Three phase partitioning, a technique which has been explored for protein separation was evaluated for extraction of turmeric oleoresin. The process consists of simultaneous addition of t-butanol and ammonium sulphate to the aqueous slurry of turmeric powder. The extraction of oleoresin was optimized with respect to the concentration of ammonium sulphate loading and the ratio of t-butanol to slurry. Pretreatment of the slurries with a commercial enzyme preparation of alpha-amylase and/or glucoamylase followed by three phase partitioning was carried out. The extraction time with this technique is lower as compared to conventional acetone extraction. (C) 2010 Elsevier Ltd. All rights reserved.