A simple and rapid method for determination of aflatoxin B₁, B₂, G₁, G₂, M₁ and M₂ in animal foodstuff by UPLC were developed.Methods Sample was extracted and diluted with methanol-water after immune affinity column chromatography purification. Components were completely separated within 5 min on a C₁₈ chromatographic column (3.0 mm×50 mm, 1.7 μm) with mobile phase of methanol-acetonitrile-water (gradient elution), and detected by a fluorescence detector. The retention time was used for qualitative analysis and peak area for quantitative analysis. Results6 kinds of aflatoxins were completely separated into aflatoxin B₁, B₂, G₁, G₂, M₁ and M2, and detection sensitivity were 0.038,0.010,0.062,0.010,0.110 and 0.016 μg/L respectively. Standard curve were linear, the recoveries were between 73.2%-94.1%, and precision between 0.2%-3.8%.Conclusion The method was simple, rapid, highly sensitive, and could be recommended for simultaneous determination of 6 kinds of aflatoxins in the animal foodstuff.