Deep graph convolutional network for small-molecule retention time prediction.

The retention time (RT) is a crucial source of data for liquid chromatography-mass spectrometry (LCMS). A model that can accurately predict the RT for each molecule would empower filtering candidates with similar spectra but differing RT in LCMS-based molecule identification. Recent research shows that graph neural networks (GNNs) outperform traditional machine learning algorithms in RT prediction. However, all of these models use relatively shallow GNNs. This study for the first time investigates how depth affects GNNs' performance on RT prediction. The results demonstrate that a notable improvement can be achieved by pushing the depth of GNNs to 16 layers by the adoption of residual connection. Additionally, we also find that graph convolutional network (GCN) model benefits from the edge information. The developed deep graph convolutional network, DeepGCN-RT, significantly outperforms the previous state-of-the-art method and achieves the lowest mean absolute percentage error (MAPE) of 3.3% and the lowest mean absolute error (MAE) of 26.55 s on the SMRT test set. We also finetune DeepGCN-RT on seven datasets with various chromatographic conditions. The mean MAE of the seven datasets largely decreases 30% compared to previous state-of-the-art method. On the RIKEN-PlaSMA dataset, we also test the effectiveness of DeepGCN-RT in assisting molecular structure identification. By 30% lessening the number of potential structures, DeepGCN-RT is able to improve top-1 accuracy by about 11%.

Discovery of contaminants with antagonistic activity against retinoic acid receptor in house dust.

Retinoic acid receptors (RARs) control reproduction and development in vertebrates, but little attention has been paid to anthropogenic chemicals exhibiting RAR agoniztic/antagonistic activity. Here we applied a His-RARα pull-down assay combined with high-resolution mass spectrometry to identify chemicals with RARα activity in house dust. After screening, a total of 540 peaks were retained as potential RARα ligands. The mass spectra of 14 chemicals matched with those in the database, of which triphenyl phosphate, galaxolidone, di(2-ethylhexyl) phthalate (DEHP), tris(2-ethylhexyl) phosphate (TEHP), and tris(2-butoxyethyl) phosphate were confirmed by their standards. While one chemical in the sample matched with monophenyl phosphate in the MS/MS database, its retention time was much higher than that of monophenyl phosphate standard, suggesting that it may be an in-source fragment. Its parent ion was finally identified to be m/z 399.2663 using a similarity analysis among chromatographic peaks of hundreds of ions at the same retention time in MS1 spectrum, and bis(2-ethylhexyl) phenyl phosphate (BEHPP) was identified. BEHPP, DEHP, and TEHP were for the first time identified to be RARα antagonists with IC50 values of 6556, 6600, and 2538 nM, respectively. This study improved structural annotation and filled the knowledge gap regarding widespread environmental contaminants with RAR antagonistic activity.

Tris(1,3-dichloro-2-propyl)phosphate Induces Mass Mortality of Crucian Carp (Carassius carassius) Embryos in Taihu Lake.

Global fishery resources have been declining for decades, leading some fisheries to collapse. Although the decline is partly due to man-made chemical contamination, causal chemicals have been identified in only a few cases. We conducted consecutive 3-year investigations of embryonic mortality in Taihu Lake, China, including heavily contaminated northern areas, including Zhushan (ZS), Meiliang (ML), and Gonghu (GH), and the less polluted southeastern Suzhou (SZ). In 2016, 65.8% of crucian carp (Carassius carassius) embryos collected from ZS died before hatching, a substantially higher mortality rate than those observed in ML (21.7%), GH (15.2%), and SZ (2.2%). In 2017, the embryonic mortality rates were 38.8% in ZS, 1.3% in ML, 6.9% in GH, and 3.5% in SZ, and these rates strongly correlated with the concentrations of tris(1,3-dichloro-2-propyl)phosphate (TDCIPP): 104.2, 1.8, 4.6, and 4.1 ng/g lipid weight (lw) in embryos from ZS, ML, GH, and SZ, respectively. In 2018, embryonic mortality decreased to 4.0% in ZS and 1.2% in GH, consistent with decreases in embryonic TDCIPP concentrations to 17.1 and 1.5 ng/g lw, respectively. Moreover, the TDCIPP concentrations in dead embryos (70.5-216.8 ng/g lw) were much higher than those in live embryos (1.2-10.5 ng/g lw). Embryonic mortality was also observed in well-controlled laboratory experiments in which wild crucian carp were exposed to TDCIPP at concentrations similar to those measured in embryos collected from Taihu Lake, thus confirming TDCIPP as a causal factor in mass crucian carp embryo mortality in Taihu Lake. TDCIPP thus poses a threat to the sustainability of fisheries worldwide, given the high worldwide production volume of this chemical and its embryonic lethal toxicity.

