A simple and sensitive method for the determination of 5-hydroxymethylfurfural (HMF) in coffee, honey, beer, Coke, and urine by high-performance liquid chromatography (HPLC) is presented. This method is based on the formation of the 2,4-dinitrophenylhydrazone of HMF and subsequent polymer monolith microextraction (PMME) of this derivative. A poly(methacrylic acid-co-ethylene glycol dimethacrylate) (MAA-co-EGDMA) monolithic capillary column was selected as the extraction medium. Several parameters affecting the derivatization of HMF with 2,4-dinitrophenylhydrazine (DNPH) followed by extraction of the derivative were optimized. The procedure is simple and offers high sensitivity and specificity since the derivative of HMF is well preconcentrated by PMME with poly(MAA-co-EGDMA) monolith and well separated from the other components of the samples under examination. The recoveries in coffee, honey, beer, Coke, and urine samples were in the range of 83.9-110.8% spiked at different levels with HMF. The inter- and intraday precisions were less than 10%. The LOD (S/N = 3) and LOQ (S/N = 10) for HMF were 1.0 ng/mL and 3.4 ng/mL, respectively.