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The degradation of fluorotelomer-based acrylate polymers (FTACPs) has been hypothesized to serve as a source of the environmental contaminants, perfluoroalkyl carboxylates (PFCAs). Studies have relied on indirect measurement of presumed degradation products to evaluate the environmental fate of FTACPs; however, this approach leaves a degree of uncertainty. The present study describes the development of a quantitative matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry method as the first direct analysis method for FTACPs. The model FTACP used in this study was poly(8:2 FTAC-co-HDA), a copolymer of 8:2 fluorotelomer acrylate (8:2 FTAC) and hexadecyl acrylate (HDA). Instead of relying on an internal standard polymer, the intensities of 40 poly(8:2 FTAC-co-HDA) signals (911-4612 Da) were normalized to the signal intensity of a matrix-sodium cluster (659 Da). We termed this value the normalized polymer response (P(N)). By using the same dithranol solution for the sample preparation of poly(8:2 FTAC-co-HDA) standards, calibration curves with coefficient of determinations (R(2)) typically >0.98 were produced. When poly(8:2 FTAC-co-HDA) samples were prepared with the same dithranol solution as the poly(8:2 FTAC-co-HDA) standards, quantification to within 25% of the theoretical concentration was achieved. This approach minimized the sample-to-sample variability that typically plagues MALDI-TOF, and is the first method developed to directly quantify FTACPs.

Fluorotelomer-based acrylate polymers (FTACPs) are a class of side-chain fluorinated polymers used for a variety of commercial applications. The degradation of FTACPs through ester hydrolysis, cleavage of the polymer backbone, or both could serve as a significant source of perfluoroalkyl carboxylates (PFCAs). The biodegradation of FTACPs was evaluated in a soil-plant microcosm over 5.5 months in the absence/presence of wastewater treatment plant (WWTP) biosolids using a unique FTACP determined to be a homopolymer of 8:2 fluorotelomer acrylate (8:2 FTAC). Although structurally different from commercial FTACPs, the unique FTACP possesses 8:2 fluorotelomer side chain appendages bound to the polymer backbone via ester moieties. Liberation and subsequent biodegradation of the 8:2 fluorotelomer appendages was indirectly determined by monitoring for PFCAs of varying chain lengths (C6-C9) and known fluorotelomer intermediates by liquid chromatography tandem mass spectrometry (LC-MS/MS). A FTACP biodegradation half-life range of 8-111 years was inferred from the 8:2 fluorotelomer alcohol (8:2 FTOH) equivalent of the unique FTACP and the increase of degradation products. The progress of FTACP biodegradation was also directly monitored qualitatively using matrix-assisted laser desorption/ionization (MALDI-TOF) time-of-flight mass spectrometry. The combination of indirect and direct analysis indicated that the model FTACP biodegraded predominantly to perfluorooctanoate (PFOA) in soils and at a significantly higher rate in the presence of a plant and WWTP biosolids.

The biotransformation of fluorotelomer-based compounds such as fluorotelomer alcohols (FTOHs) and polyfluoroalkyl phosphate esters (PAPs) are sources of exposure to perfluorinated carboxylates (PFCAs), leading in part to the observation of significant concentrations of PFCAs in human blood. The biotransformation of FTOHs and PAPs yield intermediate metabolites that have been observed to covalently modify proteins. In the current investigation, the extent of covalent protein binding in Sprague-Dawley rats upon exposure to 8:2 FTOH and the 6:2 polyfluoroalkyl phosphate diester (6:2 diPAP) was quantified. The animals were administered a single dose of 8:2 FTOH or 6:2 diPAP at 100 mg/kg by oral gavage to monitor biotransformation and extent of protein binding within the liver, kidney, and plasma. In the 8:2 FTOH-dosed animals, perfluorooctanoate (PFOA) was produced as the primary PFCA, at 623.13 ± 59.3, 459.5 ± 171.8, and 397.3 ± 133.0 ng/g in the plasma, liver, and kidney, respectively. For the animals exposed to 6:2 diPAPs, perfluorohexanoate (PFHxA) was the primary PFCA produced, with maximum concentrations of 57.4 ± 6.5, 9.0 ± 1.2, and 25.3 ± 1.2 ng/g in the plasma, liver, and kidney, respectively. Protein binding was observed in the plasma, liver, and kidney after 8:2 FTOH and 6:2 diPAP exposure, with the most significant binding occurring in the liver (>100 nmol/g protein). This is the first study to link the exposure and in vivo biotransformation of fluorotelomer-based compounds to covalent protein binding.

The study reviews the current state of knowledge regarding the biotransformation of fluorotelomer-based compounds, with a focus on compounds that ultimately degrade to form perfluoroalkyl carboxylates (PFCAs). Most metabolism studies have been performed with either microbial systems or rats and mice, and comparatively few studies have used fish models. Furthermore, biotransformation studies thus far have predominately used the 8:2 fluorotelomer alcohol (FTOH) as the substrate. However, there have been an increasing number of studies investigating 6:2 FTOH biotransformation as a result of industry's transition to shorter-chain fluorotelomer chemistry. Studies with the 8:2 FTOH metabolism universally show the formation of perfluorooctanoate (PFOA) and, to a smaller fraction, perfluorononanoate (PFNA) and lower-chain-length PFCAs. In general, the overall yield of PFOA is low, presumably because of the multiple branches in the biotransformation pathways, including conjugation reactions in animal systems. There have been a few studies of non-FTOH biotransformation, which include polyfluoroalkyl phosphates (PAPs), 8:2 fluorotelomer acrylate (8:2 FTAC), and fluorotelomer carboxylates (FTCAs, FTUCAs). The PAPs compounds and 8:2 FTAC were shown to be direct precursors to FTOHs and thus follow similar degradation pathways., (© 2013 SETAC.)

