The aim of the present study was to develop a new procedure for isolation, enrichment, and determination of 4‐nonylphenol (4NP) from aqueous matrices (surface waters: rivers, fishponds) and sewage. An attempt was focused to find selective sorbents for nonylphenol isolation by solid‐phase extraction (SPE), solvents selection for elution of this group of compounds isolated from aqueous samples, and the optimum composition of HPLC mobile and stationary phases for final determination of 4NP. New‐generation chromatographic packings were used whose structure resembles that of natural biological systems. The results obtained were verified by employing GC‐MS as a comparative method. #This paper is dedicated to memory of Prof. Csaba Horváth.