The aim of this study was to determine whether the timing or mode of application influences the antimicrobial efficacy of a sodium hypochlorite solution against microorganisms on fresh-cut Romaine lettuce. Lettuce leaves were treated with a solution of sodium hypochlorite containing 70 mg liter(-1) free chlorine prior to slicing (WLS), while submerged in the solution (CWS), or after slicing (PCS), the last being common practice in the fresh-cut industry. Immediate antimicrobial effects were determined from analyses performed directly after treatment, while delayed effects were derived from analysis of packaged samples stored at 4 and 15°C. Experiments were performed with both uninoculated and inoculated whole leaves, using Escherichia coli O157:H7 from a type collection as an inoculum. The CWS treatment provided the greatest initial reductions in aerobic plate counts of E. coli O157:H7 populations, followed by PCS and WLS. The advantage afforded by the CWS approach was maintained over 10 days in storage at 4°C. In addition, it was noted that the visual quality attributes of the fresh-cut lettuce from the CWS approach were better than those from the other two treatments. These results collectively suggest that slicing lettuce under a sanitizer solution could provide better disinfection of fresh-cut Romaine lettuce than the current practice of washing with sanitizer after cutting has taken place.