The biotransformation of fluorotelomer based compounds yields saturated and unsaturated fluorotelomer aldehydes (FTALs and FTUALs, respectively) and carboxylic acids (FTCAs and FTUCAs, respectively) as intermediate metabolites that subsequently transform to perfluorinated carboxylic acids (PFCAs). Previous studies have demonstrated that the FTCAs and FTUCAs are 1 to 5 orders of magnitude more toxic than PFCAs after exposure to aquatic organisms. Additionally, FTUALs have demonstrated reactivity with proteins, which may be associated with toxicity through the inhibition of protein function. The purpose of this study was to carry out a comprehensive assessment of the relative toxicity between PFCAs and their intermediate precursor metabolites: the FTALs, FTUALs, FTCAs, and FTUCAs. Analytes were separately incubated with human liver epithelial (THLE-2) cells to assess how varying the functional group and the fluorinated chain length affects cell viability. For each analyte, dose-response EC50 values were calculated. The EC50 values for FTUCAs and FTCAs were similar, with values ranging from 22 ± 9 and 24 ± 9 μM for the 10:2 congeners to 1004 ± 20 and 1004 ± 24 μM for the 4:2 congeners, respectively. The EC50 values for the PFCAs ranged from 65 ± 41 (PFDA) to 1361 ± 146 (PFBA) μM. The range of toxicity between PFCAs and their acid precursors were similar. However, the comparative toxicity between the 6:2 and 8:2 congeners and their corresponding PFCA had toxicity thresholds that varied depending on the functional headgroup, where FTUALs ≥ FTALs > FTUCAs ≥ FTCAs > PFCAs. For all PFCAs and acid precursors, toxicity depended on the length of the fluorinated chain, where the longer chain lengths yielded greater bioaccumulation and enhanced toxicity, results which agreed with those previously reported. By contrast, FTALs and FTUALs were the most toxic of all the analytes examined, where toxicity was enhanced at shorter chain lengths, with EC50 values of 7 ± 1 μM (6:2 FTUAL) and 8.6 ± 0.8 μM (6:2 FTAL). DNA adducts were not detectable for the aldehyde precursors, using a quantitative long-range PCR method. Our data provide the first evidence that aldehyde intermediates have demonstrated toxicity in cellular systems that is more significant than PFCAs and their corresponding acid intermediates.

Fluorotelomer unsaturated carboxylic acids and aldehydes (FTUCAs and FTUALs) are intermediate compounds that form from the biotransformation of fluorotelomer-based compounds. Previous evidence that FTUCAs and FTUALs bind to biological nucleophiles has indicated that protein binding might give rise to toxicity resulting from protein function disruption. The current study assesses the reactivity of FTUALs and FTUCAs by probing the covalent interactions of FTUALs and FTUCAs with proteins present in rat liver microsomes and bovine blood plasma. The FTUALs exhibited significant levels of protein covalent binding, with binding levels ranging from 20.1 (±2.8)% to 71.3 (±19.5)% in microsomes and 24.0 (±1.5)% to 82.5 (±14.0)% in blood plasma. By contrast, the FTUCAs did not exhibit any apparent covalent binding. Bovine serum albumin (BSA) was extracted and isolated from the plasma after incubation of 8:2 FTUAL (5-100 μM). Electrospray ionization mass spectrometry (ESI-MS) was used to investigate the stoichiometry of 8:2 FTUAL covalently bound to BSA; three measurable FTUAL adducts were formed with BSA. To compare the percent binding results from the FTUAL microsome incubation experiments, 8:2 FTOH was incubated in microsomes to determine the protein binding associated with the biotransformation of 8:2 FTOH. Results from this study showed that the biotransformation of 8:2 FTOH yielded 26.1 (±3.0)% binding, and was statistically similar to the percent binding associated with 8:2 FTUAL exposure (p > 0.05), indicating that the binding of 8:2 FTUAL to proteins might be a primary fate in the biotransformation of 8:2 FTOH.

The hydrolysis kinetics of three polyfluorinated alkyl phosphate monoesters (monoPAPs), differing in fluorinated chain length, were measured using bovine intestinal alkaline phosphatase to catalyze the reaction. Kinetic values were also measured for analogous hydrogenated phosphate monoesters to elucidate the effects of the fluorinated chain on the rate of enzymatic hydrolysis. Michaelis constants (K(m)) were obtained by a competition kinetics technique in the presence of p-nitrophenyl phosphate (PNPP) using UV-vis spectroscopy. Compared with K(m) (PNPP), Michaelis constants for monoPAPs ranged from 0.9 to 2.1 compared with hydrogenated phosphates, which ranged from 4.0 to 13.0. Apparent bimolecular rate constants (k(cat)/K(m)) were determined by monitoring rates of product alcohol formation at low substrate concentrations using gas chromatography-mass spectrometry. The experimental values for k(cat)/K(m) averaged as 1.1 × 10(7) M(-1) s(-1) for monoPAPs compared with 3.8 × 10(5) M(-1) s(-1) for hexyl phosphate. This suggests that the electron-withdrawing nature of the fluorinated chain enhanced the alcohol leaving group ability. The results were used in a simple model to suggest that monoPAPs in a typical mammalian digestive tract would hydrolyze in approximately 100 s, supporting a previous study that showed its absence after a dosing study in rats., (Copyright © 2012 SETAC.)

