Characterization of reversed-phase liquid chromatographic columns containing positively charged functionality.

To date, the most commonly used column characterization databases do not determine the relative positive charge associated with new generation RP columns, or they fail to successfully discriminate between RP columns of purportedly low level positive and neutral characters. This paper rectifies this in that it describes a convenient and robust chromatographic procedure for the assessment of the low levels of positive charge on a range of RP columns. The low degree of positive charge was determined by their electrostatic attraction towards the negatively charged 4-n-octylbenzene sulfonic acid (4-OBSA) relative to their retention of the hydrophobic marker toluene (Tol). The new parameter (α4-OBSA/Tol) was determined for 15 commercially available RP-LC columns. When this was combined with existing Tanaka parameters it was possible to guide the chromatographer towards similar columns as "backup / equivalent phases" or dissimilar columns for exploitation in method development strategies. It should be noted that under certain chromatographic conditions the retention mechanism(s) may be too complex to allow direct location of a "backup / equivalent" column(s). The α4-OBSA/Tol results indicate that even the new generation neutral alkyl phases may exhibit a small degree of positive charge at low buffer concentrations. Mobile phases containing low % MeCN were demonstrated to promote mixed mode (anionic exchange / hydrophobic) retention whereas at high % MeCN anionic exchange retention dominated. The measure of electrostatic repulsion between positively charged columns and positively charged bases was assessed by evaluating the relative retention of a range of bases and neutral analytes. The greatest electrostatic repulsion was observed with hydrophilic bases. While there was no correlation between the positive charge associated with the phases assessed by electrostatic attraction or repulsion, the columns could be broadly divided into three subsets (i.e., significant positive character, medium to low positive character and insignificant positive character). Finally, the results were used to highlight the usefulness of the column characterization database containing the new anionic exchange retention parameter (α4-OBSA/Tol) for the selection of an equivalent column possessing a low level of positive character in the analysis of a real-life biopharmaceutical application.

Repeated Aqueous Film-Forming Foams Applications: Impacts on Polyfluoroalkyl Substances Retention in Saturated Soil.

Historical practices at firefighter-training areas involved repeated aqueous film-forming foams (AFFFs) applications, resulting in source zones characterized by high concentrations of perfluoroalkyl and polyfluoroalkyl substances (PFAS). Repeated applications of AFFF composed of 14 anionic and 23 zwitterionic perfluoroalkyl substances (PFAS) were conducted on a single one-dimensional saturated soil column to quantify PFAS retention. An electrofluorination-based (3M) Milspec AFFF, which was above the mixture's critical micelle concentration (CMC), was at application strength (3%, v/v). Retention and retardation of PFAS mass increased with each successive AFFF addition, although the PFAS concentration profiles for subsequent applications differed from the initial. Greater degree of mass retention and retardation correlated with longer PFAS carbon-fluorine chain length and charged-headgroup type and as a function of AFFF application number. Anionic PFAS were increasingly retained with each subsequent AFFF application, while zwitterionic PFAS exhibited an alternating pattern of sorption and desorption. Surfactant-surfactant adsorption and competition during repeat AFFF applications that are at concentrations above the CMC resulted in adsorbed PFAS from the first application, changing the nature of the soil surface with preferential sorption of anionic PFAS and release of zwitterionic PFAS due to competitive elution. Applying a polyparameter quantitative structure-property relationship developed to describe sorption of AFFF-derived PFAS to uncontaminated, saturated soil was attempted for our experimental conditions. The model had been derived for data where AFFF is below the apparent CMC and our experimental conditions that included the presence of mixed micelles (aggregates consisting of different kinds of surfactants that exhibit characteristics properties different from micelles composed of a single surfactant) resulted in overall PFAS mass retained by an average of 27.3% ± 2.7% (standard error) above the predicted values. The correlation was significantly improved by adding a "micelle parameter" to account for cases where the applied AFFF was above the apparent CMC. Our results highlight the importance of interactions between the AFFF components that can only be investigated by employing complex PFAS mixtures at concentrations present in actual AFFF at application strength, which are above their apparent CMC. In firefighter-training areas (AFFF source zones), competitive desorption of PFAS may result in downgradient PFAS retention when desorbed PFAS become resorbed to uncontaminated soil.

