Global Tracking of Transformation Products of Environmental Contaminants by 2H-Labeled Stable Isotope-Assisted Metabolomics

Stable isotope-assisted metabolomics (SIAM) enables global tracking of isotopic labels in nontargeted metabolomics in living organisms. However, its application in tracking transformation products (TPs, as metabolites of contaminants) of environmental contaminants is still a challenge due to limits in methodology, unmatured algorithms, and the high cost of 13C-labeled contaminants. Therefore, we developed a 2H-SIAM pipeline coupled with a highly flexible algorithm 2H-SIAM(1.0) (https://github.com/kechen1984/2H-SIAM), facilitating tracking TPs of contaminants in the environmental matrix. A detailed discussion illustrates the theory, behavior, and prospect of 2H-SIAM. We demonstrate that the proposed 2H-SIAM pipeline has unique advantages over 13C-SIAM, for example, cost-effective 2H-labeled contaminants, easy synthesis of 2H-labeled emerging contaminants, and providing more structural information. A pyrene soil degradation study further confirmed its high performance. It efficiently discarded 99% of noise signals and extracted 52 features from the nontargeted high resolution mass spectrometry (HRMS) data. Among them, 13 features were annotated as TPs of pyrene with identification confidence from Level 2a to Level 5, and 5 TPs were reported for the first time. In conclusion, the proposed 2H-SIAM pipeline is powerful in tracking potential TPs of environmental contaminants with unique advantages.