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Chemosphere 78 (2010) 10561062

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Chemosphere
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Concentrations and mass loadings of hormones, alkylphenols, and alkylphenol ethoxylates in healthcare facility wastewaters
P.M. Nagarnaik a, M.A. Mills b, B. Boulanger a,*
a b

Environmental & Water Resources Engineering Division, Zachry Department of Civil Engineering, Texas A&M University, College Station, TX 77843, United States National Risk Management Research Laboratory, Ofce of Research and Development, US Environmental Protection Agency, Cincinnati, OH 45268, United States

a r t i c l e

i n f o

a b s t r a c t
Healthcare facility wastewaters are an anticipated source of known endocrine disrupting chemicals to the environment. In this study, the composition and magnitude of eight steroid hormones, octylphenol (OP), nonylphenol (NP), 16 nonylphenol ethoxylates (NPEOs), and 10 octylphenol ethoxylates (OPEOs) in wastewater from a(n) hospital, nursing facility, assisted living facility, and independent living facility are presented. Steroid hormone concentrations were variable for each sampling location, ranging from a non-detectable concentration of 17b-ethynylestradiol in all samples to 127 ng L1 androstenedione in the hospitals wastewater composite. OP and NP were not detected in any sites samples. However, NPEOs were found at each sampling location with a maximum combined concentration of 260 lg L1 for NPEOs with a chain length between 3 and 18 units in the assisted living facility composite sample. OPEOs were only found in the hospital and nursing facilities samples with a maximum combined OPEO concentration of 13 lg L1 for OPEOs with a chain length between 2 and 12 units in hospital wastewater. The total mass loading of hormones to the municipal sewer system from each facility ranged from 2.5 mg d1 at the assisted living facility to 138 mg d1 at the hospital. The total mass loading of the alklyphenol ethoxylates (NPEO + OPEO) is considerably higher than the estimated hormone mass loadings, ranging from 1.8 g d1 at the independent living facility to 54 g d1 at the hospital facility. 2009 Elsevier Ltd. All rights reserved.

Article history: Received 20 August 2009 Accepted 16 November 2009 Available online 15 January 2010 Keywords: Healthcare Wastewater EDC Hormones Alkyphenol Mass loading

1. Introduction Recent literature has documented the presence of endocrine disrupting compounds entering the natural environment through engineered systems (Petrovic et al., 2002; Falconer, 2006; Falconer et al., 2006; Khanal et al., 2006). Steroid hormones and alkylphenols (APs) are well established EDCs that enter the environment through municipal and industrial wastewater. These compounds have been reported in wastewater inuents (Belgiorno et al., 2007; Loyo-Rosales et al., 2007a) and efuents (Khanal et al., 2006; Ying, 2006; Belgiorno et al., 2007; Gonzlez et al., 2007; Isidori et al., 2007; Loyo-Rosales et al., 2007a; van der Linden et al., 2008); surface water (Berryman et al., 2004; Khanal et al., 2006; Ying, 2006; Li et al., 2007; Schlsener and Bester, 2008; van der Linden et al., 2008); and groundwater (Ying et al., 2002a; Kolodziej et al., 2003; Swartz et al., 2006; Belgiorno et al., 2007). Their presence in treated efuents leaving wastewater treatment systems contribute to the adverse physiological effects noted in sh downstream from wastewater efuent discharges (Ying et al., 2002a,b; Ying, 2006; Zoller, 2006).

* Corresponding author. Tel.: +1 979 845 9782; fax: +1 979 862 1542. E-mail address: bboulanger@tamu.edu (B. Boulanger). 0045-6535/$ - see front matter 2009 Elsevier Ltd. All rights reserved. doi:10.1016/j.chemosphere.2009.11.019