Fluorotelomer unsaturated aldehydes and acids (FTUALs and FTUCAs) are intermediate metabolites that form from the biotransformation of fluorotelomer-based chemicals. FTUALs and FTUCAs have been previously suggested to contribute to the toxicity associated with human exposure to fluorotelomer compounds by covalently binding to biological nucleophiles. However, the extent of their reactivity has only been assessed with glutathione. The purpose of the present study was to assess the reactivity of these intermediate metabolites with a series of nucleophilic amino acids and model proteins. In vitro experiments were carried out in an aqueous buffer system to determine the reactivity of nucleophilic amino acids with FTUCAs and FTUALs having varying fluorinated chain lengths. Using (19)F NMR spectroscopy to monitor the disappearance of the FTUCAs and FTUAL signals and the production of a fluoride signal, reaction rate constants were determined under pseudo-first-order conditions. The FTUCAs reacted only with cysteine with the following second order rate constants: 3.63 (± 1.37) × 10(-5) min(-1) mM(-1) (4:2 FTUCA), 1.19 (± 0.91) × 10(-5) min(-1) mM(-1) (6:2 FTUCA), and 4.56 (± 0.94) × 10(-5) min(-1) mM(-1) (8:2 FTUCA). The FTUALs were significantly more reactive than any of the FTUCAs with reactivity decreasing in the following order: cysteine >> histidine > lysine >> arginine. The following second-order rate constants were obtained: 5.7 (± 4.2) × 10(-4) min(-1) mM(-1) (histidine), 4.3 (± 1.4) × 10(-4) min(-1) mM(-1) (lysine), and 1.4 (± 0.73) × 10(-4) min(-1) mM(-1) (arginine). FTUCAs and FTUALs were also reacted with model proteins to assess their potential for forming covalent adducts. Electrospray ionization mass spectrometry (ESI-MS) was used to investigate the stoichiometry of FTUCAs and FTUALs covalently bound to apomyoglobin (ApoMg) and human serum albumin (HSA). FTUCAs were not reactive, whereas two measurable FTUAL adducts were formed with both ApoMg and HSA at each of the FTUAL chain lengths (6:2, 8:2, and 10:2). This is the first study to probe the reactivity of FTUALs and FTUCAs with nucleophiles other than glutathione, further elucidating possible FTUAL and FTUCA fate within biological systems.

The perfluorophosphonates (PFPAs) and perfluorophosphinates (PFPiAs) are high production volume chemicals that have been observed in Canadian surface waters and wastewater environments. To examine whether their occurrence would result in contamination of organisms in aquatic ecosystems, juvenile rainbow trout (Oncorhynchus mykiss) were separately exposed to a mixture of C6, C8, and C10 monoalkylated PFPAs and a mixture of C6/C6, C6/C8, and C8/C8 dialkylated PFPiAs in the diet for 31 days, followed by 32 days of depuration. Tissue distribution indicated preferential partitioning to blood and liver. Depuration half-lives ranged from 3 to 43 days and increased with the number of perfluorinated carbons present in the chemical. The assimilation efficiencies (α, 7-34%) and biomagnification factors (BMFs, 0.007-0.189) calculated here for PFPAs and PFPiAs were lower than those previously observed for the perfluorocarboxylates (PFCAs) and perfluorosulfonates (PFSAs) in the same test organism. Bioaccumulation was observed to decreased in the order of PFSAs > PFCAs > PFPAs of equal perfluorocarbon chain length and was dependent on the charge of the polar headgroup. Bioaccumulation of the PFPiAs was observed to be low due to their rapid elimination via metabolism to the corresponding PFPAs. Here, we report the first observation of an in vivo cleavage of the carbon-phosphorus bond in fish, as well as, the first in vivo biotransformation of a perfluoroalkyl acid (PFAA). As was previously observed for PFCAs and PFSAs, none of the BMFs determined here for the PFPAs and PFPiAs were greater than one, which suggests PFAAs do not biomagnify from dietary exposure in juvenile rainbow trout.

Perfluorinated acids (PFAs) have elicited significant global regulatory and scientific concern due to their persistence and global pervasiveness. A source of PFAs in the environment is through degradation of fluorotelomer carboxylic acids (FTCAs) but little is known about the toxicity of these degradation products. Previous work found that FTCAs were two to three orders of magnitude more toxic to some freshwater invertebrates than their PFA counterparts and exhibited comparable chain-length-toxicity relationships. In this study, we investigated the toxicity of the 6:2, 8:2, and 10:2 saturated (FTsCA) and unsaturated (FTuCA) fluorotelomer carboxylic acids to two species of freshwater algae, Chlorella vulgaris and Pseudokirchneriella subcapitata, and the amphipod, Hyalella azteca. C. vulgaris was generally the most sensitive species, with EC₅₀s of 26.2, 31.8, 11.1, and 4.2 mg/L for the 6:2 FTsCA, 6:2 FTuCA, 8:2 FTuCA, and 10:2 FTsCA, respectively. H. azteca was most sensitive to the 8:2 FTsCA and 10:2 FTuCA, with LC₅₀s of 5.1 and 3.7 mg/L. The toxicity of the FTCAs generally increased with increasing carbon chain length, and with saturation for most of the species tested, with the exception of P. subcapitata, which did not exhibit any trend. These observations agree with chain-length-toxicity relationships previously reported for the PFCAs and support the greater toxicity of the FTCAs compared to PFCAs. However, the toxicity values are approximately 1000-fold above those detected in the environment indicating negligible risk to aquatic invertebrates., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Human biomonitoring has traditionally focused on analyzing the perfluorocarboxylates (PFCAs) and perfluorosulfonates (PFSAs), although the presence of other unidentified fluorinated chemicals has been demonstrated through total organofluorine analysis. Exposure to legacy and current commercial fluorinated chemicals was investigated by analyzing fifty human sera samples collected in 2009 from the United States for forty fluorinated analytes that included the polyfluoroalkyl phosphate diesters (diPAPs), N-ethyl perfluorooctanesulfonamidoethanol-based polyfluoroalkyl phosphate diester (SAmPAP), one fluorotelomer mercaptoalkyl phosphate diester congener (FTMAP), fluorotelomer sulfonates (FTSs), perfluorophosphonates (PFPAs), and perfluorophosphinates (PFPiAs). DiPAP concentrations (0.035-0.136 μg/L) for the more dominant congeners (6:2, 6:2/8:2, 8:2) were lower than those reported in human sera samples collected in 2004, 2005, and 2008. The SAmPAP and 6:2 FTMAP were not detected, but exposure to SAmPAP was suggested based on the detection of one of its potential degradation intermediates, N-ethyl perfluorooctanesulfonamidoacetate (N-EtFOSAA). PFPiAs were detected for the first time in human sera, with C6/C6 and C6/C8 PFPiAs as the dominant congeners, observed in >50% of the samples.

