The sewage epidemiology approach was applied to a one-year sampling campaign in the largest wastewater treatment plant (WWTP) in Belgium. The consumption of cocaine (COC), amphetamine (AMP), methylenedioxymethamphetamine (MDMA), methamphetamine (METH), methadone (MTD) and heroin (HER) was evaluated based on measured concentrations of the parent compound and/or metabolites in daily 24-hour composite influent wastewater samples. The inevitable back-calculations used in the sewage epidemiology approach were adapted to newly available information regarding the stability of the compounds in wastewater and the excretion pattern of illicit drugs. For COC, three different back-calculation approaches were evaluated. In addition, for the first time, efforts were made to calculate the number of inhabitants living in the catchment area of the WWTP in a real-time and dynamic way, based on concentrations of nitrogen, phosphorus and oxygen in the wastewater samples. Clear variations in the amount of inhabitants in the catchment area of the WWTP were observed. For COC, AMP and MDMA a significant higher weekend use was observed while for HER and MTD no significant daily variations could be found. METH consumption was negligible. Generally, the sewage epidemiology calculations were in agreement with official statistics. This manuscript shows that sewage epidemiology provides consistent and logical results and that it is a promising tool that can be used in addition to classical studies to estimate illicit drug use in populations. Therefore, efforts should be made to further optimize this approach in the future.
The simultaneous analysis of nine drugs of abuse (DOAs) and their metabolites (amphetamine, methamphetamine, methylenedioxymethamphetamine, methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine, cocaine, benzoylecgonine, ecgonine methyl ester and 6-monoacetylmorphine) in wastewater based on hydrophilic interaction liquid chromatography (HILIC) coupled to tandem mass spectrometry (MS/MS) was optimised and validated. For each analyte, the deuterated analogue was used for quantification. The separation by HILIC showed good performance for all compounds, especially for the hydrophilic compounds, which elute early (amphetamine-like stimulants) or show no retention (ecgonine methyl ester) in reversed-phase liquid chromatography. Sample preparation based on solid-phase extraction was optimised by comparing Oasis HLB and Oasis MCX sorbents for various parameters such as sample pH, amount of sorbent bed and washing solvent. The method was validated for each compound by assessing the following parameters (following International Conference on Harmonisation guidelines): specificity, limit of quantification (LOQ), linearity, accuracy, precision, recovery and matrix effects. LOQs were 2 ng/L for 6-monoacetylmorphine, ecgonine methyl ester and amphetamine and 1 ng/L for the rest of the compounds, corresponding with the lowest point in the calibration curve. Except for 6-monoacetylmorphine, all compounds were detected from 1 to 819 ng/L in influent wastewater samples (n = 12) collected from 11 different wastewater treatment plants across Belgium. The presence of ecgonine methyl ester in wastewater could be demonstrated for the first time. In the future, the new HILIC-MS/MS method will be applied to assess the use of DOAs in Belgium using the "sewage epidemiology" approach.
Daily 24 hour composite influent wastewater samples were collected from the wastewater treatment plant of Brussels-North (Belgium) for eight months to study variations in the concentrations and mass loads of nine illicit drugs and metabolites: cocaine (COC) and its metabolites benzoylecgonine (BE) and ecgonine methyl ester (EME), amphetamine (AMP), methylenedioxymethamphetamine (MDMA), methamphetamine (METH), methadone (MTD) and its metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) and the specific metabolite of heroin, 6-monoacetylmorphine (6-MAM). Samples were analyzed using a validated analytical method based on solid-phase extraction and hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry. The selected compounds could be detected and quantified in all samples. The measured concentrations were in agreement with earlier published concentrations, when available. All illicit drugs showed significant differences in mass loads between months. Daily variations were observed for MDMA, AMP and COC and metabolites, with higher mass loads observed during the weekend. This is probably related to their recreational consumption pattern and was further confirmed when holiday periods (New Year's Eve, Belgian national holiday) were in detail investigated. For METH, 6-MAM, MTD and EDDP, stable and consistent amounts were observed during the week. In all samples, the ratio of the concentration of the parent compound to its metabolite(s) was calculated to evaluate whether measured concentrations reflect human excretion of the illicit drugs. For MTD, the ratio of parent compound/metabolite was in agreement with the excretion pattern, while for COC some deviations were observed, resulting from excretion pattern uncertainties, and stability and discharge issues.
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