Abstract: Carotenes is one of the most important vitamin A precursor in human nutrition which has numerous advantages. Palm Oil Mill Effluent (POME) is wastewater which consists of carotenes in the oil and grease content. Therefore, adsorption chromatography is used to separate the carotenes from the oil and grease in POME. Several types of adsorbents, temperatures and mass loading were studied in the experiments. The 40°C and oil:adsorbent ratio of 1:5 was recommended to be the most suitable temperature and mass loading for separation of carotenes by adsorption chromatography. Silica gel also shows better quality of adsorbent in separation of carotenes in hexane fractions.