An environmentally friendly thermosensitive nanovesicle-cloud point microextraction technique has been developed with the assistant of ultrasonic waves to determine analgesic drugs with a broad range of polarity in field water and human urine. Based on thin-film hydration, the conformation of nanovesicles formed by a binary mixing system with the nonionic surfactants was evaluated using regular and cryogenic transmission electron microscopy. The multilayered nano-spherical structure was able to capture polar and nonpolar compounds simultaneously. Analgesic drugs (acetaminophen, salicylic acid, ketoprofen, diclofenac, indomethacin, ibuprofen, and mefenamic acid) were detected by ultra-performance liquid chromatography coupled to photodiode array detection. Under optimal conditions including the type and ratio of surfactants, sonication time and sonication temperature, linear calibration curves were obtained over the range of 50–8000 μg L−1. The coefficient of determination (R2) ranged from 0.9953 to 0.9995, with detection limits of 10–100 μg L−1. The relative standard deviations ranged from 3.2% to 12.7% for intraday precision (n = 5) and 2.5% to 14.1% for interday precision (n = 15). The relative recoveries obtained from one industrial wastewater sample and two field water samples ranged from 86.1% to 108.1%. In the human urine analysis, three volunteers ingested 1500 mg of acetaminophen. After 4 h, the concentration of acetaminophen in the urine was found to range from 87.0 to 197.9 mg L−1.
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