Scaling gas chromatography–mass spectrometry for profiling environmental chemicals in human cohorts

Scaling gas chromatography – mass spectrometry for profiling environmental chemicals in human cohorts Katerina Coufalikova1,2, Romana Sevcikova1,2, Akrem Jbebli1, Hana Selicova1, Elliott James Price1,2 1 RECETOX, Masaryk University, Faculty of Science, Brno, Czech Republic 2 Czech node of the Environmental Exposure Assessment Research Infrastructure (EIRENE-CZ) Acknowledgements •Maintaining measurement consistency in large-scale small molecule profiling studies is challenging •We translated a gas chromatography – mass spectrometry (GC-MS) method for profiling environmental chemicals in human blood plasma1 for routine application to population cohorts i.e., > 1000 samples References Background Implementation The daily analytical sequence (e) comprises system suitability test samples to check background (instrumental blank), retention time, signal response and mass accuracy (alkane and PCB mixtures), study QC samples to check experimental background (procedural blank) and long-term benchmarking (external QC and NIST SRM materials), and a batch of 38 study samples. Daily intervention includes injector liner and septa change, guard column trimming and MS tune and calibration. A liquid-liquid extraction procedure (a) has been fully automated via a TriPlus RSH Autosampler (b) coupled with Orbitrap Exploris GC 240 MS (c) for the sequential, online sample preparation and analysis (d). b a c d Monitoring of the retention drift of CRM alkanes (f) showed a maximum deviance of 3.3% (total runtime) across 50+ batches. Peak areas of analytes in NIST SRM 1958 showed no discernible relationships with interventional maintenance (g). Detection of analytes over 4 orders of MS response magnitude represented 5 orders of concentration – characteristic ions for cholesterol ~300 ppm (h), tetra-, pentaand hepta-chlorobiphenyls (PCBs) and dichlorodiphenyltrichloroethane (DDT), dichlorodiphenyldichloroethylene (DDE) & dichlorodiphenyldichloroethane (DDD) ~ 1ppm (i), polybrominated diphenyl ethers (PBDEs), triand hexaand octaPCBs, mirex, chlordane & nonachlor ~450 ppb (j), nonaand decaPCBs ~350 ppt (k). study 1 21 batches 3 source exchanges 2 column exchanges 0 filament replacements 2 technical services 4.5% total error rate: 86.2% injection failure 12.3% autosampler collision 1.5% human error study 2 57 batches 11 source exchanges 5 column replacements 3 filament replacements 2 technical services 3.7% total error rate: 89.0% injection failure 9.5% autosampler collision 1.5% human error Outcomes •Population scale GC-MS chemical exposure agent profiling in routine operation •Errors tracked and rate < 5% •Full SOP available2 e f study 1 study 2 study 1 study 2 study 1 study 2study 1 study 2 hg i j k This work was supported by the EU’s H2020 research and innovation programme under grant agreements No. 857560 (CETOCOEN Excellence), 874583 (ATHLETE), and 874627 (EXPANSE), by the HEU programme under grant agreement No. 101079789 (EIRENE PPP) and by RECETOX Research Infrastructure (LM2023069). The views expressed are those of the author(s) and do not necessarily reflect those of the EU or REA. The research has been conducted within the Environmental Exposure Assessment Research Infrastructure (EIRENE RI), in cooperation with the Network of Exposomics in the US (NEXUS) and within the collaborative framework of the International Human Exposome Network (IHEN; HEU programme under grant agreement No. 101137317). The authors would like to thank Moira Zanaboni & Manuela Bergna (Thermo Fisher Scientific S.p.A) for TriPlus RSH Sampling Workflow Editor Software and automation support; Dominic Roberts (Thermo Fisher Scientific UK) for the Orbitrap GC-MS Contaminants Library; Xin Zheng (Thermo Fisher Scientific US) & Giuseppe Scollo (Thermo Fisher Scientific S.p.A) for the iConnect high temperature transferline nut; Renzo Picenoni, Thomi Preiswerk & Hagen Gegner (CTC Analytics AG) for PALscript Editor and automation support; Brooklynn McNeil & Gary Miller (Columbia University, USA) and Doug Walker & Mattie Braselton (Emory University, USA) for collaboration on GC-MS method harmonisation; Delia Castilla Fernández, Kathrin Sigl & Benedikt Warth (University of Vienna, Austria) and Isabelle Boom, Hyung Elfrink, Amy Harms & Thomas Hankemeier (Leiden University, Netherlands) for collaboration on TriPlus RSH method replication. 1 Hu, X., Walker, D.I., Liang, Y. et al. (2021). A scalable workflow to characterize the human exposome. Nat Commun 12, 5575. https://doi.org/10.1038/s41467-021-25840-9 2 Biomarker Analytical Laboratories. (2024). SOP for automated on-line liquid-liquid extraction (LLE) workflow for anthropogenic profiling of human plasma samples via GC-HRMS. Zenodo. https://doi.org/10.5281/zenodo.13692108 2x ATC stations with liquid syringe tools Centrifuge Trayholder for 3x 54-2 mL vials Vortexer Cooled drawer Fast wash station Standard wash station Solvent station