The bioaccumulation and biotransformation of the 8:2 fluorotelomer acrylate (C(8) F(17) CH(2) CH(2) OC(O)CH = CH(2) , 8:2 FTAc) was investigated in rainbow trout via dietary exposure. The 8:2 FTAc is a monomer used in the manufacture of fluorinated polymers and has been widely detected in the atmosphere. The parent 8:2 FTAc and suspected intermediate and terminal metabolites were monitored in liver, blood, kidney, bile, and feces during the 5-d uptake and 8-d elimination phases using gas chromatography-mass spectrometry (GC-MS)- and liquid chromatography-tandem mass spectrometry (LC-MS/MS)- based methods. Very low levels of the 8:2 FTAc were detected in the internal tissues and feces, suggesting that the 8:2 FTAc was rapidly biotransformed in the gut or liver. Similarly, low concentrations of the 8:2 fluorotelomer alcohol (FTOH) were accumulated in the fish tissues. The 8:2 saturated fluorotelomer carboxylate (FTCA) was formed in the highest concentration, reaching steady-state tissue concentrations of approximately 1,000 to 1,400 ng/g wet weight. The 8:2 FTUCA and 7:3 FTCA were also accumulated in high levels, at levels approximately 10-fold lower than the 8:2 FTCA. Both the 7:3 FTCA and perfluorooctanoate (PFOA) showed increasing levels throughout the uptake phase and into the initial stages of the elimination phase, indicating continued formation through precursors still present in the body. Perfluorononanoate (PFNA) was formed in low nanogram per gram wet weight levels. The intermediate and terminal metabolites were also detected in the bile and feces, indicating an important elimination pathway for these compounds. In addition, the 8:2 FTOH glucuronide conjugate was measured in relatively high concentrations in the bile and feces. The results of the current study demonstrated a scenario in which a biologically labile compound is biotransformed to terminal metabolites that are much more biologically persistent., (Copyright © 2010 SETAC.)

The biotransformation of the 8:2 fluorotelomer acrylate (C(8) F(17) CH(2) CH(2) OC(O)CH = CH(2) , 8:2 fluorotelomer-based acrylate [FTAc]) was quantitatively investigated in cytosolic (S9) fractions isolated from rainbow trout stomach and liver. The in vitro studies presented in this manuscript compliment the whole body 8:2 FTAc dietary exposure study, presented as a companion paper. The S9 fractions were prepared in our laboratory, using fish that had previously been used as control animals in our in vivo study. Before 8:2 FTAc incubations, general carboxylesterase activity was determined using paranitrophenyl acetate (PNPA) as the substrate with formation of paranitrophenol monitored using an ultraviolet-vis spectrometer. In the 8:2 FTAc incubations, the degradation of the parent compound and 8:2 fluorotelomer alcohol (FTOH) formation was monitored by gas chromatography-mass spectrometry. Incubations were performed in triplicate, over a range of concentrations encompassing two orders of magnitude, and the initial rate of 8:2 FTOH or paranitrophenol formation was determined. Enzyme kinetic parameters were determined by plotting the initial rate versus concentration, using nonlinear regression analysis. The maximum initial velocities of the enzyme-catalyzed reaction (V(max)) in the PNPA incubations were 614 ± 18 nmol/min/mg and 147 ± 16 nmol/min/mg for the liver and stomach fractions, respectively. These values are much faster than other phase I and II metabolism reactions. The calculated intrinsic clearance rates (CL(int)) for the 8:2 FTAc incubations were 1.7 and 0.40 ml/min/mg protein, respectively. These results show that the esterase activity toward the 8:2 FTAc is only fourfold greater in the liver as compared with the stomach. These trends demonstrate the potential for considerable extrahepatic metabolism of the 8:2 FTAc before uptake into the internal tissues, ultimately limiting the overall bioaccumulation., (Copyright © 2010 SETAC.)

