Application of the Ultrasonic-Assisted Extraction and Dispersive Liquid-Liquid Microextraction for the Analysis of AFB1 in Egg

A fast and simple sample preparation method based on dispersive liquid-liquid microextraction (DLLME) and ultrasonic-assisted extraction (UAE) was proposed in order to determine aflatoxin B-1 (AFB(1)) in eggs. In this study, AFB(1) was first extracted from matrixes of egg samples with acetonitrile/water 80:20 (V/V), then its eluents were utilized as dispersant of the followed UAE-DLLME for further purification and concentration of AFB(1). All parameters that could affect the effectiveness of this method were investigated and optimized such as types and volumes of extractant and dispersive solvents, volume of water, pH of extracted solution, salt addition, ultrasonic time, ultrasonic temperature, and centrifugation time. Quantification method was HPLC-UV which does not require AFB(1) derivatization in samples. Under the optimum UAE-DLLME conditions (240 mu L CHCl3 as extraction solvent, 1.2 mL acetonitrile 80% extract as disperser solvent, water volume 3 mL, sample pH 5.6, ultrasonic time 2, and ultrasonic temperature 35 degrees C) recovered result were between 91 and 94%. The limits of detection and quantification were 0.12 and 0.32 mu g kg(-1), respectively. Real samples were analyzed by the proposed UAE-DLLME method and then compared with confirmative immunoaffinity column chromatography (IAC) clean-up. The proposed UAE-DLLME method is economical, simple, high enrichment factor, quick, and environmental friendly.