Development and validation of an accurate and rapid LC-ESI-MS/MS method for the simultaneous quantification of aflatoxin B1, B2, G1 and G2 in lotus seeds

An accurate and rapid LC-ESI-MS/MS analytical method was developed and validated for the simultaneous determination of aflatoxin B₁, B₂, G₁ and G₂ in lotus seeds. The samples were firstly extracted with methanol-water solution (80:20, v/v), and then cleaned up by immunoaffinity columns. The mass spectrometer was operated in the positive ionization electrospray (ESI+) mode using multiple reaction monitoring (MRM) for analysis of four aflatoxins. The transitions of m/z 313 → 285 (AFB₁, CE 33 eV), m/z 315 → 259 (AFB₂, CE 37 eV), m/z 329 → 243 (AFG₁, CE 37 eV) and m/z 331 → 257 (AFG₂, CE 37 eV) were used to quantify these four aflatoxins, respectively. The limits of detection (LODs) of aflatoxin B₁, B₂, G₁ and G₂ were 0.007, 0.005, 0.003 and 0.005 μg kg^?1 based on a signal-to-noise ratio of 3:1, respectively. The limits of quantification (LOQs) of aflatoxin B₁, B₂, G₁ and G₂ were 0.02, 0.015, 0.01 and 0.015 μg kg^?1 based on a signal-to-noise ratio of 10:1, respectively. Recoveries for samples of spiked lotus seeds were all above 66% with relative standard deviation all below 15% for all compounds. Nineteen out of twenty batches of lotus seeds collected from different drug stores or markets in China were found to be contaminated with aflatoxins at different levels ranging from 0.02 to 688.4 μg kg^?1.