Results of the studies for determining fractions of organic contaminants in a pretreated petrochemical wastewater flowing into a pilot Aerated Submerged Fixed-Bed Biofilm Reactor (ASFBBR) are presented and discussed. The method of chemical oxygen demand (COD) fractionation consisted of physical tests and biological assays. It was found that the main part of the total COD in the petrochemical, pretreated wastewater was soluble organic substance with average value of 57.6%. The fractions of particulate and colloidal organic matter were found to be 31.8% and 10.6%, respectively. About 40% of COD in the influent was determined as readily biodegradable COD. The inert fraction of the soluble organic matter in the petrochemical wastewater constituted about 60% of the influent colloidal and soluble COD. Determination of degree of hydrolysis (DH) of the colloidal fraction of COD was also included in the paper. The estimated value of DH was about 62%. Values of the assayed COD fractions were compared with the same parameters obtained for municipal wastewater by other authors.