Science of the Total Environment

(1)

Occurrence and fate of select psychoactive pharmaceuticals and antihypertensives in two wastewater treatment plants in New York State, USA

Bikram Subedi

^a

, Kurunthachalam Kannan

^a,b,

⁎

aWadsworth Center, New York State Department of Health, and Department of Environmental Health Sciences, School of Public Health, State University of New York at Albany, Empire State Plaza, P.O. Box 509, Albany, NY 12201-0509, United States

bBiochemistry Department, Faculty of Science and Experimental Biochemistry Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589, Saudi Arabia

H I G H L I G H T S

•16 psychoactive pharmaceuticals were found at 0.98–1220 ng/L in wastewater inﬂuent.

•Over 50% of total mass of 8 psychoactives were found sorbed to particulate matter.

•Inﬂux of psychoactives in WWTPs ranged from 0.91 to 347 mg/d/1000 inhabitants.

•Environmental emission of psychoactives ranged from 0.01 to 316 mg/d/1000 inhabitants.

G R A P H I C A L A B S T R A C T

a b s t r a c t a r t i c l e i n f o

Article history:

Received 24 November 2014 Received in revised form 5 January 2015 Accepted 27 January 2015

Available online xxxx Editor: Adrian Covaci Keywords:

Pharmaceuticals Psychoactives Metabolites Fate

Removal efﬁciency Sludge

Suspended particulate matter Wastewater treatment plant

The fates of psychoactive pharmaceuticals, including two antischizophrenics, six sedative–hypnotic–anxiolytics, four antidepressants, four antihypertensives, and their select metabolites, were determined in two wastewater treatment plants (WWTPs) in the Albany area of New York. All target psychoactive pharmaceuticals and their metabolites were found at a mean concentration that ranged from 0.98 (quetiapine) to 1220 ng/L (atenolol) in wastewater and from 0.26 (lorazepam) to 1490 ng/g dry weight (sertraline) in sludge. In this study, the fraction of psychoactive pharmaceuticals that was sorbed to suspended particulate matter (SPM) was calculated for the first time. Over 50% of the total mass of aripiprazole, norquetiapine, norsertraline, citalopram, desmethyl citalopram, propranolol, verapamil, and norverapamil was found sorbed to SPM in the influent. The mass loadings, i.e., influx, of target psychoactive pharmaceuticals in WWTPs ranged from 0.91 (diazepam) to 347 mg/d/

1000 inhabitants (atenolol), whereas the environmental emissions ranged from 0.01 (dehydro-aripiprazole) to 316 mg/d/1000 inhabitants (atenolol). The highest calculated removal efﬁciencies were found for antischizophrenics (quetiapine = 88%; aripiprazole = 71%). However, the removal of some psychoactive pharmaceuticals through adsorption onto sludge was minimal (b1% of the initial mass load), which suggests that bio-degradation and/or chemical-transformation are the dominant mechanisms of removal of these pharmaceuticals in WWTPs.

⁎ Corresponding author at: Wadsworth Center, Empire State Plaza, P.O. Box 509, Albany, NY 12201-0509, United States.

E-mail address:[email protected](K. Kannan).

Contents lists available atScienceDirect

Science of the Total Environment

j o u r n a l h o m e p a g e :w w w . e l s e v i e r . c o m / l o c a t e / s c i t o t e n v

(2)

1. Introduction

Neurological disorders have emerged as a signiﬁcant public health concern. The total global burden of diseases that result from neuropsy- chiatric disorders is projected to be the second most frequent by 2020 (Menken et al., 2000). Globally, psychoactive drugs were among the most prescribed classes of pharmaceuticals in 2008, with over 30 billion doses prescribed daily in that year (INCB, 2010). In the USA, the total usage of psychoactive pharmaceuticals increased from 1998 to 2008 by 78% (Gallini et al., 2013), and psychoactives were the top-selling class of prescription medications, with $14.6 billion in sales in 2009 (IMS, 2009).

Most psychoactive pharmaceuticals undergo metabolism in the human body and excrete either unchanged, as metabolites, or as conjugates (Calisto and Esteves, 2009). Metabolites of select psychoactive pharmaceuticals are biologically active, and the conjugates can be further transformed to corresponding parent compounds in wastewater treatment processes (Calisto and Esteves, 2009). Psychoactive pharmaceuticals, their metabolites, and conjugates can enter the environment through the discharge of efﬂuents from wastewater treatment plants (WWTPs) (Ternes, 1998), efﬂuents from pharmaceutical industries (Ruhoy and Daughton, 2008), use of treated wastewater for irrigation (Pedersen et al., 2005), and the application of biosolids in agriculture (Kinney et al., 2008; Subedi et al., 2013). Psychoactive pharmaceuticals were reported to occur in wastewater, surface water, and drinking water at ng/L toμg/L levels and in sludge/biosolids at ng/g toμg/g levels (Benotti et al., 2009; Kolpin et al., 2002; Subedi et al., 2013; Wick et al., 2009; Yuan et al., 2013).

Studies have reported that exposure of aquatic organisms to psychoactive pharmaceuticals affect reproduction (Brooks et al., 2003), endocrine function (van der Ven et al., 2006), or photosynthesis (Escher et al., 2006). A mixture of psychoactive pharmaceuticals venlafaxine (VLF),ﬂuoxetine, and carbamazepine (CBZ) at environmentally rele- vant concentrations induced autism-like gene expression in fathead minnows (Thomas and Klaper, 2012). A recent study showed that exposure ofﬁsh and benthic invertebrates to psychoactive drugs altered the behavioral responses (Brodin et al., 2014; Rosi-Marshall et al., 2015). To date, however, very little is known about the ecotoxicologic effects of psychoactive pharmaceuticals in aquatic ecosystems (Alonso et al., 2010; Petersen et al., 2014).

Psychoactive pharmaceuticals, such as CBZ and VLF, are recalcitrant, and their removal efﬁciencies in WWTPs were estimated atb20%

(Lajeunesse et al., 2012; Zhang et al., 2008). Estimation of removal rates of pharmaceuticals in the aqueous phase alone can under- or over-estimate the actual removal (Petrovic and Barceló, 2007). In the USA, ~240 kg of biosolids are produced for every million liter of domes- tic wastewater treated (Kinney et al., 2008). The US Environmental Pro- tection Agency (EPA) estimated that ~ 50% of the annual production (8 × 10⁶tons in 2006) of biosolids is land-applied (USEPA, 2006).

Despite the sorption of psychoactive pharmaceuticals to suspended particulate matter (SPM), studies on the fate of these chemicals have focused only on the analysis infiltered wastewater (Lajeunesse et al., 2012; Radjenovic et al., 2009) with the assumption that pharmaceuticals partition mainly to the aqueous phase.Radjenovic et al. (2009) reported mean emission of 7.1, 41.1, and 3.9 g/d of CBZ, atenolol (ATN), and propanolol (PPN), respectively, based on the concentrations measured infiltered wastewater and treated sludge from nine WWTPs, with an average inflow of 42,000 m³/d.

In this study, psychoactive pharmaceuticals, including two antischizophrenics [aripiprazole (APPZ) and quetiapine (QTP)], six sedative– hypnotic–anxiolytics [lorazepam (LZP), alprazolam (APZ), diazepam (DZP), oxazepam (OxZP), nordiazepam (NDZP), and CBZ], four antidepressants [VLF, bupropion (BPP), sertraline (STL), and citalopram (CTP)], four antihypertensives [ANL, PPN, diltiazem (DTZ), and verapamil (VPM)], and their select metabolites, were analyzed in wastewater influent, primary effluent,final effluent, SPM, and treated sludge from

two WWTPs in the Albany area of New York. In addition to psychoactive pharmaceuticals, one antiplatelet [clopidogrel (CPG)], one antihistamine [diphenhydramine (DPH)], and their metabolites were analyzed.

