Illicit drugs and pharmaceuticals in swimming pool waters

Illicit drugs and pharmaceuticals in swimming pool waters

Science of the Total Environment 635 (2018) 956–963 Contents lists available at ScienceDirect Science of the Total Environment journal homepage: www...

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Science of the Total Environment 635 (2018) 956–963

Contents lists available at ScienceDirect

Science of the Total Environment journal homepage: www.elsevier.com/locate/scitotenv

Illicit drugs and pharmaceuticals in swimming pool waters G. Fantuzzi a,⁎, G. Aggazzotti a, E. Righi a, G. Predieri a, S. Castiglioni b, F. Riva b, E. Zuccato b a b

Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia, Via G Campi, 287, 41125 Modena, Italy Department of Environmental Health Sciences, “Mario Negri” Institute for Pharmacological Research, Via La Masa 19, 20156 Milan, Italy

H I G H L I G H T S

G R A P H I C A L

A B S T R A C T

• Four illicit drugs and eleven pharmaceuticals were found in swimming pools. • Ibuprofen was found in all the investigated pool waters. • Human Risk Assessment (HRA) was performed for different classes of age • No health risk was found for humans via swimming pool waters

a r t i c l e

i n f o

Article history: Received 8 January 2018 Received in revised form 15 March 2018 Accepted 5 April 2018 Available online xxxx Editor: Adrian Covaci Keywords: Illicit drugs Pharmaceuticals Indoor swimming pools Swimming pool water Human risk assessment

a b s t r a c t The occurrence of illicit drugs (cocaine, opioids, amphetamines and cannabis derivatives), some of their metabolites and 48 pharmaceuticals, was investigated in pool and source waters in ten Italian indoor swimming pools. The samples were analyzed by highperformance liquid chromatography-tandem mass spectrometry (HPLC-MS/ MS), after solid phase extraction (SPE). Cocaine and its metabolites were found in nine swimming pools, at concentrations from 0.3 to 4.2 ng/L for cocaine, 1.1 to 48.7 ng/L for norcocaine, 0.7 to 21.4 ng/L for benzoylecgonine and 0.1 to 7.3 ng/L for norbenzoylecgonine. Opioids, amphetamines and cannabis derivatives were never detected. The most frequent pharmaceuticals were anti-inflammatory drugs: ibuprofen was found in all the pool waters, with a maximum 197 ng/L and ketoprofen was detected in 9/10 samples (maximum 127 ng/L). Among anticonvulsants, carbamazepine and its metabolite, 10,11-dihydro-10,11dihydroxycarbamazepine, were frequent in swimming pool water (8/10 samples) at concentrations up to 62 ng/L. The cardiovascular drug valsartan was also found frequently (8/10 samples), but at lower concentrations (up to 9 ng/L). Other pharmaceuticals were detected occasionally and at lower concentrations (atenolol, enalapril, paracetamol, hydroclorothiazide, irbesartan and dehydro-erythromycin). Carbamazepine, irbesartan and dehydroerythromycin were detected at very low levels (up to 5 ng/L) in only one of the four source water samples. A quantitative risk assessment showed that the health risk for humans to these substance in swimming pool waters was generally negligible, even for vulnerable subpopulations such as children and adolescents. © 2018 Elsevier B.V. All rights reserved.

⁎ Corresponding author at: Dipartimento di Scienze Biomediche, Metaboliche e Neuroscienze - Sezione di Sanità pubblica, Università degli Studi di Modena e Reggio Emilia Via Giuseppe Campi, 287 41125 Modena, Italy. E-mail addresses: [email protected] (G. Fantuzzi), [email protected] (G. Aggazzotti), [email protected] (E. Righi), [email protected] (G. Predieri), [email protected] (S. Castiglioni), [email protected] (F. Riva), [email protected] (E. Zuccato).

https://doi.org/10.1016/j.scitotenv.2018.04.155 0048-9697/© 2018 Elsevier B.V. All rights reserved.

