Diacylglycerol oil is an edible oil with taste and usability characteristics comparable to naturally occurring oils. The objective of this review is to examine literature on diacylglycerol oil to assess its safety-in-use. Feeding rats with unheated or heated diacylglycerol oil at levels up to 5.5% in diet for 90 days did not cause any toxic effects. In chronic studies, dietary administration of diacylglycerol oil (up to 5.3%) to rats for 2 years or at 9.5% to Beagle dogs for 1 year had no adverse effects. Genotoxicity studies of unheated and heated diacylglycerol oil did not reveal any genotoxic effects. Carcinogenicity studies in rodents demonstrate that diacylglycerol oil is non-carcinogenic. In a two-generation reproductive and developmental toxicity study, gavage administration of diacylglycerol oil at dose levels of 5.0 ml/kg body weight/day did not reveal any adverse effects. In several human clinical investigations, administration of diacylglycerol oil at levels up to 0.5 g/kg body weight/day for up to 1 year did not cause adverse effects. Collectively, there is sufficient qualitative and quantitative scientific evidence available from animal and human studies suggesting that intake of diacylglycerol oil is safe for human consumption when used in a manner similar to other edible oils.