The water accommodated fractions (WAFs) of nine oils in seawater have been studied. The oils range from light condensate to heavy crude, and include one highly biodegraded oil and one very wax rich oil. This study has identified large variations in the chemical composition of WAFs, depending on oil type, temperature, and mixing time. Experiments at different temperatures (2-13 degrees C) showed that it takes longer time to reach equilibrium at the lowest temperatures, and that this varies for the different oil types. Oils with higher pour point (wax rich oils) need a longer time to establish WAF in equilibrium than oils with lower pour points (naphthenic oils). At 13 degrees C a mixing time of 48h, as recommended in standard procedures, seems to be sufficient for asphalthenic and paraffinic oils. The results demonstrated that for WAF prepared from an unknown oil, or at lower temperatures, different mixing times should be tested. Since the WAF often is used in toxicity testing, the toxicity might be underestimated if the mixing time is too short.