Occurrence of per- and polyfluoroalkyl substances (PFASs) in wastewater of major cities across China in 2014 and 2016.

China produced and consumed a large amount of per- and polyfluoroalkyl substances (PFASs). whose persistency and possible toxicity to organisms have raised public health concerns. Analyzing influent wastewater could help to assess the composition and mass load of PFASs discharged into a wastewater treatment plant (WWTP) from its catchment. In this study, we analyzed 27 PFASs in wastewater samples collected from 42 WWTPs across China in 2014 and 2016. Results indicated that perfluorobutanoic acid (PFBA), perfluorohexanoic acid (PFHxA), perfluoroheptanoic acid (PFHpA), perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid (PFOS) were the most common PFASs in wastewater. Population normalized mass loads of PFOA and PFOS were higher in Eastern China than in the other three regions, possibly due to their higher usage. Although the concentrations of PFASs in Central and West areas were showed lower than in East area, Wuhan (in Central area) and Lanzhou (in West area) were hotspots of PFASs pollution because of their industry structure. Population density and per capita Gross Domestic Product (GDP) have positive correlations with the concentration of PFBA, PFOA, PFHxA, and ∑PFASs in wastewater. The estimated annual release of PFASs to WWTPs in our study is much lower than the total emission to the environment. Our results suggest that although there was some reduction in the production volume, certain legacy PFASs were still released into wastewater and their composition and concentration vary among WWTPs.

Urinary Concentrations of Bisphenols in the Australian Population and Their Association with the Per Capita Mass Loads in Wastewater.

Health concerns and related regulation of bisphenol A (BPA) in some countries have led to an increase in the production and use of unregulated and poorly understood BPA analogues, including bisphenol S (BPS), bisphenol F (BPF), bisphenol B (BPB), and bisphenol AF (BPAF). To assess the temporal trends of human exposure to BPA analogues, urine and wastewater samples were collected from South East Queensland, Australia between 2012 and 2017 and analyzed for five bisphenols using validated isotope dilution liquid chromatography tandem mass spectrometry methods. BPA and BPS were the predominant bisphenols detected in both urine and wastewater samples, with median concentrations of 2.5 and 0.64 µg/L in urine and 0.94 and 1.1 µg/L in wastewater, respectively. BPB, BPF, and BPAF had low detection frequencies in both urine and wastewater samples. Concentrations of BPA in both urine and wastewater decreased over the sampling period, whereas concentrations of BPS increased, suggesting that BPS has become a BPA replacement. The contributions of urinary excretion to wastewater were calculated by the ratio of daily per capita urinary excretion to wastewater-based mass loads of bisphenols. Urinary BPA and BPS contributed to less than 1% of the load found in wastewater, indicating that much of the BPA and BPS originates from other sources.

Per- and poly-fluoroalkyl substances (PFASs) in follicular fluid from women experiencing infertility in Australia.

Per- and poly-fluoroalkyl substances (PFASs) have been widely used and detected in human matrices. Evidence that PFAS exposure may be associated with adverse human reproductive health effects exists, however, data is limited. The use of a human matrix such as follicular fluid to determine chemical exposure, along with reproductive data will be used to investigate if there is a relationship between PFAS exposure and human fertility. OBJECTIVE: This study aims to: (1) assess if associations exist between PFAS concentrations and/or age and fertilisation rate (as determined in follicular fluid of women in Australia who received assisted reproductive treatment (ART)); and (2) assess if associations exist between PFAS concentrations and infertility aetiology. METHODS: Follicular fluids were originally collected from participants who underwent fully stimulated ART treatment cycles at an in vitro fertilisation (IVF) clinic in the period 2006-2009 and 2010-11 in Queensland, Australia. The samples were available for analysis of 32 PFASs including perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), perfluorohexane sulfonate (PFHxS), and perfluorononanoic acid (PFNA) using high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). 97 samples were matched with limited demographic data (age and fertilisation rate) and five infertility factors (three known female factors): 1) endometriosis, 2) polycystic ovarian syndrome (PCOS), and 3) genital tract infections - tubal/pelvic inflammation disease; as well as 4) male factor, and 5) idiopathic or unknown from either males or females. SPSS was used for linear regression analysis. RESULTS: PFASs were detected in all follicular fluid samples with the mean concentrations of PFOS and PFOA, 4.9, and 2.4 ng/ml, respectively. A lower fertilisation rate was observed at higher age when age was added as a covariate, but there was no relationship between PFAS concentrations and fertilisation rate. There were few statistically significant associations between PFAS concentrations in follicular fluid and infertility factors. Log-transformed PFHxS concentrations were lower in females with endometriosis (factor 1) than in women who had reported 'male factors' as a reason of infertility, while PFHpA was higher in women who had infertile due to female factors (factor 1-3) compared to those who had infertile due to male factor. CONCLUSION: PFASs were detected in follicular fluid of Australian women who had been treated at an IVF clinic. PFAS exposure found in follicular fluids is linked to increased risk of some infertility factors, and increased age was associated with decreased fertilisation rate in our data. But there was no relationship between PFAS and ferlitisation rate. Further large-scale investigations of PFAS and health effects including infertility are warranted.

