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The opportunistic fungal human and plant pathogen Aspergillus terreus (A. terreus) can be isolated from sea water, soil or decaying organic matter such as rotting leaves and fruits. While growing on fruits A. terreus produces secondary metabolites such as terrein, which may ease its penetration into plant tissues. In addition, biological activities of terrein may support competition against other microorganisms. In summary, terrein is a small polyketide that reduces germination of seedlings, induces lesions on fruit surfaces but also shows moderate antifungal activity. With this manuscript we provide a fruit infection protocol with Aspergillus terreus with subsequent determination of terrein production rates on infected fruits using an HPLC-based quantification approach.