Determination of Organophosphate Ester Metabolites in Seafood Species by QuEChERS-SPE Followed by LC-HRMS.

Organophosphate triesters are compounds widely used in industries and are ubiquitous in the environment, where they can be transformed into organophosphate diesters. Some organophosphate diesters are also used by industry. Several studies suggest organophosphate diesters can have toxic effects for reproduction, and hazardous and mutagenic properties. Due to the impact these compounds can have on marine biota and human beings through the consumption of fish and shellfish, it is necessary to study their presence in widely consumed seafood species. We therefore developed an analytical method for determining six of the most common organophosphate diesters in seafood. The procedure is based on the Quick, Easy, Cheap, Effective, Rugged and Safe extraction method and a solid phase extraction clean-up, followed by liquid chromatography coupled to high-resolution mass spectrometry. The method was optimised and validated for seafood with different lipid content, providing satisfactory relative recoveries (from 89 to 138%) and limits of detection (1.0-50 ng g-1 dry weight), as well as repeatability values (RSD% (n = 5, 100 ng g-1 (dry weight)) lower than 15%. Eight seafood species were analysed using this method and two organophosphate diesters were detected and quantified in all the samples, demonstrating the suitability of the method.

A simple, fast method for the analysis of 20 contaminants of emerging concern in river water using large-volume direct injection liquid chromatography-tandem mass spectrometry.

A fast and sensitive method for the determination of a structurally and physico-chemically diverse group of contaminants of emerging concern (CEC) based on large-volume direct injection liquid chromatography-tandem mass spectrometry was developed. The method can be used to determine 20 CECs belonging to different pollutant families (pharmaceuticals, personal care products, and pesticides) in river water at nanogram per liter. A single analytical run is required and the positive and negative ionization modes can be used simultaneously. Because of the large-volume injections of samples and the high sensitivity of the current mass spectrometers, the method has no need of a preconcentration step. The analytes are quantitated with matrix-matched calibration curves. The estimated limits of detection were in the range 0.1-5 ng L-1. The accuracy of the method was in the range 86-114%, and the precision, expressed as a relative standard deviation (RSD %), was below 18% for all the analytes (n = 5, at 5, 10, and 25 ng L-1). The method was applied to water samples taken from different points along the lower course of the Ebro River, Spain. A total of 12 out of the 20 target analytes were detected, and the ones at higher concentrations were caffeine and the pharmaceuticals paracetamol and ibuprofen (184.8 ng L-1, 63.3 ng L-1, and 23.3 ng L-1, respectively).

Determination of benzothiazoles in seafood species by subcritical water extraction followed by solid-phase microextraction-gas chromatography-tandem mass spectrometry: estimating the dietary intake.

Benzothiazoles are high production volume chemicals widely used in many industrial and household applications. However, information on their occurrence in aquatic organisms is very limited, although a high level of bioaccumulation is expected. In this study and for the first time, a method was developed involving subcritical water extraction followed by solid-phase microextraction coupled to gas chromatography-ion trap-tandem mass spectrometry for the determination of five benzothiazoles in seafood. The repeatability and reproducibility of the method were under 21% (%RSD, n = 5, 100 ng g-1 (dw)), while method detection limits and method quantification limits were between 0.5 and 10 ng g-1 (dw) and 1 and 50 ng g-1 (dw), respectively. Ten widely consumed fish and shellfish species from the county of Tarragona (Catalonia, Spain) were selected in order to estimate dietary exposure and to assess the human health risks. The most frequently determined compounds were benzothiazole and 2-(methylthio)-benzothiazole, with squid being the species which showed the highest level of benzothiazole (82 ng g-1 (dw)). In terms of human exposure, the current concentrations of benzothiazoles found in fish and shellfish could not be compared to threshold values because of the lack of toxicological data.

Pressurized liquid extraction followed by liquid chromatography with tandem mass spectrometry to determine pharmaceuticals in mussels.

An analytical method based on pressurized liquid extraction and solid-phase extraction with a mixed-mode Oasis(®) MAX sorbent as cleanup, followed by liquid chromatography with electrospray ionization and tandem mass spectrometry was developed and validated for the determination of seven widely used pharmaceuticals in mussel species. The optimization of the pressurized liquid extraction and the solid-phase extraction parameters is described. The method provided extraction recoveries ranging from 61 to 90%, and limits of detection ranging from 2 to 50 ng/g (dry weight). The repeatability and reproducibility of the method, expressed as relative standard deviation, were lower than 15 and 19%, respectively. The method was successfully applied to the analysis of mussel samples from different locations. The analyses showed that salicylic acid was present in mussels at concentrations up to 177 ng/g (dry weight).

