The dependence of microbial killing on chloride ions present in solutions undergoing iontophoresis is addressed. A 400-,uA current was applied to vials containing synthetic urine or saline, and the production of chlorine-based substances (CBSs) was detected by the N,N-diethyl-p-phenylene diamine colorimetric method.It was found that as the time of current application increased, the'total concentration of CBSs also increased.The iontophoretic current converted (through oxidation) chloride ions present in the solutions into CBSs such as free chlorine, chlorine dioxide, chlorite, monochloramine, and dichloramine (the last two were produced by iontophoresis only when nitrogenous substances were present in the solution). Two of the CBSs (free Cl and C102), when they were separately added back to microbial suspensions (approximately 3 x 105 CFU/ml) at the same concentrations at which they were detected in either 0.46% (wt/vol) NaCl solution or synthetic urine iontophoresed for 4 h at 400 ,LA, reduced or eliminated bacterial genera and a fungus. However, when free Cl and C102 were jointly added back to microbial suspensions, bacterial and fungal killing was synergistic and more rapid and complete than when these chlorine-based biocides were added separately. Therefore, iontophoresis of solutions containing chloride ions produces chlorine-based biocides that are responsible for the antimicrobial effect of iontophoresis.For many years urinary catheters have been identified as both the leading cause of nosocomial-induced urinary tract infections (UTIs) and the most common predisposing factor in fatal gram-negative sepsis in hospitals (13,24). Therefore, reducing the threat of nosocomial UTIs from catheter usage has been an important goal for our laboratory. We have designed a catheter-power supply system that delivers an iontophoretic current of 400 ,A to the urinary bladder through platinum electrodes (7). Previous research has demonstrated that this current will eliminate microbial populations only when it is passed through solutions containing chloride ions (11). We have also previously shown that platinum electrodes have the best durability and microbial killing efficiency and that iontophoresis for 4 h at 400 ,uA in synthetic urine generates a biocidal effect against selected fungi and gram-positive and gram-negative bacteria for more than 6 h after iontophoresis ceases (9, 10). Furthermore, bacterial killing efficiency was found to be directly related to increasing microamperage (up to 400 VA) and to chloride ion concentration, while it was found to be inversely related to the number of bacteria present in the solution undergoing iontophoresis (12). Preliminary work suggested that the reason for biocidal activity was the production of chlorine-containing oxidants. Therefore, the aim of the study described here was to both identify and quantify the various iontophoretically produced chlorine-based substances (CBSs) such as free chlorine (i.e., Cl2, OCI-, and HOCI), chlorine dioxide (CG02), chlorite (CdO2-), monochlo-...