A targeted and non-targeted discovery screening approach for poly-and per-fluoroalkyl substances in model environmental biota samples.

A comprehensive analytical approach for targeted and non-targeted discovery screening of per- and polyfluoroalkyl substances (PFAS) was developed and applied to model complex environmental biotic samples. Samples were extracted by formic acid-acetonitrile solution and cleaned up and fractionated by SPE (WAX). Target PFAS quantification was performed by ultra-high performance liquid chromatography interfaced with a triple quadrupole mass spectrometer (UPLC-QqQ-MS/MS). Non-targeted analysis (NTA) PFAS screening was performed with UPLC coupled with a quadrupole-Exactive orbitrap high resolution mass spectrometer (UPLC-Q-Exactive-HRMS). An iterative exclusion (IE) approach was applied to data acquisition for NTA suspect screening to increase the potential for unknown PFAS discovery with MS/MS. A complex workflow in Compound Discoverer was set up to automate data processing of the PFAS suspects search. New mass lists and MS/MS databases, which included a large number of PFAS, were set up and introduced into the search for high-throughput structure identification using HRMS techniques. The integrated targeted-NTA method successfully analyzed for legacy and alternative PFAS in model environmental biota samples, namely polar bear liver and bird egg samples. Targeted analysis provided unequivocal identification of well known/established PFAS (mainly perfluoroalkyl acids) with quantification at very low levels. The NTA suspect screening was able to determine a broader range of PFAS. The data analysis method offered high-confidence annotations for PFAS despite lacking available authentic standards. Overall, the analytical coverage of PFAS was greater and elucidated other PFAS present in these model apex predators.

Bottom-up proteomics analysis for adduction of the broad-spectrum herbicide atrazine to histone.

Histones are the major proteinaceous components of chromatin in eukaryotic cells and an important part of the epigenome. The broad-spectrum herbicide atrazine (2-chloro-4-[ethylamino]-6-[isopropylamino]-1, 3, 5-triazine) and its metabolites are known to form protein adducts, but the formation of atrazine-histone adducts has not been studied. In this study, a bottom-up proteomics analysis method was optimized and applied to identify histone adduction by atrazine in vitro. Whole histones of calf thymus or human histone H3.3 were incubated with atrazine. After solvent-based protein precipitation, the protein was digested by trypsin/Glu-C and the resulting peptides were analyzed by high-resolution mass spectrometry using an ultra-high-performance liquid chromatograph interfaced with a quadrupole Exactive-Orbitrap mass spectrometer. The resulting tryptic/Glu-C peptide of DTNLCAIHAK from calf thymus histone H3.1 or human histone H3.3 was identified with an accurate mass shift of +179.117 Da in atrazine incubated samples. It is deduced that a chemical group with an elemental composition of C8H13N5 (179.1171 Da) from atrazine adducted with calf thymus histone H3.1 or human histone H3.3. It was confirmed by MS/MS analysis that the adduction position was at its cysteine 110 residue. Time- and concentration-dependent assays also confirmed the non-enzymatic covalent modification of histone H3.3 by atrazine in vitro. Thus, the potential exists that atrazine adduction may lead to the alteration of histones that subsequently disturbs their normal function.

Biotransformation of bisphenol-A bis(diphenyl phosphate): In vitro, in silico, and (non-) target analysis for metabolites in rat and bird liver microsomal models.

