pubs.acs.org/journal/estlcu Letter The Potential of Sewage Sludge to Predict and Evaluate the Human Chemical Exposome Ruben Gil-Solsona, Maria-Christina Nika, Mariona Bustamante, Cristina M. Villanueva, Maria Foraster, Marta Cosin-Tomás, Nikiforos Alygizakis, Maria Dolores Gómez-Roig, Elisa Llurba-Olive, Jordi Sunyer, Nikolaos S. Thomaidis, Payam Dadvand, and Pablo Gago-Ferrero* Cite This: Environ. Sci. Technol. Lett. 2021, 8, 1077−1084 Read Online ACCESS * See https://pubs.acs.org/sharingguidelines for options on how to legitimately share published articles. Metrics & More Article Recommendations sı Supporting Information Downloaded via 92.177.142.130 on January 27, 2023 at 11:52:11 (UTC). ABSTRACT: Chemicals are part of our daily lives, and we are exposed to numerous chemicals through multiple pathways. Relevant scientific evidence contributing to the regulation of hazardous chemicals require a holistic approach to assess simultaneous exposure to multiple compounds. Biomonitoring provides an accurate estimation of exposure to chemicals through very complex and costly sampling campaigns. Finding efficient proxies to predict the risk of chemical exposure in humans is an urgent need to cover large areas and populations at a reasonable cost. We conducted an exploratory study to characterize the human chemical exposome in maternal blood and placenta samples of a population-based birth cohort in Barcelona (2018−2021). Ultimate HRMS-based approaches were applied including wide-scope target, suspect, and nontarget screening. Forty-two chemicals were identified including pesticides, personal care products, or industrial compounds, among others, in the range of ng/mL and ng/g. In parallel, sewage sludge from the wastewater treatment plants serving the residence areas of the studied population were also screened, showing correlations with the type and concentrations of chemicals found in humans. Our findings were suggestive for the potential use of sewage sludge as a proxy of the human exposure and its application in early warning systems to prevent bioaccumulation of hazardous chemicals. ■ INTRODUCTION The presence of chemicals in our everyday environment plays a Therefore, scientific evidence is essential to obtain a holistic understanding of both the presence of new chemicals and their prominent role in the development of complex diseases, potential effects on human health. especially chronic ones.1−3 Over the last decades, there has Understanding the chemicals we are exposed to in our been a considerable increase in cases of allergies, infertility, workplace and daily lives remains one of the main challenges impaired brain development in children, various types of cancer, regulators face when addressing chemicals of concern. or even neurological disorders that can be related to environ- Biomonitoring provides an accurate method to estimate the mental exposures.4−8 However, it is challenging to develop exposure and bioaccumulation of organic chemicals through the compelling evidence that certain chemicals present in the direct measurement of the chemicals in the relevant biofluids environment cause specific adverse health effects. Furthermore, the increasing addition of new chemicals in the market could be Received: October 19, 2021 worsening these situations. One of the principal factors of this Revised: November 5, 2021 difficulty is the lack of knowledge on the presence and Accepted: November 8, 2021 bioaccumulation of organic chemicals; the chemical exposome, Published: November 12, 2021 defined as the totality of environmental chemical exposures from birth onward,9 is far from being completely understood. © 2021 American Chemical Society https://doi.org/10.1021/acs.estlett.1c00848 1077 Environ. Sci. Technol. Lett. 2021, 8, 1077−1084 Environmental Science & Technology Letters pubs.acs.org/journal/estlcu Letter and tissues. The analytical strategies (based on target screening) of 200 mg from the fetal villi of one of these biopsies was that have been commonly applied in human biomonitoring are analyzed. Additional details of the cohort and the mother−child biased because they only consider a very limited portion of the pairs can be found in the