This is thefirst study to describe the fate of psychoactive pharmaceuticals including influx assessment as well as the fraction sorbed to the sewage sludge and SPM in WWTPs in the USA. The fraction of each of the target pharmaceuticals sorbed to SPM was calculated and utilized in the estimation of mass loadings and emissions of these chemicals from WWTPs. The removal efficiencies of pharmaceuticals through wastewater treatment processes also were calculated.

2. Material and methods 2.1. Reagents and chemicals

Standard stock solutions (100 or 1000μg/mL) of individual pharmaceuticals and their corresponding isotopically-labeled standards were purchased from commercial vendors, as described elsewhere (Subedi et al., 2013). Purity of all of the standards was≥95%. Formic acid (98.2%) was from Sigma-Aldrich (St. Louis, MO, USA). Ultrapure water was prepared with the milli-Q ultrapure system (Barnstead Interna- tional, Dubuque, IA, USA). All standard stock solutions were stored at

−20 °C.

2.2. Sample collection and preparation

Information on the collection of wastewater samples and sludge has been described earlier (Subedi and Kannan, 2014). Briefly, 24-h composite wastewater samples, including raw wastewater (influent), primary treated wastewater (primary effluent), secondary treated wastewater (effluent orfinal effluent), and sludge were collected over a seven-day period, from July 12 to 18, 2013, consecutively from two WWTPs in the Albany area of New York. The two plants are denoted as WWTPA(population served ~ 15,000) and WWTPB (population served ~100,000), with a treatment capacity of 2.5 and 35 MGD, respectively. Both WWTPs used activated biological treatment. The activated sludge samples (3.9% solid, determined gravimetrically) were collected for seven days in WWTPA; however, activated sludge and dewatered sludge samples were collected from WWTPBfor four consecutive days within the sampling week. The activated sludge samples from both WWTPs were the combined sludge produced after primary and secondary treatments (dewatered and thickened). More detailed information on WWTPs is provided elsewhere (Sinclair and Kannan, 2006) and Table S1. All samples were collected in certified pre-cleaned amber glass jars with Teflon-faced caps, shipped to the laboratory, and stored in a refrigerator at 4 °C until extraction.

The detailed procedure for the extraction of wastewater, SPM, and sludge has been described elsewhere (Subedi and Kannan, 2014).

Briefly, wastewater samples (100 mL) were centrifuged at 5000 ×gfor 10 min, and the supernatant wasfiltered through a glassfiberfilter (37 mm, pore size, 1μm; GE Osmonics Inc., Minnetonka, MN) to sepa- rate SPM. Thefiltered wastewater was spiked with a mixture of labeled internal standards (25 ng), mixed well, allowed to equilibrate for

~ 30 min at room temperature, and extracted by passage through Oasis® HLB 6 cm³(200 mg; Waters, Milford, MA) solid phase extraction (SPE) cartridges. Prior to use, the cartridges were conditioned with 5 mL of methanol and 5 mL of milli-Q water, and wastewater samples were loaded at ~ 1 mL/min. Cartridges were allowed to dry for

~ 30 min under vacuum and then eluted with 6 mL of methanol followed by 3 mL of a mixture of acetone, methanol, and ethyl acetate (2:2:1 v/v/v). Cartridges were also eluted with 3 mL of methanol con- taining 5% ammonia. The eluents were combined and concentrated to

~100μL under a gentle stream of nitrogen at 35 °C using a TurboVap®

Evaporator (Zymark, Inc., Hopkinton, MA). Theﬁnal volume of the extract was adjusted to one milliliter with methanol in an amber glass vial, and 10μL of the extract was injected into HPLC–MS/MS.

(3)

Freeze-dried sludge (0.1 g) was spiked with a mixture of internal standards (25 ng) prior to extraction. The entire contents of SPM obtain- ed by centrifugation andfiltration of 100 mL of wastewater were trans- ferred into a clean, pre-weighed polypropylene tube along with a pre- weighed glass-fiberfilter, freeze-dried, and analyzed. Spiked sludge samples were vortex-mixed for 1 min and extracted with 6 mL of methanol:water mixture (5:3 v/v) using an ultrasonic bath (Branson®

Ultrasonics 3510R-DTH; Danbury, CT) for 30 min. Extracts were centrifuged at 4500 ×gfor 5 min (Eppendorf Centrifuge 5804, Hamburg, Germany), the supernatant was collected in a polypropylene tube, and the extraction was repeated with 6 mL of methanol. The extracts were combined and concentrated to ~1 mL under a gentle stream of nitrogen.

The concentrated extract was diluted with milli-Q water to ~12 mL and puriﬁed by passage through Oasis® HLB (6 cm³, 200 mg) cartridges, as described above for wastewater samples. Theﬁnal volume of the extract was one milliliter, and 10μL of the extract were injected into HPLC–MS/

MS for analysis.

2.3. Analysis

Target chemicals were analyzed using an API 2000 electrospray tri- ple quadrupole mass spectrometer (ESI-MS/MS; Applied Biosystems, Foster City, CA, USA), interfaced with an Agilent 1100 Series HPLC system (Agilent Technologies, Santa Clara, CA, USA). The analytes were separated using a Hypersil Gold® column (150 mm × 2.1 mm, 3μm) (Thermo Scientiﬁc, Chelmsford, MA, USA). Methanol and water (0.1%

formic acid) were used as mobile phases; a description of the mobile phase gradientﬂow is presented in Table S2. Target analytes were determined by multiple-reaction monitoring (MRM) in the positive ion- ization mode. Detailed information on the MS/MS transitions is provided in the Supporting Information (Table S3) (Subedi et al., 2013).

Briefly, analyte peak identification was based on the retention time (±0.05 min) and the ratio of qualitative to quantitative transition-ion responses (±20%). The quantitation of pharmaceuticals was based on the isotope dilution method. The calibrations curves were prepared by plotting concentration-dependent response factor of each target analyte (peak area of analyte divided by peak area of internal standard) versus the response-dependent concentration factor (concentrations of analyte divided by concentration of internal standard). However, metabolites were quantified using the internal standards of the corresponding parent compound (due to the lack of labeled standards available for the metabolites), and BPP was quantified using VLF-D8. The regression coefficients (r²) for seven- to nine-point calibration standards calculated by equal weighting quadratic regression were≥0.99 for all target analytes. The limits of quantitation (LOQs) and limits of detection (LODs) were determined as a minimum concentration of analytes in sample extracts that provide a signal to noise ratio≥10 and≥3, respectively. LODs of the target pharmaceuticals and their metabolites in wastewater, SPM, and sludge samples were in the ranges of 0.05– 10 ng/L, 0.05–5.0 ng/g, and 0.1–10 ng/g, respectively. LOQs of the target pharmaceuticals and their metabolites in wastewater, SPM, and sludge samples were ranged from 0.1–20 ng/L, 0.1–20 ng/g, and 0.5–20 ng/g, respectively. The continuing calibration verification standards injected before and after every batch (n= 21) of sample analysis showed recoveries at 100 ± 30%. A method blank was analyzed with every batch of samples. The concentrations of target pharmaceuticals and their metabolites in method blanks were below the corresponding LOQ. The concentrations of the target chemicals in SPM and sludge are reported on a dry-weight basis unless stated otherwise.

One sample was selected randomly for matrix spike (MS) and matrix spike duplicate (MSD) analyses with each batch of samples analyzed.

