Introduction: Pistachio is an important export product in some countries, especially Iran. Pistachios are with the highest risk of aflatoxin contamination between agricultural products. Aflatoxin B1 is the most potent carcinogenic aflatoxins. Aflatoxins have an important role in the etiology of cancer. Undoubtedly, the major problem is, Aspergillus flavus contamination. Aflatoxin production and find suitable ways to prevent its production. The aim of this study was to determine the effect of electron irradiation on aflatoxin B1 production in pistachio that was inoculated with Aspergillus flavus.Methods: Experimental study was performed. aflatoxigenic Aspergillus flavus spores (106 spores/ml) were inoculated into 10 pistachio samples. Contaminated samples were exposed to the electron beam radiations at doses of 1, 3, 5 and 7 kGy. The samples were stored at a room temperature for 0, 30, 60, 90, 180, 210 days and were compared with the control samples for the aflatoxin production by Kruskal Wallis test using SPSS version 16. Aflatoxin B1 was measured by ELISA kit. The test was duplicated for each samples.Results: The mean concentration of aflatoxin in the samples was 59.97 ng/g. There was no difference among the production of aflatoxin in different doses on days 0, 30, 60, 90 (p<0.05), but on days 180 and 210 was borderline significant (p=0.06).Conclusion: Exposure of the pistachio samples by electron beam could inhibit the growth and alsoprevent aflatoxin production.