The chemistry of ant alarm pheromones is incredibly diverse with identified alarm pheromones spanning many structural families, including terpenoids, alcohols, aldehydes, ketones, esters, and nitrogen heterocycles. The compounds are highly volatile, a necessity of quick but transient information transfer. We employed two analytical techniques, solid phase microextraction (SPME) and purge and trap, to collect and analyze headspace contents about workers exhibiting alarmed behavior. These techniques also eliminate the need for solvent, which complicates analysis of highly volatile compounds using the gas chromatograph-mass spectrometer (GC-MS). Using these techniques, we identified a substituted pyrazine as a component of the fire ant alarm pheromone. We tested this chemical in a behavioral bioassay with S. invicta workers and we found no significant difference between the alarm response to the headspace volatiles collected above shaken ants and the pyrazine at a concentration of 100 ng/mL, supplying a headspace concentration of 0.50 pg/mL. Additionally, compared to the response to headspace volatiles collected above unshaken ants, we found a significant alarm response to concentrations as low as 1 ng/mL, with a corresponding headspace concentration below detection limits (less than 0.03 pg/mL). We are continuing to define the alarm pheromone source and chemistry.