Evaluation of reduced graphene oxide-based nanomaterial as dispersive solid phase extraction sorbent for isolation and purification of aflatoxins from poultry feed, combined with UHPLC-MS/MS analysis.

Poultry feed comprises cereals and their by-products and is vulnerable to aflatoxins contamination. This study utilised reduced graphene oxide-titanium dioxide (rGO-TiO 2 ) nanomaterial as a dispersive solid phase extraction (d-SPE) adsorbent to extract, enrich and purify aflatoxins (aflatoxin B1, aflatoxin B2, aflatoxin G1 and aflatoxin G2). The synthesis of rGO-TiO 2 nanomaterials through hydrothermal process and characterisation by transmission electron microscopy, scanning electron microscopy, Brunauer-Emmett-Teller (BET) and X-ray diffraction reveals that the nanomaterials have a single-layer structure embedded with TiO 2 nanoparticles. The matrix-spiked technique was employed for the extraction process, optimisation of d-SPE, and analytical method validation. The most appropriate extraction solvent was acetonitrile/water/formic acid (79/20/1, v/v/v), with 30 min of extraction time assisted by ultra-sonication. The optimised d-SPE parameters were: 50 mg of rGO-TiO 2 as sorbent amount, 2% methanol as the sample loading solvent, 30 min as adsorption time, and absolute ethanol as the washing reagent. The d-SPE method exhibited good desorption efficiency with 3 mL of acetonitrile/formic acid (99/1, v/v) and 20 min desorption time. After validation, the UHPLC-MS/MS analytical method has an acceptable range of specificity, linearity ( R 2  ≥ 0.999), sensitivity (LOQ 0.04-0.1 µg kg -1 ), recoveries (74-105% at three matrix-spiked levels) and precision (RSD 1.5-9.6%). Poultry feed samples ( n  = 12) were pretreated by this method to extract, enrich and analyse aflatoxins, which were detected in all poultry feed samples. The contamination levels were within the permissible limits.