SI. existing universe of chemicals.10−13 Advances in high-resolution Sewage sludge was collected from the only two WWTPs, from mass spectrometry (HRMS) have opened windows of September 2019 to March 2020, which serve the areas of the opportunity for the identification of previously unknown whole Barcelona metropolitan area. Sludge samples (n = 6) chemicals of concern. The emergence of large chemical consisting of 250 g were collected each 4 months, as sewage repositories, containing MS/HRMS data (e.g., Massbank,14 sludge composition undergoes very small variations over time, Massbank of North America, 15 Metlin, 16 HMDB, 17 freeze dried, and stored at −20 °C until extraction. mzCloud18), as well as tools for wide-scope suspect screening Chemicals and Analysis. Details about the chemicals and (e.g., NORMAN DSFP 19 ) or automatic annotation of reagents and extensive descriptions of the sample treatment unknowns (e.g., SIRIUS4,20 MetFrag21), have provided an protocols are provided in the SI. Briefly, aliquots of plasma and unprecedented possibility to perform large-scale investigations. serum (150 μL) were deproteinized with acetonitrile (ACN), In the framework of projects and initiatives related to human and the supernatant was collected for analysis. Placenta samples exposure (such as HEALS,22 HBM4 EU,23 EXPOsOMICS,24 (200 mg) were extracted with a mixture of ACN, citric acid Project VIVA,25 ATHLETE,26 or FIGO27), few studies have buffer, and zirconium beads in a tissuelyser. Then, a SPE cleanup successfully applied these strategies, identifying potential step with mixed mode cartridges was carried out as described chemical exposures in human samples.28−31 elsewhere.37,38 Sewage sludge (1 g) was extracted with a digital However, human biomonitoring campaigns are very complex sonifier using ACN and water, and further cleanup and since they (I) deal with logistical constraints, (II) use invasive preconcentration steps were conducted through SPE. Proce- techniques and imply ethical issues, and (III) impose dural blanks (n = 5 for each matrix) were processed following the considerable economic burden. These factors make it difficult same protocols, in order to assess potential contaminations. to cover large geographical areas and reach large populations. Instrumental analysis was carried out using a MaXis Impact Finding efficient proxies to predict the risk of chemical exposure Q-ToF instrument (Bruker Daltonics) and a Q-Exactive Q- in humans is an urgent need to cover a significant fraction of the Orbitrap instrument (Thermo Scientific) coupled to UHPLC. population and specific environments. In this context, sewage Acquisition was performed using both data-independent and epidemiology, currently used to estimate the behavior of a large- data-dependent modes (DIA and DDA, respectively). The scale population regarding the consumption of specific scheme shown in Figure S1 summarizes the followed workflow, substances (illicit drugs,32 pharmaceuticals,33 and plastisizers34) and full details can be also consulted in the SI. or to predict COVID-19 outbreaks,35 can play an important role. Target, Suspect, and Nontarget Screening Strategies. In this work, we conducted an exploratory study using Wide-scope target analysis was performed for more than 2300 advanced HRMS-based strategies (wide-scope target, suspect, organic chemicals, including pharmaceuticals, illicit drugs, and nontarget) to extensively characterize the human chemical doping compounds, herbicides, surfactants, and other industrial exposome in maternal blood (N = 10) and placenta samples (N = chemicals as well as transformation products following the 9) from 19 individuals available from the population-based BiSC procedure described elsewhere.39 The complete list of chemicals (Barcelona Life Cohort Study, http://projectebisc.org) moth- as well as the analytical evaluation procedure is available in the er−child cohort. In parallel, sewage sludge (the residue of aforementioned publication. wastewater treatment) from the wastewater treatment plants For extended suspect screening (SI), DIA data from a Q- (WWTP) serving the residence areas of the studied population Exactive instrument was