Healthcare facilities are a potentially important source of hormones and APs to municipal sewer systems (Ying et al., 2002a,b; Ying, 2006; van der Linden et al., 2008). Hormones are released to healthcare facility wastewaters through patient excretions and disposal of pharmaceuticals. Alkylphenol ethoxylates (APEOs) are widely used as surfactants in domestic detergents (Koh et al., 2005) and are anticipated to be present at high amounts in healthcare facilities due to extensive laundering and cleaning practices, although existing data is not readily available. The primary aim of this study is to provide a preliminary assessment of the concentration range and magnitude of environmental loadings of natural and synthetic steroid hormones, APs, and APEOs from different types of healthcare facilities. The concentration and daily mass loading of three androgens (androstenedione, dihydrotestosterone, and testosterone); four estrogens (estriol, estrone, 17a-ethynylestradiol, and 17b-estradiol); progesterone; two alkylphenols (OP and NP); and 28 APEOs were analyzed and reported for four healthcare facilities. These four facilities include a hospital, a multi-care nursing facility, an independent living facility, and an assisted living facility located in a municipality in Central Texas. The reported data provide insight on the relative contributions and magnitudes of steroid hormones, APEOs and APs released to municipal wastewater systems from healthcare facilities.

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2. Experimental 2.1. Healthcare facility sampling sites Four different healthcare facilities in a single municipality in Texas were selected as sampling sites for this study. The sampling sites were selected based on the (1) importance and relevance to the study objective; (2) accessibility of the sampling locations onsite; (3) ability to collect wastewater samples representing only the facilitys contribution (no upstream sources, no combined storm water ows); (4) ability to collect the facilitys wastewater just above the point where the facility wastewater enters the municipal sewer; (5) historical consistency of wastewater ow at the site; and (6) range of healthcare services provided. Please note that all facility names and bed counts are withheld to protect the location identities. In addition, while presented ow information within the text is approximated, actual ow information was used to determine the estimated daily mass loadings presented in the paper. Pre-treatment at the facilities was limited to grease traps and metal recovery only at the hospital location. A description of each sampled facility including the number of beds, wastewater ow rates, and services provided is presented in Table 1. 2.2. Sampling protocols

The same pumping system was used to split the hospital samples, however, the three individual composites from the hospital sampling location were rst combined in a 35 L glass vessel prior to sub-sampling. Three 1 L sub-samples from each sites composite were collected in silanized 1 L amber glass bottles for hormone analysis. All hormone analysis samples were shipped overnight in ice packed coolers to the US Environmental Protection Agencys (EPA) National Risk Management Research Laboratorys (NRMRL) in Cincinnati, OH for analysis. Four 1 L sub-samples from each facilitys composite were also collected in non-silanized 1 L amber glass bottles for analysis of long-chain and short-chain APEOs, NP, and OP. Three of the 1 L samples were preserved with formaldehyde (1% vol:vol) for analysis of long-chain APEOs. The remaining 1 L samples were preserved with concentrated sulfuric acid to a pH of two (2) for analysis of short-chained APEOs, NP, and OP. All preserved AP and APEO samples were shipped overnight in ice packed coolers to the US EPA Region 5 Central Regional Laboratory (CRL) in Chicago, IL for analysis. Each shipped cooler sent to NRMRL or CRL contained a eld blank consisting of DI water in a 1 L appropriately treated amber glass bottles. Once received at the respective laboratories, samples were removed from the coolers and stored at 4 C until extraction. All samples were extracted within a week of receipt. 2.4. Analysis

All samples were collected from the four sampled facilities on a single day using ISCO automated composite samplers following the municipalitys normal wastewater sampling protocol. At each site samples were collected from sewer manholes located just up-line from where the facility wastewater enters the sewer. Individual autosamplers were deployed to collect the composite samples out of the multi-care nursing, assisted living, and independent living facilities, while three autosamplers were deployed to collect composite samples out of the hospital facility. Autosamplers were deployed to all sampling sites and sampling locations on April 14th, 2008. The autosamplers were programmed to collect 100 mL of sample every 15 min for 24 h into one 10 L polypropylene bottle that was chilled on ice during collection. The composite samplers were picked up 24 h after deployment and transported to the municipalitys wet laboratory. 2.3. Sample handling In the laboratory, composite samples collected from the multicare nursing, assisted living, and independent living facility were split while completely mixing into 1 L sub-samples using a peristaltic pump. The sub-samples were pumped into pre-labeled amber glass bottles. Silicone tubing was used in the pump head and teon-lined polyethylene tubing was used to convey the sub-samples. New tubing was used to split samples from each site.

