Three minor peptaibols, trichokonins (TKs)-Ia, Ib, and IX, were obtained from the culture broth of Trichoderma koningii Oudemans. Primary structures of these peptaibols were elucidated by ion-spray ionization mass spectrometry (ISI-MS) including the collision-induced dissociation (CID) technique together with two-dimensional nuclear Overhauser enhancement spectroscopy (NOESY).