Determination of pharmaceuticals in bivalves using QuEChERS extraction and liquid chromatography-tandem mass spectrometry.

A method for the quantitative determination of seven pharmaceuticals in bivalves was developed by QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) extraction, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with electrospray ionization. Both the European Standard Method EN 15662 and the AOAC Official Method 2007.01 for QuEChERS were tested. In addition, several clean-up strategies were evaluated in order to clean the matrix previous to the LC-MS/MS analyses. Dispersive solid-phase extraction with silica gel and modification of the chromatographic separation were the clean-up strategies that gave the best results. The optimized method was validated in mussels (Mytilus galloprovincialis) and allowed the determination of pharmaceuticals at nanograms per gram levels (dry weight (d.w.)). Limits of quantification ranged from 5 to 100 ng/g. Apparent recoveries ranged from 35 to 77%. The application of this method to bivalves revealed the presence of salicylic acid at concentrations up to 103 ng/g (d.w.).

Exposure of the population of Catalonia (Spain) to musk fragrances through seafood consumption: Risk assessment.

The occurrence of ten synthetic musks in samples of 10 widely consumed fish and shellfish species from Tarragona (Catalonia, Spain) was determined. The most used nitro and polycyclic musks, as well as a well-known transformation product in tissues, were analyzed. Furthermore, the human health risks derived from the musk exposure through seafood consumption were characterized. None of the nitro musks were detected in any of the analyzed samples. In contrast, most of the polycyclic musks were found, being galaxolide (HHCB) and tonalide (AHTN) present in all the samples. HHCB was the greatest contributor, with maximum levels in sardine and mackerel (367 and 304 ng g(-1) (d.w.) (dry weight), respectively). The highest exposure to individual musks was estimated for HHCB and HHCB-Lactone, with average values of 19.7 and 6.8 ng kg(-1)bw day(-1), respectively, in adults. A notably lower mean exposure was calculated for AHTN, cashmeran (DPMI) and traseolide (ATII), being ranged between 1.1 and 3.7 ng kg(-1)bw day(-1). The current concentrations of musks in fish and shellfish should not mean human health risks for the adult population living in Tarragona. However, a continuous monitoring would be desirable to assure that the exposure does not follow increasing temporal trends.

Headspace stir bar sorptive extraction followed by thermal desorption and gas chromatography with mass spectrometry to determine musk fragrances in sludge samples without sample pretreatment.

A direct, simple and solvent-free method based on headspace stir bar sorptive extraction and thermal desorption gas chromatography with mass spectroscopy was developed to determine 13 musk fragrances (six polycyclic musks, three nitro musks and four macrocyclic musks) in sludge without sample treatment. The optimal headspace stir bar sorptive extraction conditions were achieved when a polydimethylsiloxane stir bar was exposed for 45 min in the headspace of a 10 mL vial filled with 100 mg of sludge mixed with 0.2 mL of water stirred at 750 rpm at 80°C. The stir bar was then desorbed in the thermal desorption gas chromatography and mass spectrometry system, obtaining limits of detection between 5 and 30 ng/g. The method applicability was tested with sewage sludge from two urban wastewater treatment plants and from a potable water treatment plant. Results showed galaxolide and tonalide to be the most abundant musk fragrances found in wastewater treatment plants with maximal concentrations of 9240 and 7500 ng/g, respectively. Maximum concentration levels between 35 and 635 ng/g were found for musk ketone, musk moskene, traseolide, phantolide and celestolide in this kind of samples. Concentrations below the limits of quantitation of phantolide, galaxolide, tonalide and musk ketone were found in sludge from a potable water treatment plant.

Determination of sedative hypnotics in sewage sludge by pressurized liquid extraction with high-performance liquid chromatography and tandem mass spectrometry.