G. Fantuzzi et al. / Science of the Total Environment 635 (2018) 956–963

1. Introduction There is growing concern about the presence of emerging chemical contaminants in swimming pools and similar environments. Chemicals in pool water can come from a number of sources, namely the source water (polluted in origin), deliberate additions such as disinfectants, and bathers who continuously release organic matter mainly through sweat and urine (WHO, 2006). Most of the studies on the chemical risk in swimming pools are focused on disinfection by-products (DBPs) in pool water and on the mechanisms of their formation as a result of the reaction between the chlorine used as a disinfectant and the organic substances present in the waters (Zwiener et al., 2007; Richardson et al., 2010). Some hundreds of DBPs have been identified in chlorinated water of swimming pools, some being of particular importance to human health because they are toxic and/or have suspected carcinogenic or mutagenic activity (Richardson et al., 2012). Other chemical contaminants recently found in swimming pool waters include sunscreen agents, personal care products, pharmaceuticals and other chemicals. However, up to now, the occurrence of pharmaceuticals in swimming pool waters has been scarcely investigated. (Zwiener et al., 2007; Wang et al., 2013; Bottoni et al., 2014; Teo et al., 2016). Weng et al. (2014) studied 32 pharmaceuticals and personal care products in three swimming pools in USA and detected N, N-diethylm-toluamide (DEET), caffeine, and tri(2-chloroethyl)-phosphate (TCEP) in pool water. DEET, a commonly used active ingredient in commercial insect repellents, was found at very different levels. In pool A (in Georgia, USA) the concentration of DEET was highest 721 ± 3.7 ng/L in winter and 2087 ± 32 ng/L in summer, while in pool C (in Indiana, USA) DEET was found at about 200 ng/L. The authors examined the kinetics of decay of five chemicals (naproxen, ibuprofen, caffeine, DEET, and acetaminophen) identified as being the most likely to accumulate in pools. Acetaminophen and naproxen were susceptible to chlorination; N90% of both compounds degraded within the first 6 h of chlorine exposure while DEET, caffeine, and ibuprofen reacted more slowly to chlorination and N80% of these three compounds were still detectable after 24 h. Ekowati et al. (2016) observed atenolol, carbamazepine, hydrochlorothiazide, metronidazole, ofloxacin, sulphamethoxazole, acetaminophen, ibuprofen, ketoprofen and phenazone in the waters of 17 swimming pools in Catalonia, Spain. The highest concentration was found for the diuretic hydrochlorothiazide (904 ng/L), while the most frequently detected pharmaceutical was carbamazepine, observed in 53% of the pool waters. In Australia, Teo et al., 2016 investigated the occurrence and daily variability of 30 pharmaceuticals and personal care products in pool waters. Ibuprofen was above the LOQ in seven swimming pools, ranging from 16 to 83 ng/L. The main sources of these chemicals and/or their metabolites in pool waters are swimmers who eliminate organic substances, urine and sweat during their sport activities (WHO, 2006). Furthermore, when pharmaceuticals are used topically (anti-inflammatory drugs, for instance), they too can be released into pool waters from the skin. Once these substances have entered the water, their fate will be largely determined by reaction and transfer processes within the pool system. If not completely removed by chlorination, these chemicals continuously accumulate and increase in pool waters with time, due to the widespread practice of recirculating pool water continuously with little or no replacement for months or years. Bathers can release other chemicals such as drugs of abuse and/or their metabolites in urine. To date, there is no information in the literature, to our knowledge, about illicit drugs in swimming pool waters, although these chemicals are continuously found in other aquatic environments. Previous studies in Europe and Italy reported cocaine, opioids, amphetamines, cannabis and their metabolites in waste, surface and ground waters and also in drinking waters (Zuccato et al.,

