Combining High-Resolution Mass Spectrometry and Chemiluminescence Analysis to Characterize the Composition and Fate of Total <i>N</i>-Nitrosamines in Wastewater Treatment Plants

Monitoring the prevalence and persistence of <i>N</i>-nitrosamines and their precursors in wastewater treatment plants (WWTPs) and effluent-receiving aquatic compartments is a priority for utilities practicing wastewater recycling or exploiting wastewater-impacted source waters. In this work, we developed an analytical framework that combines liquid chromatography-high-resolution mass spectrometry (LC-HRMS) with acidic triiodide-chemiluminescence analysis to characterize the composition and fate of total <i>N</i>-nitrosamines (TONO) and their precursors along the treatment trains of eight WWTPs in New York. Through the parallel application of LC-HRMS and chemiluminescence methods, the TONO scores for 41 <i>N</i>-nitrosamines containing structurally diverse substituents on their amine nitrogen were derived based on their solid-phase extraction recoveries and conversion efficiencies to nitric oxide. Correcting the compositional analysis of TONO using the TONO scores of target <i>N</i>-nitrosamines refined the assessment of the reduction or accumulation of TONO and their precursors across treatment steps in WWTPs. Nontargeted analysis prioritized seven additional <i>N</i>-nitrosamines for confirmation by reference standards, including three previously uncharacterized species: <i>N</i>-nitroso-<i>tert</i>-butylphenylamine, <i>N</i>-nitroso-2-pyrrolidinmethanol, and <i>N</i>-nitrosodesloratadine, although they only served as minor components of TONO. Overall, our study establishes an adaptable methodological framework for advancing the quantitative and qualitative analysis of specific and unknown components of TONO across water treatment and reuse scenarios.