This work deals with different extraction media and reaction mixtures on photosystem II activity of Spinach chloroplasts. The photoreduction rate of ferricyanide and DPIP by intact chloroplasts which extracted with four kinds of extraction media; S-Tris-N pH 7.2, 8.0, S-Tricine-N pH 7.2, 8.0, was measured in five kinds of reaction mixtures; S-Tris-N pH 7.2, 8.0, S-Tricine-N pH 7.2, 8.0, 0.05 M-Tris pH 7.2. The extraction medium which shows the highest photoreduction rate was S-Tris-N at pH 7.2 and S-Tricine-N at pH 8.0. As to the reaciton mixture, S-Tricine-N pH 8.0 showed the highest rate. On the complex effects of extraction media and reaction mixtures, the highest photordeuction rate of Hill oxidant by intact chloroplasts was obtained by S-Tris-N pH 7.2 extraction medium and S-Tricine-N pH 8.0 reaction mixture. The second highest activity was obtained by S-Tricine-N pH 8.0 extraction medium and reaction mixture.