Article Cite This: Environ. Sci. Technol. XXXX, XXX, XXX−XXX
pubs.acs.org/est
Elevated Concentrations of Bisphenols, Benzophenones, and Antimicrobials in Pantyhose Collected from Six Countries Adela Jing Li† and Kurunthachalam Kannan*,†,‡ †
Environ. Sci. Technol. Downloaded from pubs.acs.org by UNIV OF SOUTH DAKOTA on 08/26/18. For personal use only.
Wadsworth 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, New York 12201-0509, United States ‡ Biochemistry Department, Faculty of Science and Experimental Biochemistry Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia S Supporting Information *
ABSTRACT: Pantyhose, a skin-tight item of clothing made of synthetic fibers and worn by women in many countries, is a source of exposure to several endocrine-disrupting chemicals. Little is known regarding the occurrence of and dermal exposure to chemicals present in pantyhose. In this study, concentrations and profiles of 23 endocrine-disrupting chemicals, including bisphenols, benzophenones, chlorophenols, parabens, and triclocarban (TCC), were determined in 74 pantyhose samples collected from 6 countries. Pantyhose samples were analyzed by two extraction methods: complete dissolution and ultrasonic extraction. Dissolution of the fabric in 1,1,1,3,3,3-hexafluoro-2propanol/chloroform yielded concentrations of several target chemicals that were up to 286 times higher than in the ultrasonic extraction. Bisphenol S (BPS) and bisphenol A (BPA) were found in 100% and 96% of the samples at median concentrations of 1430 and 14.3 ng/g, respectively. Several brands of pantyhose contained BPS, bisphenol F (BPF), benzophenone-1 (BP-1), ethyl-paraben (EtP), and TCC at concentrations of milligrams per gram. Benzophenone-3 (BP-3), 4-hydroxy benzoic acid (4-HB), and methyl- (MeP) and propyl-parabens (PrP) were found in ≥85% of the samples at median concentrations on the order of several tens to hundreds of nanograms per gram of fabric. Pantyhose made in Japan and China with 21−50% Spandex contained the highest concentrations of BPS (2.2 mg/g), BP-1 (2.4 mg/g), and EtP (88 μg/g). Calculated dermal exposure doses to BPS, BP1, and EtP by women via pantyhose were as high as 45 900, 50 600, and 1800 picograms per kilogram of body weight per day, respectively.
■
disorders, obesity, and type-2 diabetes;18,19 BP-3 has been associated with decreased thyroid hormone levels;20 and BP-1 has been linked to endometriosis.21 Parabens have been shown to stimulate the proliferation of tumor cells,22 and urinary concentrations of methyl- (MeP) and ethyl-parabens (EtP) have been associated with reduced fecundity.23 Exposure to CPs has been associated with carcinogenicity, oxidative stress, apoptosis, and teratogenicity.11 Exposure to TCS and TCC has been associated with adverse birth outcomes and decreased gestational age at birth and body weight and length in neonates.24 Despite rising concerns over exposure to EDCs, the sources of human exposures are still not fully understood. Recent studies suggest that textiles, especially those made of synthetic fibers such as nylon, polyester (polyethylene terephthalate), polypropylene, and Spandex contain several EDCs.25−30 Textile production involves the use of hundreds to thousands
INTRODUCTION Bisphenols (BPs; used in the production of epoxy resins and polycarbonate plastics), benzophenones (BzPs; used as sunscreen agents and photostabilizers), parabens (esters of phydroxybenzoic acid; preservatives in personal care products, pharmaceuticals, and food products), chlorophenols (CPs; preservatives for wood and vegetable fibers and intermediates in the production of pharmaceuticals and dyes), triclosan (TCS), and triclocarban (TCC) (antimicrobials) are used in a variety of consumer products. Bisphenol A (BPA), bisphenol S (BPS), benzophenone-3 (BP-3), 4-hydroxy benzoic acid (4HB), 2,4,6-trichlorophenol (2,4,6-TCP), TCS, and TCC are “high-production-volume chemicals.”1 These chemicals have been found in indoor air; indoor dust; aquatic environments; food; and human blood, urine, and breast milk.2−13 Owing to their estrogenic activity, as demonstrated in several in vitro and in vivo studies, these chemicals are referred to as endocrine disruptors,14−17 and therefore, human exposure is a matter of concern. Exposure to endocrine-disrupting chemicals (EDCs) has been linked to adverse health outcomes in populations. Exposure to BPA has been associated with male reproductive © XXXX American Chemical Society
