Environmental Exposure and Breath Analysis: Development of Innovative Methodological Approaches for Health Risk Assessment and Early Diagnosis

The exposome encompasses the totality of environmental, occupational and lifestyle exposures that interact with the human organism throughout the lifetime, influencing biological pathways and disease susceptibility. Understanding its molecular signa- tures represents one of the most ambitious challenges in contemporary biomedical and environmental health research. Within this conceptual framework, exhaled breath emerges as a promising non-invasive matrix capable of capturing dynamic molecular fingerprints that reflect both external exposures and internal metabolic responses. This study focuses on the development and validation of methodological approaches for the characterization of exhaled breath, with the aim of assessing its relevance for exposome-oriented research and its applicability to health-risk assessment and early disease detection. A breathomics approach was employed to investigate the volatile fraction of the human metabolome as a sensitive indicator of systemic processes linking exposure, metabolism and the onset of pathophysiological alterations. To achieve this objective, analytical workflows based on thermal desorption/gas chro- matography-mass spectrometry (TD/GC-MS) were optimized and combined with advanced multivariate and machine-learning techniques for the qualitative analysis of volatile organic compounds (VOCs) in human breath across different case studies. Moreover, the performance and practical application of an innovative, sensor-based breath analysis prototype were evaluated. The findings demonstrate collectively that exhaled breath can serve as a functional interface between the external and internal domains of the exposome, providing valuable insights into exposure-related metabolic perturbations. The methodological and analytical frameworks established through this work reinforce the integration of exposomic science and breath analysis, paving the way for precision-prevention strategies and translational applications in environmental and occupational health.