This study compared the effects of separating carotene with column chromatography to select the better absorbing material between magnesia and alumina. The dried powder of magnesia and alumina as the absorbent separately were analyzed to detach the carotene from spinach leaves. Two methods both showed three bands:carotene, chlorophyll a, chlorophyll b. But each band, especially the carotene band, was more compacted, orderliness, and the edge showed clearer in magnesia experiment than in alumina experiment. Therefore, the magnesia is the better than the alumina as a kind of absorbing material for column chromatograph to separate carotene.