Triphenyl phosphate delayed pubertal timing and induced decline of ovarian reserve in mice as an estrogen receptor antagonist.

Although concerns have been raised about the adverse effects of triphenyl phosphate (TPhP) on female fertility, its risk to ovarian functioning remains unknown. In this study, female C57BL/6 mice at postnatal day 21 were exposed on a daily basis to TPhP dose of 2, 10, and 50 mg/kg for 40 days. A significant delay in pubertal timing was observed in the mice exposed to 50 mg/kg of TPhP. An estrogen-responsive reporter transgenic mice assay demonstrated that TPhP significantly downregulated the estrogen receptor (ER) signaling by 45.1% in the whole body in the 50 mg/kg group, and by 14.7-43.7% in the uterus for all exposure groups compared with the control. This strong antagonistic activity of TPhP toward ER explained the delay in pubertal timing. A significant reduction in the number of follicles in all stages was observed in mice after being exposed to TPhP for 40 days at concentrations of 10 and 50 mg/kg, resulting in a decline of the ovarian reserve. The elevation of the follicle-stimulating hormone concentration may have contributed to this phenomenon, as controlled by the antagonistic activity of TPhP toward ER in the brain. The toxic effects of TPhP on ovarian functioning highlight this chemical as a potential risk factor for female fertility.

Identification of Three Novel Chloroalkyl Organophosphate Triesters in House Dust Using Halogenation-Guided Nontarget Screening Combined with Suspect Screening.

Several haloalkyl organophosphate triester (OPTE) flame retardants have been restricted in some countries due to their potential health risks, but the usage of alternative haloalkyl OPTEs is of concern. In this study, we comprehensively screened for haloalkyl OPTEs in house dust using high-resolution mass spectrometry. Through halogenation-guided nontarget screening, a rare chloroalkyl OPTE, diethylene glycol bis(bis(2-chloroisopropyl)phosphate) (DEGBBCPP), was unequivocally identified (Level 1) in house dust of Beijing, North China. In addition, by screening a suspect list of 61 haloalkyl OPTEs from the EPA's CompTox Chemicals Dashboard, we tentatively identified diethylene glycol bis(bis(2-chloroethyl)phosphate) (DEGBBCEP) and ethylene bis[bis(2-chloroethyl)phosphate] (EBBCEP) (Level 2). DEGBBCPP was detected in all 45 house dust samples, and the median concentration was 98.4 ng/g (13.6-6217 ng/g), that is, approximately one-half that of tris(1,3-dichloro-2-propyl) phosphate, a traditional high-production chloroalkyl OPTE. The detection frequencies of DEGBBCEP and EBBCEP were 96% and 98%, respectively, but at relatively low median concentrations of 10.6 ng/g (from not detected to 152 ng/g) and 3.79 ng/g (from not detected to 130 ng/g), respectively. In standard house dust SRM2585, DEGBBCEP and EBBCEP were detected at 160 ± 15.7 and 1897 ± 38.8 ng/g, respectively, but DEGBBCPP was not detected. Future studies should evaluate the potential adverse health effects of these emerging flame retardants.

Nontarget Discovery of 11 Aryl Organophosphate Triesters in House Dust Using High-Resolution Mass Spectrometry.

There is growing interest in the identification of novel aryl organophosphate triester (OPTE) congeners that may exist in the environment. In this study, we discovered 11 novel aryl OPTEs in north China house dust using a characteristic aryl phosphate fragment-guided high-resolution mass spectrometry method with data-independent acquisition. Tripentylated triphenyl phosphate (TPeTPhP), dicresyl phenyl phosphate (DCrPP), diisodecylphenyl phosphate (DIDPP), butoxyethoxyethyl octyl phenyl phosphate (BEEOPP), dioctyl nonylphenyl phosphate (DONPP), propoxypropyl bis(diphenyl phosphate) (PPBDPhP), octyl nonylphenyl phenyl phosphate (ONPPP), and (saturated mono-oxygen butoxybutyl) butoxyethoxyethyl phenyl phosphate (MBBPP) were detected in 84-100% of 45 samples. ONPPP was present in the highest median concentration of 69.0 ng/g, followed by DONPP (68.7 ng/g), DIDPP (50.3 ng/g), BEEOPP (42.5 ng/g), DCrPP (33.7 ng/g), PPBDPhP (25.0 ng/g), TPeTPhP (9.28 ng/g), and MBBPP (4.80 ng/g). Seven novel aryl OPTEs were also detected in standard house dust SRM2585, and the concentration of DIDPP (4375 ± 660 ng/g) was 4-fold higher than that (1048 ± 44.5 ng/g) of triphenyl phosphate, a typical aryl OPTE. The discovery of these novel OPTEs has significantly enriched our understanding of the aryl OPTEs present in house dust.

