.12%）were the most abundant.After treatment with Fr.1：1，the surface of the bacteria became visibly broken，extracellular alkaline phosphatase content increased，and electrical conductivity rose，while the levels of adenosine triphosphate and superoxide dismutase first rose and then fell.In conclusion，Fr.1:1 could damage the structure of the cell wall and membrane of Listeria monocytogenes，resulting in increased membrane permeability and thus release of cell contents，affecting bacterial metabolism，and causing oxidative damage to the plasma membrane，all leading to inhibition of cell growth.