There are many skin antiseptics commercially available. Although their antibacterial activity has often been well studied [1], their potential effectiveness on skin remains poorly documented. To date, in-vivo protocols designed for the testing of the antimicrobial ef®cacy of antiseptics cannot use, for ethical reasons, pathogenic bacteria or new formulations whose toxicity in human subjects is unknown. An`ex-vivo' test was recently developed to overcome these problems. Freshly excised human skin from abdominal or breast reduction was placed in a diffusion cell containing a maintenance medium in the recipient compartment. A bacterial inoculum was then applied to the stratum corneum and, after a drying step, antiseptic formulations were evaluated for their antimicrobial activity. Several micro-organisms were investigated: ± Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), Enterococcus faecalis, vancomycinresistant Ent. faecium (VRE), S. epidermidis, Pseudomonas aeruginosa and Escherichia coli ± with several biocides ± para-chloro-meta-xylenol (PCMX, active compound of Dettol), povidone iodine, triclosan (in isopropanol) and chlorhexidine. Results from the ex-vivo test were compared with results obtained in suspension and glass-carrier tests. The bactericidal activity of the biocides depended upon the test performed and results were generally signi®cantly different from one method to the other. All biocides tested in the suspension test achieved >4 log 10 reduction in viable bacterial concentrations, apart from povidone iodine tested against Ent. faecalis and VRE. The antibacterial activity of biocides tested in the glass-carrier test was signi®cantly lower than in the suspension test, with the exception of triclosan in isopropanol, which was as effective in both suspension and glass-carrier test. In the ex-vivo test, triclosan in isopropanol achieved a log 10 reduction in viable bacterial concentration of 1.105±1.771 (with the exception of P. aeruginosa with 0.758 log 10 reduction). PCMX, povidone iodine and chlorhexidine achieved log 10 reductions in viable bacterial concentration of 0.303±0.901. Chlorhexidine tested against P. aeruginosa produced a 1.94 log 10 reduction in concentration. These results con®rm previous observations about the need for testing the antimicrobial activity of antiseptics on skin surface to determine their in-situ ef®cacy and encourage further the use of the ex-vivo protocol.