Target pharmaceuticals and their corresponding internal standards were each spiked at 10 and 25 ng, respectively, and were passed through the entire analytical procedure. The average relative recoveries of target pharmaceuticals from wastewater, sludge, and SPM were 74 ± 32%, 81 ± 26%, and 84 ± 34%, respectively (Table S2).

2.4. Calculations

The fraction of the total mass of analytes sorbed to SPM, removal ef- ﬁciency of pharmaceuticals through the treatment processes, mass loadings in WWTPs, and emission from WWTPs were calculated using the following equations (Eqs.(1)–(4)), as reported bySubedi and Kannan (2014)andJelic et al. (2012).

PSPM¼

CSPMMSPM

V_W

C_SPMM_SPM V_W

þC_W

100 ð1Þ

Removal efficiencyð Þ ¼%

C_iF 100 100−PSPM

− C_eF 100 100−PSPM

þðC_STSPÞ

CiF 100 100−PSPM

100

ð2Þ

Mass load¼C_iF 100 100−P_SPM

1 10⁶

1000 Population

ð3Þ

Emission=1000 people¼ ðC_EFÞ 100 100−P_SPM

þðC_STSPÞ

1000 Population

1 10⁶

ð4Þ

where P_SPMis the fraction of the total mass of analytes sorbed to SPM (%), CSPMis the concentration of analytes in SPM (ng/g), MSPMis the mass of SPM analyzed (g), VWis the volume of wastewater (L) used to obtain M_SPM, C_Wis the concentration of analyte in wastewater (ng/L), Ciis the concentration of analyte in wastewater influent (ng/L), Ceis the concentration of analyte in wastewater effluent (ng/L), PSPMiis the fraction of the total load of analyte sorbed to SPM (%) in wastewater influent, PSPMeis the fraction of the total load of analyte sorbed to SPM (%) in wastewater effluent, mass load is the amount of individual pharmaceutical introduced into WWTP (mg/d/1000 inhabitants), F is the daily flow of wastewater influent (L/d) over a 24-h period, Cs is the concentration of analyte in sludge (ng/g wet weight), TSP is the total sludge production (g/d wet weight), population is the number of inhabitants served by the WWTP, and emission/1000 inhabitants is the quantity of pharmaceutical discharged through wastewater effluent, SPM, and sludge (mg/d/1000 inhabitants).

3. Results and discussion

3.1. Psychoactive pharmaceuticals in wastewater

APPZ and QTP were found in 64% and 100%, respectively, of inﬂuent samples (n= 14; seven samples from each plant) (Table 1and Fig. S1).

The mean concentration of APPZ found in influent in our study was ~6 times lower than that reported for wastewater from a psychiatric hospital in China (Yuan et al., 2013); however, APPZ was not reported in wastewater from a centralized municipal WWTP earlier (Table 1). Sim- ilarly, the mean concentration of QTP in influent (19.9 ng/L) was 4.2 times higher than that reported from centralized municipal WWTPs in China and 176 times lower than that reported for wastewater from a psychiatric hospital (Yuan et al., 2013). APPZ and QTP were among the top anti-schizophrenics prescribed in the USA in 2008 (Gallini et al., 2013). QTP undergoes extensive metabolization in the human body and is excreted predominantly as metabolites (92% in urine and feces) (Sheehan et al., 2010). The mean concentration of nor-quetiapine (NQTP), one of the active metabolites of QTP, was 3.4 times higher than the concentration of QTP in influents. This is thefirst study to report the concentrations of NQTP in wastewater.

(4)

LZP, APZ, and DZP were found in all inﬂuent samples (df: 100%) at mean concentrations of 18.2, 6.06, and 3.58 ng/L, respectively; the average consumption of these three drugs was 2420, 2650, and 7060 kg,

respectively, in the USA in 2010–2012 (INCB, 2013). Benzodiazepines are metabolized extensively in the human body and form pharmacologically inactive glucuronides, which are excreted through urine (Calisto

Table 1

Concentrations of select psychoactive pharmaceuticals and their metabolites in wastewater (ng/L) and sludge (ng/g dry wt) from two centralized wastewater treatment plants in Albany area, New York, USA, in July 2013.

Analytes WWTPA WWTPB Literature

Inﬂuent Primary efﬂuent

Efﬂuent *Sludge (n = 7)

Inﬂuent Primary efﬂuent

Efﬂuent *Sludge (n = 4)

Inﬂuent Efﬂuent Sludge

Mean/df (%) Range

Antischizophrenics

APPZ 5.58/57 3.21/100 10.3/86 115/100 5.43/71 1.53/41 1.69/57 16.8/100 ND^o ND^o 6.68/94^r

ND-22.4 1.27–5.70 ND-20.9 21.0–267 ND-14.1 ND-5.28 ND-5.93 13.0–21.1 bLOQ-23.9^r

DAPPZ ND ND ND 1.49/100 ND ND ND 2.93/100 – – 1.87/75^r

0.59–2.40 0.65–6.26 bLOQ-3.81^r

QTP 24.4/100 17.5/100 0.98/57 17.8/100 15.5/100 12.5/100 4.60/41 21.1/100 4–6^o ND^o 5.41/94^r

11.0–43.6 6.94–40.0 ND-3.43 7.41–25.3 8.29–20.9 0.65–18.7 ND-29.3 19.4–22.1 bLOQ-17.3^r

NQTP 71.0/100 69.4/100 74.3/100 133/100 66.1/100 60.3/86 82.3/100 196/100 – – 11.9/75^r

39.5–159 50.9–94.8 25.0–131 69.3–260 34.0–131 ND-102 42.9–144 99.1–267 bLOQ-31.6^r

Sedatives–hypnotics–anxiolytics

LZP 20.3/43 16.2/100 64.2/100 0.26/14 16.2/100 27.1/86 78.4/100 ND 74^e 140/100^f 11.6^r

9.14–33.0 4.86–28.7 37.3–85.5 ND-1.80 6.11–34.3 ND-45.9 42.8–114 51–82^e 30–160^f,j

APZ 6.24/100 4.85/86 6.20/100 0.61/71 5.89/100 4.53/100 4.59/100 0.28/50 ND^f,o ND^f,o 10.6/38^r

3.09–12.6 ND-8.84 5.01–8.21 ND-1.08 3.47–8.77 3.01–6.37 3.10–6.20 ND-0.62 bLOQ-14.1^r

AHA 21.4/100 5.56/43 9.2/100 0.37/14 17.4/100 12.4/71 12.7/86 1.43/50 – – –

5.86–28.5 ND-21.3 7.25–12.4 ND-2.06 5.93–26.0 ND-28.2 ND-46.6 ND-4.39

DZP 3.38/100 1.62/57 1.73/71 ND 3.79/100 3.01/86 2.58/100 0.48/50 17^e 1.4/100^m 3.3^r

2.29–4.57 bLOQ-4.70 bLOQ-3.45 2.29–9.78 bLOQ-8.21 1.28–4.00 ND-1.43 13–19^e ND-6.2^g,m 2.03–23^l,r

OxZP 8.43/100 6.76/100 9.87/100 0.86/43 6.52/100 7.28/100 7.72/100 1.60/50 480/100^h 320/100^h 4.62–13.1^r 2.30–16.9 3.56–10.1 7.09–14.4 ND-1.91 4.06–9.30 2.77–10.4 4.38–10.8 ND-3.81 up to 860^h 18.9–630^g,h,j

NDZP 5.30/100 4.29/100 4.53/100 0.96–57 4.04/100 3.82/100 3.69/100 1.08/50 – – 3.82^r