Several studies have shown that fluorotelomer-based compounds can be metabolized to poly- and perfluorinated carboxylates, such as perfluorooctanoate (PFOA). Research has predominately focused on the 8:2 fluorotelomer alcohol (8:2 FTOH), however, the biotransformation pathway is not well understood. Specifically, there is uncertainty regarding the biological fate of the 8:2 fluorotelomer unsaturated carboxylate (FTUCA) and 7:3 fluorotelomer saturated carboxylate (FTCA). The objective of this study was to further elucidate the pathway for 8:2 FTOH biotransformation through dosing rainbow trout with three 8:2 FTOH metabolism intermediates: the 7:3 FTCA (CF(3)(CF(2))(6)CH(2)CH(2)COO(-)), 8:2 FTCA (CF(3)(CF(2))(7)CH(2)COO(-)), and 8:2 FTUCA (CF(3)(CF(2))(6)CF horizontal lineCHCOO(-)). This study represents the first investigation of these three labile intermediate metabolites in an in vivo system. The parent compounds were dosed via the diet and the parent compounds and intermediates were monitored in the blood and liver during the 7-day uptake phase and 10-day elimination phase. Exposure to the 7:3 FTCA did not result in the formation and accumulation of PFOA, but resulted in low levels of the 7:3 FTUCA and perfluoroheptanoate, a novel finding. PFOA was formed in the 8:2 FTCA and 8:2 FTUCA dosing. In addition, the 7:3 FTCA was formed during exposure to both the 8:2 FTCA and 8:2 FTUCA. Elimination half-lives were 5.1 d (95% confidence interval: 3.1-14 d) for 7:3 FTCA, 1.2 d (1.1-1.3 d) for 8:2 FTCA, and 0.39 d (0.31-0.53 d) for 8:2 FTUCA. The observed differences in the elimination half-life may be the result of differences in either the depuration or metabolism rate. Based on the findings of this study, and reported analogous literature pathways, we proposed a "beta-like-oxidation" pathway for PFOA formation proceeding from the 8:2 FTUCA > 7:3 beta-keto acid > 7:2 ketone > PFOA. Alternatively PFOA could be formed directly through the beta-oxidation of the 7:3 beta-keto acid.

Saturated and unsaturated fluorotelomer carboxylic acids (fluorotelomer acids: FTAs) represent important intermediates in the degradation of fluorotelomer alcohols to perfluorinated carboxylic acids (PFCAs). Recent studies have detected FTAs at low concentrations (ng/L) in precipitation and surface waters; however, information regarding chronic toxicity is lacking. The present study assessed the chronic toxicity of the 8:2 saturated fluorotelomer carboxylic acid (8:2 FTCA) to Chironomus dilutus and the 10:2 saturated and unsaturated fluorotelomer carboxylic acids (10:2 FTCA and 10:2 fluorotelomer unsaturated carboxylic acid [FTuCA]) to Daphnia magna in separate life-cycle tests. In D. magna tests the FTCA was consistently more toxic than the FTuCA. Lethal concentrations (LC50s) were 150 and >60 microg/L for FTuCA and FTCA, respectively. Reproduction was significantly reduced relative to the controls, with respective median effective concentrations (EC50s) for time to first brood and mean number of offspring/female of 287 and 214 microg/L for FTuCA and 50 and 48 microg/L for FTCA. In tests with C. dilutus, EC50s for survival and growth at 20 d were 2,610 and 1,250 microg/L. Total emergence and time to first emergence, the most sensitive endpoints, yielded EC50s of 440 and 890 microg/L. Few adults emerged and no reproduction occurred at the two highest concentrations (600 and 1540 microg/L). Mean number of eggs/female was not affected. These results represent the first chronic toxicity data for FTCAs and additional evidence that FTCAs are more toxic than some PFCAs. While the results indicate that current environmental concentrations of FTAs likely pose negligible risk to aquatic biota, additional quantification of FTAs in surface waters and assessment of their toxicity is needed before meaningful assessments of potential risks to aquatic biota are possible., (Copyright (c) 2010 SETAC.)

Perfluorocarboxylic acids (PFCAs) can be found from the hydrolysis of perfluoroacyl fluorides and chlorides, which can be produced in three separate ways in the atmosphere. Alternatively, PFCAs can be formed directly in the gas phase through reaction of perfluoroacyl peroxy radicals or perfluorinated aldehyde hydrates. All five mechanisms have been elucidated using smog chamber techniques. Yields of the PFCAs from this process vary from less than 10% to greater than 100%, depending on the mechanism. The formation of perfluorosulfonic acids in the atmosphere can also occur, though the mechanism has not been entirely elucidated. A large number of compounds have been confirmed as perfluorinated acid precursors, including CFC-replacement compounds, anesthetics, fluorotelomer compounds, and perfluorosulfonamides. Levels of some of these compounds have been measured in the atmosphere, but concentration for the majority have yet to be detected. It is clear that atmospheric oxidation of volatile precursors contributes to the overall burden of PFAs, though the extent to which this occurs is compound and environment dependent and is difficult to assess accurately.

Relative rate techniques were used to measure the rate constants k[Cl + C(4)F(9)CH(2)CH(2)OC(O)CH=CH(2)] = (2.21 +/- 0.16) x 10(-10) and k[OH + C(4)F(9)CH(2)CH(2)OC(O)CH=CH(2)] = (1.13 +/- 0.12) x 10(-11) cm(3) molecule(-1) s(-1) in 700 Torr of N(2) or air diluent at 296 K. The atmospheric lifetime of C(4)F(9)CH(2)CH(2)OC(O)CHCH(2) (4:2 FTAc) is determined by its reaction with OH radicals and is approximately 1 day. The chlorine-atom-initiated oxidation of 4:2 FTAc in 700 Torr of air at 296 K gives C(4)F(9)CH(2)C(O)H in molar yields of 18% and 10% in the absence and presence of NO, respectively. The OH-radical-initiated oxidation of 4:2 FTAc in 700 Torr of air in the presence of NO gives HCHO in a molar yield of (102 +/- 7)%, with C(4)F(9)CH(2)CH(2)OC(O)C(O)H (4:2 fluorotelomer glyoxylate) as the expected coproduct. The atmospheric fate of the 4:2 fluorotelomer glyoxylate will be photolysis and reaction with OH radicals, which will lead to the formation of C(4)F(9)CH(2)C(O)H and ultimately perfluorinated carboxylic acids. The atmospheric oxidation of fluorotelomer acrylates is a potential source of perfluorinated carboxylic acids in remote locations.