Increased production and usage of organophosphate esters (OPEs) as flame retardants and plasticizers has trended towards larger and 'novel' (oligomeric) OPEs, although there is a dearth of understanding of the environmental fate, stability, toxicokinetics, biotransformation and bioaccumulation of novel OPEs in exposed biota. The present study characterized in vitro biotransformation of the novel OPE bisphenol-A bis(diphenyl phosphate) (BPADP) using Wistar-Han rat and herring gull liver based microsomal assays. Hypothesized target metabolites bisphenol-A (BPA) and diphenyl phosphate (DPHP) and other metabolites were investigated by applying a lines of evidence approach. In silico modelling predicted both BPA and DPHP as rat metabolites of BPADP, these metabolites were quantified via UHPLC-QQQ-MS/MS. Additional non-target metabolites were determined by UHPLC-Q-Exactive-Orbitrap-HRMS/MS and identified by Compound Discoverer software. Mean BPADP depletion of 44 ± 10% was quantified with 3.9% and 2.6% conversion to BPA and DPHP, respectively, in the rat assay. BPADP metabolism was much slower when compared to the well-studied OPE, triphenyl phosphate (TPHP). BPADP depletion in gull liver assays was far slower relative to the rat. Additional non-target metabolites identified included two Phase I, O-dealkylation products, five Phase I oxidation products and one Phase II glutathione adduct, demonstrating agreement between lines of in vitro and in silico evidence. Lines of evidence suggest that BPADP is biologically persistent in exposed mammals or birds. These findings add to the understanding of BPADP stability and biotransformation, and perhaps of other novel OPEs, which are factors highly applicable to hazard assessments of exposure, persistence and bioaccumulation in biota.

Identification and characterization of serum albumin covalent adduct formed with atrazine by liquid chromatography mass spectrometry.

The present study developed an analytical technique to investigate the possible covalent adduct formation of albumin with the herbicide atrazine, and to characterize the protein modifications in vitro using liquid chromatography separation coupled with high resolution time-of-flight mass spectrometry (LC-TOF-MS). Tandem mass spectrum analysis (MS/MS) with collision induced dissociation (CID) revealed the specific sites of rat, human and bovine serum albumin adduct with atrazine. The formation of b-ion, y-ion series in MS/MS showed a covalent adduct with an addition mass of 179.1 Da located on Cys-34 of serum albumin from rats, human and bovine. This clearly indicated that the chemical group C8H13N5 forms an adduct with Cys-34 despite the sequences differences between of rat, human and bovine serum albumin. To confirm the method reliability, concentration-dependent and time-dependent formation of adducts between serum albumins and atrazine were also investigated. Our results confirmed that atrazine can directly react with Cys-34 of serum albumin and form covalent adducts without prior metabolism.

Side-chain fluorinated polymer surfactants in biosolids from wastewater treatment plants.

High concentrations of the main components in Scotchgard™ fabric protector products (pre-2002 and post-2002; side-chain fluorinated polymer surfactants, S1 and S2, respectively) were detected in biosolids samples from twenty pan-Canadian wastewater treatment plants (WWTPs). Based on mass spectrometric analysis, S1 and S2 can be named as side-chain perfluorooctane sulfonamide-urethane polymer and side-chain perfluorobutane sulfonamide-urethane polymer, respectively. S1 (with C8F17 side-chain) concentrations ranged from 1.08-105 ng/g d.w. and S2 (with C4F9 side-chain) concentrations ranged from 37.5-2051 ng/g d.w., which were much higher than that of other commonly monitored perfluoroalkyl substances (PFAS). S1 and S2 concentrations were significantly correlated (p < 0.001; r2 = 0.6142) indicating similar source origins. A negative linear correlation was observed (p < 0.05) between concentrations of S1 (or S2) with the volume of WWTP treated wastewater per day per person (m3/person/day). The total concentration of 22 other PFAS ranged from 4.93 to 92.6 ng/g d.w., and approximately thirty times lower than S1 and S2 concentrations. The calculated elemental fluorine concentrations of Æ©FS1&S2 were generally much higher than the sum of the other PFAS. PFAS concentrations in biosolids are likely underestimated without consideration of S1 and S2.

Uptake, Deposition, and Metabolism of Triphenyl Phosphate in Embryonated Eggs and Chicks of Japanese Quail (Coturnix japonica).

