HPLC Analysis of AFB1 Degradation

AFB₁ in the control groups and samples was extracted three times using chloroform with an equal volume before the solvent evaporated under N₂ at room temperature [23 (link),26 (link),57 (link),58 (link)]. Dimethyl sulfoxide (DMSO) (50 µL) was used to dissolve the dried extracts, and 20 µL of the mixture was injected in UltiMateTM3000 HPLC (Thermo Scientific, Bremen, Germany). HPLC analysis was conducted with a C18 Polaris column (250 mm × 4.6 mm i.d., 5 µm) in a mobile phase of water and methanol in a 1:1 ratio (v/v). The flow rate was set as 1 mL min⁻¹, and a UV/VIS detector (Thermo Scientific, Germany) was used for absorbance measurements at a wavelength of 360 nm. The column temperature was set to 35 °C for detection. The software Chromeleon v6.8 was used for data analysis. The rate of AFB₁ degradation was determined and calculated with (1 − AFB₁ peak area in treatment/AFB₁ peak area in control) × 100%.