Laboratory studies were conducted to identify the new and old milled rice by single grain among mixed rice using the ELISA plate and multi-channel adjustable micropipette method. The separated single grain were injected with minute amount of chemical reagent, and the reaction of reagent solution or grain were observed and recorded. Two chemical solutions were used in this studies. The first solution is contained Brom-Thymol Blue and Phenol Red, which were dissolved in NaOH, and the color reaction of grain or solution were to determine the freshness of milled rice. The new grain solution showed violet color, while the old grain turned into yellow. The second solution is contained Brom-Thymol Blue and Methyl Red, whice were dissolved in ethyl alcohol, new grain solution showed green color, while the old grain solution turned into yellow to orange. This method could be determined the rate of new and old milled rices among the mixed rice grains.