In this work, a simple and miniaturized solid‐phase extraction device was constructed by connecting a commercial nylon needle filter to a syringe, which was applied for extracting 1‐hydroxypyrene from a urine sample via hydrophobic and hydrogen bond interactions. The nylon membrane in the needle filter acted as the solid‐phase extraction adsorbent, meanwhile, it filtered the particles in the urine sample. To obtain high extraction efficiency, key parameters influencing extraction recovery were investigated. The entire pretreatment process was accomplished within 5 min under the optimal conditions. By coupling high‐performance liquid chromatography–ultraviolet, a rapid, low‐cost, and convenient nylon needle filter‐based method was established for the analysis of 1‐hydroxypyrene in a complex urine matrix. Within the linearity range of 0.2–1000 μg/L, the method exhibited a satisfactory correlation coefficient (R = 0.9999). The limit of detection was 0.06 μg/L, and the recoveries from urine sample spiked with three concentrations (5, 20, and 100 μg/L) ranged from 105.8% to 113.1% with the relative standard deviations less than 6.7% (intra‐day, n = 6) and 8.9% (inter‐day, n = 4). Finally, the proposed method was successfully applied for detecting 1‐hydroxypyrene in urine samples from college students, smokers, gas station workers, and chip factory workers. The detected concentration in actual urine samples ranged from 0.46 to 5.26 μg/L. Taken together, this simple and cost‐effective nylon needle filter‐based solid‐phase extraction device showed an excellent application potential for pretreating hydrophobic analytes from aqueous samples.