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2000; Castiglioni et al., 2006; Zuccato et al., 2008; van der Aa et al., 2013; Mendoza et al., 2014). Pharmaceuticals and illicit drugs, documented in chlorinated drinking water at concentrations in the order of ng/L, can further react with chlorine and DBPs already formed and present, becoming in turn additional by-products, with greater toxicity than the original compounds (González-Mariño et al., 2012). Besides other chemicals in pool waters, such as disinfection byproducts (DBPs) from chlorine-based water treatment, swimmers may be exposed to pharmaceuticals and illicit drugs through ingestion of small amounts of water while swimming or by skin contact and even by inhalation of aerosols; the amount of water ingested depends on various factors including experience, age, skill and type of activity (Aggazzotti et al., 1995; WHO, 2006). In view of the potential hazards of pharmaceuticals and illicit drugs and the scarcity of data in the literature on their levels in swimming pool waters, this study investigated for the first time the occurrence of these substances and some of their metabolites in pool and source waters in a sample of Italian indoor swimming pools. The aim was to quantify pharmaceuticals and illicit drugs actual amounts in pool waters and to link them to the different water treatments such as disinfection and filtration. Another aim was to assess the potential exposure for swimmers, taking into account sensitive subpopulations. 2. Material and methods 2.1. Sample collection The study was done in ten public indoor swimming pools in the Emilia-Romagna Region (Northern Italy). One indoor swimming pool from the major cities of the Region (ten cities) was sampled. Each pool was visited once and information about the pool water treatments and disinfection procedures was collected. All sampling sessions were carried out in May 2016 in order to obtain waters that had been in the pools a long time, as the complete draining and filling with fresh water is generally done at the end of the summer. The time since the last complete change of water was collected. During each session, source (only in four swimming pools) and pool waters were sampled according to the EPA sampling procedure (USEPA, 2016). A grab sample of source water was collected as close as possible to the point where source water was added to the recirculating pool water. Individual samples of pool water (100 mL) were collected for trihalomethanes (THMs) determination at three different points, at a depth of 20 cm, and 40 cm from the edge of the pool. Samples for the determination of illicit drugs and pharmaceuticals were collected in 1000 mL polypropylene bottles, and were frozen and stored at −20 °C until analysis. 2.2. Analysis of swimming pool water physico-chemical parameters Temperature, pH, free and total chlorine, were analyzed in water samples on the poolside. Water temperature was taken with a digital thermometer, pH and oxidation reduction potential (ORP) were measured with a portable pH meter (pH 110 EUTECH Ins, USA). Free and combined chlorine were evaluated at the poolside using a colorimetric method based on N,N-diethyl-p-phenylendiamine (DPD) (PC compact – Aqualityc, USA). THMs, considered the most representative of the DBPs, were examined in order to link any by-products deriving from chlorination to other chemicals in the waters (WHO, 2006). Triplicate water samples for the determination of THMs (chloroform, bromodichloromethane, dibromochloromethane and bromoform) were collected in screwcapped glass vials (40 cm3) with Teflon faced septa. Five mg of sodium thiosulfate were added to the vials to quench residual chlorine reactions. The analysis were done within a few days, according to the EPA sampling procedure (USEPA, 2016).

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Table 1 Pharmaceuticals and illicit drugs in swimming pool water grouped by class. Pharmaceuticals Antibiotics Amoxicillin Cefazoline Ciprofloxacin Clarithromycin Erythromycin Dehydroerythromycin Lincomycin Ofloxacin Spiramycin Sulfamethoxazole Vancomicin Anticancer Cyclophosphamide Methotrexate Tamoxifen Anti-inflammatory and analgesics Diclofenac Ibuprofen Ketoprofen Naproxene Paracetamol Antihypertensive Enalapril Enalaprilat Irbesartan Losartan Ramipril Ramiprilat Valsartan

Bronchodilator Salbutamol Cardiovascular Atenolol CNS-drugs Carbamazepine 10,11-dihydro-10-hydroxycarbamazepine Demetildiazepam Diazepam Paroxetine Diuretics Furosemide Hydrocholorotiazide Estrogens 17-β estradiol Estrone 17-α ethynylestradiol Gastrointestinal Omeprazole Sulphide Lansoprazol Ranitidine Lipid regulators Atorvastatine Bezafibrate Clofibric Acid Gemfibrozil Rosuvastatin Simvastatine Hypoglycemic Metformine

Illicit drugs Cocaine and metabolites Cocaine Norcocaine Benzoylecgonine Norbenzoylecgonine Cocaethylene Amphetamine like substances Amphetamine Methamphetamine 3,4 MDMA (Ecstasy) New psycoactive substances Ketamine Mephedrone

Opioids Morphine 6-acethyl morphine Morphine 3D-glucuronide Morphine 6D-glucuronide Oxycodone Hydrocodone Codeine Methadone EDDP (methadone metabolites) Cannabinoids Tetrahydrocannabinol (THC) OH-THC THC-COOH

The levels of THMs in water samples were analyzed using a static head space gas chromatographic technique (SHS-GC) according to previous studies (Aggazzotti et al., 1995; Fantuzzi et al., 2010; Righi et al., 2014). A Varian 3380 gas chromatograph equipped with a 63Ni electron-capture detector (ECD) and a Vocol capillary column (30 m ×

0.53 mm I.D., film thickness 3.0 μm (Supelco) was used. Quantitative and qualitative analyses of THMs were managed using Dionex Chromeleon 6.0 software. Calibration was done according to the external standard method. THMs standards were prepared with pure analysis reagents (Supelco) in specific concentrations of methanol. A working standard solution (1–8 μg/L) was prepared every day in synthetic water (chloride 100 mg/L, nitrate 20 mg/L and sulfate 150 mg/L). The limit of detection (LOD) for each THM was 0.1 μg/L. Aliquots of 5 cm3 from the water sample, external standard and control blank (vials with THMfree water) were each placed in a 10 cm3 glass vial, sealed, and heated to a specific temperature (1 h at 37 °C). After incubation, 100 μL of the head-space sample was injected directly into the GC using a gas-tight syringe (Hamilton).

