By comparing the recoveries of different solid phase extraction columns for aflatoxin B1(AFB1) clean-up,the silicone-diatomite-neutral alumina mixed column was chosen,and the analytical method of Reversed Phase High Performance Liquid Chromatography(RP-HPLC) was developed for determination of aflatoxin B1 in raw materials of beer(barley malt,barley,rice and wheat malt).The results showed that the linear range of detection of AFB1 ranged from 0.5 μg/kg to 50 μg/kg.Standard curve correlation coefficient(r) was 0.9997.The detection limit was 0.15 μg/kg.The average recovery ranged from 80.1% to 109.2%,and the relative standard deviation(RSD) ranged from 0.23% to 4.29%.The method was simple,accurate and cost saving.It was also proposed to be used for the quality control of AFB1 in raw materials of beer.