The toxicokinetics of triphenyl phosphate (TPHP) in vivo including the uptake, deposition, and biotransformation into the metabolite diphenyl phosphate (DPHP) is presently reported in embryonated eggs and chicks of Japanese quail. Quail were dosed with TPHP at 3 concentrations by air cell egg injection on embryonic day 0, followed by daily oral dosing after chicks hatched (5 d). Vehicle-only exposed controls were also used. In dosed eggs, only 33% of the TPHP remained 2 d after injection (no hepatic development); after 10 d (post-hepatogenesis), only 2% remained. The estimated TPHP half-lives in the eggs ranged from 1.1 to 1.8 d for the 3 dosed groups. In all exposed eggs and chicks, DPHP significantly increased with dose (0.001 < p < 0.044). It appears that DPHP is an important metabolite in quail, making up 41 to 74% of all metabolites formed in embryonated eggs. In chicks, at medium and high doses, DPHP concentrations significantly exceeded those of TPHP (p ≤ 0.007), making up 67 and 76% of the total burden, respectively. Our findings suggest that rapid TPHP metabolism occurred in chicks and embryonated quail eggs but that this may vary with the age of the embryonated egg and the stage of embryo development, which should be considered when evaluating concentrations of TPHP and DPHP measured in eggs of wild birds. Environ Toxicol Chem 2020;39:565-573. © 2019 Her Majesty the Queen in Right of Canada. Environmental Toxicology and Chemistry © 2019 SETAC.

In vitro metabolic activation of triphenyl phosphate leading to the formation of glutathione conjugates by rat liver microsomes.

The present study investigated the metabolism of the flame retardant and plasticizer chemical, triphenyl phosphate (TPHP), in a rat liver microsome-based in vitro assay with glutathione (GSH) in order to elucidate metabolic pathways leading to formation of conjugates. A highly sensitive and efficient method was developed for the detection and characterization of GSH reactive metabolites using LC-Q-TOF-MS/MS both in the negative and positive electrospray ionization modes. Seven GSH conjugates formed as a result of microsomal incubation, which were identified as S-conjugates based on MS/MS spectra, and confirmed by subsequent time-dependent incubation assays. With the exception of hydrolysis reactions leading to formation of a diester metabolite, diphenyl phosphate (DPHP), the results demonstrated that Phase I epoxidation on phenyl ring of TPHP leading to mono- and di-hydroxylated TPHP metabolites, which can further conjugate with GSH. Depending on hydroxylated TPHP formation, an o-hydroquinone intermediate formed in vitro via Phase I metabolism, and the o-benzoquinone form reacted with GSH and also formed GSH conjugates. The present study showed that via hydroxylated TPHP Phase I formation that GSH conjugates are important Phase II metabolites for TPHP metabolism in vitro. Some GSH conjugates may be valuable candidate biomarkers for monitoring TPHP exposure in biota.

Covalent binding of the organophosphate insecticide profenofos to tyrosine on α- and ß-tubulin proteins.

Organophosphorus (OP) compounds can bind covalently to many types of proteins and form protein adducts. These protein adducts can indicate the exposure to and neurotoxicity of OPs. In the present work, we studied adduction of tubulin with the OP insecticide profenofos in vitro and optimized the method for detection of adducted peptides. Porcine tubulin was incubated with profenofos and was then digested with trypsin, followed by mass spectrometric identification of the profenofos-modified tubulin and binding sites. With solvent-assisted digestion (80% acetonitrile in digestion solution), the protein was digested for peptide identification, especially for some peptides with low mass. The MALDI-TOF-MS and LC-ESI-TOF-MS analysis results showed that profenofos bound covalently to Tyr83 in porcine α-tubulin (TGTY*83R) and to Tyr281 in porcine ß-tubulin (GSQQY*281R) with a mass increase of 166.02 Da from the original peptide fragments of porcine tubulin proteins. Tyrosine adduct sites were also confirmed by MALDI-TOF/TOF-MS analysis. This result may partially explain the neurotoxicity of profenofos at low doses and prolonged periods of exposure.

A mixed-mode chromatographic separation method for the analysis of dialkyl phosphates.