This paper describes a method for the determination of eight sedative hypnotics (benzodiazepines and barbiturates) in sewage sludge using pressurized liquid extraction and liquid chromatography with tandem mass spectrometry. Pressurized liquid extraction operating conditions were optimized and maximum recoveries were reached using methanol under the following operational conditions: 100ºC, 1500 psi, extraction time of 5 min, one extraction cycle, flush volume of 60% and purge time of 120 s. Pressurized liquid extraction recoveries were higher than 88% for all the compounds except for carbamazepine (55%). The repeatability and reproducibility between days, expressed as relative standard deviation (n = 5), were lower than 6 and 10%, respectively. The detection limits for all compounds were lower than 12.5 µg/kg of dry weight. The method was applied to determine benzodiazepines and barbiturates in sewage sludge from urban sewage treatment plants, and carbamazepine showed the highest concentration (7.9-18.9 µg/kg dry weight).

Comparison of the presence of high production volume chemicals in farmed and wild fish highly consumed in catalonia and their risk assessment.

The decline in fish populations and the depletion of marine resources have sparked concerns about sustainable fish production, driving the innovation of new aquaculture methods. While some argue that wild fish are healthier than farmed fish due to less exposure to contaminants and pathogens, wild fish can accumulate contaminants from more contaminated water sources. The slower growth of wild fish and their longer exposure to the environment may contribute to higher pollutant levels in fish tissues. In this study, we focus on 25 contaminants considered as high production volume chemicals (HPVCs), such as organophosphate esters (OPEs), benzothiazoles (BTs), benzosulfonamides (BSAs) and phthalates (PAEs). The compounds were extracted from the edible part of the fish using the QuEChERS method and analysed by gas chromatography-tandem mass spectrometry. A total of 74 samples were analysed from three of the most commonly consumed species in Catalonia, Spain (turbot, sea bass and sea bream). Two samples of each species were collected each month, one form farmed and one from wild origin. In general, the compounds were found in all the samples in a wide concentrations range, although no significant differences were observed between the mean concentration of wild and farmed samples. Although similar mean concentrations for the OPEs, BTs and BSAs were found between farmed and wild origin samples, PAEs were more frequently detected in farmed samples. Di-n-octyl phthalate and diethyl phthalate showed the highest concentrations in all fish samples, with values up to 19505 and 17605 ng g-1 (d.w.), in sea bass and sea bream, respectively. Di-(2-ethylexyl)-adipate proved to be the most relevant carcinogenic compound, with no associated health risk. Despite the detection of the studied HPVCs, no health risk was associated with the consumption of these three fish species.

Optimization of the ultrasound-assisted extraction method for determining high production volume chemicals in fish liver and skin samples.

The aquatic ecosystem is one of the most delicate environments, housing a diverse range of organisms, including fish, all of which are exposed to a wide variety of pollutants. The accumulation of these harmful substances in fish, which are part of the human diet, presents a significant health risk to humans. In our study, we have optimized an extraction technique to determine the presence of 25 high production volume chemicals in liver and skin samples taken from commonly consumed fish species. We have employed ultrasound-assisted extraction in conjunction with gas chromatography tandem mass spectrometry to achieve this goal. Apparent recoveries of the method ranged from 50% to 111% for both sample types with some exceptions such as most of the benzosulfonamides and benzothiazole. Additionally, the method's detection and quantification limits varied from 0.1 to 1.7 ng g-1 (dry weight, d.w) and 0.2-4.5 ng g-1 (d.w), respectively. Our investigation focused on three frequently consumed fish species in Tarragona: sea bass, sea bream, and turbot. Almost all of the samples we analysed contained traces of contaminants, with phthalates being the most commonly detected. The highest concentrations were observed for diethyl phthalate, with levels peaking at 8350 ng g-1 (d.w.). Organophosphate esters, such as triethyl phosphate and tributyl phosphate, also showed notable presence, with peak concentrations of 93.6 and 34.0 ng g-1 (d.w.), respectively.

Wastewater-based epidemiology to assess pharmaceutical consumption. Spanish perspective.