Target and Suspect Screening Integrated with Machine Learning to Discover Per- and Polyfluoroalkyl Substance Source Fingerprints.

This study elucidates per- and polyfluoroalkyl substance (PFAS) fingerprints for specific PFAS source types. Ninety-two samples were collected from aqueous film-forming foam impacted groundwater (AFFF-GW), landfill leachate, biosolids leachate, municipal wastewater treatment plant effluent (WWTP), and wastewater effluent from the pulp and paper and power generation industries. High-resolution mass spectrometry operated with electrospray ionization in negative mode was used to quantify up to 50 target PFASs and screen and semi-quantify up to 2,266 suspect PFASs in each sample. Machine learning classifiers were used to identify PFASs that were diagnostic of each source type. Four C5-C7 perfluoroalkyl acids and one suspect PFAS (trihydrogen-substituted fluoroethernonanoic acid) were diagnostic of AFFF-GW. Two target PFASs (5:3 and 6:2 fluorotelomer carboxylic acids) and two suspect PFASs (4:2 fluorotelomer-thia-acetic acid and N-methylperfluoropropane sulfonamido acetic acid) were diagnostic of landfill leachate. Biosolids leachates were best classified along with landfill leachates and N-methyl and N-ethyl perfluorooctane sulfonamido acetic acid assisted in that classification. WWTP, pulp and paper, and power generation samples contained few target PFASs, but fipronil (a fluorinated insecticide) was diagnostic of WWTP samples. Our results provide PFAS fingerprints for known sources and identify target and suspect PFASs that can be used for source allocation.

Ecotoxicity and Accumulation of Perfluorononanoic Acid in the Fathead Minnow (Pimephales promelas) and an Approach to Developing Protective Thresholds in the Aquatic Environment Through Species Sensitivity Distribution.

Per- and polyfluoroalkyl substances (PFAS) are ubiquitous in the environment. Locations where PFAS-containing aqueous film-forming foam (AFFF) has been used or accidentally released have resulted in persistently high concentrations of PFAS, including in surface water that may be adjacent to release sites. Perfluorooctane sulfonic acid (PFOS) is most frequently measured near AFFF release sites; however, other PFAS are being quantified more frequently and, of those, perfluorononanoic acid (PFNA) is common. The goal of our study was to fill data gaps on PFNA toxicity to freshwater fish using the fathead minnow (Pimephales promelas). We aimed to understand how PFNA may impact apical endpoints following a 42-day exposure to mature fish and a 21-day exposure to second-generation larval fish. Exposure concentrations were 0, 124, 250, 500, and 1000 µg/L for both adult (F0) and larval (F1) generations. The most sensitive endpoint measured was development in the F1 generation at concentrations of ≥250 µg/L. The 10% and 20% effective concentration of the tested population for the F1 biomass endpoint was 100.3 and 129.5 µg/L, respectively. These data were collated with toxicity values from the primary literature on aquatic organisms exposed to PFNA for subchronic or chronic durations. A species sensitivity distribution was developed to estimate a screening-level threshold for PFNA. The resulting hazard concentration protective of 95% of the freshwater aquatic species was 55 µg PFNA/L. Although this value is likely protective of aquatic organisms exposed to PFNA, it is prudent to consider that organisms experience multiple stressors (including many PFAS) simultaneously; an approach to understand screening-level thresholds for PFAS mixtures remains an uncertainty within the field of ecological risk assessment. Environ Toxicol Chem 2023;42:2229-2236. © 2023 SETAC.

Spatial and temporal variability of per- and polyfluoroalkyl substances (PFAS) in environmental media of a small pond: Toward an improved understanding of PFAS bioaccumulation in fish.