1.68–9.73 2.56–7.23 3.05–6.56 ND-2.29 2.63–5.25 2.37–6.12 3.29–4.08 ND-2.71 1.27–8.71^r

CBZ 145/100 301/86 310/100 83.1/100 241/100 238/100 268/100 118/100 757/100ⁱ 713/100ⁱ 23.3/100^r

61–347 ND-564 150–731 28.6–189 109–588 112–501 91.3–631 57.3–191 6–1032^b,d,e,i 6–961^b,d,i 17–245^i,r Antidepressants

VLF 415/100 471/100 480/100 129/100 336/100 359/100 339/100 84.2/100 1343/100ⁱ 1087/100ⁱ 8.94/94^r

169–609 309–702 389–553 76.5–162 194–407 245–451 209–431 66.6–106 40–1769^f,i,k 60–2190^f,i,k,n 0.97–499^i,r

BPP 110/100 75.0/100 67.4/100 23.7/100 147/100 73.8/100 34.1/100 12.5/100 191^k 104–500^k,n –

17.1–231 12.8–182 18.2–264 8.23–46.2 25.2–378 39.3–108 7.31–89.2 7.07–19.9

STL 80.8/100 58.5/100 62.8/100 1490/100 43.1/100 47/100 24.5/100 862/100 20/100ⁱ 12/100ⁱ 56.7/100^r

46.7–114 33.4–73.0 27.9–88.3 976–1993 31.6–52.5 23.5–70.4 15.7–49.8 788–961 7.6–34^i,k 5.7–70^i,k,n 11.3–2117^c,i,l,r

NSTL 71.1/100 56.8/100 54.4/86 688/100 65.3/100 33.3/100 16.4/41 394/100 20/100ⁱ 15/100ⁱ 117/100^r

23.7–137 19.7–119 ND-78.6 269–1192 11.0–183 11.7–50.6 ND-83.6 175–716 15–30î,k 12–50î,n 24.3–279î,r

CTP 133/100 221/100 280/100 283/100 59.4/100 127/100 150/100 170/100 236/100ⁱ 173ⁱ 26.8/94^r

77.7–170 180–271 205–414 170–429 35.1–146 47.6–211 104–215 131–230 144–326î,k 86–500î,k,n 10.7–1381î,r

DCTP 55.4/71 68.0/100 118/100 222/100 12.8/29 35.6/71 79.3/86 130/100 133^k 111^k 41.5^r

ND-126 32.5–135 35.9–310 36.8–434 ND-55.3 ND-105 bLOQ-183 33.2–223

Antihypertensions

ANL 1220/100 1040/100 594/100 23.0/100 606/100 568/100 426/100 27.6/100 7801^b 2772^b ND^c

377–2000 537–1390 382–754 9.33–40.8 377–904 363–722 299–852 17.7–46.4 916–11239^b,e 118–5910^b,j

PPN 24.5/100 46.5/100 82.2/100 83.3/100 24.1/100 64.5/100 74.2/100 49.7/100 183^e 93/100^a 53.7/100^r

3.04–53.8 23.2–82.9 29.8–225 35.0–137 2.99–97.5 12.1–125 28.2–153 13.9–115 72–309^e 16–284^a,j 22.7–849^c,l,r

DTZ 105/100 NA 194/100 61.9/100 168/100 NA NA 48.5/100 6–19^d 6–13^d 2.59/44^r

15.3–215 179–218 38.2–134 75.0–354 35.9–68.7 bLOQ-3.91^r

DAD 483/100 361/100 294/100 179/100 473/100 295/100 327/100 84.9/100 – – 27.5/100^r

125–1090 188–613 163–368 23.2–503 109–1840 118–618 138–1000 69.7–104 1.70–159^r

VPM 18.5/100 21.1/100 49.2/100 218/100 7.30/100 23.5/100 38.5/100 170/100 3100^h 510^h 3.66/94^r

5.34–42.8 6.76–33.5 28.8–63.2 112–380 1.79–17.4 8.82–42.9 25.3–58.6 145–219 bLOQ-551^c,l,r

NVP 14.8/100 19.3/100 20.3/100 385/100 8.88/100 15.2/100 8.29/100 175/100 – – 7.43/94^r

4.22–33.5 9.36–42.5 2.63–39.9 246–509 1.11–24.1 2.53–28.1 1.87–18.3 92.4–249 bLOQ-458^c,r

Antiplatelet

CPG 35.5/100 16.3/100 21.8/100 32.4/100 31.4/100 28.4/100 23.7/100 29.0/100 124^e – 8.51/81^r

7.39–92.8 14.0–23.0 13.9–30.1 19.1–66.4 22.9–38.3 14.7–40.7 18.2–39.9 19.0–34.8 106–133^e bLOQ-20.8^r

CPGC 160/100 129/100 194/100 3.24/100 124/100 166/100 116/100 2.26/100 – – 3.74/100^r

43.5–278 75.0–193 166–249 1.59–5.19 67.8–166 64.8–168 49.5–170 1.33–3.25 1.19–12.6^r

Antihistamine

DPH 462/100 609/100 194/86 444/100 227/100 287/100 85.7/100 293/100 160–600^p 586^q 87.2/100^r

286–615 421–764 bLOQ-704 351–621 105–390 14.4–575 5.63–426 225–339 87.2–5957^l,r

DPMA 3.7/86 3.65/29 4.5/29 ND 3.79/29 11.4/57 4.14/14 ND – – –

bLOQ-5.52 ND-9.55 bLOQ-12.9 ND-9.45 ND-32.7 ND-16.5

(5)

and Esteves, 2009). The glucuronide conjugates are transformed to the corresponding parent compounds byβ-glucuronidase enzyme produced by fecal bacteria, e.g.,Escherichia coli. OxZP and NDZP were found in all influent samples at mean concentrations of 7.47 and 4.67 ng/L, respectively. The measured concentrations of OxZP and NDZP in wastewater represent both the parent molecule and the metabolite of benzodiazepines (Hass et al., 2012).α-hydroxy alprazolam (AHA), a metabolite of APZ, was found for thefirst time in wastewater influents at mean concentrations of 19.4 ng/L (df: 100%). CBZ is one of the co-prescribed pharmaceuticals, along with antischizophrenics and anxiolytics (Kim et al., 2007), and is found at the highest concentration (194 ng/L; df: 100%) in wastewater among several schizophrenics and sedative–hypnotics–anxiolytics analyzed in this study (Table 1).

VLF, a selective serotonin and norepinephrine re-uptake inhibitor (SSNRI), was found at the highest mean concentration (376 ng/L) in all influents, followed by BPP (a dopamine and norepinephrine inhibitor) and selective serotonin re-uptake inhibitors (SSRI: CTP and STL) (Table 1).Metcalfe et al. (2010)reported the occurrence of VLF at the highest concentrations among all antidepressants analyzed (N500 ng/L) in wastewater effluents from two WWTPs employing conventional activated sludge and tertiary treatment followed by UV- disinfection in Canada. The parent compounds and the metabolites of antidepressants are excreted predominantly as conjugates. The mean concentration of norsertraline (NSTL), a pharmacologically active metabolite of STL, was similar to that of STL in influents. However, the mean concentration of CTP was ~ 3 times higher than its metabolite, N-desmethyl citalopram (DCTP: 34.1 ng/L; df: 50%) in influents.