Three different analytical techniques were used for the analysis of steroid hormones, long-chain APEOs, APs, and short-chain APEOs. All analyses were performed using laboratory standard operating procedures developed in accordance with EPAs quality assurance program. 2.4.1. Hormone sample extraction and analysis All hormone samples were extracted and analyzed according to NRMRLs standard operating procedure (SOP) for the analysis of steroid hormones in aqueous samples (Revision 4r). This SOP details the extraction and analysis of eight hormones: 17b-estradiol, estrone, estriol, 17a-ethynylestradiol, testosterone, androstenedione, progesterone, and dihydrotestosterone. The SOP used to analyze the hormones is similar to the procedure previously reported by Esperazna et al. ( 2004). The primary difference between the recently updated SOP and the Esperanza et al. method is that the new SOP incorporates deuterated compounds (d4-estrone, d3-testosterone, and d4-17a-ethynylestradiol) as method surrogates. The acceptable surrogate recovery was between 60% and 140%. 2.4.2. Long-chain APEO extraction and analysis CRLs SOP MS006 V1 Laboratory for analysis of nonylphenol polyethoxylates (NPnEO, 3 6 n 6 18) and octylphenol polyethoxylates (OPnEO, 2 6 n 6 12) in wastewater samples using selected

Table 1 Descriptions for each healthcare facility sampled in this study. Facility name Hospital NAICS name and code General medical and surgical hospital, 622110 Nursing care facility; 623110 Continuing care retirement communities. 623311 Homes for elderly (NAICS: 623312) Number of beds 375 Facility wastewater owrate (L d1) 500 000 Description of facility services In patient general hospital care, coronary care, surgical, and intensive care; outpatient cardiac, orthopedic, obstetrics and gynecology, pulmonary, surgical, gastrointestinal and respiratory care Nursing care, physical rehabilitation, Alzheimers care, and hospice Custodial non-medical care, as well as personalized medical care including medication management services, Alzheimers care, and short-term skilled nursing care Short-term custodial non-medical care services and limited healthcare services in the area of onsite therapy, wellness, and rehabilitation providers

Nursing care Assisted living Independent living

300 225

100 000 50 000

225

70 000

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ion recording liquid chromatography/mass spectrometry (LC/MS) was used to analyze all long-chain APEO samples. Because this method has not been previously published, a full description of the method is provided here. One liter sub-samples were received in the laboratory and immediately ltered using a syringe driven Millex HV PVDF lter unit with a 0.45 lm lter size. The ltered samples were then analyzed using a Waters 2659 liquid chromatograph coupled to a Waters single quadrapole ZQ mass spectrometer. Analytes were chromatographically separated using an Atlantis MS C18 150 mm 2.1 mm 3 lm column at a ow rate of 0.75 mL min1. The injection volume was 50 lL. A gradient elution was used with acetonitrile (A), water (B), and 100 lM ammonium acetate (C). C was held constant at 5% of the total ow throughout the gradient elution between A and B. The remaining linear gradient conditions were 95%B held from 0 2 min; 65% B at 5 min; 55% B at 10 min; 20% B at 15 min; 0% B at 35 min; back to 95% B at 40 min. Target analytes were quantitated in single ion recording (SIR) mode on the Waters ZQ mass spectrometer operated in electrospray positive ion mode under atmospheric pressure. The mass spectrometer instrument conditions included: 3.5 kV capillary voltage; 120 C source temperature; 300 C desolvation temperature; 500 L h1 desolvation gas ow; 50 L h1 cone gas ow; 0.1 s inter-scan delay; and a 0.1 s dwell time. The cone voltage varied for each analyte and ranged from 20 V for the shorter APEO chains (<4 units) to 50 V for the longer chain analytes. A total of 27 SIR experiments were carried out to determine individual concentrations of 16 NPEOs (NPnEO, 3 6 n 6 18) and 11 OPEOs (OPnEO, 2 6 n 6 12) for each run, where n indicates the chain length of the ethoxylate group. Individual APEOs quantication was based upon the response of the primary ion, m/z = [MNH3]+ (parent compound molecular weight + ammonium adduct) for each individual APEO compared to a ve point external calibration curve. The presence of a secondary ion was used for target compound verication. The secondary conrmation ion was 359.2 for OP6EO; 315.2 for OP5EO; 271.4 for NP4EO and OP4EO; 227.4 for NP3EO and OP3EO; 183.4 for OP2EO; and 133.3 for NP5EO through NP18EO and OP7EO through OP12EO. As an additional quality control measure, n-NP2EO was used as a method surrogate (primary ion 326.4; secondary ion 309.2). QuanLynx was used to process concentration data for each analyte. Matrix spike samples were prepared for each analytical batch and were spiked with NPnEO (3 > n > 18), OPnEO (2 > n > 12), and n-NP2EO to achieve a concentration of 100 parts-per-billion (ppb) of each analyte in the sub-sample. The acceptable range of matrix spike recovery was dened to be between 60% and 140%. DI water laboratory prepared blanks and eld blanks were also analyzed for each analytical batch. 2.4.3. Alkylphenol and short-chain alkylphenol ethoxylates extraction and analysis ASTM D7065-06 Standard test method for determination of nonylphenol, bisphenol A, p-tert-octylphenol, nonylphenol monoethoxylate and nonylphenol diethoxylate in environmental waters by gas chromatography mass spectrometry was used to analyze all AP and short-chain APEOs (ASTM Standard D7065-06, 2003). The method is described briey here. Approximately 300 mL of aqueous sample was combined with 300 mL of methylene chloride in a continuous liquidliquid extractor to extract the analytes from the aqueous sample. After 18 h the DCM phase was collected and dried using sodium sulfate. The extract was concentrated to a volume of 0.5 mL and analyzed by GC/MS operated in select ion monitoring mode. The four target compounds reported in this study were quantitated using the primary ion response and external calibration. Acenaphthene-d10 and phenanthrene-d10 were used as internal standards.