2.3. Analysis of illicit drugs and pharmaceuticals in swimming pool water The most used illicit drugs (cocaine, opioids, amphetamines and cannabis) and their metabolic residues, and pharmaceuticals aheady found in significant concentrations in the aquatic environment in Italy in previous studies were investigated (Zuccato et al., 2005: Riva et al., 2015). Among pharmaceuticals, antibiotics (n. 11), anticancers drugs (n. 3), non-steroidal anti-inflammatory drugs (n. 5), antihypertensive drugs (n. 7), bronchodilators (n. 1), cardiovascular drugs (n. 1), central nervous system – drugs (n. 5), diuretics (n. 2), estrogen and hormones (n. 3), gastrointestinal drugs (n. 4), lipid regulators (n. 6), and hypoglycemic drugs (n. 1) were investigated (Table 1). Samples were analyzed by high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS), after solid-phase extraction (SPE). The methods were adapted from previous publications (Castiglioni et al., 2005; Castiglioni et al., 2006; Riva et al., 2015). Briefly, water samples (200 mL) were acidified to pH 2.0 with 37% HCl, spiked with labeled internal standards (10–100 ng/L) and solidphase extracted using mixed reverse-phase cation exchange cartridges (Oasis-MCX, Waters Corp., Milford, MA). Cartridges were then vacuum-dried and eluted with 2 mL of methanol and 2 mL of a 2% ammonia solution in methanol. The eluates were pooled in glass tubes, dried under a gentle nitrogen stream, redissolved in 100 μL MilliQ water and transferred into glass vials for instrumental analysis. For analysis a 1200 Series pump system (Agilent Technologies, CA, USA) coupled to a triple quadrupole mass spectrometer (AB SCIEX QqQ 5500, Ontario, Canada) were used. Compounds were quantified by selected reaction monitoring (SRM) using both the positive and negative ionisation modes. Quantification was done by isotope dilution for each compound using the most abundant precursor/product ion transition (quantifier ion) and comparing the area with its corresponding deuterated analog (IS). Retention times were also compared with reference standards to identify the compounds.

Table 2 Main characteristics of ten indoor swimming pools: water treatments and water quality. Name

A B C D E F G H I J

Treatment

Physical and chemical parameters

Filtration

Disinfection

Water source

t° (°C)

pH

ORP^ mV

Free chlorine mg/L

Total chlorine (mg/L)

THMs (μg/L)

Sand filter Sand filter + GAC⁎ Sand filter + GAC⁎ Sand filter + GAC⁎ Sand filter + GAC⁎

Sodium hypoclorite Trichloroisocyanuric acid Sodium hypoclorite Calcium hypoclorite Calcium hypoclorite Calcium hypoclorite Calcium hypoclorite Sodium hypoclorite Trichloroisocyanuric acid Sodium hypoclorite

Tap water Tap water Tap water Tap water Tap water Tap water Tap water Tap water Tap water Tap water

26.5 28.1 28.0 28.6 28.0 28.6 28.9 27.7 28.9 28.5

7.65 7.18 7.60 6.86 7.30 7.29 7.33 7.38 7.41 7.30

579 620 678 680 665 686 616 706 609 609

1.20 1.54 0.70 1.05 1.38 1.31 1.07 1.29 2.05 1.62

1.57 2.11 1.30 1.51 1.89 1.82 1.48 2.03 2.46 2.40

35.3 45.4 31.7 35.5 22.9 28.1 43.8 55.8 33.2 61.7

Sand filter Sand filter Sand filter + GAC⁎ Sand filter Sand filter

^ ORP: oxidation reduction potential. ⁎ GAC: granular active carbon.