2-Ethylhexyl Diphenyl Phosphate and Its Hydroxylated Metabolites are Anti-androgenic and Cause Adverse Reproductive Outcomes in Male Japanese Medaka (Oryzias latipes).

Although high concentrations of 2-ethylhexyl diphenyl phosphate (EHDPP) have been detected in wild fish, its reproductive toxicity in fish remains unclear. In this study, we for the first time observed that EHDPP elicited androgen receptor (AR) antagonistic activity with a 50% inhibitory concentration of 37.5 µM. 2-Ethyl-5-hydroxyhexyl diphenyl phosphate was proved to be the dominant metabolite of EHDPP in Japanese medaka and elicited 3.1-fold stronger AR antagonistic activity than that of EHDPP. Medaka larvae (0-day post hatching) were exposed to EHDPP for 100 days, and intersex was observed in males from all exposure groups with significantly increased incidence (13.5-48.6%). 17ß-E2 was promoted at 104 ng/L, and androgens were suppressed at 434 ng/L, which account for the intersex incidence in the high-exposure groups but do not explain the significant incidence of intersex in the 29.9 ng/L exposure group. The AR antagonistic activity of EHDPP and its metabolites must therefore play a key role in intersex incidence. EHDPP also significantly (p < 0.05) repressed reproductive behaviors of males in the 434 ng/L group and decreased fertility in high-exposure groups compared with the control. All the adverse outcomes were observed under environmentally relevant concentrations, implying that EHDPP poses an ecological risk for wild fish populations.

Nontargeted identification of per- and polyfluoroalkyl substances in human follicular fluid and their blood-follicle transfer.

The female reproductive toxicity of per- and polyfluoroalkyl substances (PFAS) has raised concerns, but knowledge about their human preconception exposure is limited. In this study, 15 emerging PFAS were identified in follicular fluid samples from healthy women by using high-resolution mass spectrometry, and Cl-substituted perfluoroalkyl ether sulfonates (Cl-PFESAs) including 4:2, 5:2, 6:2, and 8:2 Cl-PFESAs, 4:4 C8 perfluoroalkyl ether sulfonate (PFESA), C8 perfluoroalkyl ether carboxylate (PFECA), and C8 polyether PFECA (Po-PFECA) were detected in over 50% of 28 follicular fluid samples. Ten legacy PFAS were also detected, and the geometric mean concentration of PFOS was the highest (4.82 ng/mL), followed by PFOA (4.60 ng/mL), 6:2 Cl-PFESA (1.09 ng/mL), PFHxS (0.515 ng/mL), PFNA (0.498 ng/mL), and C8 PFECA (0.367 ng/mL). The blood-follicle transfer efficiencies for PFCAs decreased with increasing chain length (0.96 for PFHpA, 0.56 for PFTriDA), and the transfer efficiencies of C8 PFECA (0.78) was significantly higher than that of PFOA (0.76). The transfer efficiencies of 4:2 Cl-PFESA (0.73), 6:2 Cl-PFESA (0.75) and 8:2 Cl-PFESA (0.91) were significantly higher than that (0.70) of PFOS (p = 0.028, 0.026 and 0.002, respectively). This study constitutes the first report of the human oocyte exposure to emerging PFAS and their blood-follicle transfer abilities.

Triphenyl phosphate modulated saturation of phospholipids: Induction of endoplasmic reticulum stress and inflammation.

Although triphenyl phosphate (TPHP) has been reported to disrupt lipid metabolism, the effect of TPHP on lipid saturation remains unexplored. In this study, a lipidomic analysis demonstrated decreases in the levels of poly-unsaturated phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylserine (PS) in RAW264.7 murine macrophage cells exposed to 10 µM TPHP. The expression of the gene encoding lysophosphatidylcholine acyltransferase 3 (Lpcat3) was significantly downregulated by 0.76 ± 0.03 and 0.70 ± 0.08-fold in 10 and 20 µM TPHP exposure groups, relative to the control group. This finding explains the observed decrease in lipid saturation. Correspondingly, exposure to 10 and 20 µM TPHP induced endoplasmic reticulum (ER) stress and inflammatory responses, which have been linked to metabolic dysfunction such as insulin resistance and hypertriglyceridemia. Therefore, TPHP may pose a risk to human health by promoting metabolic diseases.

