Linoleoyl benzotriazole was synthesized from linoleic acid and benzotriazole by means of dicyclohexylcarbodiimide coupling method and characterized by FT-IR and 1H NMR.Linoleoyl insulin was prepared by chemical modification of insulin with linoleoyl benzotriazole without using any specific protecting group.MALDI-TOF-MS analysis showed that the modification of insulin was successful and the monoacylated insulin was the major product.A little diacylated insulin was observed and no triacylated insulin was detected.