Analysis of Volatile Compounds Emitted by Filamentous Fungi Using Solid-Phase Microextraction-Gas Chromatography/Mass Spectrometry

Here, we describe a solid-phase microextraction-gas chromatography/mass spectrometry (SPME-GC/MS) analytical approach that identifies and analyzes volatile compounds in the headspace above a live fungal culture. This approach is a sensitive, solvent-free, robust technique; most importantly from a practical standpoint, this approach is noninvasive and requires minimal sample handling. Aliquots of liquid fungal cultures are placed into vials equipped with inert septa and equilibrated at a constant temperature, and headspace gases are sampled using an SPME fiber inserted through the septum into the headspace above the fungal culture for a standardized period of time. The outer polymer coating of a fused silica fiber absorbs volatiles from the headspace; the volatiles are then desorbed in the hot GC inlet and chromatographed in the usual manner. The separated compounds are subsequently identified by mass spectrometry. All steps in volatile profiling of a single sample from volatile sorption on a fiber to obtaining a list of volatiles can take as little as 15 min or can be extended to several hours if longer sorption is required for compounds present at very low levels and/or have low rates of diffusion.