The characterization of components within actinomycin complexes may often be complicated by the lack of material and standards of known actinomycins. Mass spectrometry-mass spectrometry can be employed both as a separatory device and as a means of structural analysis. This technique has been applied to an actinomycin complex obtained from a previously unidentified Streptomyces strain. The method involved initial work on a known material, in this case actinomycin D, and application to the unknown material. Three major components within the unknown complex were characterized as actinomycins D, F(8), and F(9).