Efficient Validation Strategies in Environmental Analytical Chemistry: A Focus on Organic Micropollutants in Water Samples

This article critically reviews analytical method validation and quality control applied to the environmental chemistry field. The review focuses on the determination of organic micropollutants (OMPs), specifically emerging contaminants and pesticides, in the aquatic environment. The analytical technique considered is (gas and liquid) chromatography coupled to mass spectrometry (MS), including high-resolution MS for wide-scope screening purposes. An analysis of current research practices outlined in the literature has been performed, and key issues and analytical challenges are identified and critically discussed. It is worth emphasizing the lack of specific guidelines applied to environmental analytical chemistry and the minimal regulation of OMPs in waters, which greatly affect method development and performance, requirements for method validation, and the subsequent application to samples. Finally, a proposal is made for method validation and data reporting, which can be understood as starting points for further discussion with specialists in environmental analytical chemistry.D.F.-S. acknowledges the Ministerio de Universidades in Spain for his Margarita Salas postdoctoral grant (Ref. MGS/2021/15). The authors from University Jaume I acknowledge financial support from Generalitat Valenciana (Research Group of Excellence Prometeo 2019/040) and Ministerio de Ciencia e Innovación (Ref. PID2021-127451OB-I00). J.B.Q. acknowledges funding provided by Xunta de Galicia (Ref. ED431C 2021/06), the Spanish Agencia Estatal de Investigación – MCIN/AEI/10.13039/501100011033 (Ref. PID2020-117686RB-C32).S

Analytical research of pesticide biomarkers in wastewater with application to study spatial differences in human exposure

Wastewater-based epidemiology (WBE) relies on the assessment and interpretation of levels of biomarkers in wastewater originating from a well-defined community. It has provided unique information on spatial and temporal trends of licit and illicit drug consumption, and has also the potential to give complementary information on human exposure to chemicals. Here, we focus on the accurate quantification of pesticide biomarkers (i.e., predominantly urinary metabolites) in influent wastewater at the ng L−1 level to be used for WBE. In the present study, an advanced analytical methodology has been developed based on ultra-high-pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS), for the simultaneous determination of 11 specific human biomarkers of triazines, urea herbicides, pyrethroids and organophosphates in urban wastewater. The sample treatment consisted of solid-phase extraction using Oasis HLB cartridges. Direct injection of the samples was also tested for all compounds, as a simple and rapid way to determine these compounds without sample manipulation (i.e., minimizing potential analytical errors). However, if extraction recoveries are satisfactory, SPE is the preferred approach that allow reaching lower concertation levels. Six isotopically labelled internal standards were evaluated and used to correct for matrix effects. Due to the difficulties associated with this type of analysis, special emphasis has been placed on the analytical challenges encountered. The satisfactory validated methodology was applied to urban wastewater samples collected from different locations across Europe revealing the presence of 2,6-EA, 3,4-DCA, 3-PBA and 4-HSA i.e, metabolites of metolachlor-s, urea herbicides, pyrethroids and chlorpropham, respectively. Preliminary data reported in this paper illustrate the applicability of this analytical approach for assessing human exposure to pesticides through WBE.Funding for open access charge: CRUE-Universitat Jaume

Analytical investigation of cannabis biomarkers in raw urban wastewater to refine consumption estimates