transformed from a proprietary (∗.raw) was also collected and screened using compatible method- to generic (∗.mzML) format and uploaded in the DSFP ologies. As such, this study is a proof of concept for the platform19 along with calculated retention time indexes of evaluation of the potential use of sewage sludge as a proxy of the calibrant substances.40 The screening included 7586 chemicals human chemical exposome and its application in early warning (EXPHRMSMSAVAL list) with a wide range of uses and systems to prevent the bioaccumulation of hazardous chemicals, physicochemical properties (full list available in the SI without the need of conducting extensive biomonitoring spreadsheet). Experimental MS/MS information for all campaigns. The similarity of human and sewage sludge samples substances was available. The identification of the chemicals was evaluated in terms of presence/absence of xenobiotics and was first based on mass accuracy, isotopic fit, and an advanced overall matrix composition using nontarget approaches. ■ LC retention time prediction model combined with the inspection of MS/MS fragments. Then, tentative identifications MATERIALS AND METHODS for exogenous compounds were verified or discarded manually Study Population and Sample Collection. Recruitment checking the DDA data. Other metadata linked with commercial occurred from October 2019 until March 2020 in three main indicators was used in some cases as described elsewhere.41,42 tertiary hospitals of Barcelona, Spain. Exogenous chemicals For nontarget screening, DDA data from Q-Exactive was enter the human body, are distributed by blood, and finally reach transformed to generic format (∗.mzML) and data mined in different tissues, where they can be heterogeneously bioaccumu- MZMine2.43 Peak picking was performed only for those features lated depending on tissue, species, and chemical.36 The pilot with one MS/MS spectrum acquired in, at least, one sample of study included 10 blood samples (serum and plasma) from the whole data set (in order to avoid the inclusion of mothers obtained at the time of delivery and nine placentas from instrumental noise as much as possible). The feature list was a total of 19 pregnant women. Maternal blood was collected in then exported to SIRIUS4.20 Both molecular formula and different tubes and centrifuged to obtain serum and plasma. compound annotations were obtained, based on the information Placental biopsies (0.4 mm × 0.4 mm × full thickness) were on accurate mass, isotopic pattern, and MS/MS spectra. taken from two different quadrants, rinsed in saline, snap frozen Chemicals with observed negative mass defect, suggesting in liquid nitrogen, and stored at −80 °C until analysis. A fraction halogenated species and therefore of anthropogenic origin, and 1078 https://doi.org/10.1021/acs.estlett.1c00848 Environ. Sci. Technol. Lett. 2021, 8, 1077−1084 Environmental Science & Technology Letters pubs.acs.org/journal/estlcu Letter Table 1. Details on 43 Chemicals Identified in Human Samples Confidence Serum: Detection (%)/ Placenta: Detection (%)/ Sludge: Detection (%)/ Identity Common use Strategy levela Conc. range (ng/mL) Conc. range (ng/g) Conc. range (ng/g) Tris(chloropropyl) Flame retardant Nontarget Level 1 100/(0.2−15) 22/(n.d. − 1.4) 100/(3.6−13) phosphate 1,7-Dimethylxanthine Food component Suspect Level 2 50/(n.d. − 0.05) 0 (−) 50/(n.d. − 2.2) Theobromine Food component Target Level 1 100/(0.01−25) 78/(n.d. − 542) 17/(n.d. − 2.6) Theophylline Food component Target Level 1 90/(n.d. − 3.8) 89/(n.d. − 540) 33/(n.d. − 2.2) Lenticin Food component Suspect Level 2 100/(3.7−13) 78/(n.d. - 211) 83/(n.d.