Many species of frogs of the genus Litoria secrete bioactive peptides from their skin glands. These peptides are normally host-defence compounds and may have one, or more of the following activities; smooth muscle contraction, analgesic, antimicrobial, antiviral, lymphocyte proliferator (immunomodulator) and neuronal nitric oxide synthase (nNOS) inactivation. Two frog species of the Litoria rubella Group that have been studied before, namely, Litoria electrica and Litoria rubella, are different from other species of the genus Litoria in that they produce small peptides that show neither membrane, lymphocyte nor nNOS activity. In this study we have used electrospray mass spectrometry together with Edman sequencing to identify eight skin peptides of the third member of this Group, Litoria dentata: surprisingly, none of these peptides show activity in our biological screening program. However, two major peptides (FPWL-NH2 and FPWP-NH2) from L. electrica and L. rubella are opioids at the micromolar concentration. Copyright © 2009 John Wiley & Sons, Ltd.