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Table 3 Analytical performance of the method (recoveries and sensitivity) and parameters used for quantification (labeled standards, calibration curves ranges and coefficients of correlation). Chemicals

Labeled standard

Linearity range (μg/L)

Coefficient of correlation (r2)

Recovery ± SD (%)

LOD (ng/L)

LOQ (ng/L)

Pharmaceuticals Atenolol Carbamazepine 10,11-Dihydro-10-hydroxycarbamazepine Dehydroerythromycin Enalapril Hydrochlorothiazide Ibuprofen Irbesartan Ketoprofen Paracetamol Valsartan

Atenolol-d7 Carbamazepine-d10 Carbamazepine-d10 Carbamazepine-d10 Ramipril-d5 Ibuprofen-d3 Ibuprofen-d3 Valsartan-d3 Ibuprofen-d3 Salbutamol-d3 Valsartan-d3

0–50 0–50 0–50 0–50 0–50 0–50 0–50 0–50 0–50 0–50 0–50

0.99993 0.99980 0.99962 0.99922 0.99984 0.99951 0.99992 0.99961 0.99997 0.99925 0.99956

81 ± 8.7 93 ± 3.6 116 ± 5.9 71 ± 2.6 101 ± 3.4 122 ± 7.1 98 ± 8.0 82 ± 8.9 101 ± 10.3 100 ± 0.4 76 ± 8.3

0.01 0.03 0.11 0.51 0.03 0.07 0.59 0.05 2.5 0.46 0.11

0.03 0.01 0.35 1.7 0.1 0.2 2.0 0.2 8.2 1.5 0.4

Illicit drugs Cocaine Norcocaine Benzoylecgonine Norbenzoylecgonine

Cocaine-d3 Cocaine-d3 Benzoylecgonine-d3 Norbenzoylecgonine-d3

0–15 0–15 0–15 0–15

0.99989 0.99936 0.99929 0.99970

96 ± 5 112 ± 7 107 ± 9 85 ± 5

0.02 0.02 0.018 0.018

0.07 0.08 0.06 0.06

The detection limits (LOD) and quantification limits (LOQ) were calculated from chromatograms of swimming pool samples; the LOD was the concentration with a signal/noise ratio of 3 and the LOQ was the concentration with a signal/noise ratio of 10.

risk for swimmers due to accidental ingestion of the water can be considered negligible even for lifetime exposure. The DWGL for a compound was calculated as reported in Eq. 1: DWGL ½μg=kg day ¼ ADI  BW  P  103 =V

2.4. Risk assessment for human health Human Risk Assessment (HRA) was conducted for children (3 years old), teens (14 years old) and adults, dividing each category by sex. The weights used in the assessment were obtained from Cacciari et al. (2006) considering the 50th percentile; for the average amount of water swallowed we considered the data reported in Dufour et al. (2006). A worst case scenario was designed taking the maximum concentration of each contaminant measured in waters and assuming that the subjects swam daily. HRA was first assessed considering the exposure to each single contaminant, and concentrations were compared with Drinking Water Guideline Levels (DWGLs) (AwwaRF, 2008; Environment Protection and Heritage Council and Natural Resource Management Ministerial Council, 2008); these are theoretical concentrations below which the

ð1Þ

where ADI is the Acceptable Daily Intake, i.e., the dose that can be ingested daily over a lifetime with a negligible risk of adverse effects; BW is the body weight considered; P is the fraction of the substance ingested in water (we took this as 100%); V is the daily volume of water drunk (data reported in Dufour et al., 2006). These levels were calculated according to the guidelines, as described elsewhere (Riva et al., 2018). The ADI used to calculate DWGL for THM were also collected from the guidelines (WHO, 2005). The ratios between the concentrations measured and the DWGLs are called Hazard Quotients (HQs). HRA was later assessed for co-exposure to all contaminants and was measured following the Hazard Index (HI) approach, in which the HQs of single contaminants are summed to obtain a single risk value. Generally, if the HQ and HI are below 1 the risk

Table 4 Concentrations of pharmaceuticals and illicit drugs in ten indoor swimming pool waters and four source waters (mean and range). Swimming pools

Pharmaceutical compounds Ibuprofen Ketoprofen Carbamazepine 10,11-Dihydro-10-hydroxycarbamazepine Valsartan Atenolol Enalapril Paracetamol Hydrochlorothiazide Irbesartan Dehydroerythromycin Illicit drugs of abuse Cocaine Norcocaine Benzoylecgonine Norbenzoylecgonine

Source waters

Frequency of determination

Mean ± SD

Range ng/L

10/10 9/10 8/10 8/10 8/10 4/10 3/10 2/10 1/10 1/10 1/10

64.58 ± 54.54 75.35 ± 45.91 1.06 ± 1.79 12.59 ± 20.68 4.01 ± 3.58 0.13 ± 0.11 0.65 ± 0.31 2.10 ± 0.72

