The test possibility of L-ascorbic acid(AsA) assay by ascorbate oxidase(A.O) solution obtained from cucumber was estimated. The results obtained were summarized as follow. The difference of absorbance before and after oxidation of AsA by A.O solution was proportional to the AsA concentration. Iso- AsA compared to same concentration of AsA was 97% response. Therefore, the 0 specificity on Iso- AsA a was deficient. On the sucrose, glucose, fructose were coexisted from 5 times, 500 times of AsA concentration, none of them were affected by any level of AsA concentration. In the case of EDTA, it was not nearly affected that EDTA was coexisted 50 times of AsA concentration, but EDTA Inhibited considerably from 500 times of AsA concentration. The effect of citric acid on the AsA concentration was not in the same level of AsA, however, it was slightly inhibited from 5 times, 50 times of AsA concentration and 500 times of AsA inhibited remarkably As known L-AsA was catalycally autooxidated by Cu2+ and Fe2+. They were not nearly affected In the above 0.01M level on the Ash assay. However, Fe3+ ion was neither nearly affected in the 0.001M level nor In the above 0.01M level on the AsA assay.