- 13) Caffeine Food component Target Level 1 100/(0.6−45) 78/(n.d. − 840) 100/(0.9−4.8) Endothall Herbicide Target Level 1 50/(n.d. − 0.3) 0 (−) 83/(n.d. − 16) Denatonium Industrial Suspect Level 1 0 (−) 44/(n.d. − 44) 67/(n.d. − 2.3) Benzotriazole Industrial Suspect Level 1 0 (−) 44/(n.d. − 3.2) 100/(0.4−0.6) Dibenzylamine Industrial Suspect Level 2 0 (−) 44/(n.d. − 99) 50/(n.d. − 24) Tributyl phosphate Industrial Suspect Level 2 100/(0.2−1.2) 11/(n.d. − 0.4) 100/(0.6−10) Triphenyl phosphine Industrial Suspect Level 1 0 (−) 70/(n.d. − 8.3) 100/(0.3−0.5) oxide Triphenyl phosphate Industrial Suspect Level 1 100/(0.6−1.6) 0 (−) 10/(n.d. − 0.01) 2-hydroxybenzothiazole Industrial Suspect Level 1 0 (−) 78/(n.d. − 34) 100/(0.2−1.2) 2-benzothiazolesulfonic Industrial Nontarget Level 1 0 (−) 78/(n.d. − 2.3) 84/(n.d. − 0.02) acid DEET PCP/Pesticide Target Level 1 40/(n.d. − 0.4) 0 (−) 100/(0.9−13) N,N- PCP Suspect Level 2 90/(n.d. − 0.8) 44/(n.d. − 0.2) 100/(0.1−1.2) Dimethyldodecylamine- N-oxide Panthenol PCP Suspect Level 1 0 (−) 44/(n.d. − 21) 17/(n.d. − 1.4) Bis-(2-ethylhexyl)amine PCP Suspect Level 2 60/(n.d. − 27) 22/(n.d. − 48) 100/(>1000) N-(2-hydroxyethyl)- PCP Suspect Level 2 0 (−) 100/(0.03−1.5) 100/(0.6−14) Tetradecanamide Lauryl diethanolamine PCP Suspect Level 1 90/(n.d. − 5.2) 67/(n.d. − 3.2) 100/(0.4−0.5) Benzododecinium PCP Target Level 1 60/(n.d. − 86) 67/(n.d. − 118) 100/(326−664) PFOS PFAS Suspect Level 1 100/(0.08−0.3) 55/(n.d. − 34) 100/(4.1−10) PFHpS PFAS Suspect Level 1 0 (−) 33/(n.d. − 2.9) 0 (−) PFBS PFAS Suspect Level 1 0 (−) 11/(n.d. − 0.8) 0 (−) 6:2 FTS PFAS Suspect Level 1 0 (−) 11/(n.d. − 1.4) 50/(n.d. − 0.2) PFHxS PFAS Suspect Level 1 20/(n.d. − 0.02) 0 (−) 0 (−) Bupivacaine PhAC Suspect Level 2 0 (−) 100/(0.8−126) 0 (−) Ciprofloxacin PhAC Suspect Level 1 0 (−) 11/(n.d. − 7.3) 0 (−) Amoxicillin PhAC Suspect Level 1 0 (−) 11/(n.d. − 6.6) 0 (−) Indole-3-acetic acid Plant hormone Suspect Level 1 100/(2.8−8.9) 78/(n.d. − 1.4) 100/(0.5−2.6) Dibutyl phthalate Plastic additive Target Level 2 80/(n.d. − 9.6) 100/(11.1−23) 100/(27−63) Monobuthyl phthalate Plastic additive Suspect Level 2 30/(n.d. − 2.5) 89/(n.d. − 10) 100/(9.2−15) 4-Ethoxyethylbenzoate Plastic additive Suspect Level 2 100/(11−69) 100/(51.5−186) 100/(70.3−96) 1,3-Diphenylguanidine Plastic additive Suspect Level 2 70/(n.d. − 28) 0 (−) 100/(16−79) Nicotine Stimulant Suspect Level 1 0 (−) 67/(n.d. − 15) 100/(2.4−70) Cotinine Stimulant TP Target Level 1 20/(n.d. − 56) 33/(n.d. − 7.5) 67/(n.d. − 587) Acesulfame Sweetener Target Level 1 20/(n.d. − 0.5) 11/(n.d. − 8.1) 0 (−) Methylsulfonyl-1,3- Unknown Nontarget Level 3 0/(−) 22/(n.d. − 6.2) 0 (−) thiazol-2-ylamine 1-Benzothiophene-2- Unknown Nontarget Level 3 0 (−) 89/(n.d. − 19) 17/(n.d. − 0.1) Sulfonamide Benzophenone UV filter Suspect Level 1 10/(n.d. − 0.07) 78/(n.d. − 3.4) 100/(0.2−7.7) a 41 According to Schymanski et al. . PCP, personal care products; PhAC, pharmaceutical active compounds; PFAS, per- and polyfluoroalkyl substances; n.d., not detected. not previously identified in the target and suspect analysis were Concentrations for the newly confirmed chemicals and the further explored in a nontarget manner (SI), using different tentatively identified ones were estimated45,46 (SI-7). spectral online databases (e.g., MassBank,14 MoNa,15 or Metlin16) and automatic annotation tools (e.g., SIRIUS20). In all cases, three procedural blanks for serum, placenta, and ■ RESULTS AND DISCUSSION Extensive Screening of Organic Chemicals in Blood sludge were carefully assessed to avoid false positives. Reference and Placenta. In total, 42 chemicals were identified using the standards were purchased for the confirmation of most three various workflows (target, suspect, and nontarget analysis) tentatively identified compounds, including all listed in Table in the selected serum and placenta samples (18 in common, 25 1 with a level of confidence 1. When the standard was not in serum, 35 in placenta). Out of those chemicals, 28 were available, chemicals remained with a lower confidence level.44 confirmed with reference standards, and 14 remained as 1079 https://doi.org/10.1021/acs.estlett.1c00848 Environ. Sci. Technol. Lett. 2021, 8, 1077−1084 Environmental Science & Technology Letters pubs.acs.org/journal/estlcu Letter Figure 1. (A) Correlation between the mean concentration levels found in sludge and serum (A1) and placenta (B1). Stimulants (Caffeine, Theobromine, and Theophylline) were excluded due to their recent intake by human individuals before sample collection. (B) Venn diagram representing all features (combining positive and negative ionization modes) in common between human matrices and sludge. Only features with available MS/MS data were considered. tentatively