Seven Sesamum indicum (sesame) honey samples were collected from Hooghly district of West Bengal, India and analyzed for polyphenol and flavonoid content along with their invitro free radical scavenging activities. Antibacterial activity and stimulatory effect on multiplication of probiotic bacteria were evaluated. Antioxidant markers like IC50 value for DPPH (1, 1-diphenyl-2-picrylhydrazyl) and FRAP (ferric reducing antioxidant power) of sesame honey were positively correlated to its polyphenolic content (28.9±0.6mg GAE/100g) and color intensity (r ranges between 0.872 and 0.931). Four flavonoids viz., apigenin, quercetin, myricetin, rutin have been identified along with one cinnamic acid derivative (ferulic acid) and two sesame lignans (sesamin and episesamin) by High performance liquid chromatography which can be used as a tool for authentication of sesame honey. Antibacterial activity of sesame honey against some enteropathogenic bacteria Escherichia coli, Vibrio cholerae, S. Typhi, S. Typhimurium were studied. Minimum inhibitory concentration was found to be lowest against S. Typhi (12.5% w/v) and S. Typhimurium (12.5% w/v). Plasmid DNA degradation by sesame honey administration evinced its molecular level of action. Interestingly, it was also found that sesame honey exhibited significant growth promoting property of probiotic strains such as Lactobacillus acidophilus and Bifidobacterium bifidum. © 2014 Elsevier Ltd.