Perfluorinated acids (PFAs) and their precursors (PFA-precursors) exist in the environment as linear and multiple branched isomers. These isomers are hypothesized to have different biological properties, but no isomer-specific data are currently available. The present study is the first in a two-part project examining PFA isomer-specific uptake, tissue distribution, and elimination in a rodent model. Seven male Sprague-Dawley rats were administered a single gavage dose of approximately 500 microg/kg body weight perfluorooctane sulfonate (C(8)F(17)SO(3)(-), PFOS), perfluorooctanoic acid (C(7)F(15)CO(2)H, PFOA), and perfluorononanoic acid (C(8)F(17)CO(2)H, PFNA) and 30 microg/kg body weight perfluorohexane sulfonate (C(6)F(13)SO(3)(-), PFHxS). Over the subsequent 38 d, urine, feces, and tail-vein blood samples were collected intermittently, while larger blood volumes and tissues were collected on days 3 and 38 for isomer analysis by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). For all PFAs, branched isomers generally had lower blood depuration half-lives than the corresponding linear isomer. The most remarkable exception was for the PFOS isomer containing an alpha-perfluoromethyl branch (1m-PFOS), which was threefold more persistent than linear PFOS, possibly due to steric shielding of the hydrophilic sulfonate moiety. For perfluoromonomethyl-branched isomers of PFOS, a structure-property relationship was observed whereby branching toward the sulfonate end of the perfluoroalkyl chain resulted in increased half-lives. For PFHxS, PFOA, and PFOS, preferential elimination of branched isomers occurred primarily via urine, whereas for PFNA preferential elimination of the isopropyl isomer occurred via both urine and feces. Changes in the blood isomer profiles over time and their inverse correlation to isomer elimination patterns in urine, feces, or both provided unequivocal evidence of significant isomer-specific biological handling. Source assignment based on PFA isomer profiles in biota must therefore be conducted with caution, because isomer profiles are unlikely to be conserved in biological samples.

Two major industrial synthetic pathways have been used to produce perfluorinated acids (PFAs) or their precursors: Telomerization and electrochemical fluorination (ECF). Products of telomer and ECF origin can be distinguished by structural isomer profiles. A mixture of linear and branched perfluoroalkyl isomers is associated with ECF. Telomer products characteristically consist of a single perfluoroalkyl geometry, typically linear. In biota, it is unclear if the isomer profile is conserved relative to the exposure medium and hence whether PFA isomer profiles in organisms are useful for distinguishing environmental PFA sources. A companion study suggested isomer-specific disposition following a single oral gavage exposure to rats. To confirm these findings under a more realistic subchronic feeding scenario, male and female rats were administered PFA isomers by diet for 12 weeks, followed by a 12-week depuration period. The diet contained 500 ng/g each of ECF perfluorooctanoate (PFOA, approximately 80% n-PFOA), ECF perfluorooctane sulfonate (PFOS, approximately 70% n-PFOS), and linear and isopropyl perfluorononanoate (n- and iso-PFNA). Blood sampling during the exposure phase revealed preferential accumulation of n-PFOA and n-PFNA compared to most branched isomers. Female rats depurated all isomers faster than males. Both sexes eliminated most branched perfluorocarboxylate isomers more rapidly than the n-isomer. Elimination rates of the major branched PFOS isomers were not statistically different from n-PFOS. Two minor isomers of ECF PFOA and one branched PFOS isomer had longer elimination half-lives than the n-isomers. Although extrapolation of these pharmacokinetics trends in rats to humans and wildlife requires careful consideration of dosage level and species-specific physiology, cumulative evidence suggests that perfluorocarboxylate isomer profiles in biota may not be suitable for quantifying the relative contributions of telomer and ECF sources.

In an effort to reduce volatile organic compounds (VOCs) in paint solvents, replacements containing non-VOC compounds have been proposed. One such compound is 4-chlorobenzotrifluoride (CBTF), for which environmental fate studies have not been conducted. The objective of the present study was to determine the products of the atmospheric oxidation of CBTF and the aqueous fate of these products. Smog chamber experiments were performed to measure the kinetics and mechanism of atmospheric oxidation. A rate constant of 2.22 (+/-0.30) x 10(-13) cm3 molecule(-1) s(-1) was determined for the reaction of hydroxyl radicals with CBTF in 700 Torr of air at 296 K. Using offline sampling and gas chromatography coupled to mass spectroscopic analysis, it was determined that 2-chloro-5-trifluoromethylphenol (o-CTFP) was the primary product of CBTF atmospheric oxidation. Aqueous photolysis of o-CTFP in deionized water proceeded at a rate of 1.3 (+/-0.1) x 10(-4) s(-1), corresponding to a half-life of 1.5 +/- 0.1 h and a quantum yield of 6.6 (+/-0.4) x 10(-4). The mechanism of photolysis was investigated using liquid chromatography coupled to tandem mass spectrometry, which suggested that degradation of o-CTFP occurred via photonucleophilic displacement of chlorine, followed by photoinduced hydrolysis of the trifluoromethyl group to yield 3,4-dihydroxybenzoic acid (an approved food additive considered to be nontoxic).