identified. Of the 42 reported chemicals, nine were our study area, and its presence has been previously reported in identified via wide-scope target analysis and 29 as a part of the both human (at similar levels, ≈3.2 ng/mL in maternal blood)52 suspect analysis, including pharmaceuticals, food-derived and environmental samples including air and drinking water.53 chemicals, herbicides, insect repellents, and a variety of High levels of the disinfectant Benzododecinium (up to 118 ng/ industrial chemicals. Additionally, features with a negative g (placenta) and 86 ng/mL (serum)) were detected in greater mass defect and Cl and Br isotope information, indicator of an than 60% of the samples. This chemical is present in products exogenous origin and potentially hazardous, were prioritized for such as Sterinol, widely used as disinfectant before and during further identification via nontarget analysis. Despite this the wake of the COVID-19 pandemic46 and have been found to restrictive criterion, four additional chemicals not included in cross skin with intensive usage, reaching levels in plasma of up to any of the previous list were tentatively identified including 141.2 ng/L after an intensive five day usage.54 Additional tris(chloropropyl)phosphate, 2-benzothiazolesulfonic acid, chemicals used in personal care products were also present in methylsulfonyl-1,3-thiazol-2-ylamine, and 1-benzothiophene-2- human matrices (Table 1) including N,N-dimethyldodecyl- sulfonamide, classified as flame retardants (and other industrial amine-N-oxide, bis(2-ethylhexyl)amine, N-(2-hydroxyethyl)- uses). For all the identified chemicals, their identity, common tetradecanamide, and lauryl diethanolamine. Furthermore, we uses, types of strategy that led to their identification, confidence detected various plastic additives such as 1,3-diphenylguanidine, levels, frequencies of detection, and (semi)quantitative concen- dibutyl phthalate, and its metabolite monobutyl phthalate. tration ranges are summarized in Table 1. Limits of detection Additionally, high levels (up to 186 ng/g in placenta and 69 ng/ (LODs) were in the range of 0.01−1.7 ng/mL in serum and mL in serum) were determined for 4-ethoxyethylbenzoate, 0.01−4.9 ng/g in placenta. The analytical evidence that led to which is used in food contact plastics (e.g., polypropylene). Also, the identifications are provided in Table S2. The results a wide variety of industrial compounds such as benzotriazole, obtained in plasma and serum were identical in qualitative and dibenzylamine, tributyl phosphate, triphenylphospine oxide, quantitative terms. Therefore, in this study, we show only serum benzothiazole derivatives, or the flame retardant tris- results. (chloropropyl)phosphate were identified. Six PFAS (for which The presence of the exogenous chemicals in biofluids and various routes of exposure have been recognized55,56) were tissues could be explained by multiple types of exposure, mainly found at low levels in both matrices, except for PFOS, where inhalation, ingestion, or dermal contact.47,48 In exceptional concentrations up to 34 ng/g were detected in the placenta. cases, such as for some drugs, it may be by injection, as is the case Significant differences were observed in the distribution of the with Bupivacaine (used as epidural and intradural anesthesia).49 chemicals in these matrices. In general, compounds with high This chemical was found in all placental samples (since all the LogP (Table S2) showed higher levels in placenta compared to mothers used local anesthesia during delivery). Nicotine- serum, indicating bioaccumulation. On the contrary, polar detected levels (up to 15 ng/g in placenta) and its main compounds found in serum at low concentrations were not metabolite Cotinine (up to 56 ng/mL in serum and 7.5 ng/g in detected in the placenta (e.g., dimethylxanthine, 1,3-diphenyl- placenta) showed a certain active or passive exposure to tobacco guanidine, endothall, or DEET). for 67% of the individuals. This percentage is in line with Human Samples and Sewage Sludge: Similarities and previous studies that in some cases have reported higher levels of Differences in Their Chemical Profiles. Chemicals are