The mixture was incubated at 45 C for 120 min with regular shaking at 120 rpm followed by centrifugation (8545g, 30 C) for 15 min (Sigma, 3 K 30, Osterode am Harz, Germany) yielding three distinct phases (i) an oil phase, (ii) creamy phase and (iii) aqueous phase (Latif et al., 2008).