2.4.4. Potential estrogenicity calculation The potential estrogenicity of each facility composite wastewater sample was calculated using the following method. First, individual APEO and hormone concentrations in a given sample were converted from units of lg L1 to lM and ng L1 to lM, respectively. Second, because the highest potential estrogenicity of the samples will occur when the APEOs degrade to their AP parent, each APEO lM concentration was transformed into a potential alklyphenol concentration using a 1:1 M conversion. The resulting individual potential alkylphenol concentrations in a sample were summed to give a total potential alkylphenol value. Following this calculation, the values for total potential nonylphenol and total potential octylphenol were calculated. Finally, the total potential nonylphenol, total potential octylphenol, and individual estrogen concentrations for each composite sample were multiplied by the relative potency of each analyte in the yeast screen (YES) assay. Relative potencies used in the calculation were found in the literature (Routledge and Sumpter, 1996; Beck et al., 2006) 2.4.5. Mass loading The average daily and monthly ow rates from each facility were obtained from the municipality where the samples were acquired. The total mass loading from each facility was calculated by multiplying individual analyte concentrations in each facility composite by the average daily ow for the facility. The estimated mass loadings are reported as ng d1 for the hormone analytes and g d1 for the APEO analytes. 3. Results and discussion 3.1. Hormone concentrations in facility composite samples Seven out of the eight analyzed steroid hormones were detected in at least one of the facility composite samples. Table 2 reports the individual mean steroid hormone concentrations for each location. Androstenedione and progesterone were found in wastewater from all of the healthcare facilities with a concentration range from 9 ng L1 progesterone in the independent living facility composite to 127 ng L1 androstenedione in the hospitals composite sample. Overall, the hospital composite sample had the highest measured concentration of each analyte, except for 17b-estradiol. The maximum concentration 17b-estradiol was nominally higher in the nursing care facility. Perhaps surprisingly, the synthetic 17a-ethynylestradiol was not observed in any of the facility composites. Deuterated surrogate recoveries for d4-estrone, d3-testosterone, and d4-17a-ethynylestradiol were within 60140% for the hospital, nursing care, and independent living facility composite samples. The mean deuterated surrogate recoveries for the assisted living facility composite were lower than the acceptable range, with the minimum mean recovery obtained for d4-17a-ethynylestradiol (27% 1%), d4-estrone (30 9%), and d3-testosterone (58 7%). All of the concentrations reported in this study are corrected based on the surrogate recovery performance. Because the surrogate recoveries from the assisted living facility were lower than 60%, the steroid hormone concentrations from this facility are not reported in order to maintain the data quality even though a signal was present in the sample. 3.2. AP and APEO concentrations in facility composite samples The concentration of individual APs and APEOs measured in healthcare facility wastewater are presented in Table 3. NP and OP were not detected in the matrix spiked sample, indicating signicant matrix interference for these two compounds. For NP1EO

