Filamentous fungi have long been recognized to be a rich source of secondary metabolites with potential medicinal applications. The recent genomic sequencing of several Aspergillus species has revealed that many secondary metabolite gene clusters are apparently silent under standard laboratory conditions. Several successful approaches have been utilized to upregulate these genes and unearth the corresponding natural products. A straightforward, reliable method to purify and characterize new metabolites therefore should be useful. Details are provided herein on the cultivation of Aspergillus nidulans and the LC/MS analysis of the metabolic profile. Following is an explanation of silica gel chromatography, HPLC, and preparative TLC. Finally, the NMR characterization of previously unknown A. nidulans metabolites is detailed.