ANL, an antihypertensive, was found at the highest concentration (913 ng/L, df: 100%) among all pharmaceuticals analyzed in inﬂuents;

this concentration was 2 and 9 times lower than those reported in Spain (Dolar et al., 2012) and Korea (Behera et al., 2011), respectively (Table 1). However, Dolar et al. studied the removal of pharmaceuticals from municipal wastewater through a membrane bioreactor coupled to reverse osmosis in Spain (Dolar et al., 2012) whereas Behera et al. mon- itored pharmaceuticals in Korean WWTPs employing conventional treatment process including primary treatment and activated sludge process (Behera et al., 2011). The mean concentrations of antiplatelet (CPG) and antihistamine (DPH) analyzed in inﬂuents were 3.8 times lower than those reported in Spain (Dolar et al., 2012) but similar to those reported in an earlier study in the USA (Table 1) (Du et al., 2014). Clopidogrel carboxylic acid (CPGC, a metabolite of CPG) was not reported in wastewater before; however, in this study, this metabolite was found at concentrations 4.2 times higher than those of CPG

in influents (df: 100%). Similarly, 2-(diphenylmethoxy) acetic acid (DPMA: metabolite of DPH) was found for thefirst time in influents at 3.76 ng/L (df: 57%) in our study.

3.2. Psychoactive pharmaceuticals in sludge

APPZ and DAPPZ were consistently detected in all sludge samples (Table 1). APPZ is excreted from human bodies mainly through feces, as a metabolite (37%) as well as an unchanged parent molecule (~18%) (Sheehan et al., 2010). It can be presumed that pharmaceuticals found in feces often partition to solid particles, such as SPM, rather than to the aqueous phase (Metcalfe et al., 2010). Similarly, QTP is excreted predominantly as a metabolite (72.5%) in urine and (19.5%) through feces (Sheehan et al., 2010). NQTP was detected in all efﬂuent and sludge samples at concentrations of 28 and 8.2 times higher, respectively, than the concentrations of QTP. In dewatered sludge, the mean concentrations of ANL, DTZ, BPP, CBZ, and NDZP were 3.3 times lower than those found in activated sludge (Table S5).

The sorption coefﬁcients (Kd) of psychoactive pharmaceuticals were determined based on the assumption that the analytes are in equilib- rium between the aqueous phase and the particulate phase as reported byRadjenovic et al. (2009). The concentrations measured in inﬂuent (ng/L) and SPM (ng/kg dw) were applied in the calculation of Kd (L/kg) (Table S5). The highest Kd values were found for VPM (12000 L/kg) and its metabolite norverapamil, NVP (14300 L/kg) and some antidepressants (STL: 3460 L/kg and NSTL: 5680 L/kg). The Kd

values of APPZ and QTP were 1930 and 430 L/kg, respectively. The reported Kdvalues for STL, NSTL, and VLF, based on the measured concentrations in sludge and effluent (Lajeunesse et al., 2012) were similar to those found in our study. The sorption coefficients of PPN and CBZ were ~ 3.5 times higher than those reported in a study from Spain (Radjenovic et al., 2009). Removal by sorption is considered negligible for substances having log Kdvalues below two and considered significant for substances having log Kdvalues above four (Deegan et al., 2011). The sorption of antischizophrenics (APPZ and QTP), antidepressants (STL and NSTL), antihypertensive metabolite (NVPM), and antihistamine (DPH) was significant (KdN4) (Table S5); however, the sorption of LZP, ANL, and CPGC (log Kd= 1.21–1.73) was low.

3.3. Sorption of psychoactive drugs on SPM

Based on the measured concentrations of psychoactive pharmaceuticals in SPM and the aqueous phase (Eq.(1)), the fraction of drugs found Notes to Table 1:

Italics refer to metabolites. df: detection frequency (%), *sludge corresponds to the combined sludge produced after primary and secondary treatments; ND: non-detect; LOQ: limit of quantitation; aripiprazole (APPZ); dehydro-aripiprazole (DAPPZ); quetiapine (QTP); norquetiapine (NQTP); lorazepam (LZP); alprazolam (APZ);α-hydroxy alprazolam (AHA); diazepam (DZP); oxazepam (OxZP); nordiazepam (NDZP); carbamazepine (CBZ); venlafaxine (VLF); bupropion (BPP); sertraline (STL); norsertraline (NSTL); citalopram (CTP); N-desmethyl citalopram (DCTP); atenolol (ANL); propranolol (PPN); diltiazem (DTZ); desacetyl diltiazem (DAD); verapamil (VPM); norverapamil (NVP); clopidogrel (CPG); clopidogrel carboxylic acid (CPGC); diphenhydramine (DPH); 2-(diphenylmethoxy) acetic acid (DPMA).

aIn efﬂuent fromﬁve WWTPs in the UK (Ashton et al., 2004).

b In efﬂuent fromﬁve WWTPs in South Korea (Behera et al., 2011).

c In composite biosolid samples from 96 WWTPs in the USA (Chari and Halden, 2012).

d In wastewater from 4 WWTPs in Korea (Choi et al., 2008).

e In a pilot scale WWTP in Spain (Dolar et al., 2012).

f In 3 WWTPs in Spain (Gracia-Lor et al., 2012).

g In 6 WWTPs in Berlin, Germany (Hass et al., 2012).

h In 11 WWTPs in Germany (Hummel et al., 2006).

i In 5 WWTPs in Canada (Lajeunesse et al., 2012).

j In a WWTP in Spain (Lopez-Serna et al., 2010).

k In a WWTP in Canada (Metcalfe et al., 2010).

l In seven sewage sludge samples in France (Peysson and Vulliet, 2013).

mIn eleven WWTPs in Korea (Ryu et al., 2011).

n In a WWTP in the USA (Schultz et al., 2010).

o In WWTPs in China (Yuan et al., 2013).

p In municipal wastewater in the USA (Du et al., 2014).

q In a WWTP in the USA (Bartelt-Hunt et al., 2009).

r In 16 WWTPs in Korea (Subedi et al., 2013).

(6)

in SPM ranged from 0.68% (ANL) to 80.2% (VPM) (Table 2). Many studies that examined the fate of psychoactive pharmaceuticals in WWTPs based their measurements onﬁltered wastewater (Lajeunesse et al., 2012;

Metcalfe et al., 2010; Radjenovic et al., 2009). We found, however, that the fraction of APPZ, NQTP, STL, NSTL, CTP, DCTP, PPN, VPM, and NVP sorbed to SPM wasN50% of the total mass. This suggests that studies that determine mass loadings of psychoactive pharmaceuticals in WWTPs based only on aqueous phase concentrations can underesti- mate the total loadings by up to 80%. Nevertheless, the fraction of select sedative–hypnotics–anxiolytics (APZ, DZP, NDZP, and CBZ), antidepressants (VLF and BPP), and antihypertensives (ANL and DTZ) sorbed to SPM wasb11%, which suggests that these pharmaceuticals are present predominantly in the aqueous fractions.

3.4. Removal of pharmaceuticals from WWTPs

The removal efficiencies of psychoactive pharmaceuticals through wastewater treatment were calculated based on PSPMcorrected mean concentrations in influent and effluent (Eq.(2)) (Fig. 1). The average removal efficiencies of psychoactive pharmaceuticals and their metabolites in two WWTPs ranged from 0% (VLF) to 71% (APPZ) through the primary treatment and from 0.3% (DCTP) to 87% (QTP) through the final (primary and secondary) treatment. Select pharmaceuticals and their metabolites, including LZP, CBZ, NSTL, PPN, CPGC, and DPMA,

showed a negative removal efﬁciency in WWTPs. Microbial transformation of conjugated forms of drugs during the wastewater treatment processes can increase the residue levels of parent drugs in waste streams (Calisto and Esteves, 2009).Gracia-Lor et al. (2012)reported a negative removal for LZP in WWTPs. Negative removal efﬁciency for CBZ was at- tributed to cleavage of hydroxylated carbamazepine metabolite to CBZ by microbial activity in WWTPs (Miao et al., 2005). APPZ, QTP, and NQTP were removed in WWTPs at 68%, 87%, and 31%, respectively.

