BACKGROUND: Decreasing the microbial skin burden reduces the risk of surgical site infection (SSI). The present study compares the activity of an innovative 2% chlorhexidine gluconate (CHG)-impregnated preoperative skin preparation cloth (PC) with a standard application procedure with a 4% CHG surgical skin preparation (SP). METHODS: A paired, randomized, parallel phase III study was conducted adhering to the Food and Drug Administration (FDA) design criteria for evaluating preoperative skin preparations. Subjects' left and right sides of the inguinal and abdominal skin sites (n = 30) were randomized to either PC or SP treatment. Following baseline cultures, PC sites were prepped for 3 minutes, and SP sites were prepped for 4 minutes. Skin site cultures were obtained at 10 minutes, 30 minutes, and 6 hours postpreparation. Bacterial recovery was expressed as log(10) colony-forming units (cfu)/cm(2) for baseline and postapplication microbial recovery. RESULTS: Mean microbial baseline for the abdominal and inguinal skin sites were as follows: PC = 3.36 cfu/cm(2) and 6.15 cfu/cm(2); SP = 3.51 cfu/cm(2) and 6.16 cfu/cm(2), respectively. Log(10) reduction for PC abdominal and inguinal prepped sites at 10 minutes, 30 minutes, and 6 hours postpreparation were 2.50, 2.33, and 2.54; 3.45, 3.50, and 3.64, respectively. Log(10) reductions for SP abdominal and inguinal prepped sites at 10 minutes, 30 minutes, and 6 hours were 2.18, 2.19, and 2.77; 2.78, 2.63, and 3.15, respectively. CONCLUSION: Microbial reductions from abdominal-inguinal PC prepped sites were significantly reduced (P < .05) compared with baseline, exceeding the FDA log-reduction criteria for a preoperative topical skin preparation. Compared with baseline, microbial reductions at the SP-prepped abdominal-inguinal sites were significant (P < .05). SP abdominal-prepped sites met the FDA log-reduction criteria; inguinal sites, however, failed to meet expected FDA log-reduction criteria at 10 minutes postpreparation. The PC-treated inguinal sites at 10 minutes, 30 minutes, and 6 hours post-skin preparation demonstrated significantly greater microbial reductions than did the SP-treated inguinal sites (P < .01).