Prenatal phthalate exposure, oxidative stress-related genetic vulnerability and early life neurodevelopment: A birth cohort study.

Prenatal phthalate chemicals may have adverse effects on brain development by various mechanisms including oxidant damage. However, birth cohort findings have been conflicting. This study aimed to (i) investigate the interplay between maternal prenatal phthalate levels, infant genetic vulnerability to oxidative stress, and child neurodevelopment and (ii) examine combined putative oxidant exposures. In a population-based birth cohort of 1064 women with prenatal recruitment in Victoria, Australia, maternal urine was collected at 36 weeks of pregnancy and phthalate metabolite concentrations measured. An unweighted genetic score for oxidative stress was made using a candidate gene approach. Cognition was assessed using the BAYLEY-III at two years (n = 678). Parents completed questionnaires for doctor diagnosed autism spectrum disorder (ASD) (1.4 %), ASD traits (4.9 %) and child inattention/hyperactivity (n = 791). Analyses included multiple linear and logistic regression. Higher prenatal phthalate levels and a higher oxidative stress genetic score were each associated with subsequent ASD. Several oxidative stress-related SNPs modified the association between prenatal phthalates and ASD and other outcomes. Consistent patterns were evident across gene score-phthalate combinations for cognition, ASD, ASD traits and inattention/hyperactivity. Other putative oxidant factors such as prenatal smoking further increased risk. Prenatal phthalate levels and infant oxidative stress-related genetic vulnerability are associated with adverse neurodevelopment. Combined exposures are important. Current recommendations and regulation on maternal phthalate exposure during pregnancy require re-evaluation.

Predictors with regard to ingestion, inhalation and dermal absorption of estimated phthalate daily intakes in pregnant women: The Barwon infant study.

Human exposure to phthalate chemicals, used in consumer product plastics, occurs throughout the day. Phthalate levels in pregnant women are associated with offspring health effects including obesity and neurodevelopmental problems. Knowledge of predictors of exposure is necessary in order to effectively reduce phthalate exposure. The present study aims to identify predictors of phthalate levels in Australian pregnant women from the Barwon Infant study birth cohort. Maternal urine samples from 841 women were analyzed for phthalate metabolites. Maternal diet and food preparation practices, use of volatile household products, household characteristics and personal care product use were assessed with questionnaires. All maternal urine contained phthalate metabolites. Maternal prenatal high-fat milk consumption was associated with higher benzyl butyl phthalate (BBzP) (p < 0.001), and bis(2-ethylhexyl) phthalate (DEHP) (p = 0.0023). Higher phthalate levels were associated with consumption of tinned food (fish and tomatoes). Diethyl phthalate (DEP) levels were significantly higher when women reported using air freshener (35% increase, p = 0.01), aerosols (40% increase, p = 0.005), hair treatment chemicals (28% increase, p = 0.031), and chlorine (34% increase, p = 0.009) compared to no use. Maternal phthalate levels did not vary by reported plastic avoidance during pregnancy. The study showed that phthalate exposure is ubiquitous and increased by multiple factors. Future intervention studies to reduce phthalate levels among pregnant women will need to take into account the variety of sources identified in this study.

