Abstract: A molecularly imprinted polymer (MIP) able to bind 4- nitrophenol (4-NP) was prepared using noncovalent molecular imprinting methods and evaluated as a selective sorbent in molecularly imprinted solid-phase extraction (MISPE) on-line coupled to a reversed-phase HPLC. It has been shown that the conditions chosen for washing the MIP and for eluting the analyte in the MISPE process are extremely important for ensuring good selectivity and recovery. River water samples, spiked with the 11 Environmental Protection Agency phenolic compounds at microgram per Liter levels, were preconcentrated on-line using this MIP, and 4-NP was selectively extracted. The humic acid interference was simultaneously reduced considerably. The MTP was also compared with a commercially available highly cross-linked polymer (LiChrolut EN) and the former yielded cleaner extracts