A method of pre-column derivatization ultra-performance liquid chromatography with fluorescence detector was developed for simultaneous determination of aflatoxins (AFB₁, AFB₂, AFG₁ and AFG₂) in beers.Methods The aflatoxins were extracted from beer samples by acetonitrile and cleaned up with multifunctional purification column to remove the matrix interference. The purified solution was concentrated and dried with nitrogen. After derivatized by trifluoroacetic acid, the aflatoxins were separated by reverse liquid chromatography (T3) with gradient elution and detected by fluorescence detector (FLD). Results The linear ranges of each compound were wide, and relative coefficients were between 0.999 2 and 0.999 6. The recoveries were within 90.47%-108.17%, and the relative standard deviations (RSDs) were between 0.97%-8.13%. The limits of detected (LODs) of 4 aflatoxins were 0.05,0.02,0.08 and 0.02 μg/kg in beers, respectively.Conclusion The established method was sensitive, selective, accurate and simple for the simultaneous determination of aflatoxins in beers.