16.1–197.0 12.6–127.0 0.1–5.4 0.7–62.2 0.7–8.7 0.03–0.2 0.4–1.0 1.6–2.6 0.3 0.2 2.4

9/10 9/10 9/10 9/10

1.29 ± 1.31 13.05 ± 15.32 4.56 ± 6.56 2.38 ± 2.44

0.3–4.2 1.1–48.7 0.7–21.4 0.1–7.3

Frequency of determination

Mean ± SD

Range ng/L

2/4

0.82 ± 0.62

0.4–1.3

1/4 1/4

3.3 1.0

1/4 1/4

0.1 0.7

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posed by exposure to these contaminants in swimming pools can be considered negligible.

3. Results 3.1. Main characteristics of swimming pools

2.5. Statistical analyses All statistical analyses were carried out using the Statistical Package for Social Sciences (SPSS version 18.0 for Windows, Chicago, IL, USA). As environmental data were not normally distributed, non-parametric tests were applied. The Kruskal Wallis test was used to evaluate differences in mean concentrations of pharmaceuticals and drugs of abuse. Correlations between chemical-physical parameters and/or pharmaceuticals and drugs of abuse levels were evaluated with Spearman's rank correlation coefficient. Statistical significance for all statistical tests was set at p b 0.05.

Table 2 reports the main characteristics of the disinfection treatments of the ten indoor swimming pools and some physical and chemical parameters. Four swimming pools were disinfected with sodium hypochlorite, four with calcium hypochlorite and two with trichloroisocyanuric acid. Sand filters with granular active carbon (GAC) were used in five swimming pools (50%). Water temperature ranged from 26.6 °C to 28.9 °C (28.2 ± 0.7 °C), and pH from 6.86 to 7.65. ORP ranged from 579 to 706 mV with a mean of 644 ± 42.86 mV. Free and total chlorine in water ranged from respectively 0.7 to 2.0 mg/L and 1.3 to 2.5 mg/L. Among THMs, chloroform, BDCM and DBCM were found in swimming pool waters at concentrations

Pharmaceucals 250

200

150

100

50

0 A

B

C

D

E

10,11-dihydro-10,11-hydroxycarbamazepine

F Ketoprofen

G

H

Ibuprofen

I

J

Atenolol

Valsartan

H

I

Illicit drugs 60

50

40

30

20

10

0 A

B

C Cocaine

D Norcocaine

E

F

Benzoylecgonine

G

J

Norbezoylecgonine

Fig. 1. Concentrations (ng/L) of the most abundant pharmaceuticals and illicit drugs in the investigated swimming pools waters.

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ranging respectively from 19.5 μg/L to 54.8 μg/L, 2.8 μg/L to 7.9 μg/L and 0.3 μg/L to 3.7 μg/L. The mean level of THMs was 39.34 ± 12.25 μg/L, with a maximum of 61.7 μg/L. Tap water was the only source water and the last complete filling with water had been done nine months before, in September 2015, in all the swimming pools investigated. 3.2. Illicit drugs and pharmaceuticals in swimming pool water The main performance parameters of the method are reported in Table 3. Recoveries were higher than 70% with standard deviations lower than 10% and LOQs for the method were in the low ng/L or high pg/L range. Among illicit drugs, only cocaine and its metabolites were detected above the LOQ in swimming pool waters (Table 4). Cocaine and its metabolites were found in nine pools, at concentrations from 0.3 to 4.2 ng/L for cocaine, 1.1 to 48.7 ng/L for norcocaine, 0.7 to 21.4 ng/L for benzoylecgonine and 0.1 to 7.3 ng/L for norbenzoylecgonine. Opioids, amphetamines and cannabis derivatives were never detected. Norcocaine and benzoylecgonine were found in one source water sample at ng/L levels. Eleven pharmaceuticals were detected at levels above their LOQs (Table 4). The most frequently detected pharmaceuticals were antiinflammatory drugs. Ibuprofen was present in all the pool waters investigated, with maximum 197 ng/L and ketoprofen was detected in 9/10 samples (maximum 127 ng/L). The anticonvulsant carbamazepine and its metabolite 10,11-dihydro-10,11-dihydroxy-carbamazepine were frequent in pool water samples (8/10 samples) at concentrations up to 62 ng/L. The cardiovascular drug valsartan was also found frequently (8/10 samples), but at lower concentrations (up to 9 ng/L). The other pharmaceuticals were found occasionally (1–4 samples) and at lower concentrations (i.e. atenolol, enalapril, paracetamol, hydroclorothiazide, irbesartan and dehydroerythromycin). In the four source water samples investigated only carbamazepine, irbesartan and dehydroerythromycin were detected at very low levels (up to 5 ng/L). Fig. 1 shows concentrations (ng/L) of the most abundant pharmaceuticals and cocaine and its metabolites in swimming pool waters. No significant correlations were observed between the substances investigated and physical and chemical parameters. Disinfection procedures were also not related to the presence of pharmaceuticals and illicit drugs. Instead, when sand filters + granular active carbon (GAC) were used for water treatment, drug levels were lower than in swimming pools where only sand filters were used. Significant differences were observed for illicit drugs applying a Mann-Whitney test