Temporal trends in perfluoroalkyl compounds (PFCs) were investigated in liver samples from two seabird species, thick-billed murres (Uria lomvia) and northern fulmars (Fulmaris glacialis), from Prince Leopold Island in the Canadian Arctic. Thick-billed murre samples were from 1975, 1993, and 2004, whereas northern fulmars were from 1975, 1987, 1993, and 2003. Between 8 and 10 individuals were analyzed per year. Analytes included C7-C15 perfluorinated carboxylates (PFCAs) and their suspected precursors, the 8:2 & 10:2 fluorotelomer saturated and unsaturated carboxylates (FTCAs, FTUCAs), C6, C8 (perfluorooctane sulfonate, PFOS), C10 sulfonates, and perfluorooctane sulfonamide (PFOSA). Liver samples were homogenized, liquid-liquid extracted with methyl tert-butyl ether, cleaned-up using hexafluoropropanol, and analyzed by LC-MS/ MS. Overall, concentrations in seabirds were lower than those in other marine animals that occupy similar or higher trophic positions. In contrast to most other wildlife samples, PFC profiles were dominated by the PFCAs which comprised 81% and 93% of total PFC profiles in the 2004 thick-billed murre and 2003 northern fulmar samples, respectively. As well, the PFCA profiles were mainly comprised of the C11-C15 PFCAs, which appears to be unique among other wildlife species. PFC concentrations were found to increase significantly from 1975 to 2003/2004. Doubling times in thick-billed murres ranged from 2.3 yrs for perfluoropentadecanoate (PFPA) to 9.9 yrs for perfluorododecanoate (PFDoA), and from 2.5 yrs for PFPA to 11.7 yrs for perfluorodecanoate (PFDA) in northern fulmars. PFCA concentration increases in thick-billed murres were significant for both time periods (1975-1993, 1993-2004), but in northern fulmars appeared to remain steady after 1993. Differences in the temporal trends observed may be the result of differing migratory patterns of the seabirds. Finally, the detection of the 8:2 and 10:2 FTUCAs in seabirds is suggestive of fluorotelomer alcohols as a source of some PFCAs.

Perfluorosulfonates (PFSAs) and perfluorocarboxylates (PFCAs) have been hypothesized to reach remote locations such as the Canadian Arctic either indirectly as volatile precursor chemicals that undergo atmospheric transport and subsequent degradation, or directly via oceanic and atmospheric transport of the PFSAs and PFCAs themselves. Water, sediment, and air samples were collected from three Arctic lakes (Amituk, Char, and Resolute) on Cornwallis Island, Nunavut, Canada. Samples were analyzed for PFSAs and PFCAs, precursor chemicals including the fluorotelomer alcohols (FTOHs) and polyfluorinated sulfonamides (FSAs), and precursor degradation products such as the fluorotelomer unsaturated carboxylates (FTUCAs). PFSAs and PFCAs were detected in water and sediment of all three Arctic lakes (concentrations ranged from nondetect to 69 ng/L and nondetect to 85 ng/g dry weight, respectively). FTOHs and FSAs were observed in air samples (mean concentrations ranged from 2.8 to 29 pg/m3), and confirm that volatile precursors are reaching Arctic latitudes. The observation of degradation products, including FTUCAs observed in sediment and atmospheric particles, and N-ethyl perfluorooctanesulfonamide (NEtFOSA) and perfluorooctanesulfonamide (PFOSA) in air samples, indicate that degradation of the FTOHs and FSAs is occurring in the Arctic environment. PFSAs and PFCAs were also observed on atmospheric particles (mean concentrations ranged from < 0.1 to 5.9 pg/m3). In addition, results of this study also indicate that local perfluoroalkyl contamination of Resolute Lake, which is located downstream of an airport wastewater input, has occurred.

Perfluorinated acids (PFAs) are ubiquitously found in water and biota, including remote regions such as the High Arctic. Under environmental conditions, PFAs exist mainly as anions and are not expected to be subject to long-range atmospheric transport in the gas phase. Fluorinated telomer alcohols (FTOHs) are volatile and can be atmospherically oxidized to form perfluorocarboxylic acids. Analogously, fluorosulfamido alcohols can be oxidized to form perfluorooctane sulfonate (PFOS). High Arctic ice caps experience contamination solely from atmospheric sources. By examining concentrations of PFAs in ice cap samples, it is possible to determine atmospheric fluxes to the Arctic. Ice samples were collected from high Arctic ice caps in the spring of 2005 and 2006. Samples were concentrated using solid-phase extraction and analyzed by LC-MS-MS. PFAs were observed in all samples, dating from 1996 to 2005. Concentrations were in the low-mid pg L(-1) range and exhibited seasonality, with maximum concentrations in the spring-summer. The presence of perfluorodecanoic acid (PFDA) and perfluoroundecanoic acid (PFUnA) on the ice cap was indicative of atmospheric oxidation as a source. Ratios of PFAs to sodium concentrations were highly variable, signifying PFA concentrations on the ice cap were unrelated to marine chemistry. Fluxes of the PFAs were estimated to the area north of 65 degrees N for the 2005 season, which ranged from 114 to 587 kg year(-1) for perfluorooctanoic acid (PFOA), 73 to 860 kg year(-1) for perfluorononanoic acid (PFNA), 16 to 84 kg year(-1) for PFDA, 26 to 62 kg year(-1) for PFUnA, and 18 to 48 kg year(-1) for PFOS. The PFOA and PFNA fluxes agreed with FTOH modeling estimations. A decrease in PFOS concentrations through time was observed, suggesting a fast response to changes in production. These data suggest that atmospheric oxidation of volatile precursors is a primary source of PFAs to the Arctic.