nicotine in placenta (up to 108 ng/g)50 and plasma (up to 19.6 retained in sewage sludge according to their chemical ng/mL of nicotine and 97.1 ng/mL of cotinine)51 of smokers. abundance, bioaccumulation potential, and environmental Low levels (up to 0.4 ng/mL) of the insect repellent N,N- persistence.57 We hypothesize that the presence of xenobiotics diethyl-3-methylbenzamide (DEET) were also determined in in urban sludge could be indicative of their bioaccumulation in 40% of the serum samples. This chemical is commonly used in humans. Under this premise, a comparison of the evaluated 1080 https://doi.org/10.1021/acs.estlett.1c00848 Environ. Sci. Technol. Lett. 2021, 8, 1077−1084 Environmental Science & Technology Letters pubs.acs.org/journal/estlcu Letter human samples and sewage sludge from WWTP serving the area rich structure (as mentioned before), it can act as a proxy to the of residence of those individuals (collected during the same human body. Moreover, the concentrations of organic chemicals period) was performed in terms of prevalence and levels of undergo very small variations over time (it is equivalent to an xenobiotics, physicochemical properties, and overall chemical integrated sample of several days).58,59 Nevertheless, excretion is composition (feature distribution). not the only way for chemicals to reach urban WWTPs. Minor More than 80% of the compounds identified in the human industrial effluents and other inputs also influence the samples (serum or placenta) were present in the sludge (LODs composition of sludge. The addition of those compounds with were in the range 0.09−11 ng/g). The overlap was 67% in the a nonstrictly urban origin will help to obtain a more case of placenta and 55% for serum. Chemicals not detected in comprehensive overview of what chemicals a community may sludge (Acesulfame, Bupivacaine, Ciprofloxacin, Amoxicillin, be exposed to through other pathways and evaluate potential methylsulfonyl-1,3-thiazol-2-ylamine, PFAS) were all deter- future exposures. Our results are reinforced by studies based on mined at very low levels (0.02−8 ng/g) and low frequency of data extracted from the literature. A set of chemicals detected in appearance (mainly less than 30%) in the human samples. Also, the USA representative of biological specimens from humans those chemicals were mainly highly biodegradable except the (performed by the Centers for Disease Control and PFASs and Acesulfame, whose LODs in sludge were not Prevention60) and municipal sewage sludge (U.S. Environ- sufficient for such low levels. mental Protection Agency’s national sewage sludge surveys61), A certain correlation (R2 > 0.8) was observed between the revealed important (70%) overlap (out of the 52 compounds levels found in the human samples and those found in the sludge, evaluated using basic target strategies).57 as can be observed in Figure 1A (R2 > 0.6 if excluding As in any study focused on the analysis of organic chemicals, Benzododecinium, the chemical present at the highest levels). the covered chemical space is limited. In this work, we Although the number of compounds evaluated is small and, in implemented methodologies for a large coverage of semipolar some cases, the reported levels are semiquantitative, the graphic and polar compounds, combining a minimally invasive sample reflects the potential link between the concentrations in sludge treatment with different instrumental techniques. However, to and those found in humans. obtain a comprehensive picture including highly lipophilic Obviously, sewage sludge may contain a large number of compounds, additional analysis using gas chromatography (GC) chemicals that were not detected in human samples. Those coupled to HRMS would be necessary. However, the observed compounds, especially the ones with high toxicity potential, co-occurrence of chemicals in sludge and humans in our poor biodegradability, and high concentrations, would require experiments, as well as because of data crossing from different an additional assessment in larger cohorts, since they are national monitoring studies, strongly suggests that wide-scope candidates to bioaccumulate and result in potential hazardous analysis