While reversed-phase (RP) liquid chromatography can separate a wide range of analytes, for strongly acidic compounds such as environmentally relevant dialkyl phosphates (DAPs), this remains a challenge because they have low affinity for standard RP columns or they exhibit inferior peak shapes. Mixed-mode chromatographic (MMC) columns, which contain both RP and ion-exchange functionalities, can address these resolution problems. However, using current MMC separation approaches, analyte peaks are relatively broad as compared to conventional RP chromatography. Herein we present an enhanced MMC-based UHPLC/ESI-MS method for the analysis of DAPs. In contrast to commonly available MMC-based methods, we applied the MMC Luna® Omega PS C18 column that was conditioned by 0.1% formic acid and equilibrated with the initial mobile phase before sample injection. This conditioning step tremendously improved the retention and separation of the DAPs, especially for those with high water solubility and shorter carbon chain lengths. Using water/methanol (95 v/5 v) and ammonium acetate in methanol as the mobile phases, nine DAPs could be baseline resolved with very sharp peaks, including the shorter-chain dimethyl phosphate, diethyl phosphate and bis(2-chloroethyl) phosphate. Other columns were examined to facilitate method optimization, and to identify stationary phases with the ability to separate DAPs as well as to elucidate the retention and separation mechanisms. With this novel UHPLC and post-column dication ion-pairing ESI-MS/MS method, instrumental detection limits as low as 0.01 ng/mL level were achieved. Representing other strongly acidic analytes, the short-chain perfluoroalkyl acid, perfluorobutyl sulfonic acid could also be analyzed with this method.

Side-chain fluorinated polymer surfactants in aquatic sediment and biosolid-augmented agricultural soil from the Great Lakes basin of North America.

Side-chain fluorinated polymer surfactants are the main components of fabric protector sprays and used extensively on furniture and textiles. The composition of these commercial protector products has changed, but there is currently a total dearth of information on these novel fluorinated polymers in the environment. Using a developed analytical approach, two complementary studies examined the distribution of Scotchgard™ fabric protector components in aquatic sediment and in agricultural soils where wastewater treatment plant (WWTP) sourced biosolid application occurred, and in samples from sites in the Laurentian Great Lakes basin of North America. The main components in the pre- and post-2002 Scotchgard™ fabric protectors were identified by MS/MS and Q-TOF-MS to contain a perfluorooctane sulfonamide (S1) and perfluorobutane sulfonamide (S2) based side-chain, respectively, and bonded to a polymer backbone. In fifteen sediment samples collected in 2012-2013 from western Lake Erie and Saginaw Bay (Lake Huron), S1 was in all sediment samples (0.18 to 461.59ng/g dry weight (d.w.)); S2 was in 80% of the sediment samples (<0.03 to 24.08ng/g d.w.). Thirteen soil samples were collected (2015) from a biosolid applied and two non-biosolid applied farm field sites in southern Ontario (Canada). S1 was detected in 100% of the soil samples from biosolid-augmented agricultural sites (mean 236.36ng/g d.w.; range 41.87 to 622.46ng/g d.w.), and at concentrations much greater than in the aquatic sediment samples. The concentration of S1 and S2 in soil and sediment samples were also much greater than the total concentration of other per-and poly-fluoroalkyl substances (PFASs) that were measured. The ratio of S1 concentration versus ∑22PFAS concentration was up to 1616 in sediment samples from Lake Erie. This results helps to explain why known PFASs account for low percentages of the total extractable organic fluorine (EOF) content in sediment.

Exploring adduct formation between human serum albumin and eleven organophosphate ester flame retardants and plasticizers using MALDI-TOF/TOF and LC-Q/TOF.

