Freeze-Thaw Sample Preparation Method Improves Detection of Volatile Compounds in Insects Using Headspace Solid-Phase Microextraction.

Headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS) is commonly used in analyzing insect volatiles. To improve the detection of volatiles in insects, a freeze-thaw method was applied to insect samples before the HS-SPME-GC-MS analysis. Insect samples were first frozen at -80 °C for 10 min and then thawed at 25 °C for 5 min before SPME extraction was performed. The freeze-thaw method clearly improved the detection of volatile compounds for all six tested insect species, including red imported fire ants, Solenopsis invicta Buren, black imported fire ants, Solenopsis richteri Forel, little black ants, Monomorium minimum (Buckley), pharaoh ants, Monomorium pharaonis (Linnaeus), eastern subterranean termites, Reticulitermes flavipes (Kollar), and spotted lady beetles, Coleomegilla maculate De Geer. This method helped identify various volatile compounds in the tested insects which have never been reported previously. This improved method may facilitate the identification of insect derived volatiles such as insect semiochemicals.