of sludge can be used to obtain information on chemical effects. exposure in human populations. ■ Apart from the xenobiotics identified through target and nontarget strategies, ≈3500 additional common features were determined out a total of 6500 (54%). It is noteworthy that only ASSOCIATED CONTENT features with MS/MS spectra available after acquisition in DDA * Supporting Information sı were considered to avoid the inclusion of instrumental noise. The Supporting Information is available free of charge at More than 2000 features were common in sludge, serum, and https://pubs.acs.org/doi/10.1021/acs.estlett.1c00848. placenta, 1000 more between placenta and sludge, and 300 between serum and sludge. This reveals a certain similarity Summary of study population and sample collection between the matrices, as can be seen in the Venn diagram (Section SI-1), summary of chemicals and materials (Figure 1B). These common features include potential xeno- (Section SI-2), detailed information regarding the sample biotics, for which the identity is still unknown, and mainly treatment (Section SI-3), full details on instrumental endogenous biomolecules (mostly lipid-like and protein-like, as analysis (Section SI-4), table containing chromatographic it can be observed in the van Krevelen diagram (Figure S2)) gradients (Table S1), figure with instrumental and data among the evaluated matrices. treatment workflow (Figure S1), detailed information Perspectives on Use of Sewage Sludge as a Proxy to regarding suspect screening data processing (Section SI- the Human Body to Explore Human Exposure to Organic 5), nontarget screening (Section SI-6), semiquantification Chemicals. The experimental results obtained in this proof of strategy (Section SI-7), van Krevelen diagrams creation concept study showed that the screening of sludge can be useful (Section SI-8), and table with analytical evidence leading to obtain information on the risk of certain substances to to chemical identifications (Table S2) (PDF) bioaccumulate (or pseudobioaccumulate) in humans. Optimiz- Complete list of chemicals evaluated in the suspect ing this approach could allow covering a large part of the analysis (XLSX) population at a small cost, limiting complex biomonitoring ■ campaigns to cases with specific objectives. The existing nonexperimental approaches usually consider the persistence, AUTHOR INFORMATION general bioaccumulation potential, and toxicity. Using sewage epidemiology (understood as the analysis of sludge), we add to Corresponding Author the equation the current chemical production rates, con- Pablo Gago-Ferrero − Department of Environmental sumption, and, above all, the individual behavior of chemicals Chemistry, Institute of Environmental Assessment and Water in real world biological systems. Research − Severo Ochoa Excellence Center (IDAEA), We hypothesize that sewage sludge can offer a clearer picture Spanish Council of Scientific Research (CSIC), Barcelona of the bioaccumulation of persistent chemicals than other 08034, Spain; orcid.org/0000-0002-5987-0399; matrices such as wastewater since, due to its carbon- and lipid- Phone: +34 934 00 61 00; Email: pablo.gago@idaea.csic.es 1081 https://doi.org/10.1021/acs.estlett.1c00848 Environ. Sci. Technol. Lett. 2021, 8, 1077−1084 Environmental Science & Technology Letters pubs.acs.org/journal/estlcu Letter Authors Agreement No. 785994 (AIR-NB), and the Health Effects Ruben Gil-Solsona − Department of Environmental Chemistry, Institute (HEI) with Grant Agreement No. 4959-RFPA15-1/ Institute of Environmental Assessment and Water Research − 18-1 (FRONTIER). IDAEA-CSIC and ISGlobal are Centres of Severo Ochoa Excellence Center (IDAEA), Spanish Council of Excellence Severo Ochoa (Spanish Ministry of Science and Scientific Research (CSIC), Barcelona 08034, Spain Innovation). ■ Maria-Christina Nika − Laboratory of Analytical Chemistry, Department of Chemistry, National and Kapodistrian REFERENCES University of Athens, Athens 15771, Greece (1) Cui, Y.; Balshaw, D. M.; Kwok, R. K.; Thompson, C. L.; Collman, Mariona Bustamante − ISGlobal, Barcelona 08003, Spain; G. W.; Birnbaum, L. S. 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