Miniaturized vortex assisted-dispersive molecularly imprinted polymer micro-solid phase extraction and HPLC-MS/MS for assessing trace aflatoxins in cultured fish

A dispersive micro-solid phase extraction approach using a molecularly imprinted polymer as an adsorbent has been developed for pre-concentrating aflatoxins from cultured fish. Aflatoxins were first isolated from fish muscle and liver by an ultrasound assisted extraction procedure using a 60 : 40 acetonitrile/0.1 M KH2PO4 aqueous buffer (pH 6.0) mixture. Polymeric adsorbent beads were synthesized using 5,7-dimethoxycoumarin as a dummy template, methacrylic acid as a functional monomer, divinylbenzene as a crosslinker, and 2,2'-azobisisobutyronitrile as an initiator. Parameters affecting the steps of extraction procedure, including the sample (fish extract) pH, adsorption stirring speed and time, desorption stirring speed and time, elution solvent ratio, and polymer capacity, were investigated and optimized. The limit of detection was found to vary from 0.29 to 0.61 μg kg-1 for the several aflatoxins. The proposed method was shown to be accurate and precise. Intraday and inter-day relative standard deviations were lower than 20%, and intraday and inter-day analytical recoveries were within the 80-100% range. The prepared adsorbent in the dispersive micro-solid phase extraction format was re-usable, and the pre-concentration procedure was found to be simple, rapid and highly selective and sensitive to identify/quantify AFs in fish.