The average removal efﬁciencies of antidepressants and their metabolites in WWTPs (27%) were similar to those reported in two Canadian WWTPs (Metcalfe et al., 2010). The secondary biological treatment using activated sludge enhanced the removal of QTP (49%), STL (16%), NVP (26%), and DPH (50%) (Fig. 1).

3.5. Mass loading and emission of psychoactive drugs through WWTPs

Mass loadings (mg/d/1000 inhabitants) of psychoactive pharmaceuticals were calculated using the concentration of psychoactive pharmaceuticals determined in wastewater inﬂuent, including the fraction sorbed to SPM (Eq.(3)) (Table 2). The mass loadings of psychoactive pharmaceuticals ranged from 0.91 (DZP) to 347 mg/d/1000 inhabitants (ANL). The mass load of psychoactive pharmaceuticals in the WWTPA

was 3.5 (PPN) to 16 (STL) times higher than in the WWTP_B. The average mass loadings of antidepressants and antihypertensives were similar;

however, the average mass loading of sedative–hypnotic–anxiolytic Table 2

The fraction of pharmaceutical sorbed, average mass load, and emission of pharmaceuticals and their metabolites in two centralized wastewater treatment plants in Albany area, New York, USA.

Analytes PSPM

(%)^a

Mass load (mg/d/1000 inhabitants)

Environmental emission (mg/d/1000 inhabitants) Antischizophrenics

Aripiprazole (APPZ) 68.3 6.46 3.82

Dehydro-aripiprazole(DAPPZ) NA 0.01

Quetiapine (QTP) 22.9 7.41 2.27

Norquetiapine(NQTP) 50.4 36.4 59.5

Sedatives–hypnotics–anxiolytics

Lorazepam (LZP) 34.7 7.74 47.7

Alprazolam (APZ) NA 1.66 3.36

α-Hydroxy alprazolam (AHA) NA 5.60 7.44

Diazepam (DZP) NA 0.92 1.48

Oxazepam(OxZP) 48.2 4.84 6.56

Nordazepam(NDZP) NA 1.38 2.60

Carbamazepine (CBZ) 4.13 43.8 186

Antidepressants

Venlafaxine (VLF) 10.8 122 255

Bupropion (BPP) 1.03 31.0 30.8

Sertraline (STL) 71.5 78.9 38.0

Norsertraline(NSTL) 68.5 76.6 71.7

Citalopram (CTP) 60.9 72.2 131

Desmethyl citalopram (DCTP) 69.0 30.7 61.6

Antihypertensions

Atenolol (ANL) 0.68 347 316

Propranolol (PPN) 56.1 12.6 50.7

Diltiazem (DTZ) 8.93 34.5 45.8

Desacetyl diltiazem(DAD) 13.5 143 217

Verapamil (VPM) 80.2 18.8 30.0

Norverapamil(NVP) 71.5 10.6 11.2

Antiplatelet

Clopidogrel (CPG) 19.5 12.3 15.1

Clopidogrel carboxylic acid (CPGC)

0.72 41.8 94.3

Antihistamine

Diphenhydramine (DPH) 42.4 184 116

2(Diphenylmethoxy)acetic acid (DPMA)

NA 1.00 2.77

aAverage PSPMin inﬂuents of two WWTPs; metabolites are italicized.

Removal Efficiency (%)

-100 -75 -50 -25 0 25 50 75 100

Analytes

DPMA DPH CPGC CPG NVP VPM DAD DTZ PPN ANL DCTP CLP NSTL STL BPP VLF CBZ NDZP OxZP DZP AHA APZ LZP NQTP QTP AAPZ Primary Treatment

Total Treatments Antischizophrenic

Antisedative-hypnotic-anxiolytic

Antidepressant

Antihypertensive

Antiplatalet Antihistamine

Fig. 1.The average removal efﬁciencies (%) of psychoactive pharmaceuticals from two WWTPs: aripiprazole (APPZ); dehydro-aripiprazole (DAPPZ), quetiapine (QTP);

norquetiapine (NQTP); lorazepam (LZP); alprazolam (APZ);α-hydroxy alprazolam (AHA); diazepam (DZP); oxazepam (OxZP); nordiazepam (NDZP); carbamazepine (CBZ); venlafaxine (VLF); bupropion (BPP); sertraline (STL); norsertraline (NSTL);

citalopram (CTP); N-desmethyl citalopram (DCTP); atenolol (ANL); propranolol (PPN);

diltiazem (DTZ); desacetyl diltiazem (DAD); verapamil (VPM); norverapamil (NVP);

clopidogrel (CPG); clopidogrel carboxylic acid (CPGC); diphenhydramine (DPH); 2- (diphenylmethoxy) acetic acid (DPMA). Total treatments refer to the primary treatment as well as secondary treatment.

(7)

and antischizophrenic was approximately 7.3 and 4.1 times lower than that found for antidepressants, respectively.

The emission of psychoactive pharmaceuticals from WWTPs to the environment was calculated based on the concentration in effluents, fraction sorbed to SPM in effluent, and volume of activated sludge produced (Eq.(4)). The environmental discharge of psychoactive pharmaceuticals from WWTPs, on average, ranged from 0.01 (dehydro- aripiprazole) to 316 (ANL) mg/d/1000 inhabitants (Table 2). The average emission rates of antidepressants and antihypertensives analyzed in this study were 2.6–6.8 times higher than those for antischizophrenics and sedative–hypnotic–anxiolytics. The calculated per-capita emission of CBZ, ANL, and PPN in this study were 2.1–5.1 times higher than that reported for a WWTP from Spain that serves a population of 277,000 (Radjenovic et al., 2009); however, the fraction of pharmaceuticals sorbed to SPM was not calculated in the latter study. The environmental emission of psychoactive pharmaceuticals we determined here can be a lower bound estimate because the peaks of prescription of psychoactive pharmaceuticals are reported in May to early June (Skegg et al., 1986; Tansella and Micciolo, 1992). Moreover, seasonal enhance- ment of removal of pharmaceuticals in WWTPs was reported in sum- mer (Lajeunesse et al., 2012; Sui et al., 2011), that can result in decreased concentration of pharmaceuticals in effluent.

QTP, STL, VPM, and their metabolites were found emitted at≤17.3%

of their mass load from WWTPB(Fig. 2). However, APZ and its metabolite AHA were discharged at 77.8% and 73.1%, respectively, of the mass load from WWTPB. The removal of some psychoactive pharmaceuticals through sorption to sludge was found minimal (b1% of initial mass) (Fig. S2), which suggests that biodegradation and/or chemical transformation can be the dominant mechanism of removal for these drugs (Fig. 2) (Meakins et al., 1994; Radjenovic et al., 2009). Further studies are required to monitor psychoactive pharmaceuticals, pharmacoactive metabolites, and potential transformed products in receiving waters.

Acknowledgments

Authors would like to thank the WWTP facilities, Mr. Anthony DeJulio, and Mr. Jingchuan Xue for assistance with the sample collection. This study was funded by a grant (1U38EH000464-01) from the Centers for Disease Control and Prevention (CDC, Atlanta, GA) to Wadsworth Center, New York State Department of Health. Its contents

are solely the responsibility of the authors and do not necessarily represent the ofﬁcial views of the CDC.

