Amphibian skin secretions, ansmg from specialised granular glands, are often complex peptidomes containing many components of entirely novel structure and unique site-substituted analogues of known peptide families. Following the discovery that the granular gland transcriptome is present in such secretions in a PCR amenable form, we have designed a strategy for peptide structural characterisation involving the integration of "shotgun" cloning of peptide precursor encoding cDNAs, deduction of putative bioactive peptide structures and confirmation of these using tandem MSIMS sequencing. Here, we illustrate this strategy by means of the elucidation of the primary structures of antimicrobial peptides from the defensive skin secretions of the Chinese smelly frog, Odorrana livida. Synthetic replicates of the peptides were found to possess antimicrobial activity, anticancer cell activity and trypsin inhibition activity. These peptides are thus of widespread occurrence in the skin secretions of Asian ranid frogs and in those of the Odorrana group and they are particularly well-represented and of diverse structure in some species. The integration of the molecular analytical technologies described can thus provide 'a means of rapid structural characterisation of novel peptides from complex natural libraries in the absence of systematic on-line database data.