Organophosphate (OP) and organophosphate ester (OPE) adducts of albumin are valuable biomarkers for retrospective verification of exposure. In the present study, our goal was to determine whether OPE flame retardants (OPE FRs) and OPE plasticizers can covalently bind to human serum albumin (HSA), which would allow the resulting adducts to be used to evaluate exposure. Eleven OPE FRs and plasticizers were examined in a HSA-adduct in vitro assay. Pure HSA was incubated with the target OPEs, as well as with an OP insecticide (profenofos) positive control. After enzymatic cleavage with pepsin or Glu-C, the digested albumin was analyzed by matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS) and liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-ToF-MS). Under optimized HSA assay conditions, tyrosine adducts were formed at Y411 and Y148/Y150 with a characteristic mass shift for phosphorylation (Δm/z 166) for the profenofos positive control. However, no such phosphorylated peptides were detected for the 11 target OPEs. This negative result suggests that these OPEs have very different affinities from the OP insecticide. They are less reactive or they may specifically interact with other proteins.

Organophosphate pesticide method development and presence of chlorpyrifos in the feet of nearctic-neotropical migratory songbirds from Canada that over-winter in Central America agricultural areas.

Recent modeling analysis suggests that numerous birds may be at risk of acute poisoning in insecticide-treated fields. Although the majority of avian field studies on pesticides have focused on treated seed, granule, insect or vegetation (oral exposure) ingestion, dermal exposure is an important exposure route when birds come into contact with deposited pesticides on foliage and other surfaces. Some nearctic-neotropical migratory songbirds are likely exposed to pesticides on their non-breeding habitats and include treated crops, plantations or farmlands. In the present study, we developed a method for four environmentally-relevant organophosphate (OP) pesticides (fenthion, fenamiphos, chlorpyrifos and diazinon) in the feet of migratory songbirds (i.e. Common yellowthroat, Gray catbird, Indigo bunting, America redstart, Northern waterthrush, Northern parula, and an additional 12 species of warblers). A total of 190 specimens of the 18 species of songbirds were sampled from available window-killed birds (spring of 2007 and 2011) in downtown Toronto, Canada. The species that were available most likely over-wintered in Mexican/Central American crops such as citrus, coffee and cacao. The feet of the dead birds were sampled and where OP foot exposure likely occurred during over-wintering foraging on pesticide-treated crops. Chlorpyrifos was the only measurable OP (pg mg feet weight(-1)) and in the 2011-collected feet of Black throated blue warbler (0.5), Tennessee warbler (1.0), Northern parula (1.2), Northern waterthrush (0.6), Common yellowthroat (1.0) and the Blue winged warbler (0.9). Dermal contact with OP pesticides during over-wintering in agricultural areas resulted in low levels of chlorpyrifos and long time retention on the feet of a subset of songbirds.

A New Fluorinated Surfactant Contaminant in Biota: Perfluorobutane Sulfonamide in Several Fish Species.

Environmental contamination and regulation of longer-chain perfluoroalkyl substances (PFASs) such as perfluorooctanesulfonate (PFOS) has given rise to the increased use of shorter-chain PFASs as alternatives in new products, although confirmation of their presence in the environment remains limited. In this study, the PFAS alternative, perfluoro-1-butane-sulfonamide (FBSA), was identified for the first time in biota in homogenate samples of fish by liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-ToF-MS) and quantified by ultra high performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS). In one flounder (Platichthys flesus) muscle sample from the Western Scheldt, The Netherlands, FBSA concentration was at 80.12 ng/g wet weight (w.w.) and was exceeded only by PFOS. FBSA was also detected in 32 out of 33 samples of freshwater fish collected (2009-2010) from water bodies across Canada. In lake trout (Salvelinus namaycush) from northern Canada (e.g., Lake Kusawa (Yukon Territory), Great Bear Lake (Northwest Territories and in the Arctic), and Lake Athabasca (northern Alberta)), the concentrations of FBSA ranged from below method detection limit (<0.01 ng/g w.w) to 0.44 ng/g w.w. and were much lower than those reported for lake trout from the more urbanized and industrialized Laurentian Great Lakes sites (3.17 ± 1.53 ng/g w.w.). In three species of fish purchased from a supermarket in Ottawa (ON, Canada), FBSA concentrations were the lowest of all fish and ranged from < MLOD to 0.29 ng/g w.w. and 0.03 to 0.76 ng/g w.w. in muscle and liver, respectively. FBSA is a bioaccumulative contaminant in fish in Canada and possibly in The Netherlands. It is likely sourced from new alternative perfluorobutane-based products, as well as other shorter chain perfluoroalkyl-based products.

