The present work is the first attempt to use countercurrent chromatography for xenobiotics extraction and separation from complex natural matrices without sample pretreatment. A procedure has been developed for the direct extraction and separation of some PAHs from a sewage sludge medium being in fact a concentrated suspension. The sewage sludge and aqueous solutions containing the trace amounts (0.01–10 μg/mL) of PAHs were used as the stationary phase in the column whereas organic solvents (n-heptane, dichloromethane) or their mixture were used as mobile phase.