Per- and polyfluoroalkyl substances (PFAS) are highly fluorinated compounds with many industrial applications, for instance as ingredients in fire-suppressing aqueous film-forming foams (AFFF). Several PFAS have been demonstrated to be persistent, bioaccumulative and toxic. This study better characterizes the bioaccumulation of PFAS in freshwater fish through a spatial and temporal analysis of surface water and sediment from a stormwater pond in a former Naval air station (NAS) with historic AFFF use. We sampled environmental media from four locations twice per week for five weeks and sampled fish at the end of the sampling effort. The primary PFAS identified in surface water, sediment, and biota were perfluorooctane sulfonate (PFOS) and perfluorohexane sulfonate (PFHxS) followed by perfluorooctanoic acid (PFOA) in environmental media and perfluoroheptane sulfonate (PFHpS) in biota. We observed significant temporal variability in surface water concentrations at the pond headwaters following stochastic events such as heavy rainfall for many compounds, particularly PFHxS. Sediment concentrations varied most across sampling locations. In fish, liver tissue presented the highest concentrations for all compounds except PFHxS, which was highest in muscle tissue, suggesting the influence of fine-scale aqueous PFAS fluctuations on tissue distribution. Calculated log bioaccumulation factors (BAFs) ranged from 0.13 to 2.30 for perfluoroalkyl carboxylates (PFCA) and 0.29-4.05 for perfluoroalkane sulfonates (PFSA) and fluctuated greatly with aqueous concentrations. The variability of PFAS concentrations in environmental media necessitates more frequent sampling efforts in field-based studies to better characterize PFAS contamination in aquatic ecosystems as well as exercising caution when considering single time-point BAFs due to uncertainty of system dynamics.

Practical Semiquantification Strategy for Estimating Suspect Per- and Polyfluoroalkyl Substance (PFAS) Concentrations.

Semiquantitation of suspect per- and polyfluoroalkyl substances (PFAS) in complex mixtures is challenging due to the increasing number of suspect PFAS. Traditional 1:1 matching strategies require selecting calibrants (target-surrogate standard pairs) based on head group, fluorinated chain length, and retention time, which is time-consuming and requires expert knowledge. Lack of uniformity in calibrant selection for estimating suspect concentrations among different laboratories makes comparing reported suspect concentrations difficult. In this study, a practical approach whereby the area counts for 50 anionic and 5 zwitterionic/cationic target PFAS were ratioed to the average area of their respective stable-isotope labeled surrogates to create "average PFAS calibration curves" for suspects detected in negative- and positive-ionization mode liquid chromatography quadrupole time-of-flight mass spectrometry. The calibration curves were fitted with log-log and weighted linear regression models. The two models were evaluated for their accuracy and prediction interval in predicting the target PFAS concentrations. The average PFAS calibration curves were then used to estimate the suspect PFAS concentration in a well-characterized aqueous film-forming foam. Weighted linear regression resulted in more target PFAS that fell within 70-130% of their known standard value and narrower prediction intervals than the log-log transformation approach. The summed suspect PFAS concentrations calculated by weighted linear regression and log-log transformation were within 8 and 16% of those estimated by a 1:1 matching strategy. The average PFAS calibration curve can be easily expanded and can be applied to any suspect PFAS even if the confidence in the suspect structure is low or unknown.

Role of Mineral-Organic Interactions in PFAS Retention by AFFF-Impacted Soil.

A comprehensive, generalized approach to predict the retention of per- and polyfluoroalkyl substances (PFAS) from aqueous film-forming foam (AFFF) by a soil matrix as a function of PFAS molecular and soil physiochemical properties was developed. An AFFF with 34 major PFAS (12 anions and 22 zwitterions) was added to uncontaminated soil in one-dimensional saturated column experiments and PFAS mass retained was measured. PFAS mass retention was described using an exhaustive statistical approach to generate a poly-parameter quantitative structure-property relationship (ppQSPR). The relevant predictive properties were PFAS molar mass, mass fluorine, number of nitrogens in the PFAS molecule, poorly crystalline Fe oxides, organic carbon, and specific (BET-N2) surface area. The retention of anionic PFAS was nearly independent of soil properties and largely a function of molecular hydrophobicity, with the size of the fluorinated side chain as the main predictor. Retention of nitrogen-containing zwitterionic PFAS was related to poorly crystalline metal oxides and organic carbon content. Knowledge of the extent to which a suite of PFAS may respond to variations in soil matrix properties, as developed here, paves the way for the development of reactive transport algorithms with the ability to capture PFAS dynamics in source zones over extended time frames.