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(norcocaine p = 0.016; benzoylecgonine norbenzoylecgonine p = 0.036) (Fig. 2).

p

=

0.032;

3.3. Risk assessment for human health HRA for the single substances and for co-exposure are reported in Table 5, and show negligible risks for swimmers of any age or sex. The HQs for pharmaceuticals were below 0.001 for almost all the subjects, with the sole exception of ketoprofen for children where HQs were 0.004 and 0.003 respectively for boys and girls. The HQs for cocaine and its metabolites were generally higher than those of pharmaceuticals, ranging up to 0.146, but were all far below the threshould of 1. The HQs for THMs ranged from b0.001 to 0.011. being in the same range as the other emerging pollutants. The cumulative risk calculated as HI were also all below 1, the highest being 0.260 for boys three years old and 0.173 for girls three years old. 4. Discussion This study, the first in Italy, demonstrated the presence of illicit drugs and pharmaceuticals in Italian swimming pool waters at levels up to the hundreds of ng/L. Pharmaceuticals were detectable in several swimming pools and anti-inflammatory agents, anticonvulsants and beta-blockers were the most widespread. In general, pharmaceutical concentrations were in accordance with the reports in the literature. In a study in Spain 32 pharmaceuticals were evaluated and only eleven were detected above their LOD. The most prevalent were antibiotics and anti-inflammatory agents and analgesics, and their concentrations in pool waters were in general lower than 100 ng/L. The most frequently detected pharmaceutical was carbamazepine, which was observed in more than half the water samples (53%, 27/51) (Ekowati et al., 2016). Teo et al. (2016) observed concentrations of ibuprofen in eight chlorinated pools in the range of 16–83 ng/L, but no other pharmaceuticals were found up to the LOD. Regarding illicit drugs opioids, amphetamines and cannabis derivatives were not detected in the present study (Table 4), while cocaine and its main metabolites were found in almost all the samples up to 49 ng/L. These chemicals are present in pool waters as a consequence of swimmers releasing them in urine and sweat, or skin when pharmaceuticals are used topically, as in source waters they were observed at very low concentrations and in only a few samples.

ng/L 20

*

18 16 14 12 10 8 6

*

4

*

2 0 cocaine

norcocaine sand filter

benzoylecgonine

norbenzoylecgonine

sand filter + granular acve carbon

Fig. 2. Median concentrations of cocaine and its metabolites in swimming pool waters according to the use of sand filters or sand filters + granular active carbon. *p b 0.05.

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Table 5 Hazard Quotients (HQs) for emerging contaminants in swimming pool waters, and Hazard Indexes (HIs) for each class of swimmers. Swimmers

Boys 3 years

Girls 3 years

Boys 14 years

Girls 14 years

Men

Woman

Weight (kg)

15

15

55

54

70

55

0.045

0.030

0.045

0.030

0.022

0.012

Water swallowed (L)

Hazard quotients Pharmaceuticals Atenolol Carbamazepine 10.11-dihydro-10-hydroxycarbamazepine Ibuprofen Hydroclorothyazide Irbesartan Ketoprofen Paracetamol Valsartan

