A new Wastewater-Based Epidemiology workflow to estimate community wide non-communicable disease prevalence using pharmaceutical proxy data

This manuscript introduces a new wastewater-based epidemiology workflow for estimation of non-communicable diseases (NCDs) prevalence by using wastewater-based epidemiology (WBE) and pharmaceuticals/their metabolites as proxies for NCDs prevalence. 83 targets were selected (54 parent pharmaceuticals and 29 metabolites). Three critical aspects were tested: (i) Solid-Phase Extraction - Ultra-Performance Liquid Chromatography and Tandem Mass Spectrometry (SPE-UHPLC-MS/MS) method performance, (ii) biomarker stability under variable storage conditions (during sampling and long-term storage) and (iii) accounting for human metabolism in WBE back-calculations. High stability of most analytes was observed under tested storage conditions. A few exceptions include diazepam, dihydroketoprofen and 5-hydroxy-lansoprazole. Analyte recoveries varied between 75% and 125% for most analytes. MDLs ranged from 0.2 ng L^-1 to 5.6 ng L^-1, while MQLs from 0.2 ng L^-1 to 16.8 ng L^-1. The overall average method accuracy and precision were: 99.5% and 4.0% respectively. A fully validated method was tested using community wastewater in the Southwest of England to estimate pharmaceutical usage, test metabolism correction factors established and compare results with prescription data. The new WBE method for NCD approximation allowed for the estimation of the daily usage/intake of 69 NCD targets with a standardized approach and a consistent reporting format.