Wastewater-based epidemiology (WBE) is a well-established approach that can provide objective and real-time data on the consumption of substances such as pharmaceuticals. However, most of the studies reported so far compares consumption data obtained using WBE with those derived from prescription data from public health systems, which is often incomplete and might represent a source of uncertainty. This study aims to compare the measured pharmaceutical consumption back calculated with the WBE approach with consumption derived from dispensed pharmaceuticals in two regions of Spain, managed by two different Health Systems. To do so, a group of 17 pharmaceuticals, including the most representative ones of every therapeutic family, were monitored in influent wastewater (IWW) samples collected over a week campaign in spring 2022 at four different wastewater treatment plants (WWTPs) in Spain: two WWTPs in Madrid city (center of Spain) and two WWTPs in Catalonia (Northeast of Spain). Population-normalized daily loads (PNDL) revealed that the patterns of pharmaceutical occurrence in the different WWTPs are very similar, being acetaminophen, 4-acetamidoantipyrine and valsartan the pharmaceuticals with the highest PNDL values: 17162 ± 1457 mg day-1 1000 inh-1 for acetaminophen, 2365 ± 696 and 2429 ± 263 mg day-1 1000 inh-1 for 4-acetamidoantipyrine, 2006 ± 541 and 2041 ± 352 mg day-1 1000 inh-1 for valsartan. Pharmaceutical PNLDs were then transformed into measured pharmaceutical consumption (MC) and compared with dispensed consumption (DC) data obtained from the pharmacies in the catchment area where the WWTPs are located. A ratio MC/DC within 0.8 to 1.2 was obtained for 11 out of the 17 studied pharmaceuticals. Highlighting a match in all the cardiovascular system pharmaceuticals, with the exception of losartan (1.29-1.39 ratio) and valsartan (1.35-1.43) in all WWTPs. In summary, the degree of correlation between MC/DC is higher than those previously reported comparing with the prescribed pharmaceutical consumption.

An automated headspace solid-phasemicroextraction followed by gas chromatography­mass spectrometry method to determine macrocyclic musk fragrances in wastewater samples.

A fully automated method has been developed for determining eight macrocyclic musk fragrances in wastewater samples. The method is based on headspace solid-phase microextraction (HS-SPME) followed by gas chromatography­mass spectrometry (GC-MS). Five different fibres (PDMS 7 µm, PDMS 30 µm, PDMS 100 µm, PDMS/DVB 65 µm and PA 85 µm) were tested. The best conditions were achieved when a PDMS/DVB 65 µm fibre was exposed for 45 min in the headspace of 10 mL water samples at 100 °C. Method detection limits were found in the low ng L−1 range between 0.75 and 5 ng L−1 depending on the target analytes. Moreover, under optimized conditions, the method gave good levels of intra-day and inter-day repeatabilities in wastewater samples with relative standard deviations (n =5, 1,000 ng L−1) less than 9 and 14 %, respectively. The applicability of the method was tested with influent and effluent urban wastewater samples from different wastewater treatment plants (WWTPs). The analysis of influent urban wastewater revealed the presence of most of the target macrocyclic musks with, most notably, the maximum concentration of ambrettolide being obtained in WWTP A (4.36 µg L−1) and WWTP B (12.29 µg L−1), respectively. The analysis of effluent urban wastewater showed a decrease in target analyte concentrations, with exaltone and ambrettolide being the most abundant compounds with concentrations varying between below method quantification limit (

Performance of zwitterionic hydrophilic interaction LC for the determination of iodinated X-ray contrast agents.

This study compares the separation performance of a group of iodinated X-ray contrast media on four different columns. The first three were two stationary phases (SPs) modified with C18 and a polar-embedded SP (polar amide group bonded to an alkyl chain), all of which worked under RP-LC mode. The fourth was a zwitterionic sulphoalkylbetaine SP, working under the hydrophilic interaction LC (HILIC) mode. After the optimisation of the different parameters, the zwitterionic column displayed the best separation, which also overcomes the problems encountered when these analytes were separated under RP-LC. Moreover, when HILIC is coupled to MS/MS, sensitivity is enhanced. However, when sewage samples were analysed by SPE followed by the optimal HILIC-MS/MS, the sensitivity of the method was affected due to the high matrix effect, which had to be solved by dilution of the extract. Finally, the method was preliminarily validated with sewage and the figures of merit were comparable to those of the SPE-RP-LC-MS/MS.

Determination of benzothiazoles, benzotriazoles and benzenesulfonamides in seafood using quick, easy, cheap, effective, rugged and safe extraction followed by gas chromatography - tandem mass spectrometry: Method development and risk assessment.

