Australian isolates ofFusariumspecies were grown on potato dextrose agar. Trichothecenes produced by these species were extracted by ethyl acetate followed by methanol and a silica gel column was used to clean‐up the extract. The extracted samples were derivatized by acetylation with trifluoroacetic anhydride and the derivatives analysed by gas chromatography/mass spectrometry (GC/MS). Multiple ion detection was used to trace ions characteristic of the trichothecenes expected to be present. Quantitation of those found was based on a known mass of pentabromophenol that was added as an internal standard. Eight species ofFusarium(nineteen strains) were surveyed, of which three species,F. acuminatum, F. equisetiandF. sporotrichioidesproduced the trichothecenes scirpentriol, diacetoxyscirpenol, neosolaniol, HT‐2 toxin, T‐2 toxin, T‐2 tetraol and deoxynivalenol. Wheat samples were inoculated with four different species ofFusarium, F. acuminatum, F. equiseti, F. graminearumandF. sporotrichioidesand in these samples diacetoxyscirpenol, neosolaniol, HT‐2 toxin and T‐2 toxin were found.