Wastewater analysis of Δ9-tetrahydrocannabinol (THC) biomarkers can provide essential information on trends in cannabis consumption. Although analysis is mostly focused on the aqueous phase, previous studies have illustrated the need of improving the measurements of raw influent wastewater (IWW) considering also suspended solids. This is important for cannabis biomarkers, because a substantial part of them is expected to be found in the suspended solids due to their more lipophilic character compared with other metabolites/drugs included in these types of studies. However, it remains open to which extent trend estimates might be affected by solely analysing the liquid phase. To investigate this aspect, robust analytical methodologies are required to measure both the liquid and solid phases of IWW. In this work, we firstly tested liquid-liquid extraction (LLE) for THC and its major metabolites (THC-OH, and THC-COOH). Using LLE, no filtration or centrifugation step was required for raw IWW analysis, and the three analytes were extracted from both the liquid and the solid phase simultaneously. In parallel, the raw IWW was centrifuged and the obtained solid and liquid phases were analyzed separately: the liquid phase by both LLE and solid phase extraction (SPE) for comparison of data, and the suspended solids by solid-liquid extraction (SLE). The separate analysis of both phases in a number of samples revealed that a significant amount of cannabis biomarkers (ranging from 42 to 90%) was found in the suspended solids. In addition, the total amount of cannabis biomarkers obtained by analysing raw IWW on the one hand, and by separate analysis of the liquid and the solid phases, on the other hand, was in good agreement. Data from this study show that the sole analysis of the liquid phase would lead to a notable underestimation of cannabis biomarkers concentrations in IWW

Analytical investigation of cannabis biomarkers in raw urban wastewater to refine consumption estimates

Wastewater analysis of Δ9-tetrahydrocannabinol (THC) biomarkers can provide essential information on trends in cannabis consumption. Although analysis is mostly focused on the aqueous phase, previous studies have illustrated the need of improving the measurements of raw influent wastewater (IWW) considering also suspended solids. This is important for cannabis biomarkers, because a substantial part of them is expected to be found in the suspended solids due to their more lipophilic character compared with other metabolites/drugs included in these types of studies. However, it remains open to which extent trend estimates might be affected by solely analysing the liquid phase. To investigate this aspect, robust analytical methodologies are required to measure both the liquid and solid phases of IWW. In this work, we firstly tested liquid-liquid extraction (LLE) for THC and its major metabolites (THCsingle bondOH, and THCsingle bondCOOH). Using LLE, no filtration or centrifugation step was required for raw IWW analysis, and the three analytes were extracted from both the liquid and the solid phase simultaneously. In parallel, the raw IWW was centrifuged and the obtained solid and liquid phases were analyzed separately: the liquid phase by both LLE and solid phase extraction (SPE) for comparison of data, and the suspended solids by solid-liquid extraction (SLE). The separate analysis of both phases in a number of samples revealed that a significant amount of cannabis biomarkers (ranging from 42 to 90%) was found in the suspended solids. In addition, the total amount of cannabis biomarkers obtained by analysing raw IWW on the one hand, and by separate analysis of the liquid and the solid phases, on the other hand, was in good agreement. Data from this study show that the sole analysis of the liquid phase would lead to a notable underestimation of cannabis biomarkers concentrations in IWW

Spatial and temporal assessment of crack cocaine use in 13 European cities through wastewater-based epidemiology

Already in early 2000s, concerns have been growing in the EU about increasing use of cocaine and it is estimated that below 1% of the population administer the drug by smoking crack cocaine. New available data suggests an increase in the use of crack cocaine and an increase in the number of crack cocaine users entering treatment has been reported in several European countries. Robust estimations of crack cocaine use are however not available yet. The use of crack cocaine has long been associated with severe adverse socio-economic conditions as well as mental health problems, such as suicide ideation and depression. The aim of this study was to assess spatial trends in population-normalized mass loads of crack cocaine biomarkers (i.e., anhydroecgonine and anhydroecgonine methyl ester) in 13 European cities in six countries (the Netherlands, Belgium, Ireland, Portugal, Spain and Italy). Furthermore, temporal trends over a five-year period were evaluated through the analysis of historic samples collected in the Netherlands. Finally, the stability of the crack cocaine biomarkers in wastewater was investigated through batch experiments. The samples were analyzed with a new developed and validated hydrophilic interaction liquid chromatography coupled to mass spectrometry method. Targeted crack cocaine biomarkers were found in all cities. Also, crack cocaine biomarker was detected in wastewater from 2017 to 2021 in the Netherlands, but no significance between the years were found. With respect to biomarker in-sample stability, AEME was found to be stable in wastewater. This study assessed crack cocaine use for the first time on a broad scale, both temporal and in cities across Europe, with wastewater-based epidemiology and it shows the importance of wastewater analysis to monitor community loads of crack cocaine use