The invention relates to a method for determining the fluorescent luminosity for the immunity affinity column of aflatoxicosis in rice, which comprises such procedures as extracting, cleaning, determining and calculating, etc.; and is characterized in that: the extracting procedure is to fetch 10.0g sample, add 2.0-2.5g NaCl and 60-80% methanol-water solution 50.omL, and extract by ultrasonic for 10-20 min; in the cleaning procedure, when cleaning the affinity column, first the affinity column is showered by a tween -20 / PBS solution, then by a methanol: water =2:8 solution, finally by water. The invention has the advantages that the recovery rate is more near to the actual value, the accuracy is higher, the reagent is reduced, the inspection cost is reduced, the environmental pollution is reduced, and the calculation is convenient.