Recently, wastewater-based epidemiology (WBE), has been established as a useful tool to verify public health status. In brief, a wastewater sample is collected at the influent of a wastewater treatment plant and analysed for wide range chemical substances including endogeneously formed human products that can be related to aspects of public health. This approach has been successfully validated and applied to monitor substances of abuse in large scale studies across the globe and to measure the concentrations of several pharmaceuticals, metabolites, and personal care products1, and most recently to SARS-CoV-2. However, there also exists the possibility to adapt this approach for the analysis of a spectrum of endogenous biomarkers for stress and disease (e.g., proteins, lipids, and nucleosides), specifically, proteins, to evaluate the state of public health. The aim of this work is to detect and quanitify C-reactive protein (CRP), one of the positive acute-phase response proteins2, in influent wastewater to investigate public health within populations. The challenges of analysing endogenous biomarkers such as CRP in wastewater include the dilute nature of the material and the diverse complexity of the organic matter in the influent that can interfere with the methods3. Hyphenated mass spectrometry (MS) demonstrated accurate and reproducible quantification of some of these endogenous biomarkers in plasma samples, by the analysis of peptides produced via enzymatic digestion of protein targets, which is known as bottom-up (shotgun) proteomics4. In our initial study, one of the signature peptide of CRP showed better recovery after digestion as well as quantification in influent samples. This demonstrates the development of an LC-MS/MS method to identify and quantify CRP in wastewater at low concentration levels. This method has a potential for WBE to investigate community-wide public health status. References: 1. Kasprzyk-Hordern, B., Kondakal, V. V. R. & Baker, D. R. Journal of Chromatography A Enantiomeric analysis of drugs of abuse in wastewater by chiral liquid chromatography coupled with tandem mass spectrometry. 2010, 1217, 4575-4586. 2. Ironson, G., Banerjee, N., Fitch, C. & Krause, N. Social Science & Medicine Positive emotional well-being, health Behaviors, and inflammation measured by C-Reactive protein. 2018, 197, 235-243. 3. Avella, A. C., Görner, T. & de Donato, P. Science of The Total Environment The pitfalls of protein quantification in wastewater treatment studies. 2010, 408, 4906-4909. 4. Kuhn, E. et al. PROTEOMICS Quantification of C‐reactive protein in the serum of patients with rheumatoid arthritis using multiple reaction monitoring mass spectrometry and 13C‐labeled peptide standards. 2004, 4, 1175-1186. Acknowledgement: This work is a part of the Wastewater Fingerprinting for Public Health Assessment (ENTRUST) project funded by Wessex Water and EPSRC IAA (grant no. EP/R51164X/1).
Original languageEnglish
Publication statusPublished - 3 May 2021
EventSETAC Europe 31st Annual meeting: Global Challenges. An emergency for Environmental Sciences - Virtual conference
Duration: 3 May 20216 May 2021


ConferenceSETAC Europe 31st Annual meeting
Abbreviated titleSociety of Environmental Toxicology and Chemistry Europe 31st Annual meeting
Internet address


  • Wastewater-based epidemiology
  • Proteomics
  • Biomarkers
  • Public Health
  • Water fingerprinting

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