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Table 2 Individual mean (standard deviation) steroid hormone analyte concentrations measured in each facilitys 24 h composite wastewater sample. All concentrations are given as ng L1. All units (ng L1) Androgens Compound Androstenedione Dihydrotestosterone Testosterone Estriol Estrone 17a-Ethynylestradiol 17b-Estradiol Progesterone Hospital 127(11) 42(42) 19(21) 17(22) 42(19) nd() 4(14) 31(8) Nursing care 81(5) 32(5) 14(2) 4(23) 25(4) nd() 5(8) 10(3) Assisted livinga p np p np p np np p Independent living 60(26) 12(38) 14(13) nd() 11(14) nd() 3(17) 9(15)

Estrogens

Progestagens

p: Signal present in sample, not reported due to surragate recoveries <60%. np: Signal not present during analysis, not reported due to surrogate recoveries <60%. nd: Not detected, acceptable surrogate recovery. a Concentration data from the assisted living facility composite sample demonstrated recoveries less than 60% for each of the three method surrogates. Therefore, the data is not quantitatively reported.

and NP2EO, the monoethoyxlate and diethoxylate surrogate recoveries did not meet QA/QC limits, therefore, NP1EO and NP2EO are not reported although no presence of either compound was evident during analysis. A range of NPEO and OPEO analytes with longer chain lengths were detected. For discussion purposes, all NPEO and OPEO chain P P lengths are summed to give a (NPnEO, 3 < n < 18) and (OPnEO, 2 < n < 12) concentration value, respectively. The resulting P NPEOs concentration are an order of magnitude higher than P the OPEOs concentration for each facility composite. Overall, P the assisted living facility composite had the highest NPEO con1 centrations (258 lg L ), followed by the hospital composite (111 lg L1), the independent living facility composite (26 lg L1), and the nursing facility composite (19 lg L1). The hospital com-

P posite had the highest concentration of OPEOs (13 lg L1) folP OPEOs lowed by multi-care nursing care facility (2 lg L1). were not detected in the assisted living or independent living facility composites. The higher incidence and magnitude of NPEO occurrence compared to OPEOs in this study appear to be supported by production estimates. Approximately 80% of the total APEOs used as surfactants are the NPEOs (Ying et al., 2002b; Goel et al., 2003; Ying, 2006). The presence of all longer-chain APEOs and lack OP and NP is also of interest. The high amount of long-chain APEOs dominating the healthcare facility wastewaters sampled in this study signies a source signature. The presence of NPEOs distributed in chain lengths from eight to three indicate that breakdown of the APEO chain length reductions begin to occur at the source.

Table 3 Individual alkylphenol and alkylphenol ethoxylate analyte concentrations measured in each facilitys 24 h composite wastewater sample. All concentrations are given as lg L1. All units (lg L1) Alkyl phenols NPEOs Compound Nonylphenol (NP) Octylphenol (OP) NP3EO NP4EO NP5EO NP6EO NP7EO NP8EO NP9EO NP10EO NP11EO NP12EO NP13EO NP14EO NP15EO NP16EO NP17EO NP18EO OP2EO OP3EO OP4EO OP5EO OP6EO OP7EO OP8EO OP9EO OP10EO OP11EO OP12EO Hospital nd nd 2 4 11 21 33 38 nr nr nr nr nr nr nr nr nr 1 nd nd 4 nd nd nd 6 nr nr nr 3 Nursing care nd nd nd 2 3 4 5 5 nr nr nr nr nr nr nd nd nd nd nd nd nd nd nd nd 2 nr nd nd nd Assisted living nd nd 3 16 27 51 73 86 nr nr nr nr nr nr nr nr nr 2 nd nd nd nd nd nd nd nd nd nd nd Independent living nd nd 1 2 3 4 7 8 nd nr nr nr nr nr nr nr nr 0 nd nd nd nd nd nd nd nd nd nd nd

OPEOs

Quantiable data is center justied to make the table easier to read. nr: Signal present in sample, but concentration not reported due surrogate recovery <60%. nd: Analyte signal not detected during analysis, acceptable surrogate recovery.