Concentrations of phthalate metabolites in Australian urine samples and their contribution to the per capita loads in wastewater.

Exposure to phthalates is a public health concern. In this study, we collected both urine and wastewater samples from 2012 to 2017 and analysed for 14 phthalate metabolites to assess human exposure to phthalates in Southeast Queensland (SEQ), and for associations between phthalate metabolites in urine and wastewater samples. Twenty-four pooled urine samples were prepared from 2400 individual specimens every two years (stratified by age, gender and collection year). Wastewater samples were collected from the three major wastewater treatment plants (WWTPs) representing locations in the SEQ region including a regional city, part of the state capital city and a third major urban WWTP in the region. Over the period, decreases for most phthalate metabolites, i.e. mono-butyl phthalate (MBP), mono-isobutyl phthalate (MiBP), monobenzyl phthalate (MBzP), monocyclohexyl phthalate (MCHP), mono(3-carboxypropyl) phthalate (MCPP), mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), mono(2-ethyl-5-oxohexyl) phthalate (MEOHP), and monomethyl phthalate (MMP), but an increase in monoethyl phthalate (MEP, particularly in young children) were observed in urine. In general, temporal changes were smaller in urine pools representing older age groups. We also found substantial variation in per capita mass loads of phthalate metabolites between samples from the three WWTPs with generally higher concentrations of most phthalates in the metropolitan areas. Per capita mass loads of most phthalate metabolites in wastewater were higher than would be expected from the per-capita excretion in urine, suggesting there are additional sources contributing to the majority of the observed phthalate metabolites in wastewater. For MEHHP and MEOHP we estimate that the urinary excretion accounts for a substantial fraction (average about 50%) of the mass load observed in the wastewater hence wastewater data may provide useful for monitoring trends in exposure.

Machine learning combined with non-targeted LC-HRMS analysis for a risk warning system of chemical hazards in drinking water: A proof of concept.

Guaranteeing clean drinking water to the global population is becoming more challenging, because of the cases of water scarcity across the globe, growing population, and increased chemical footprint of this population. Existing targeted strategies for hazard monitoring in drinking water are not adequate to handle such diverse and multidimensional stressors. In the current study, we have developed, validated, and tested a machine learning algorithm based on the data produced via non-targeted liquid chromatography coupled with high resolution mass spectrometry (LC-HRMS) for the identification of potential chemical hazards in drinking water. The machine learning algorithm consisted of a composite statistical model including an unsupervised component (i.e. principal component analysis PCA) and a supervised one (i.e. partial least square discrimination analysis PLS-DA). This model was trained using a training set of 20 drinking water samples previously tested via conventional suspect screening. The developed model was validated using a validation set of 20 drinking water samples of which 4 were spiked with 15 labeled standards at four different concentration levels. The model successfully detected all of the added analytes in the four spiked samples without producing any cases of false detection. The same validation set was processed via conventional trend analysis in order to cross validate the composite model. The results of cross validation showed that even though the conventional trend analysis approach produced a false positive detection rate of ≤5% the composite model outperformed that approach by producing zero cases of false detection. Additionally, the validated model went through an additional test with 42 extra drinking water samples from the same source for an unbiased examination of the model. Finally, the potentials and limitations of this approach were further discussed.

Multi-residue screening of non-polar hazardous chemicals in green turtle blood from different foraging regions of the Great Barrier Reef.

