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<bold>Background:</bold> Pleural mesothelioma (PM) is an aggressive cancer with poor prognosis. Volatile organic compounds (VOCs) in exhaled breath are promising non-invasive biomarkers, but their relation to tumour pathophysiology remains unclear. Establishing a baseline VOC profile from murine PM cell lines may provide translational insights into tumour metabolism and allows to bridge in vitro findings with in vivo breath-based diagnostics. <bold>Aim:</bold> Identify VOCs released by murine PM cell lines and explore their biochemical origin. <bold>Methods:</bold> VOCs of the headspace of murine PM cell lines (AB1-HA, AE17) and medium controls were collected and analysed via gas chromatography quadrupole time-of-flight mass spectrometry (GC-qTOF-MS). VOCs significantly higher in tumour samples compared to medium only samples were further investigated via pathway analysis. <bold>Results:</bold> <fig><object-id>erj;66/suppl_69/PA5305/F1</object-id><object-id>F1</object-id><object-id>F1</object-id><graphic></graphic></fig> In AE17 and AB1-HA cells, a ketone was significantly elevated compared to the medium. In AB1-HA cells, ketones, alcohols, aromatic hydrocarbons, and alkanes were significantly higher. <bold>Conclusion:</bold> PM cell lines exhibit distinct VOC profiles, suggesting tumour-specific metabolic changes. Elevated ketones, alkanes, and alcohols indicate altered lipid metabolism, while aromatic hydrocarbons may reflect oxidative stress. These findings provide mechanistic insights into PM metabolism and lay the foundation for in vivo validation in PM mouse models, advancing breath-based diagnostics for PM.