Sensitive Metal Oxide-Clay Nanocomposite Colorimetric Sensor Development for Aflatoxin Detection in Foods: Corn and...

The work reports on zinc oxide bentonite nanocomposite (ZnOBt) chemical route synthesis, characterization, and investigation of curcumin (Cur) functionalization for a label-free colorimetric detection of total aflatoxins (AFs) in foods. XRD of ZnO nanoparticles (NPs) confirmed the wurtzite structure (2θ = 36.2°) and that of ZnOBt showed the intercalated interlayer composite phase. The Debye-Scherrer relation calculated the crystallite size as 20 nm (ZnO) and 24.4 nm (ZnOBt). Surface morphology by SEM exhibited flower-like hexagonal, rod-shaped ZnO NPs on the bentonite surface. Colorimetric reaction involved two-stage redox reactions between ZnOBt and dye Cur followed by AFs phenolic group and Zn(Cur)OBt. Cur gets oxidized at its diketone moiety in the presence of ZnOBt to form a red colored complex Zn(Cur)OBt, which further scavenge protons from AFs phenolic group, and gets oxidized to AFs-Zn(Cur)OBt (yellow). Binding of AFs-Zn(Cur)OBt is characterized by FT-IR ascribed to C-H bending (1966.615 cm^-1), O-H stretching (3256.974 cm^-1), and C=O stretching (1647.362 cm^-1). ¹H NMR chemical shifts (δ) (ppm) showed an increase in proton at the aliphatic region (0 to 4.4) while removal of proton in ether at 4.4 to 6 regions. Job plot calculation using UV-Vis data resulted in a higher total AF binding coefficient of Zn(Cur)OBt (K _a = 3.77 × 10⁶ mol^-1 L) compared to Zn(Cur)O (K _a = 0.645 × 10⁶ mol^-1 L) as well as a molar ratio of 1:1 by the Benesi-Hildebrand plot equation. Corn and almond food samples showed the total AFs LOD of 2.74 and 4.34 ppb, respectively. The results are validated with standard LC/MS-MS in compliance with MRL value as per the regulatory standard (EU).The NP-based method is facile and rapid and hence can be utilized for onsite detection of total AFs in foods.