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Table 4 Calculated contribution of each individual analyte to the potential estrogenicity of each facility wastewater determined by literature values (Routledge and Sumpter, 1996; Beck et al., 2006) for the yeast estrogen screen (YES) assay response. Potential estrogenicity is presented as lM 17b-estradiol (E2) equivalents for each analyte. All units (lM) Relative potency Hospital Sample conc. 17b-Estradiol (E2) 17a-Ethinylestradiol Estrone Estriol NP OP 1 1.25 2.5E01 5.9E03 1.5E03 7.0E03 1.5E05 0 1.6E04 5.9E05 0.21 0.01 E2 equiv 1.5E05 0 3.9E05 3.5E07 3.2E04 1.0E04 Nursing care Sample conc. 1.8E05 0 9.2E05 1.4E05 3.8E02 3.6E03 E2 equiv. 1.8E05 0 2.3E05 8.2E08 5.7E05 2.5E05 Assisted living Sample conc. 0 0 0 0 0.51 0 E2 equiv. 0 0 0 0 7.6E04 0 Independent living Sample conc. 1.1E05 0 4.1E05 0 5.0E02 0 E2 equiv. 1.1E05 0 1.0E05 0 7.5E05 0

3.3. Estimation of the wastewaters potential estrogenicity The potential estrogenicity for each facilitys wastewater was calculated for discussion purposes based upon YES assay published literature values for relative potency (Routledge and Sumpter, 1996; Beck et al., 2006). Estrogenicity was benchmarked against 17b-estradiol and individual mean steroid hormone concentrations were used in the calculation. The YES assay relative potency of each estrogen measured in the sample is presented in Table 4 along with the relative potency for OP and NP. Because relative estrogenic potency information for APEO with a chain length greater than two (2) was not found in the literature during our review, we converted all of the APEOs present in the sample to either NP or OP rst on a molar basis (outlined in the methods section). Because the healthcare facility wastewater is sent through the municipal sewer system and out into the environment, converting all APEOs to their OP or NP parent is consistent with what is observed for the fate of NPEOs during anaerobic and aerobic biotransormation under ideal conditions (Giger et al., 1984; Ejlertsson et al., 1999; Langford et al., 2005; Zhang et al., 2008). The potential estrogenicity of the hospital, nursing, assisted living, and independent living facility wastewaters were calculated to be 130, 34, 207, and 26 ng L1 as 17b-estradiol, respectively. The calculated contribution of the anticipated NP and OP to the potential estrogenicity is greater than 65% of the total potential estrogenicity for all facilities. The high calculated estrogenicities and the high contribution of AP can be explained because APEOs are present at part-per-billion concentrations in the samples compared to the part-per-trillion concentrations of hormones. Since the relative potencies of the alkylphenols in the YES assay are three orders of magnitude lower than 17b-estradiol, the three order of magnitude

difference in concentration makes up for the difference in relative potency. The calculated estrogenicity of the samples based upon sample chemistry alone most likely is an over estimate of the actual estrogenicity that would be measured through a bioassay. The actual estrogenicity of complex samples is impacted by many variables, including the presence of other analytes that are antagonists of the estrogen receptor (Beck et al., 2006). This calculation also assumes that no loss of either alkyphenol or hormone analytes occurs due to sorption, which is not the case because these compounds are known to sorb to surfaces (Johnson et al., 1998; Ying et al., 2002b; Andreu et al., 2007; Navarro et al., 2009). However, if estrogenicity is an end point of concern, we argue that the total potential for estrogenicity of a sample needs to be taken into consideration when making risk management decisions. Based upon our interpretation, NPEOs present in the sampled healthcare facility wastewaters contribute to the high calculated value of the samples potential estrogenicity. A reduction of potential estrogenicity at this site could be realized by replacing NPEO use with other compounds as a precautionary source reduction measure. 3.4. Mass loading estimates The estimated mass loadings of individual hormones and APEOs from each healthcare facility into the municipal sewer system are reported in Figs. 1 and 2, respectively. Steroid hormone mass loadings from the healthcare facilities are three orders of magnitude lower in comparison to the APEO loadings. The estimated total mass ux of hormones ranged from 92 mg d1 at the hospital to 2 mg d1 at the assisted living facility. The overall androgen contribution to the total mass loading of hormones was more than 65% at

Fig. 1. Estimated mass loading (ng d1) of analyzed hormones leaving each healthcare facility in wastewater. Individual analyte mass loadings are shown within each bar (or directly below each bar). The analyte signier is given above each bar and the key is provided at the bottom of the gure. Data for the assisted living facility is not provided due to a lack of reportable concentration data (<60% surrogate recovery) at that site.

