Biodegradation has proved to be a versatile technique to remediate benzene, toluene, ethyl benzene and xylene mixtures in contaminated soil and groundwater. In this study, a mixed microbial culture obtained from a wastewater treatment plant was used to degrade liquid phase ethyl benzene and xylene, at initial concentrations varying between 15 to 75 mg/l. Experiments were conducted according to the 2k−1 fractional factorial design to identify the main and interaction effects of parameters and their influence on biodegradation of individual ethyl benzene and xylene compounds in mixtures. The removal efficiencies of these compounds varied between 2 to 90% depending on the concentration of other compounds and also on their interaction effects. A statistical interpretation of the results was done based on the Fishers variance ratio (F) and probability (P) values. Though all the main effects were found significant (P < 0.05) at the 5% confidence level, the interactions between benzene and toluene and benzene and xylene concentrations were also found to be statistically significant and play a major role in affecting the total ethyl benzene and xylene removal.