Determination of organophosphate flame retardants and plasticizers in lipid-rich matrices using dispersive solid-phase extraction as a sample cleanup step and ultra-high performance liquid chromatography with atmospheric pressure chemical ionization mass spectrometry.

A fast, robust and highly sensitive analysis method for determination of trace levels of organophosphate ester (OPE) flame retardants and plasticizers in lipid-rich samples was presently developed, and based on ultra-high performance liquid chromatography-tandem mass spectrometry coupled to a positive atmospheric pressure chemical ionization source (UHPLC-MS/MS-APCI(+)). The target OPEs in the sample were extracted from the biota samples, such as egg and liver, by ultrasonic extraction, and cleaned up further by dispersive solid phase extraction (d-ESP). As a result, background contamination was largely reduced. Different dispersive ESP sorbents were tested and primary secondary amine (PSA) bonded silica sorbents showed the best recoveries for these target OPEs. The recoveries obtained were in the range 54-113% (RSD<17%), with method limits of quantification (MLOQs) ranging between 0.06 and 0.29ng/g in egg, and 0.05 and 0.50ng/g w.w. in liver sample. The matrix effects (MEs) associated with using APCI(+) and ESI(+) sources were investigated. APCI(+) showed much less ion suppression than ESI(+) for the determination of these OPEs. For egg and liver samples, the APCI(+) ME values ranged from 40% to 94%, while ESI(+) ME values ranged from 0% to 36%. Although APCI(+) was used for the determination of OPEs, the ionization mechanism might mainly be a thermospray ionization process. This UHPLC-MS/MS-APCI(+) method showed good response linearity for calibration (R2>0.99). The proposed method was applied to real environmental bird egg and fish samples, where several OPE were quantifiable and different OPE patterns was observed between samples.

Perfluoroalkyl sulfonates and carboxylic acids in liver, muscle and adipose tissues of black-footed albatross (Phoebastria nigripes) from Midway Island, North Pacific Ocean.

The Great Pacific Garbage Patch (GPGP) is a gyre of marine plastic debris in the North Pacific Ocean, and nearby is Midway Atoll which is a focal point for ecological damage. This study investigated 13 C4-C16 perfluorinated carboxylic acids (PFCAs), four (C4, C6, C8 and C10) perfluorinated sulfonates and perfluoro-4-ethylcyclohexane sulfonate [collectively perfluoroalkyl acids (PFAAs)] in black-footed albatross tissues (collected in 2011) from Midway Atoll. Of the 18 PFCAs and PFSAs monitored, most were detectable in the liver, muscle and adipose tissues. The concentrations of PFCAs and PFSAs were higher than those in most seabirds from the arctic environment, but lower than those in most of fish-eating water birds collected in the U.S. mainland. The concentrations of the PFAAs in the albatross livers were 7-fold higher than those in Laysan albatross liver samples from the same location reported in 1994. The concentration ranges of PFOS were 22.91-70.48, 3.01-6.59 and 0.53-8.35 ng g(-1) wet weight (ww), respectively, in the liver, muscle and adipose. In the liver samples PFOS was dominant, followed by longer chain PFUdA (8.04-18.70 ng g(-1) ww), PFTrDA, and then PFNA, PFDA and PFDoA. Short chain PFBA, PFPeA, PFBS and PFODA were below limit of quantification. C8-C13 PFCAs showed much higher composition compared to those found in other wildlife where PFOS typically predominated. The concentrations of PFUdA in all 8 individual albatross muscle samples were even higher than those of PFOS. This phenomenon may be attributable to GPGP as a pollution source as well as PFAA physicochemical properties.

Hexabromocyclododecane Flame Retardant Isomers in Sediments from Detroit River and Lake Erie of the Laurentian Great Lakes of North America.