Received: June 8, 2018 Revised: August 1, 2018 Accepted: August 10, 2018
A
DOI: 10.1021/acs.est.8b03129 Environ. Sci. Technol. XXXX, XXX, XXX−XXX
Article
Environmental Science & Technology of chemicals to enhance the desired properties of finished fabric and garment products. Approximately 2000 chemicals have been reported to be used in textiles to obtain preferred properties, such as wrinkle, stain, soil, and UV resistance; antifading; waterproofing; softening; and microbial, fungal, and antistatic protection.28−31 Our recent studies showed the presence of BPs, BzPs, benzothiazoles, and benzotriazoles in infant clothing,25,26 and brominated and phosphate flame retardants in curtains at notable concentrations.32 Among infant clothing, socks made of polyester and Spandex had the highest concentrations of BPA (∼13 300 ng/g) and BPS (∼394 ng/g).25 It has been reported that the production of skin-tight activewear involves the use of chemicals to enhance the desired performance.33 Pantyhose is a skin-tight clothing worn widely by women in many countries. Sales of pantyhose in the United States exceeded those of other kinds of stockings since 1970.34 A variety of synthetic fibers are used in the production of pantyhose, and, thus, they can contain EDCs. Despite this, little is known about the occurrence and profiles of EDCs in pantyhose. In this study, pantyhose collected from six countries were analyzed to determine the concentrations of 23 EDCs, including BPs, BzPs, CPs, parabens, and TCC. On the basis of the measured concentrations, an assessment of dermal exposure to EDCs was performed for women who wear pantyhose.
included traditional ultrasonic extraction25 and complete dissolution.32 Briefly, 0.2 g of the garment was cut into small pieces with clean, stainless-steel scissors and placed in 15 mL polypropylene (PP) tubes; 40 ng each of isotope-labeled internal standard (IS) mixture (21 ISs for each of the 23 target analytes; see the Supporting Information) was then added. For ultrasonic extraction, 5 mL of a mixture of acetone and dichloromethane (v/v, 1:4) was used. After ultrasonication for 30 min (3510R-DTH, Branson Ultrasonics Corporation, Danbury, CT) and centrifugation at 3000 rpm for 10 min (Centrifuge 5804, Eppendorf, Marshall Scientific, Hampton, NH), the solvent layer was collected. The samples were extracted twice, and the solvents were combined. After evaporation to near-dryness under nitrogen (Organomation Associates Inc., West Berlin, MA), 0.5 mL of methanol was added and filtered through a 0.2 μm nylon filter (Spin-X, Costar, Corning, NY) prior to instrumental analysis. For the dissolution method of extraction, 0.2 g of pantyhose fabric was dissolved in 2 mL of 25% 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP)/chloroform (v/v) solution followed by sonication for 20 min. A total of 4 mL of toluene were added drop-wise to precipitate the dissolved polymer matrix. After sonication for 10 min and centrifugation at 3000 rpm for 10 min, the supernatant was collected. The samples were extracted twice and concentrated as described above. Instrumental analysis was performed, using a Shimadzu Prominence modular high-performance liquid chromatography (HPLC) system (Shimadzu Corporation, Kyoto, Japan) interfaced with an API 3200 electrospray triple quadrupole mass spectrometer (ESI-MS/MS; Applied Biosystems, Foster City, CA), operated in the negative-ionization mode. The details of instrumental analysis and compound specific analytical parameters are provided in