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200393 https://hdl.handle.net/2286/R.2.N.200393 http://rightsstatements.org/vocab/InC/1.0/ http://creativecommons.org/licenses/by-nc-sa/4.0 2025-05 31 pages Botham, Maeve Liebig, Juergen Haight, Kevin Barrett, The Honors College School of Life Sciences School of Molecular Sciences The volatility of cuticular hydrocarbons (CHCs) plays an important role in insect chemical communication by influencing their transmission and detection. This study investigates how the volatility of hydrocarbons is affected by (1) whether they are presented in isolation or as part of a mixture and (2) how structural modifications–such as the introduction of a cis-alkene or a longer-chain hydrocarbon–impact volatility. Solid-phase microextraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS) was used to quantify relative changes in evaporation rates under controlled conditions. Experiment 1 compared the volatility of tricosane (C23H48, abbr. C23), tetracosane (C24H50, abbr. C24), and pentacosane (C25H52, abbr. C25) hydrocarbons under isolated and mixed conditions. All three compounds showed significantly higher peak areas in the mixed treatment, indicating increased volatility, and C23 and C24 also differed in their proportional representation, suggesting that chemical context influences not only evaporation but also the relative contribution of each compound to the volatile components. Experiment 2 tested whether the addition of tricosene (C23:1, a cis-alkene) or heptacosane (C27H56, a longer-chain hydrocarbon, abbr. C27) altered the volatility of the focus CHCs (C23, C24, and C25) in the mixture. The results showed no significant differences in volatility across treatments, suggesting that these structural modifications did not measurably affect evaporation behavior under the tested conditions. These findings demonstrate that CHC volatility is modulated by chemical context and mixture interactions, rather than by molecular identity alone. This contributes to a growing understanding of how CHC mixtures function in short-range insect communication and may inform future research on signal availability. Hydrocarbons Volatility GC-MS Communication Ants Insects GC-MS Analysis of Hydrocarbon Volatility: Mixture and Structural Effects