Appendix A. Supplementary data

Supplementary data to this article can be found online athttp://dx.

doi.org/10.1016/j.scitotenv.2015.01.098.

References

Alonso, S.G., Catalá, M., Maroto, R.R., Gil, J.L.R.A., Miguel, Á.G.d., Valcárcel, Y., 2010.Pollu- tion by psychoactive pharmaceuticals in the Rivers of Madrid metropolitan area (Spain). Environ. Int. 36, 195–201.

Ashton, D., Hilton, M., Thomas, K.V., 2004.Investigating the environmental transport of human pharmaceuticals to streams in the United Kingdom. Sci. Total Environ. 333 (1–3), 167–184.

Bartelt-Hunt, S.L., Snow, D.D., Damon, T., Shockley, J., Hoagland, K., 2009.The occurrence of illicit and therapeutic pharmaceuticals in wastewater efﬂuent and surface waters in Nebraska. Environ. Pollut. 157 (3), 786–791.

Behera, S.K., Kim, H.W., Oh, J.E., Park, H.S., 2011.Occurrence and removal of antibiotics, hormones and several other pharmaceuticals in wastewater treatment plants of the largest industrial city of Korea. Sci. Total Environ. 409 (20), 4351–4360.

Benotti, M.J., Trenholm, R.A., Vanderford, B.J., Holady, J.C., Stanford, B.D., Snyder, S.A., 2009.Pharmaceuticals and endocrine disrupting compounds in U.S. drinking water.

Environ. Sci. Technol. 43 (3), 597–603.

Brodin, T., Piovano, S., Fick, J., Klaminder, J., Heynen, M., Jonsson, M., 2014.Ecological effects of pharmaceuticals in aquatic systems—impacts through behavioural alter- ations. Philos. Trans. R. Soc. Lond. Ser. B Biol. Sci. 369 (1656).

Brooks, B.W., Turner, P.K., Stanley, J.K., Weston, J.J., Glidewell, E.A., Foran, C.M., Slattery, M., La Point, T.W., Huggett, D.B., 2003.Waterborne and sediment toxicity ofﬂuoxetine to select organisms. Chemosphere 52 (1), 135–142.

Calisto, V., Esteves, V.I., 2009.Psychiatric pharmaceuticals in the environment.

Chemosphere 77 (10), 1257–1274.

Chari, B.P., Halden, R.U., 2012.Validation of mega composite sampling and nationwide mass inventories for 26 previously unmonitored contaminants in archived biosolids from the U.S National Biosolids Repository. Water Res. 46 (15), 4814–4824.

Choi, K., Kim, Y., Park, J., Park, C.K., Kim, M.Y., Kim, H.S., Kim, P., 2008.Seasonal variations of several pharmaceutical residues in surface water and sewage treatment plants of Han River, Korea. Sci. Total Environ. 402, 120–128.

Deegan, A.M., Shaik, B., Nolan, K., Urell, K., Oelgemoller, M., Tobin, J.M., rrisey, A., 2011.

Treatment options for wastewater efﬂuents from pharmaceutical companies. Int.

J. Environ. Sci. Technol. 8, 649–666.

Dolar, D., Gros, M., Rodriguez-Mozaz, S., Moreno, J., Comas, J., Rodriguez-Roda, I., Barcelo, D., 2012.Removal of emerging contaminants from municipal wastewater with an integrated membrane system, MBR-RO. J. Hazard. Mater. 239–240, 64–69.

Du, B., Price, A.E., Scott, W.C., Kristofco, L.A., Ramirez, A.J., Chambliss, C.K., Yelderman, J.C., Brooks, B.W., 2014.Comparison of contaminants of emerging concern removal, discharge, and water quality hazards among centralized and on-site wastewater treatment system efﬂuents receiving common wastewater inﬂuent. Sci. Total Environ.

466–467, 976–984.

Influent

Primary Effluent

Combined Sludge Primary Clarifier

Effluent

Primary Sludge

Secondary Treatment Secondary Clarifier

Secondary Sludge

River

?

7850 (74,100) 491 (1450) 113,000 (109,000)

52,900 (65,800)

12.4 (115) 0.17 (0.84) 508 (232) 100 (103)

Degradation Transformation

?

Ash

?

Incineration

Discharge 1800 (34,200)

377 (1030) 34,900 (29,000) 14,800 (49,200)

383 (10,700) 382 (1060) 4200 (18,900)

4130 (1020)

Fig. 2.A schematic showing the fate of select antipsychotic pharmaceuticals through a WWTP serving a population of approximately 100,000 in Albany area, New York, USA. The total amount (mg) of select antipsychotic pharmaceuticals per day at different stages of wastewater treatments is shown in the order of quetiapine (QTP, an anti-schizophrenic), alprazolam (APZ, an anti-anxiolytic), sertraline (STL, an antidepressant), and verapamil (VPM, an antihypertensive). The values in parenthesis represent the respective metabolites: norquetiapine (NQTP, a metabolite of QTP),α-hydroxy alprazolam (AHA, a metabolite of APZ), norsertraline (NSTL, a metabolite of STL), and norverapamil (NVP, a metabolite of VPM).

(8)

Escher, B.I., Bramaz, N., Richter, M., Lienert, J., 2006.Comparative ecotoxicological hazard assessment of beta-blockers and their human metabolites using a mode-of-action- based test battery and a QSAR approach. Environ. Sci. Technol. 40 (23), 7402–7408.

Gallini, A., Donohue, J.M., Huskamp, H.A., 2013.Diffusion of antipsychotics in the US And French markets, 1998–2008. Psychiatr. Serv. 64 (7), 680–687.

Gracia-Lor, E., Sancho, J.V., Serrano, R., Hernandez, F., 2012.Occurrence and removal of pharmaceuticals in wastewater treatment plants at the Spanish Mediterranean area of Valencia. Chemosphere 87 (5), 453–462.

Hass, U., Duennbier, U., Massmann, G., 2012.Occurrence and distribution of psychoactive compounds and their metabolites in the urban water cycle of Berlin (Germany).

Water Res. 46 (18), 6013–6022.

Hummel, D., Lofﬂer, D., Fink, G., Ternes, T.A., 2006.Simultaneous determination of psychoactive drugs and their metabolites in aqueous matrices by liquid chromatography mass Spectrometry. Environ. Sci. Technol. 40 (23), 7321–7328.

IMS, 2009. Press Release, IMS Health reports U.S. prescription sales grew 5.1 percent in 2009 to $300.3 billion. Available online:.www.imshealth.com.

INCB, 2010. International Narcotics Control Board. Psychotic Substances 2009. Available online:http://www.incb.org/documents/Psychotropics/technical-publications/2009/

Psychotropic_Report_2009.pdf. United Nations, New York.

INCB, 2013. International Narcotics Control Board. Psychotic Substances 2012. United Nations, New York (Available online:http://www.incb.org/documents/Psychotropics/

technical-publications/2012/en/Eng_2012_PUBlication.pdf).

Jelic, A., Fatone, F., Di Fabio, S., Petrovic, M., Cecchi, F., Barcelo, D., 2012.Tracing pharmaceuticals in a municipal plant for integrated wastewater and organic solid waste treatment. Sci. Total Environ. 433, 352–361.

Kim, Y., Choi, K., Jung, J., Park, S., Kim, P.G., Park, J., 2007.Aquatic toxicity of acetamino- phen, carbamazepine, cimetidine, diltiazem and six major sulfonamides, and their potential ecological risks in Korea. Environ. Int. 33 (3), 370–375.