Screening of House Dust from Chinese Homes for Chemicals with Liver X Receptors Binding Activities and Characterization of Atherosclerotic Activity Using an in Vitro Macrophage Cell Line and ApoE-/- Mice.

BACKGROUND: Atherosclerotic cardiovascular disease has become the leading cause of death worldwide, and environmental pollutants are increasingly recognized as risk factors for atherosclerosis. Liver X receptors (LXRs) play a central role in atherosclerosis; however, LXR activity of organic pollutants and associated potential risk of atherosclerosis have not yet been characterized. OBJECTIVES: This study aimed to explore whether LXR-antagonistic chemicals are present in indoor house dust and, if so, to characterize this activity in relation to changes in macrophages in vitro and cardiovascular disease indicators in vivo in an atherosclerosis ApoE-/- mouse model. METHODS: We used a His-LXRα-pull-down assay and a nontarget high-resolution mass spectrometry method to screen house dust collected from Chinese homes for LXRα- and LXRß-antagonist activity. A chemical identified in this manner was assessed for its ability to induce cholesterol efflux and foam cell formation in RAW264.7 macrophages, to down-regulate the expression of two LXR-dependent genes, ABCA1 and ABCG1, and finally to induce atherosclerotic lesions in vivo using an ApoE-/- mouse model. RESULTS: We identified the flame retardants triphenyl phosphate (TPHP) and 2-ethylhexyl diphenyl phosphate (EHDPP) in house dust samples and demonstrated their ability to antagonize LXRs. The potency of TPHP was similar to that of the LXR-antagonist SR9238. TPHP could also inhibit cholesterol efflux and promote foam cell formation in RAW264.7 macrophages and mouse peritoneal macrophages and significantly promoted atherosclerotic lesion formation in the ApoE-/- mouse model. CONCLUSIONS: We found LXR-antagonist chemicals in environmental samples of indoor dust from Chinese homes. One of the chemicals, TPHP, was able to promote the development of atherosclerotic lesions in the ApoE-/- mouse model. These results highlight the need to assess the LXR-antagonist activities of pollutants in future environmental management programs. https://doi.org/10.1289/EHP5039.

Contribution of phthalates and phthalate monoesters from drinking water to daily intakes for the general population.

Although phthalates (PAEs) are ubiquitous in drinking water, and phthalate monoesters (MPAEs) have been recognized as the bioactive metabolites of PAEs, little information is available regarding the occurrence of MPAEs in drinking water and the contributions of PAEs and MPAEs to human exposure. In this study, the concentrations of PAEs and MPAEs in 146 samples of drinking water collected from 24 cities throughout China were determined. The mean concentrations of dimethyl phthalate (DMP), diethyl phthalate (DEP), diisobutyl phthalate (DiBP), di-n-butyl phthalate (DnBP), and di-2-ethylhexyl phthalate (DEHP) were 14.31 ±â€¯26.28, 5.905 ±â€¯11.57, 103.8 ±â€¯310.5, 595.9 ±â€¯1794, and 178.2 ±â€¯422.0 ng/L, respectively. Monomethyl phthalate (MMP), monoethyl phthalate (MEP), monoisobutyl phthalate (MiBP), mono-n-butyl phthalate (MnBP), and mono-2-ethylhexyl phthalate (MEHP) were detected in drinking water for the first time, at mean concentrations of 12.1 ±â€¯18.0, 2.4 ±â€¯5.8, 11.3 ±â€¯37.2, 36.3 ±â€¯103, and 9.9 ±â€¯18.0 ng/L, respectively. The geometric mean concentrations of MMP, MEP, MiBP, MnBP, and MEHP in urine samples collected from 1040 participants from 16 cities were 10.1, 19.3, 29.6, 47.3, and 3.63 µg/g creatinine, respectively. The concentrations of PAEs and MPAEs in drinking water and daily intakes (DIs) of PAEs from nine cities where drinking water and urine samples were simultaneously collected were used to estimate the contributions from drinking water. The percentages of DMP, DEP, DiBP, DnBP, and DEHP from drinking water accounted for DIs of 0.60%, 0.049%, 1.26%, 2.76%, and 0.56%, respectively. The percentages of MMP, MEP, MiBP, MnBP and MEHP via intake of drinking water accounted for urinary concentrations of 0.86%, 0.032%, 0.14%, 0.089%, and 0.045%, respectively.

Relationship between perfluorooctanoate and perfluorooctane sulfonate blood concentrations in the general population and routine drinking water exposure.

