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Knowledge of in-process oxygen transfer is essential to the optimum design and operation of aeration systems in activated sludge processes. In this study, non-steady state H2O2 oxygen transfer tests were performed in a laboratory scale, University of Cape Town configuration biological nutrient removal process (BNR) to measure the in-process oxygen transfer rates. Given the small quantity (about 1 ml @30% concentration) of H2O2 used in the aerobic reactor with a mixed liquor volume of 161 (total system working volume of 33.65 l), no effect on the process performance itself was expected. However, the process performance data obtained indicated results to the contrary. Use of H2O2 in measuring process oxygen transfer rates may not be suitable for BNR processes, as all the major process performance indicators (carbon, nitrogen and phosphorus removal by the system and carbon uptake and phosphorus release in the anaerobic zone) were negatively affected. Evidence in thiswork leads to the conclusion that external addition of H2O2 leads to excessive production of the hydroxyl radical. Since microorganisms do not have enzyme systems capable of acting upon this additional reactive radical, it resulted in loss of process performance. It is also possible that H2O2 could have upset the normal aerobic respiration process by introducing oxidative stress conditions on the catalase, peroxidase and superoxide dismutase enzymes that deal with other oxygen radicals generated as part of the overall process of reducing oxygen to water. Any test that interferes with the system in which the test is being performed is suspect. Therefore, in light of these results, the H2O2 method to test in-process oxygen transfer should be re-evaluated.