ADI (μg/kg day) 2000 3400 3400 2860 0,180 1071 0,100 5000 0,290

b0.001 b0.001 b0.001 b0.001 b0.001 b0.001 0.004 b0.001 b0.001

b0.001 b0.001 b0.001 b0.001 b0.001 b0.001 0.003 b0.001 b0.001

b0.001 b0.001 b0.001 b0.001 b0.001 b0.001 0.001 b0.001 b0.001

b0.001 b0.001 b0.001 b0.001 b0.001 b0.001 0.001 b0.001 b0.001

b0.001 b0.001 b0.001 b0.001 b0.001 b0.001 b0.001 b0.001 b0.001

b0.001 b0.001 b0.001 b0.001 b0.001 b0.001 b0.001 b0.001 b0.001

Illicit drugs Cocaine Norcocaine Benzoylecgonine Norbenzoylecgonine

0,001 0,001 0,001 0,001

0.012 0.146 0.064 0.022

0.008 0.097 0.043 0.015

0.003 0.040 0.018 0.006

0.002 0.027 0.012 0.004

0.001 0.015 0.007 0.002

0.001 0.011 0.005 0.002

THMs Chloroform DBCM

15,000 214,000

0,011 b0.001

0,007 b0.001

0,003 b0.001

0,002 b0.001

0,001 b0.001

b0.001 b0.001

0,260

0,173

0,071

0,048

0,027

0,019

Hazard index

In Italy it is not uncommon for pool water to remain unchanged for several months at a time or longer except for the fresh water replaced daily after evaporation or splashing by swimmers. This management approach is motivated by a number of factors, including simplicity and the costs of water replacement, heating, and treatment. However, it also offers the potential for accumulate of constituents that are stable, in terms of degradation by reaction and gas/ liquid transfer. Previous studies have demonstrated the formation of disinfection by-products due to the reaction of chlorinate-based disinfectants with certain pharmaceuticals. It was interesting to note the higher concentrations of norcocaine than the other metabolites and cocaine itself (Table 4, Fig. 2). This result is unusual, since norcocaine is a minor metabolite of cocaine and is normally found in wastewater (as a result of human excretion) at much lower levels than the other metabolites (Castiglioni et al., 2011). Nevertheless, González-Mariño et al. reported recently that chlorine in water can react with cocaine to give this metabolite (González-Mariño et al., 2012). This reaction is therefore confirmed by our study in a real case scenario - chlorination in a swimming pool. Other disinfection by-products could originate from the reaction of other pharmaceuticals with chlorine and further investigation is needed. The widespread presence of chemicals in pool waters obviously raises the question whether these concentrations pose a risk for people using indoor swimming pools. Swimmers swallow more or less water depending on variables such as age, sex, technical skill, swimming style, intensity and duration. Few data exists about the amount of water ingested during swimming. Dufour et al. (2006) calculated the average amount of water swallowed by swimmers and found considerable differences in water ingestion, depending on age (37 mL in children and 16 mL in adults) and sex (45 mL in boys, 30 mL in girls, 22 mL in men and 12 mL in women). Obviously, subjects who do not know or are learning how to swim may swallow more. These data were used to assess the human risk related to ingestion of water during swimming and no risk was found either for children or for adults. This is in line with current risk assessments related to drinking water, indicating that concentrations of pharmaceuticals in the ng/L range are very unlikely to pose any risks to human health (Houtman et al., 2014).

However, the risk of long-term exposure to active pharmaceuticals and drugs of abuse and to disinfection by-products in swimming pool waters has not been investigated up to now so health effects cannot be ruled out yet, especially with regard to vulnerable subpopulations like children. Investigation of possible additive or synergistic effects of mixtures would also be necessary for accurate assessment of the potential risk to human health, taking into account sensitive subpopulations such as babies and small children. 5. Conclusion This study investigated the concentrations of several categories of pharmaceuticals and illicit drugs in ten Italian indoor swimming pools. Cocaine and its metabolites were widespread, at concentrations up to 49 ng/L for norcocaine, while opioids, amphetamines and cannabis derivatives were never observed. Among pharmaceuticals, ibuprofen, ketoprofen, carbamazepine and its metabolite, 10,11-dihydro-10,11dihydroxy-carbamazepine and valsartan were found frequently, but at low concentrations. The human risk assessment indicated that the health risk from oral exposure to these compounds in swimming pools in this study was generally low, with hazard quotients less than one. However, further research is needed to check for any additive or synergistic effects due to disinfection by products in pool waters for accurate exposure assessment, to see whether there are any potential risks for human health, in sensitive subpopulations as well. Acknowledgements This research received financial support from the University of Modena and Reggio Emilia, Department of Biomedical, Metabolic and Neural Sciences (FAR, 81711/2015). The authors thank the managers of the indoor swimming pools involved in the study who supported our monitoring campaign. References Aggazzotti, G., Fantuzzi, G., Righi, E., Predieri, G., 1995. Environmental and biological monitoring of chloroform in indoor swimming pools. J. Chromatogr. A 710, 181–190.

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