Tables S2 and S3. Quality Assurance and Quality Control. A 10- to 14point standard calibration curve, with concentrations ranging from 0.05 to 1000 ng/mL, was used in the quantification of target analytes by an internal standard method. Samples with concentrations above the calibration range were diluted with a solvent and reanalyzed. The regression coefficients (r) were ≥0.997 for all standard calibration curves. The method limits of quantification (MLOQs) were estimated based on the lowest concentrations of the calibration standard with a signalto-noise (S/N) ratio of ≥10, the mass of the pantyhose taken for analysis, and the concentration factor (Table S4). The MLOQs of the target compounds in pantyhose ranged from 0.1 to 12.5 ng/g for both the ultrasonic and the dissolution methods of extraction (Table S4). Procedural blanks were analyzed with every 20 samples to determine contamination that arises from laboratory materials and solvents. Trace concentrations of target compounds (0.1−5.4 ng/mL) were found in procedural blanks (Table S4). The concentrations of target analytes found in procedural blanks were subtracted from measured concentrations in pantyhose samples. As a check for instrumental drift in response factors over time, a midpoint calibration standard was injected after every 20 samples. Sample-to-sample carryover of target analytes was monitored by the injection of a pure solvent (methanol) after every 10 samples. Throughout the analysis, three different types of pantyhose were selected for pre- and post-extraction matrix spike by fortifying known concentrations of target analytes (i.e., 10 and 100 ng/mL) and passing them through the entire analytical procedure.
■
MATERIALS AND METHODS Standards and Reagents. Details of the standards and reagents used in this study are provided in the Supporting Information. Sample Collection and Preparation. Between September 2017 and January 2018, we collected 36 pairs of pantyhose from Harbin, China (n = 13); Nagoya, Japan (n = 8); Seoul, Korea (n = 2); Porto and Lisbon, Portugal (n = 6); Santiago, Chile (n = 2); and Albany (New York), United States (n = 5). The samples represented various fabric types, colors, deniers (unit of linear mass density of fibers; fabric thickness), and lengths and were major brands commonly used by women. A total of 30 pantyhose (83%) samples contained the information on fabric types (50−100% nylon blended with 3−50% Spandex, 2% polyester, and 2% cotton). Among these, 90% of the pantyhose samples were composed of nylon with a mixture of Spandex (which provides the elasticity and formfitting nature). Popular colors of pantyhose were sampled, including black (43.2% of the total number of samples), khaki (36.5%), and tan (20.3%). Denier information was available for 55.6% of the samples and ranged from 5 (ultra-sheer) to 80 (opaque). A total of three lengths of hose were sampled, i.e., full (55.6% of the samples), knee (19.4%), and ankle (25%). Information on the country of manufacture was available for 92% of the samples, of which 70% were from Asian countries (China, Japan, and Korea), 18% were from the Americas (United States, Chile, and Honduras), and 12% were from Europe (Croatia, Italy, Portugal, and France). A detailed list of samples analyzed in this study is provided in Table S1. The pantyhose samples were purchased in retail stores in the cities listed above in bulk quantities with the package material (when they were sold in packages) and then sealed in polyethylene bags for storage at 4 °C in the dark until analysis. Fabric from several parts of the pantyhose was cut into small pieces to provide a representative sample for extraction. In total, 74 samples were analyzed, and 2 different extraction methods were used for each sample. The extraction methods B
DOI: 10.1021/acs.est.8b03129 Environ. Sci. Technol. XXXX, XXX, XXX−XXX
Article