Green turtles spend a large part of their lifecycle foraging in nearshore seagrass habitats, which are often in close proximity to sources of anthropogenic contaminants. As most biomonitoring studies focus on a limited number of targeted chemical groups, this study was designed to screen for a wider range of hazardous chemicals that may not have been considered in prior studies. Whole blood of sub-adult green turtles (Chelonia mydas) were sampled from three different locations, a remote, offshore 'control' site; and two coastal 'case' sites influenced by urban and agricultural activities on the Great Barrier Reef in North Queensland, Australia. In order to screen blood samples for chemicals across a wide range of KOW's, a modified QuEChER's extraction method was used. The samples were analysed using a multi-residue gas chromatography with tandem mass spectrometry system (GC-MS/MS method that allowed simultaneous quantification of polychlorinated biphenyls (PCBs), polychlorinated diphenyl ethers (PBDES), organochlorine pesticides (OCPs) and polycyclic aromatic hydrocarbons (PAHs). While PBDEs, PCBs and OCPS were below the limits of quantification, PAHs were detected in all turtle blood samples. However, PAH levels were relatively low (maximum ΣPAH = 13 ng/mL ww) and comparable to or less than those reported from other green turtles globally. The present study provides the first baseline PAH levels in blood samples from green turtles from nearshore and offshore locations in the Southern Hemisphere.

Calibration and validation of a novel passive sampling device for the time integrative monitoring of per- and polyfluoroalkyl substances (PFASs) and precursors in contaminated groundwater.

Per-and polyfluoroalkyl substances (PFASs) as key components in aqueous film forming foams (AFFF) have led to growing incidences of environmental contamination. The aim of this study was to investigate a novel diffusion based passive sampling device comprising of microporous polyethylene (PE) for the long-term time-integrative monitoring of PFASs in groundwater systems. PE passive samplers (PEs) were deployed for 83 d and calibrated at five AFFF impacted groundwater sites representing different PFASs concentration levels (ΣPFAS 0.001 to 0.1 ng mL-1). Grab samples were collected simultaneously. Linear accumulation of 12 PFASs (r2 ≥ 0.84) were observed in the PEs over 83 d and PFASs sampling rates were 2-5 mL d-1. Estimated mean half-times to equilibrium for PFASs ranged between 122 and 490 d. A separate validation study compared PEs and grab sampling during a 93 d field deployment, at seven groundwater sites near a fire fighting training ground. Seventeen PFASs were detected in PEs and fifteen in grab samples. PEs showed higher sensitivity for precursors (i.e. 4:2 FTS and FOSA). Time-weighted-average water concentrations across all validation sites for all PFASs determined from PEs were strongly correlated (r2 = 0.98) with grab samples, (within range 0.3-60 ng mL-1 PFOS). Results represent the first application of passive sampling technology for the quantitative assessment of PFASs in groundwater systems.

Evaluating internal exposure of sea turtles as model species for identifying regional chemical threats in nearshore habitats of the Great Barrier Reef.

Marine megafauna that forage in proximity to land can be exposed to a diverse mixture of chemicals that - individually or combined - have the potential to affect their health. Characterizing such complex exposure and examining associations with health still poses considerable challenges. The present study summarizes the development and application of novel approaches to identifying chemical hazards and their potential impacts on the health of coastal wildlife, using green sea turtles as model species. We used an epidemiological study approach to collect blood and keratinized scute samples from free-ranging turtles foraging in nearshore areas and an offshore control site. These were analyzed using a combination of non-targeted, effect-based and multi-chemical analytical screening approaches to assess internal exposure to a wide range of chemicals. The screening phase identified a suite of elements (essential and non-essential) as priority for further investigation. Many of these elements are not commonly analyzed in marine wildlife, illustrating that comprehensive screening is important where exposure is unknown or uncertain. In particular, cobalt was present at highly elevated concentrations, in the order of those known to elicit acute effects across other vertebrate species. Several trace elements, including cobalt, were correlated with clinical indicators of impaired turtle health. In addition, biomarkers of oxidative stress (e.g. 3-indolepropionic acid and lipid peroxidation products) identified in the blood of turtles showed significant correlations with clinical health markers (particularly alkaline phosphatase and total bilirubin), as well as with cobalt. To assist interpretation of trace element blood data in the absence of sufficient information on reptile toxicity, we established exposure reference intervals using a healthy control population. In addition, trace element exposure history was investigated by establishing temporal exposure indices using steady-state relationships between blood and scute. Overall, the data provide a strong argument for the notion that trace element exposure is having an impact on the health of coastal sea turtle populations.

