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E-101 solution is a first-in-class myeloperoxidase containing antimicrobial solution developed for topical application. The active ingredients in E-101 solution are two enzymes, porcine myeloperoxidase (pMPO) and glucose oxidase (GO) in an aqueous solution and activated by the addition of a glucose solution. Once activated, the reactive species hydrogen peroxide, hypochlorous acid/hypochlorite (HOCl/OCl-), and singlet oxygen (1O2) are generated. We evaluated the effect of whole human blood on the performance of E-101 solution compared to commercially available wound antiseptics and commonly used biocides. The wound cleansers NeutroPhase, Microcyn, and Vashe with the active HOCl/OCl- component were tested according to the USP-51 effectiveness test in the presence of 0, 1, 2 and 5% blood. Comparative time-kill studies against chlorhexidine, povidone-iodine, sodium oxychlorosene were tested in the presence of 0, 2, 5 10, and 20% blood. In the USP-51 test, E-101 solution demonstrated >2 log10 reduction against bacterial and fungal isolates in the presence of 5% blood at days 14 and 28. With the exception of NeutroPhase activity against S. aureus, all comparable wound antiseptics demonstrated <2 log10 reduction in the presence of 5% blood at days 14 and 28. Time-kill microbicidal data observed in the presence of blood demonstrated that E-101 solution was the most active biocide, followed by chlorhexidine and povidone-iodine. The presence of 2% blood completely inhibited the activity of sodium oxychlorosene. In summary, E-101 solution remained active in the presence of blood containing catalase and other substances that competitively react with 1O2 and HOCl/OCl- as a safe and effective wound antiseptic