In regions with heavily contaminated drinking water, a significant contribution of drinking water to overall human perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) exposure has been well documented. However, the relationship of PFOA/PFOS blood concentrations in the general population to routine drinking water exposure is not well characterized. This study determined the PFOA and PFOS concentrations in 166 drinking water samples across 28 cities in China. For 13 of the studied cities, PFOA and PFOS concentrations were analyzed in 847 human blood samples which were collected in parallel with the drinking water samples. The geometric mean PFOA and PFOS concentrations in drinking water were 2.5 ±â€¯6.2 ng/L and 0.7 ±â€¯11.7 ng/L, and population-weighted geometric mean blood concentrations were 2.1 ±â€¯1.2 ng/mL and 2.6 ±â€¯1.3 ng/mL, respectively. We found a significant correlation between the PFOA concentration in drinking water and blood (r = 0.87, n = 13, p < 0.001). The total daily intake of PFOA (0.24-2.13 ng/kg/day) and PFOS (0.19-1.87 ng/kg/day) were back-calculated from the blood concentrations with a one-compartment toxicokinetic model. We estimated relative source contributions (RSCs) of drinking water to total daily intake in China of 23 ±â€¯3% for PFOA and 12.7 ±â€¯5.8% for PFOS. Using the mean RSCs, we derived the health advisory values of 85 ng/L for PFOA and 47 ng/L for PFOS in China.

Urinary biomarkers for assessment of human exposure to monomeric aryl phosphate flame retardants.

While monomeric aryl organophosphate flame retardants (m-aryl-OPFRs) are used worldwide in a variety of consumer products, specific biomarkers for epidemiologic studies are lacking. To explore the potential of urinary hydroxylated metabolites of m-aryl-OPFRs as the biomarkers, we detected triphenyl phosphate (TPHP), 2-ethylhexyl diphenyl phosphate (EHDPP), and tricresyl phosphate (TCrP) in 259 whole blood samples and their 5 hydroxylated and 2 diester metabolites in the paired urine samples from the general population. 2-Ethyl-5-hydroxyhexyl diphenyl phosphate (5-OH-EHDPP), 4-hydroxyphenyl diphenyl phosphate (4-OH-TPHP), and 3-hydroxy-4-methylphenyl di-p-tolyl phosphate (3-OH-MDTP) were detected in >80% of urine samples after enzymatic hydrolysis of conjugates, and their concentrations showed significant positive correlations with the blood concentrations of their corresponding parent compounds, respectively. To characterize the temporal reliability, the m-aryl-OPFRs metabolites were also determined in urine samples repeated nine times from six volunteers over 3 months. Urinary 5-OH-EHDPP showed strong temporal reliability (creatinine-corrected intraclass correlation coefficients (ICCs), 0.77; 95% confidence interval [CI], 0.58 to 0.90), and urinary 3-OH-MDTP (creatinine-corrected ICC, 0.52; 95% CI, 0.37 to 0.87) and 4-OH-TPHP (0.56; 95% CI, 0.32 to 0.80) showed moderate-to-strong temporal reliability, while relatively weak temporal reliability was found for urinary DPHP (creatinine-corrected ICC, 0.37; 95% CI, 0.12 to 0.62). This study confirmed specific, reliable, and frequently detected biomarkers for TPHP and EHDPP and developed new biomarker of TCrP for future epidemiological research on health effects of m-aryl-OPFRs.

Screening of Chlorinated Paraffins and Unsaturated Analogues in Commercial Mixtures: Confirmation of Their Occurrences in the Atmosphere.

Characterizing the detailed compositions of chlorinated paraffins (CPs) commercial mixtures is crucial to understand their environmental sources, fates, and potential risks. In this study, dichloromethane (DCM)-enhanced UPLC-ESI-QTOFMS analysis combined with characteristic isotope chlorine peaks is applied to screen all CPs and their structural analogues in the three most commonly produced CP commercial mixtures (CP-42, CP-52, and CP-70). Mass fractions of total short-chain CPs (SCCPs), medium-chain CPs (MCCPs) and long-chain CPs (LCCPs) ranged from 0.64 to 31.9%, 0.64 to 21.8%, and 0.04 to 43.9%, respectively, in the three commercial mixtures. 113 unsaturated SCCPs, MCCPs, and LCCPs were identified in the commercial mixtures. The detailed mass percentages of saturated and unsaturated CPs with carbon numbers of 10-30, chlorine numbers of 5-28, and unsaturated degrees of 0-7 were characterized in all commercial mixtures. Occurrences of the predominant saturated and unsaturated CPs were further confirmed in air samples collected in Guangdong Province, one of the major CP production areas in China, over one year. The profiles of the detected compounds indicated that LCCPs in air samples might come mainly from the production and usage of CP-52, and unsaturated C24-29-LCCPs were specifically originated from CP-70 used in the area.