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This work describes the application of ultrasound-assisted dispersive solid-phase extraction (UA-DSPE) as a sample preparation approach for aflatoxin M1 (AFM1) and also its subsequent determination by HPLC–fluorescence detection. A Box–Behnken design in combination with response surface methodology was implemented to determine the variables affecting the extraction procedure. The effects of different variables, including type and quantity of clean-up phase, ultrasonication time, ultrasonication temperature, nature and volume of the leaching solvent, were investigated in the optimization study. Primary secondary amine (PSA) and acetonitrile were selected as the clean-up phase and the leaching solvent, respectively. The obtained optimized values were 30 mg of PSA, 10 min ultrasonication time, 32°C ultrasonication temperature and 10 mL of acetonitrile. Under the optimal conditions, the limit of detection and limit of quantification were 0.0012 and 0.0045 µg kg −1 , respectively. The recoveries of the UA-DSPE procedure ranged from 80 to 92%, with relative standard deviations lower than 10% in all cases. Eventually, this method was successfully applied to the extraction of AFM1 in milk powder samples.

Determination of Aflatoxin M1 in Milk Powder by Ultrasonic-Assisted Extraction and Dispersive Solid-Phase Clean-up