Sediments collected in 2004 from along the Detroit River (n = 19) and across all of Lake Erie (n = 18) were analyzed for isomers of the flame retardant chemical, hexabromocyclododecane (HBCD), using liquid chromatography-tandem mass spectrometry. Sediment samples had ΣHBCD concentrations ranging from not detected to 1.6 ng/g d.w. γ-HBCD (56 %-100 % of ΣHBCDs) was the predominate isomer, observed in 7 of 19 samples from the Detroit River and 6 of 18 samples from Lake Erie (all within the western basin). α-HBCD was found in 4 Detroit River and 2 Lake Erie western basin sites, while ß-HBCD was only in two Detroit River samples. High ΣHBCD concentrations (>100 ng/g d.w.) were found in two sludge samples from two Windsor, ON, wastewater treatment plants that feed into the Detroit River upstream. HBCD contamination into the Detroit River is a major input vector into Lake Erie and with an apparent sediment dilution effect moving towards the eastern basin.

Methodology and determination of tetradecabromo-1, 4-diphenoxybenzene flame retardant and breakdown by-products in sediments from the Laurentian Great Lakes.

Tetradecabromo-1,4-diphenoxybenzene (TeDB-DiPhOBz) is a brominated polyphenyl ether flame retardant (FR) that is known to photolytically degrade to produce lower brominated polybrominated-diphenoxybenzenes (PB-DiPhOBzs), which may be precursors to MeO-PB-DiPhOBzs recently reported in the Great Lakes herring gulls eggs. To our knowledge, there are no reports on TeDB-DiPhOBz or other PB-DiPhOBz by-products in any environmental sample. The present study analyzed for the presence of PB-DiPhOBzs (including TeDB-DiPhOBz) and MeO-PB-DiPhOBzs in surficial sediment from sites in Saginaw Bay in western Lake Huron (n = 7), and in comparison to southern Lake Huron (open water) (n = 5) and Lake Erie (n = 3) sediment collected in the summers of 2012 or 2013. To analyze for possible PB-DiPhOBzs (Br14­Br0), the first known analytical method was developed for extraction and cleanup of sediment samples, and analysis by HPLC­atmospheric pressure photoionization (−)-quadrupole time-of-flight-mass spectrometry. The overall recovery efficiency was optimized to on average 33­104% progressing from Br14- to Br10-PB-DiPhOBzs. Br10- to Br14-PB-DiPhOBz detection and quantification limits ranged from 0.05 to 0.15 ng g(−1) dw and 0.17 to 0.49 ng g(−1) dw, respectively. Although this is the first report, PB-DiPhOBzs (Br14­Br10) and MeO-PB-DiPhOBzs were not detectable in any sediment sample. This included a site near the mouth of the highly FR-contaminated Saginaw River, near the confined disposal facility (CDF) located in Saginaw Bay at Channel-Shelter Island, which receives dredged sediment from the Saginaw River. Our findings suggest sediments from the presently studied sites in the Great Lakes ecosystem are not a sink for TeDB-DiPhOBz and PB-DiPhOBz by-product contaminants.

Comparative hepatic in vitro depletion and metabolite formation of major perfluorooctane sulfonate precursors in Arctic polar bear, beluga whale, and ringed seal.