Fluorotelomer alcohols (FTOHs; CF(3)(CF(2))(x)C(2)H(4)OH; where x=3, 5, 7, 9) are a novel class of polyfluorinated contaminants, recently detected in the North American atmosphere, that are possible precursors to the series of perfluoroalkyl carboxylates (PFCAs) in human blood. An in vivo rat study validated earlier independent work that poly- and per-fluoroalkyl carboxylates were metabolites of FTOHs, but our detection of several novel metabolites prompted us to examine their pathways in greater detail using isolated rat hepatocytes. Using 8:2 FTOH (i.e. where x=7) as a model compound, the metabolic products formed by isolated rat hepatocytes were identified, and three synthesized intermediates were incubated separately to elucidate the metabolic pathways. For 8:2 FTOH, a major fate was direct conjugation to form the O-glucuronide and O-sulfate. Using 2,4-dinitrophenylhydrazine (DNPH) trapping, the immediate oxidation product of 8:2 FTOH was identified as 8:2 fluorotelomer aldehyde (8:2 FTAL; CF(3)(CF(2))(7)CH(2)C(H)O). 8:2 FTAL was transient and eliminated HF non-enzymatically to yield 8:2 fluorotelomer alpha,beta-unsaturated aldehyde (8:2 FTUAL; CF(3)(CF(2))(6)CFCHC(H)O) which was also short-lived and reacted GSH and perhaps other endogenous nucleophiles. Four polyfluorinated acid intermediates were also detected, including 8:2 fluorotelomer carboxylate (8:2 FTCA; CF(3)(CF(2))(7)CH(2)C(O)O(-)), 8:2 fluorotelomer alpha,beta-unsaturated carboxylate (8:2 FTUCA; CF(3)(CF(2))(6)CFCHC(O)O(-)), tetrahydroperfluorodecanoate (CF(3)(CF(2))(6)(CH(2))(2)CO(2)(-)), and dihydroperfluorodecenoate (CF(3)(CF(2))(6)CHCHCO(2)(-)). The pathways leading to 8:2 FTCA and FTUCA involve oxidation of 8:2 FTAL, however, the pathways leading to the latter two polyfluorinated acids remain inconclusive. The fate of the unsaturated metabolites, 8:2 FTUAL and FTUCA, included conjugation with GSH and dehydrofluorination to yield alpha,beta-unsaturated GSH conjugates, and GS-8:2 FTUAL which was subsequently reduced to the corresponding alcohol. Perfluorooctanoate (PFOA) and minor amounts of perfluorononanoate (PFNA) were confirmed as metabolites of 8:2 FTOH, and the respective roles of beta- and alpha-oxidation mechanisms are discussed. The analogous acids, aldehydes, and conjugated metabolites of 4:2, 6:2, and 10:2 FTOH (i.e. where x=3, 5, and 9, respectively) were also detected, and metabolite profiles among FTOHs generally differed only in the length of their perfluoroalkyl chains. Preincubation with aminobenzotriazole, but not pyrazole, inhibited the formation of metabolites from all FTOHs, suggesting that their oxidation was catalyzed by P450, not alcohol dehydrogenase.

The ubiquitous detection of perfluorooctane sulfonate (PFOS) in humans and animals has produced a need for sensitive and compound-specific analytical methods to determine the environmental distribution of fluorinated organic contaminants. A suite of potential PFOS precursors (sulfonamides) and fluorotelomer alcohols (FTOHs) were separated by gas chromatography and detected by chemical ionization mass spectrometry (GC/CI-MS). Full-scan spectra were collected in both positive and negative chemical ionization (PCI and NCI, respectively) mode to determine retention time windows and fragmentation patterns. In selected ion monitoring (SIM) mode, instrumental detection limits ranged from 0.2 to 20 pg for individual analytes, depending on ionization mode. PCI mode was preferred for routine analysis because of the simple mass spectra produced, typified by the presence of a major molecular ion [M + H]+. High-volume air samplers collected gaseous and particle-bound fluoroorganics on composite media consisting of XAD-2, polyurethane foam (PUF), and quartz-fiber filters. The combined collection efficiency for individual analytes was 87 to 136% in breakthrough experiments. Application of the method to the analysis of ambient air from urban and rural sites confirmed the presence of six novel fluorinated atmospheric contaminants at picogram per meter3 concentrations. Low concentrations of fluoroorganics were consistently detected in blanks (<4 pg m(-3)); however, this did not prevent confirmation or quantification of environmental concentrations.

This investigation was carried out to evaluate 19F NMR as an analytical tool for the measurement of trifluoroacetic acid (TFA) and other fluorinated acids in the aquatic environment. A method based upon strong anionic exchange (SAX) chromatography was also optimized for the concentration of the fluoro acids prior to NMR analysis. Extraction of the analyte from the SAX column was carried out directly in the NMR solvent in the presence of the strong organic base, DBU. The method allowed the analysis of the acid without any prior cleanup steps being involved. Optimal NMR sensitivity based upon T1 relaxation times was investigated for seven fluorinated compounds in four different NMR solvents. The use of the relaxation agent chromium acetylacetonate, Cr(acac)3, within these solvent systems was also evaluated. Results show that the optimal NMR solvent differs for each fluorinated analyte. Cr(acac)3 was shown to have pronounced effects on the limits of detection of the analyte. Generally, the optimal sensitivity condition appears to be methanol-d4/2M DBU in the presence of 4 mg/mL of Cr-(acac)3. The method was validated through spike and recovery for five fluoro acids from environmentally relevant waters. Results are presented for the analysis of TFA in Toronto rainwater, which ranged from < 16 to 850 ng/L. The NMR results were confirmed by GC-MS selected-ion monitoring of the fluoroanalide derivative.