Evaluating Neutral PFAS for Potential Dermal Absorption from the Gas Phase.

Exposures to per- and polyfluoroalkyl substances (PFAS) are of increasing concern. Assessments typically focus only on ingestion and inhalation exposure due to a lack of generally accepted approaches for estimating dermal absorption. Prior work indicates limited dermal absorption of ionic PFAS, but absorption of neutral PFAS has not been examined from the liquid vehicle or from vapor. Partitioning of semivolatile organic compounds from the gas phase to the skin surface (i.e., stratum corneum) is well known, but the potential for partitioning of neutral PFAS from the gas phase to the stratum corneum has yet to be estimated. The SPARC-estimated physicochemical properties were used to calculate transdermal permeability coefficients (kp_g) and dermal-to-inhalation (D/I) exposure ratios for two groups of neutral PFAS, including those on a U.S. Environmental Protection Agency PFAS list. 11 neutral PFAS gave calculated D/I ratios >5, indicating that direct transdermal absorption may be an important exposure pathway compared to inhalation. Data on consumer products or indoor air is needed for the 11 neutral PFAS, followed by possible biomonitoring to experimentally verify dermal absorption from air. Additional PFAS should be estimated by the protocol used here as they are identified in commercial products.

Leaching of Perfluoroalkyl Acids during Unsaturated Zone Flushing at a Field Site Impacted with Aqueous Film Forming Foam.

While several studies have focused on perfluoroalkyl acid (PFAA) leaching from soils, field studies evaluating the relationship between PFAA mass removal and porewater concentrations as the PFAA source becomes depleted are lacking. Herein, in situ water flushing was performed at a site historically impacted with AFFF to accelerate the leaching of PFAAs from unsaturated soils in a highly characterized field test cell. Porous cup suction lysimeters were used to assess the changes in PFAA porewater concentrations as a function of PFAA mass removal from the unsaturated soils, where flushing was intermittently paused to determine ambient PFAA porewater concentrations. Results showed that the fractional decreases in PFAA porewater concentrations during flushing exceeded the fractional decrease in PFAA mass removal from the soil. PFOS porewater concentrations decrease by 76% (with negligible rebound) compared to only a 7.4% decrease in overall PFOS mass removed from the unsaturated zone. Overall, the results observed herein suggest that, when considering soil impacts to groundwater, less stringent soil cleanup criteria than those that consider an equivalent relationship between mass removal and mass discharge may be appropriate. In addition, remedial approaches that remove only a modest fraction of the PFAA soil mass may be protective of underlying groundwater, particularly for perfluorinated sulfonates with at least six carbons.

Paints: A Source of Volatile PFAS in Air─Potential Implications for Inhalation Exposure.

Paints are widely used in indoor settings yet there are no data for volatile per- and polyfluoroalkyl substances (PFAS) for paints or knowledge if paints are potentially important sources of human exposure to PFAS. Different commercial paints (n = 27) were collected from local hardware stores and analyzed for volatile PFAS by gas chromatography-mass spectrometry (GC-MS), nonvolatile PFAS by liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-qTOF), and total fluorine by 19F nuclear magnetic resonance spectroscopy (NMR). Diluted paint required clean up to remove 6:2 fluorotelomer phosphate diester (diPAP), which thermally transforms into 6:2 FTOH at 280 °C (GC inlet temperature). Only 6:2 FTOH (0.9-83 µg/g) and 6:2 diPAP (0.073-58 µg/g) were found in five exterior and nine interior paints and only accounted for a maximum of 17% of total fluorine. Upon drying, 40% of the FTOH mass was lost, and the loss was verified by measurements of the cumulative FTOH mass measured in the air of a small, confined space over a 3 h period. Based on the liquid paint results, the ConsExpo model was used for potential exposure assessment and one commercial paint exceeded the chosen reference dose (5 µg/kg-day) for children and adults, indicating the potential for human exposure during painting.

Tools for Understanding and Predicting the Affinity of Per- and Polyfluoroalkyl Substances for Anion-Exchange Sorbents.