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Fig. 2. Estimated mass loading (g d1) of detected nonylphenol ethoxylates (NPEO, white bars, left side of vertical line) and octylphenol ethoxylates (OPEO, black bars, right side of vertical line) leaving each healthcare facility in wastewater efuent. Individual analyte mass loadings are shown within each bar (or directly below each bar). The signier above each bar (nEO) stands for the alkylphenol ethoxylate, where n signies the chain length ranging from 3 to 18. Only chain lengths detected in at least one site are included in this gure.

all the facilities, with androstenedione contributing more than 40% of the observed mass loading for each site. With such a small mass loading of steroid hormones, it appears unlikely that healthcare facilities are a signicantly elevated source of hormones to municipal wastewater systems. This result is partially surprising, because the hospital wastewater included wastewater from a labor and delivery unit. However, an elevated estrogen signal was not observed. The estimated total mass loading of APEOs from all facilities indicates that healthcare facilities may be a signicant point source contributor of APEOs (and thus APs) to the municipal wastewater system. The total mass loading of APEOs from all facilities was estimated to be 70 g d1. The hospital contributed approximately 80% of the total mass loading of APEOs from all facilities with 54 g d1 leaving the hospital. This nding highlights the importance of understanding mass loadings compared to only understanding the concentration of analytes in wastewater. The concentration of APEOs in the assisted living facility wastewater composite sample was more than two times higher than in the hospital wastewater composite sample. However, the hospitals daily wastewater ow (500 000 L d1) was ten times higher than the assisted living facilitys ow (50 000 L d1). In order to make risk management decisions, characterization data presented in the literature must include ow data from each source in order to determine mass loadings. 3.5. Future research Further characterization to address the temporal variation of steroid hormones, APs, and APEOs is planned to provide additional insight concerning the composition and magnitude of these known EDCs from the healthcare sector. While the amount of hormones

being released from these facilities is likely not signicant compared to domestic inputs, the intensive use of detergents in healthcare facilities could be an important source of NPs to the environment when NPEO based detergents enter the municipal system and transform to NP during conveyance and treatment. In order to determine how important healthcare facility sources are with respect to the release of NPEOs, additional sources in the municipal system will be evaluated. Other facilities with intensive laundering practices, such as hotels, commercial laundry facilities, and car washes are under investigation. The mass loading presented in this paper is a conservative estimate. Because the particulate fraction was not analyzed, any analytes retained on the lter were lost. This research also did not measure alkylphenol ethoxycarboxylates present in the sample. Alkylphenol ethoxycarboxylates have been reported in surface water and wastewater (Lara-Martn et al., 2006; Loos et al., 2007; Loyo-Rosales et al., 2007b; Koh et al., 2008) and these analytes should be included as part of future studies. While this presented research provides a preliminary assessment of several important EDCs in healthcare facility wastewater, additional source characterization is needed as regulatory and political attention continues to focus on the presence of EDCs in the environment. Our presented research provides an initial dataset municipal ofcials, regulators, and healthcare facility operators can use to identify if risk management approaches should be implemented to reduce the release of endocrine active compounds in healthcare facilities in their jurisdiction. Acknowledgements The authors wish to thank Nathan Engel grantee at US EPAs National Risk Management Research Laboratory, Yongui Shan and Ka-

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ren Koran on-site contractors to NRMRL from Pegasus Technical Services, Inc., Larry Zintek at EPAs Region 5 Central Research Laboratory, and Aditya Bhat from Texas A&M for their help during sample collection and processing. We also wish to thank members of the municipal utility (MP, TO, JG, and JH) who were integral in collecting samples and information at each facility. Funding for this project was provided through the State of Texas as part of the program of the Texas Hazardous Waste Research Center project # 067TAM0964 and the Texas Engineering Experiment Station. The United States Environmental Protection Agency through its Ofce of Research and Development provided analytical and nancial support. This paper has been subjected to the US EPAs administrative review and approved for publication. The contents do not necessarily reect the views and policies of the sponsors nor does the mention of trade names or commercial products constitute endorsement or recommendation for use. References
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