A new strategy has been developed for the determination of the amino acid sequence of apamin, a two disulfide-bond neurotoxic oligopeptide. Relative molecular mass up to about 2000 was determined using electrospray mass spectrometry. This technique shows that three basic amino acid residues are present in apamin. A hydrophobic tryptic digest fragment was sequenced using fast-atom bombardment combined with tandem mass spectrometry. But the full sequence has been directly observed using a tandem mass spectrometer: the daughter-ion mass spectrum of protonated molecules of dithiothreitol-reduced apamin gives the sequence.