Anion-exchange (AE) sorbents are gaining in popularity for the remediation of anionic per- and polyfluoroalkyl substances (PFAS) in water. However, it is unclear how hydrophobic and electrostatic interactions contribute to anionic PFAS retention. The goal of this study was to understand the effects of PFAS chain length and head group on electrostatic interactions between PFAS and an aminopropyl AE phase. Liquid chromatography-mass spectrometry (LC-MS) was used with an aminopropyl AE guard column to find relative retention times. The average electrostatic potential (EPavg) of each PFAS was calculated, which correlated positively with the PFAS chromatographic retention time, demonstrating the value of EPavg as a proxy for predicting electrostatic interactions between PFAS and the aminopropyl AE phase. The order of greatest to lowest PFAS AE affinity for an aminopropyl column based on chromatographic retention times and electrostatic interactions was n:3 fluorotelomer carboxylic acids (n:3 FtAs) > n:2 fluorotelomer carboxylic acids (n:2 FtAs) > perfluoroalkyl carboxylates (PFCAs) > perfluoroalkyl sulfonamides (FASAs) ∼ n:2 fluorotelomer sulfonates (n:2 FtSs) > perfluoroalkyl sulfonates (PFSAs). This study introduces a methodology for qualitatively characterizing electrostatic interactions between PFAS and AE phases and highlights that electrostatic interactions alone cannot explain the affinity of PFAS for AE resins in water treatment/remediation scenarios.

Enrichment of poly- and perfluoroalkyl substances (PFAS) in the surface microlayer and foam in synthetic and natural waters.

Bench-scale experiments were performed to interrogate poly- and perfluoroalkyl substance (PFAS) enrichment in the water surface microlayer (SML). In initial experiments using electrolyte-only solutions, the perfluorooctane sulfonate (PFOS) and perfluorooctane carboxylate (PFOA) enrichment in the SML were reasonably (with a factor of 2) described by the Gibbs adsorption equation coupled with a Freundlich-based interfacial adsorption model. Enrichment in the SML among perfluorinated sulfonates and perfluorinated carboxylates of varying chain lengths was proportional to their surface activity. The PFOS enrichment factor (EF), defined as the PFAS concentration in the SML divided by the concentration in the bulk water, was 18 in a 200 mg/l NaCl solution. The presence of elevated organic carbon levels in synthetic surface waters inhibited PFAS accumulation in the SML, with resulting EF values of approximately 1 for all PFAS. However, in the presence of elevated organic levels coupled with foam, PFAS enrichment in the foam was observed, with a foam EF of 25 measured for PFOS in synthetic surface waters. PFAS EF values measured in several natural surface waters without foam showed little variation among the waters tested, with PFOS EF values ranging between 6 and 10. Together, these results suggest that PFAS accumulation in the SML is largely controlled by PFAS sorption at the air-water interface for the conditions examined in this study, and the presence of foam with natural organics enhances PFAS uptake at the water surface.

Dietary Perfluorohexanoic Acid (PFHxA) Exposures in Juvenile Zebrafish Produce Subtle Behavioral Effects across Generations.

Ubiquitous anthropogenic contaminants of concern, per- and polyfluoroalkyl substances (PFAS) are frequently detected in the environment and human populations around the world. Diet is a predominate route of human exposure, and PFAS are frequently measured in food. Manufacturing trends have shifted from legacy PFAS to shorter-chain alternatives that are suggested to be safer, such as perfluorohexanoic acid (PFHxA). However, the current amount of data to support safety assessments of these alternatives is not yet sufficient. The present study investigated the effects of a 42-day dietary exposure to 1, 10, or 100 ng/g PFHxA in juvenile zebrafish. The zebrafish model was leveraged to interrogate morphometrics, fecundity, and numerous behavior endpoints across multiple generations. Dietary PFHxA exposure did not result in measurable body burden and did not affect growth, fecundity, adult social perception behavior, or associative learning. PFHxA exposure did induce abnormal adult anxiety behaviors in the F0 generation that persisted transgenerationally in the F1 and F2. Abnormal larval and juvenile behavior was observed in the F1 generation, but not in the F2. PFHxA juvenile dietary exposure induced subtle and multigenerational behavior effects that warrant further investigation of this and other alternative short-chain PFAS.