Multiplatform targeted/untargeted approaches for the identification of possible biomarkers of occupational exposure

During the last decades, exposure to chemicals in the workplace it’s shown to be associated with an increased risk of different occupational diseases [1]. Nowadays several strategies can be used to monitor the health status of workers, such as air monitoring or biomonitoring of chemicals and their metabolites, in workers’ biofluids [2]. The application of Metabolomics to the field of occupational exposure is innovative and rapidly evolving since it could correlate a metabolic phenotype with several exposure parameters. Among the analytic platforms, nuclear magnetic resonance (NMR) spectroscopy is particularly suitable for metabolomics studies, as it allows the simultaneous qualiquantitative analysis of hundreds of metabolites belonging to different chemical classes in complex real matrices. On the other hand, liquid chromatography coupled to mass spectrometry represents another important platform for target analysis with lower LOD and LOQ than NMR. Therefore, a combined approach where different platforms’ outputs were fused together could provide important advantages over traditional exposure assessment techniques. For this purpose, data fusion strategies and multivariate statistical analysis were necessary used to evaluate urinary metabolic profiles and identify key metabolites associated with exposure. The choice of biological matrix to investigate fell on the urine due to the low invasiveness of the sampling and the possibility of repeated sampling in the short term. Over the years several biological monitoring campaigns were carried out, during which urinary samples from both professionally exposed subjects (employed in various industrial sectors) and healthy volunteers were collected. Subsequently all samples have been analyzed by NMR for the evaluation of metabolic profile, and by HPLC-MS/MS, for known exposure biomarkers determination. From profiling analysis approximately 40 metabolites have been identified and quantified and then correlated with target analysis’ results. The observed subclinical metabolic alterations provided information about novel possible exposure biomarkers to be monitored to predict the onset of an occupational diseases [3]. Interestingly, some molecules were common between groups, indicating a nonspecific response of the organism, such as a redox state alteration, while others have been correlated to specific xenobiotic exposure. The described approach allows for early evaluation of occupational exposures, and it could be a useful tool that focuses attention on aspects that remain unclear still today, such as the long-term effects at concentrations of toxics below the threshold limit values, or the evaluation of exposure to mixtures of substances or again exposures to both physical and chemical agents. The scope is to promote targeted risk assessments and developing personalized protective measures for the health of specific individuals or vulnerable groups. Acknowledgements: Research was funded by Grant: BRIC ID 09/2019, ID 31/2019, ID 08/2022, from INAIL. References [1] Jakubowski M. J Environ Monit., 2012 Feb;14(2):348-52. [2] Nicholson J.K., Lindon J.C., Holmes E. Xenobiotica, 1999;29:1181–1189. [3] Giampaoli O., Sciubba F., Tranfo G., et al. Toxics 2024, 12, 182.