Environmental Science & Technology Method Performance. Of the 28 target chemicals measured, the quantification of bisphenol AP (BPAP), 2,4,5trichlorophenol (2,4,5-TCP), 2,4,6-TCP, 3,4-dihydroxy benzoic acid (3,4-DHB), and TCS was affected by the matrix effect, and, therefore, these chemicals were excluded from further data analysis. Relative recoveries of target chemicals were calculated based on the ratio of the analyte signal to that of the corresponding internal standard in spiked pantyhose samples. The relative recoveries of all target compounds extracted by traditional ultrasonication method ranged from 75.2% to 136%, except for BPS and TCC, which were present at elevated concentrations in pantyhose samples (Table S4). The relative recoveries of BPS in material 2 (a pantyhose that contained relatively lower levels of target chemicals) were 111%. The relative recoveries of target chemicals extracted by the complete dissolution method were 64% to 130% except for BPS and TCC. The relative standard deviation (RSD) from repeated analysis of samples by both of the methods of extraction was BPA (96%) > BPB (60%) > BPF (49%) (Table 1). The median concentrations of BPS, BPA, BPF, and BPB in pantyhose samples were 1430, 14.3, 8.8, and 1.7 ng/g, respectively (Table 1). BPS was the most-abundant compound found in pantyhose and was detected at concentrations as high as 2 190 000 ng/g. BPF (1 280 000 ng/g) and BPB (7230 ng/ g) were also found at very high concentrations in some pantyhose samples, whereas the highest concentration determined for BPA was 504 ng/g. BPA is being replaced with other analogues in consumer products, and a decrease in BPA concentrations in human urine with a concomitant increase in BPS, BPF, and BPB was reported recently.6,9,36
C × D × SA × Fmig × Fcontact × Fpen × T × N BW
where EXPderm is the dermal absorption dose (pg/kg-bw/day), C is the concentration of the chemicals measured in pantyhose (ng/g), D is the density of the pantyhose (mg/cm2), SA is the skin contact surface area,35 Fmig is the migration rate of chemicals to the skin per day (0.005, 1/d),27 Fcontact is the fraction of the skin contact area (1, unitless),27 Fpen is the penetration rate of chemicals into the body (0.01, unitless),27 T is the contact duration between pantyhose and skin (assumed to be 1 day), N is the mean number of events per day (assumed to be 1/day), and BW is the average body weight for a woman (67.3 kg, 15−44 years).35
■
RESULTS AND DISCUSSION Extraction of Target Chemicals. The efficiency of the traditional ultrasonic extraction and the recently developed complete dissolution method of extraction32 for target chemicals in pantyhose was evaluated for those analytes with a detection rate (DR) of >50% (Figure S1). The solvent mixture, HFIP and chloroform, used for the dissolution of pantyhose fabric yielded a homogeneous solution (Figure S2) that facilitated efficient extraction of the target compounds from pantyhose (Figure 1). Both of the DRs (i.e., BPA, BPS, bisphenol B [bisphenol B (BPB)], BP-1, BP-3, propyl-paraben [propyl-paraben (PrP)], and 4-HB) and the median C
DOI: 10.1021/acs.est.8b03129 Environ. Sci. Technol. XXXX, XXX, XXX−XXX
Article
Environmental Science & Technology Table 1. Overall Concentrations (ng/g) of Bisphenols, Benzophenones, Chlorophenols, Parabens, and Triclocarban Determined by the Complete-Dissolution Method in Pantyhose Fabric (n = 74) Collected from Six Countries analyte
detection rate (%)
mean (ng/g)
standard deviation
median (ng/g)
range (ng/g)
BPA BPS BPF BPB BPP BPZ BPAF ∑7 bisphenols BP-1 BP-2 BP-3 BP-8 4-OH-BP ∑5 benzophenones MeP EtP PrP BuP BzP HeP OH−MeP OH-EtP 4-HB ∑9 parabens TCC PCP ∑23 analytes
95.9 100 48.6 59.5 4.1 0.0 37.8
40.8 161 000 63 600 409 1.4 n.a.a 0.2 225 000 62 200 0.4 54.3 0.2 0.6 62 200 46.2 2780 10.8 0.5 0.5 0.3 3.6 1.3 104 2950 14 200 4.8 304 000
75.3 350 000 209 000 1400 3.9 n.a. 0.2 438 000 376 000 0.1 110 0.1 2.0 376 000 102 12 600 21.1 0.8 2.4 0.3 7.8 4.1 62.1 12 600 42 200 15.2 759 000
14.3 1430 8.8 1.7 0.9 n.a. 0.1 13 400 3.8 0.4 12.0 0.2 0.2 20.5 12.2 2.7 1.7 0.2 0.1 0.2 0.2 0.2 85.1 145 23.6 0.9 16 600
