This study was conducted to investigate the effect of Lentinus edodes powder on the quality characteristics and antioxidant of rice cookies. The cookies for this experiment was made by 3 main ingredients including rice powder, flour, Lentinus edodes powder. The proportion of rice powder was fixed to 50% for the total amount of ingredients. The amount of reminded 2 ingredients (flour and Lentinus edodes powder) were varied. The amount of flour was decreased up to 38%, and the addition of Lentinus edodes powder was increased to 4%, 8% and 12%. Density of the dough, hardness, total polyphenol content, DPPH free radical scavenging activity, and reducing power increased with increasing Lentinus edodes powder content. Moisture content, pH of the dough, spread factor, loss rate, leavening rate, and L values of the cookies were decreased with increasing Lentinus edodes powder content. The result of consumer acceptance test revealed that there were no significant differences in smell, taste and overall acceptability, while the preference of color decreased as the amount of Lentinus edodes powder increased. To sum up, the cookies with 12% Lentinus edodes powder can produced the best results in terms of quality and antioxidant potential.