The effect of the cyclophosphamide and its mean metabolites on extracellular polymeric substances (EPS) formation was investigated. Two lab-scale membrane bioreactors were followed in parallel (one with the cytostatic drugs, the second without). Chromatographic and spectroscopic studies (UV-Vis spectroscopy and IR spectroscopy) showed that the presence of CPs induced an increase in EPS concentration in the biological sludge, especially of soluble substances, mainly polysaccharides and proteins. Size exclusion chromatography analysis revealed that in the presence of CPs, macromolecular EPS were formed (polysaccharides corresponding to about 6 KDa and proteins to about 18 KDa). The formation of EPS seemed to be a protection mechanism. More important membrane fouling in reactor with CPs seemed to be related to the retention of an increased amount of soluble substances. (c) 2009 Elsevier Ltd. All rights reserved.