Kinney, C.A., Furlong, E.T., Kolpin, D.W., Burkhardt, M.R., Zaugg, S.D., Werner, S.L., Bossio, J.P., Benotti, M.J., 2008.Bioaccumulation of pharmaceuticals and other anthropogenic waste indicators in earthworms from agricultural soil amended with biosolid or swine manure. Environ. Sci. Technol. 42 (6), 1863–1870.

Kolpin, D.W., Furlong, E.T., Meyer, M.T., Thurman, E.M., Zaugg, S.D., Barber, L.B., Buxton, H.T., 2002.Pharmaceuticals, hormones, and other organic wastewater contaminants in U.S. streams, 1999–2000: a national reconnaissance. Environ. Sci. Technol. 36 (6), 1202–1211.

Lajeunesse, A., Smyth, S.A., Barclay, K., Sauve, S., Gagnon, C., 2012.Distribution of antidepressant residues in wastewater and biosolids following different treatment processes by municipal wastewater treatment plants in Canada. Water Res. 46 (17), 5600–5612.

Lopez-Serna, R., Perez, S., Ginebreda, A., Petrovic, M., Barcelo, D., 2010.Fully automated determination of 74 pharmaceuticals in environmental and waste waters by online solid phase extraction–liquid chromatography-electrospray–tandem mass spectrometry. Talanta 83 (2), 410–424.

Meakins, N.C., Bubb, J.M., Lester, J.N., 1994.Fate and behavior of organic micropollutants during wastewater treatment process: a review. Int. J. Environ. Pollut. 4, 27–58.

Menken, M., Munsat, T.L., Toole, J.F., 2000.The global burden of disease study: implica- tions for neurology. Arch. Neurol. 57 (3), 418–420.

Metcalfe, C.D., Chu, S., Judt, C., Li, H., Oakes, K.D., Servos, M.R., Andrews, D.M., 2010.Anti- depressants and their metabolites in municipal wastewater, and downstream exposure in an urban watershed. Environ. Toxicol. Chem. 29 (1), 79–89.

Miao, X.S., Yang, J.J., Metcalfe, C.D., 2005.Carbamazepine and its metabolites in wastewater and in biosolids in a municipal wastewater treatment plant. Environ. Sci. Technol.

39 (19), 7469–7475.

Pedersen, J.A., Soliman, M., Suffet, I.H., 2005.Human pharmaceuticals, hormones, and personal care product ingredients in runoff from agriculturalﬁelds irrigated with treated wastewater. J. Agric. Food Chem. 53 (5), 1625–1632.

Petersen, K., Heiaas, H.H., Tollefsen, K.E., 2014.Combined effects of pharmaceuticals, personal care products, biocides and organic contaminants on the growth ofSkeletonema pseudocostatum. Aquat. Toxicol. 150, 45–54.

Petrovic, M., Barceló, D., 2007.Analysis, Fate and Removal of Pharmaceuticals in the Water Cycle. Wilson & Wilson's, Amsterdam, The Netherlands.

Peysson, W., Vulliet, E., 2013.Determination of 136 pharmaceuticals and hormones in sewage sludge using quick, easy, cheap, effective, rugged and safe extraction followed by analysis with liquid chromatography–time-of-ﬂight-mass spectrometry.

J. Chromatogr. A 1290, 46–61.

Radjenovic, J., Petrovic, M., Barcelo, D., 2009.Fate and distribution of pharmaceuticals in wastewater and sewage sludge of the conventional activated sludge (CAS) and advanced membrane bioreactor (MBR) treatment. Water Res. 43 (3), 831–841.

Rosi-Marshall, E.J., Snow, D., Bartelt-Hunt, S.L., Paspalof, A., Tank, J.L., 2015.A review of ecological effects and environmental fate of illicit drugs in aquatic ecosystems.

J. Hazard. Mater. 282, 18–25.

Ruhoy, I.S., Daughton, C.G., 2008.Beyond the medicine cabinet: an analysis of where and why medications accumulate. Environ. Int. 34 (8), 1157–1169.

Ryu, J., Yoon, Y., Oh, J., 2011.Occurrence of endocrine disrupting compounds and pharmaceuticals in 11 WWTPs in Seoul, Korea. KSCE J. Civ. Eng. 15 (1), 57–64.

Schultz, M.M., Furlong, E.T., Kolpin, D.W., Werner, S.L., Schoenfuss, H.L., Barber, L.B., Blazer, V.S., Norris, D.O., Vajda, A.M., 2010.Antidepressant pharmaceuticals in two U.S.

efﬂuent-impacted streams: occurrence and fate in water and sediment, and selective uptake inﬁsh neural tissue. Environ. Sci. Technol. 44 (6), 1918–1925.

Sheehan, J.J., Sliwa, J.K., Amatniek, J.C., Grinspan, A., Canuso, C.M., 2010.Atypical antipsychotic metabolism and excretion. Curr. Drug Metab. 11 (6), 516–525.

Sinclair, E., Kannan, K., 2006.Mass loading and fate of perﬂuoroalkyl surfactants in wastewater treatment plants. Environ. Sci. Technol. 40 (5), 1408–1414.

Skegg, K., Skegg, D.C., McDonald, B.W., 1986.Is there seasonal variation in the prescribing of antidepressants in the community? J. Epidemiol. Community Health 40 (4), 285–288.

Subedi, B., Kannan, K., 2014.Mass loading and removal of select illicit drugs in two wastewater treatment plants in New York State and estimation of illicit drug usage in com- munities through wastewater analysis. Environ. Sci. Technol. 48 (12), 6661–6670.

Subedi, B., Lee, S., Moon, H.B., Kannan, K., 2013.Psychoactive pharmaceuticals in sludge and their emission from wastewater treatment facilities in Korea. Environ. Sci.

Technol. 47 (23), 13321–13329.

Sui, Q., Huang, J., Deng, S., Chen, W., Yu, G., 2011.Seasonal variation in the occurrence and removal of pharmaceuticals and personal care products in different biological wastewater treatment processes. Environ. Sci. Technol. 45 (8), 3341–3348.

Tansella, M., Micciolo, R., 1992.Trends in the prescription of antidepressants in urban and rural general practices. J. Affect. Disord. 24 (2), 117–125.

Ternes, T.A., 1998.Occurrence of drugs in German sewage treatment plants and rivers.

Water Res. 32 (11), 3245–3260.

Thomas, M.A., Klaper, R.D., 2012.Psychoactive pharmaceuticals induceﬁsh gene expression proﬁles associated with human idiopathic autism. PLoS ONE 7 (6), e32917.

USEPA, 2006.Emerging Technologies for Biosolid Management. U.S. Environmental Pro- tection Agency, Washington, DC (EPA 832-R-06-005).

van der Ven, K., Keil, D., Moens, L.N., Hummelen, P.V., van Remortel, P., Maras, M., De Coen, W., 2006.Effects of the antidepressant mianserin in zebraﬁsh: molecular markers of endocrine disruption. Chemosphere 65 (10), 1836–1845.

Wick, A., Fink, G., Joss, A., Siegrist, H., Ternes, T.A., 2009.Fate of beta blockers and psychoactive drugs in conventional wastewater treatment. Water Res. 43 (4), 1060–1074.

Yuan, S., Jiang, X., Xia, X., Zhang, H., Zheng, S., 2013.Detection, occurrence and fate of 22 psychiatric pharmaceuticals in psychiatric hospital and municipal wastewater treatment plants in Beijing, China. Chemosphere 90, 2520–2525.

Zhang, Y., Geissen, S.U., Gal, C., 2008.Carbamazepine and diclofenac: removal in wastewater treatment plants and occurrence in water bodies. Chemosphere 73 (8), 1151–1161.