Perfluorooctane sulfonate (PFOS) has been reported to be among the most concentrated persistent organic pollutants in Arctic marine wildlife. The present study examined the in vitro depletion of major PFOS precursors, N-ethyl-perfluorooctane sulfonamide (N-EtFOSA) and perfluorooctane sulfonamide (FOSA), as well as metabolite formation using an assay based on enzymatically viable liver microsomes for three top Arctic marine mammalian predators, polar bear (Ursus maritimus), beluga whale (Delphinapterus leucas), and ringed seal (Pusa hispida), and in laboratory rat (Rattus rattus) serving as a general mammalian model and positive control. Rat assays showed that N-EtFOSA (38 nM or 150 ng mL(-1)) to FOSA metabolism was >90% complete after 10 min, and at a rate of 23 pmol min(-1) mg(-1) protein. Examining all species in a full 90 min incubation assay, there was >95% N-EtFOSA depletion for the rat active control and polar bear microsomes, ∼65% for ringed seals, and negligible depletion of N-EtFOSA for beluga whale. Concomitantly, the corresponding in vitro formation of FOSA from N-EtFOSA was also quantitatively rat≈polar bear>ringed seal>>>beluga whale. A lack of enzymatic ability and/or a rate too slow to be detected likely explains the lack of N-EtFOSA to FOSA transformation for beluga whale. In the same assays, the depletion of the FOSA metabolite was insignificant (p>0.01) and with no concomitant formation of PFOS metabolite. This suggests that, in part, a source of FOSA is the biotransformation of accumulated N-EtFOSA in free-ranging Arctic ringed seal and polar bear.

In vitro metabolic formation of perfluoroalkyl sulfonamides from copolymer surfactants of pre- and post-2002 scotchgard fabric protector products.

Currently there is a scientific debate on whether fluorinated polymers (or copolymers) are a source, as a result of their degradation and subsequent formation, of perfluorinated carboxylic acids (PFCAs) and perfluorinated alkanesulfonates (PFSAs). The present study investigated whether commercially available fluorinated surfactants, such as Scotchgard fabric protector (3M Company), can be metabolically degraded, using a model microsomal in vitro assay (Wistar-Han rats liver microsomes), and with concomitant formation of PFCAs, PFASs, and/or their precursors. The results showed that the main in vitro metabolite from the pre-2002 product was perfluorooctane sulfonamide (FOSA), and coincident with the detection of the major fabric protector components, which contains the N-ethyl-perfluorooctanesulfonyl chemical moiety (C8F17SO2N(C2H5)-); the main in vitro metabolite of the post-2002 product was perfluorobutane sulfonamide (FBSA), which was coincident with the detection of the major fabric protector components, and contains the N-methyl-perfluorobutanesulfonyl chemical moiety (C4F9SO2N(CH3)-). FOSA or FBSA metabolite concentrations increased over the 0-60 min microsomal incubation period. However, concentrations of their small molecule precursors such as alkylated FOSAs or FBSAs were not detectable (

Halogenated phenolic compound determination in plasma and serum by solid phase extraction, dansylation derivatization and liquid chromatography-positive electrospray ionization-tandem quadrupole mass spectrometry.

A robust, sensitive and accurate method was developed for the simultaneous determination in plasma and serum of suite a halogenated phenolic compounds (HPCs) for which several are known to persist in the environment and analytically pure standards are available. Namely, 14 congeners of hydroxylated polybrominated diphenyl ethers (OH-PBDEs), six congeners of hydroxylated polychlorinated biphenyls (OH-PCBs), pentachlorophenol, pentabromophenol and the flame retardant tetrabromobisphenol A (TBBPA). Solid phase extraction (SPE) enriched the target compounds and cleaned up the samples as a result of efficient adsorption on a strong anion-exchange solid phase SPE cartridge (Oasis MAX). After final clean-up the HPCs were derivatized with dansyl chloride and analyzed by liquid chromatography-tandem mass spectrometry with electrospray ionization in positive mode (ESI(+)). Chromatographic separation was achieved on a Luna PFP(2) column (2mm×100mm, 3µm particle size) with mobile phases of water and acetonitrile (both containing 0.1% formic acid). The addition of the dansyl moiety to the HPCs greatly improved analyte sensitivity as the electrospray ionization efficiency was enhanced. Instrument limits of detection (ILODs) for LC-ESI(+)-MS/MS analysis of the HPCs were in the range of 0.01-0.07ng/mL and the method limits of quantification (MLOQs) were in the range of 0.02-0.15ng/g. Recovery efficiencies of the suite of HPCs ranged from 64% to 118% with relative standard deviations from 2% to 12% from fortified bovine serum samples. The method was successfully applied for HPC determination in representative polar bear and snapping turtle plasma samples.