The common use of benzothiazoles, benzotriazoles and benzenesulfonamides has led to widespread ubiquity in several environmental matrices. Their occurrence in edible fish could represent an additional exposure route for the population. The present study aims to develop a method for the simultaneous determination of these three compound families in seafood samples. Based on QuEChERS extraction, different salt combinations and clean-up strategies have been evaluated to achieve the highest recoveries while reducing the matrix effect in low and high lipidic content species. The best results were obtained with the original method salts and the lipid-selective push-through clean-up, which combined with gas chromatography-tandem mass spectrometry led to recoveries between 50 and 112% with negligible matrix effects and method detection limits between 0.15-9.50 ng g-1 dw. The application of the method to commercially available samples confirmed the presence of BTs as well as BSAs, with the latter being determined in seafood for the first time. Exposure and risk assessment calculations indicated a minor risk for the population when consuming fish.

High production volume chemicals in the most consumed seafood species in Tarragona area (Spain): Occurrence, exposure, and risk assessment.

Seafood consumption has become a potential exposure route towards high production volume chemicals (HPVs) due to the pathway of these compounds reaching the aquatic environment via industrial and domestic discharges. The present study focuses on the determination of phthalate esters (PAEs), organophosphate esters (OPEs), benzothiazoles (BTs), benzotriazoles (BTRs) and benzenesulfonamides (BSAs) in the ten most consumed fish species in Catalonia. A total of 120 commercially available seafood specimens were purchased throughout February 2019-February 2020 in three different stores (supermarket, local market, and local fishmonger) of the city of Tarragona, Spain, to cover the most typical places where seafood can be obtained. ΣOPEs, ΣBTs, ΣBSAs and ΣPAEs concentrations ranged between 5.99 and 139.45 ng g-1 w.w., 8.41-54.08 ng g-1 w.w., 8.38-304.47 ng g-1 w.w and 2.86-323.80 ng g-1 w.w., respectively. BTRs were not detected in any of the samples. PAEs and BSAs had similar contributions which combined represented nearly the 70% of detected compounds and sardine resulted as the species with the higher HPVs mean concentration. No considerable threat was posed due to the individual intake of these compounds via seafood consumption.

Seafood consumption as a source of exposure to high production volume chemicals: A comparison between Catalonia and the Canary Islands.

Seafood plays an important role in diet because of its health benefits. However, the fact that chemical compounds such as high production volume chemicals may be present in seafood means that its consumption can be a potential risk for population. To assess the occurrence of HPVs and estimate the exposure and risk associated with their consumption, specimens of the most consumed seafood species in Catalonia and the Canary Islands, Spain, were collected and analysed. Results showed higher levels of HPVs in samples from Catalonia and a prevalence of phthalate esters and benzenesulfonamides over the other target compounds in samples from both locations. Multivariate analysis showed spatial differences between the mean concentration profiles of HPVs for the samples from Catalonia and the Canary Islands. Exposures were higher for the samples from Catalonia, although the intake of HPVs via seafood was not of any real concern in either of the locations.

Determination of musk fragrances in sewage sludge by pressurized liquid extraction coupled to automated ionic liquid-based headspace single-drop microextraction followed by GC-MS/MS.

A method for the quantitative determination of ten musk fragrances extensively used in personal care products from sewage sludge was developed by using a pressurized liquid extraction (PLE) followed by an automated ionic liquid-based headspace single-drop microextraction and gas chromatography-tandem mass spectrometry. The influence of main factors on the efficiency of PLE was studied. For all musks, the highest recovery values were achieved using 1 g of pretreated sewage sludge, H(2) O/methanol (1:1) as an extraction solvent, a temperature of 80°C, a pressure of 1500 psi, an extraction time of 5 min, 2 cycles, a 100% flush volume, a purge time of 120 s, and 1 g Florisil as in-cell clean-up extraction sorbent. The use and optimization of an in-cell clean-up sorbent was necessary to remove fatty interferents of the PLE extract that make the subsequent ionic liquid-based headspace single-drop microextraction difficult. Validation parameters, namely LODs and LOQs, ranged from 0.5-1.5 to 2.5-5 ng/g, respectively. Good levels of intra- and interday repeatabilities were obtained analyzing sewage sludge samples spiked at 10 ng/g (n = 3, RSDs < 10%). The method applicability was tested with sewage sludge from different wastewater treatment plants. The analysis revealed the presence of all the polycyclic musks studied at concentrations higher than the LOQs, ranging from 6 to 530 ng/g. However, the nitro musk concentrations were below the LOQs or, in the case of musk xylene, was not detected.