Non-targeted, high resolution mass spectrometry strategy for simultaneous monitoring of xenobiotics and endogenous compounds in green sea turtles on the Great Barrier Reef.

Chemical contamination poses a threat to ecosystem, biota and human health, and identifying these hazards is a complex challenge. Traditional hazard identification relies on a priori-defined targets of limited chemical scope, and is generally inappropriate for exploratory studies such as explaining toxicological effects in environmental systems. Here we present a non-target high resolution mass spectrometry environmental monitoring study with multivariate statistical analysis to simultaneously detect biomarkers of exposure (e.g. xenobiotics) and biomarkers of effect in whole turtle blood. Borrowing the concept from clinical chemistry, a case-control sampling approach was used to investigate the potential influence of xenobiotics of anthropogenic origin on free-ranging green sea turtles (Chelonia mydas) from a remote, offshore 'control' site; and two coastal 'case' sites influenced by urban/industrial and agricultural activities, respectively, on the Great Barrier Reef in North Queensland, Australia. Multiple biomarkers of exposure, including sulfonic acids (n=9), a carbamate insecticide metabolite, and other industrial chemicals; and five biomarkers of effect (lipid peroxidation products), were detected in case sites. Additionally, two endogenous biomarkers of neuroinflammation and oxidative stress were identified, and showed moderate-to-strong correlations with clinical measures of inflammation and liver dysfunction. Our data filtering strategy overcomes limitations of traditional a priori selection of target compounds, and adds to the limited environmental xenobiotic metabolomics literature. To our knowledge this is the first case-control study of xenobiotics in marine megafauna, and demonstrates the utility of green sea turtles to link internal and external exposure, to explain potential toxicological effects in environmental systems.

Screening of organic and metal contaminants in Australian humpback dolphins (Sousa sahulensis) inhabiting an urbanised embayment.

As a marine mammal species that inhabits shallow nearshore waters, humpback dolphins are likely exposed to a wide range of pollutants from adjacent land-based activities. Increased mortality rates of Australian humpback dolphins (Sousa sahulensis) in waters off a major urbanised centre triggered investigations into the threats to these species, including their contaminant exposure. The present study utilised archived tissues from 6 stranded animals to screen for a range of pollutants (PCDD/Fs, PBDEs, PCBs, organochlorine pesticides, PAHs, organotins, essential and non-essential elements) to inform future biopsy based biomonitoring strategies. Concentrations of PCBs and DDXs in blubber of some of these animals were remarkably high, at levels near or above toxicological thresholds associated with immune- and reproductive toxicity or population declines in other marine mammals. PBDEs, PAHs, HCB, organotins, 'drins' as well as other organic pesticides were not detected, or present at relatively low concentrations. Profiles of elements were similar in epidermis compared to other tissues, and apart from some exceptions (e.g. Fe, Cr, Co, Cu) their concentrations fell within 25th-75th percentiles of cetacean baselines in four of the five animals. Non-essential elements (Al, V, Pb, Ba, Ni, Cd) were notably elevated in one specimen which may have experienced poor health or nutritional status. These data provide a first insight into the contaminant status of a rare and poorly studied population inhabiting an urbanised area. The results highlight a need for future biomonitoring of live populations, and inform on priorities in the typically limited blubber and skin sample volumes obtained through biopsies.

C3 and 2D C3 Marfey's Methods for Amino Acid Analysis in Natural Products.

We validate the improved resolution and sensitivity of the C3 Marfey's method, including an ability to resolve all Ile isomers, against an array of amino acids commonly encountered in natural products and by comparison to an existing Marfey's method. We also describe an innovative 2D C3 Marfey's method as an analytical approach for determining the regiochemistry of enantiomeric amino acid residues in natural products. The C3 and 2D C3 Marfey's methods represent valuable tools for probing and defining the stereocomplexity of hydrolytically accessible amino acid residues in natural products.