HPLC-FL Analysis of Aflatoxin B1

The analytical conditions for AFB₁ were as described by Kushiro et al. [29 (link)]. The HPLC-FL system was composed of an HPLC column Capcell pack C18 UG120 (5 μm, 250 mm × 4.6 mm i.d.; Osaka Soda, Osaka, Japan), pump LC-20AD, column heater CTO-10A, autosampler SIL-20AC, fluorescence detector RF-20A, communication bus module CBM-20A, and LabSolutions software (Shimadzu, Kyoto, Japan). The mobile phase consisted of water, methanol, and acetonitrile (6:3:1), and the flow rate was 0.3 mL min⁻¹. The column heater temperature was set at 40 °C. AFB₁ was detected at wavelengths of 365 (extraction) and 450 nm (emission). The standard AF mix was diluted to generate the calibration curve. The limit of detection (LOD) and limit of quantitation (LOQ) for the analysis were approximately 0.25 and 0.74 ng mL⁻¹, respectively, as calculated from the standard deviation of the y-intercept on the calibration curve.