Determination and occurrence of organic micropollutants in reverse osmosis treatment for advanced water reuse.

The growing demand on water resources has increased the interest in wastewater reclamation for multiple end-use applications such as indirect and direct potable reuse. In these applications, the removal of organic micropollutants is of a greater concern than in conventional wastewater treatment. This article presents a collection of data of trace organic micropollutants in an urban wastewater treatment plant (WWTP) in North East Spain using reverse osmosis (RO) membrane treatment. The RO rejection values of the organic molecules studied with a wide range of solute size and hydrophobicity were determined. Several chromatographic methods monitoring different endocrine disrupting chemicals (EDCs), pharmaceuticals and personal care products (PPCPs) were used. Results indicated that secondary effluents from this Spanish WWTP contained most of the studied organic compounds indicating incomplete removal of organics in the conventional treatment of the plant. However, the rejection of most micropollutants was high for all three RO membrane types (low energy, high rejection, fouling resistant) tested. It was observed that some selected micropollutants were less efficiently removed (e.g. the small and polar and the more hydrophobic) and the molecular weight and membrane material influenced removal efficiencies.

Automated on-fiber derivatization with headspace SPME-GC-MS-MS for the determination of primary amines in sewage sludge using pressurized hot water extraction.

An automated, environmentally friendly, simple, selective, and sensitive method was developed for the determination of ten primary aliphatic amines in sewage sludge at µg/kg dry weight (d.w.). The procedure involves a pressurized hot water extraction (PHWE) of the analytes from the solid matrix, followed by a fully automated on-fiber derivatization with 2,3,4,5-pentafluorobenzaldehyde (PFBAY) and headspace solid-phase microextraction (HS-SPME) and subsequent gas chromatography ion-trap tandem mass spectrometry (GC-IT-MS-MS) analysis. The limits of detection (LODs) of the method were between 0.5 and 45 µg/kg (d.w.) for all compounds except for ethyl-, isopropyl-, and amylamine, whose LODs were 70, 109, and 116 µg/kg (d.w.), respectively. The limits of quantification (LOQs) were between 10 and 350 µg/kg (d.w.). Repeatability and intermediate precision, expressed as RSD(%) (n=3), were lower than 18 and 21%, respectively. The method developed enabled to determine primary aliphatic amines in sludge from various urban and industrial sewage treatment plants as well as from a potable treatment plant. Most of the primary aliphatic amines were found in the sewage sludge samples analyzed corresponding to the maximum concentrations to the samples from the urban plant: for instance, isobutylamine and methylamine were found at 7728 and 12 536 µg/kg (d.w.), respectively. Amylamine was detected only in few samples but always at concentrations lower than its LOQ.

Drugs of abuse and their metabolites in waste and surface waters by liquid chromatography-tandem mass spectrometry.

A solid-phase extraction and liquid chromatography-tandem mass spectrometry (SPE/LC-MS-MS) method was developed and validated for the simultaneous determination of nicotine, five drugs of abuse (morphine, cocaine, codeine, methadone, and 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine) and four metabolites (dihydrocodeine, 6-acetylmorphine, 11-nor-carboxy-Δ(9)-tetrahydrocannabinol, and benzoylecgonine) in water samples. A Fused-Core™ particle column was used as an alternative to sub-2-µm particles in chromatographic separations to work with low backpressures and high efficiencies in short analysis times. Drugs were extracted from waste and surface water with SPE using Oasis MCX cartridges. Electrospray (ESI) in positive and negative mode and tandem MS selected reaction monitoring mode were used for identification and quantification. Calibration by linear regression analysis with deuterated internal standards was used to compensate the matrix effects. Limits of detection were found as low as 0.5-1 ng/L (surface water) and 1-50 ng/L (wastewater). The method was applied to the analysis of different kinds of samples. Wastewater from a sewage treatment plant was collected from three sampling points (after primary, secondary, and tertiary treatments) for a week. The analysis of the samples revealed a significant presence of these drugs in samples from primary treatments, where maximum concentrations of nicotine (1105 ng/L) and benzoylecgonine (3336 ng/L) were found. Most of the compounds showed values between