Abstract: The chemical composition and active constituents of Jordanian planted saffron (Crocus sativus L) were studied, to evaluate a natural preservative effect of saffron on food. High Performance Liquid Chromatography (HPLC) studies using photodiode array analyses were performed on a waters HPLC system for the separation of several ingredients from alcoholic extracts of Jordanian saffron. Eight saffron peaks were identified by comparison of their retention times with those of known reference compounds and quantified from samples of saffron as follows: picrocrocin, picrocrocin acid, kaempferol, trans-crocin, trans-crocin-3, trans-crocin-4, safranal and cis-crocin-4. In this study the antibacterial effect of ethanolic extracts from petal of saffron against the most important foodborne pathogens Salmonella typhimurium, Listeria monocytogenes, E. coli, Bacillus cereus and Staphylococcus aureus were evaluated. The Minimum Inhibitory Concentration (MIC) of extracts were evaluated by agar dilution and broth micro dilution method. The most sensitive strain to extracts was S. typhimurium and the most resistant strains were S. aureus and E. coli MICs of all extracts were estimated against all bacteria using agar dilution method. The results of this study revealed that the studied extracts might be potential sources of natural antimicrobial agents and propose their potential application in food system as a natural preservative.