Identification of Mid-Polar and Polar AhR Agonists in Cetaceans from Korean Coastal Waters: Application of Effect-Directed Analysis with Full-Scan Screening.

Major aryl hydrocarbon receptor (AhR) agonists were identified in extracts of blubber, liver, and muscle from six long-beaked common dolphins (Delphinus capensis) and one fin whale (Balaenoptera physalus) collected from Korean coastal waters using effect-directed analysis. Results of the H4IIE-luc bioassay indicated that the polar fractions of blubber and liver extracts from the fin whale exhibited relatively high AhR-mediated potencies. Based on full-scan screening with high-resolution mass spectrometry, 37 AhR agonist candidates, spanning four use categories: pharmaceuticals, pesticides, cosmetics, and natural products, were selected. Among these, five polar AhR agonists were newly identified through toxicological confirmation. Concentrations of polar AhR agonists in cetaceans were tissue-specific, with extracts of blubber and liver containing greater concentrations than muscle extracts. Polar AhR agonists with great log KOA values (>5) were found to biomagnify in the marine food chain potentially. Polar AhR agonists contributed 8.9% of the observed AhR-mediated potencies in blubber and 49% in liver. Rutaecarpine and alantolactone contributed significantly to the total AhR-mediated potencies of blubber, whereas hydrocortisone was a major AhR contributor in the liver of the fin whale. This study is the first to identify the tissue-specific accumulation of polar AhR agonists in blubber and liver extracts of cetaceans.

Use of molecular composition and compound-specific isotope analysis for source appointment of PAHs in sediments of a highly industrialized area.

This study utilized both conventional molecular analysis and compound-specific isotopic techniques to identify the sources of polycyclic aromatic hydrocarbons (PAHs) in sediments of Ulsan Bay, South Korea. The concentrations of 15 traditional and 11 emerging PAHs were determined in sediments from 21 source sites and 26 bay sites. The concentrations and compositions of traditional and emerging PAHs varied significantly, even at sites close to the source areas. The results obtained from diagnostic ratios and the positive matrix factorization model for source identification were inconsistent in adjacent source areas. The δ13C profiles of PAHs, such as phenanthrene (Phe), fluoranthene (Fl), pyrene (Py), and benz[a]anthracene (BaA) in the sediments showed distinct features depending on the surrounding sources. In urban sediments, lighter δ13CPhe values were observed (mean: -25.1‰), whereas relatively heavier values of δ13CPy were found in petroleum industry areas (mean: -23.4‰). The Bayesian isotope mixing model indicates that the predominant source of PAHs in Ulsan Bay sediments was the petroleum industry (45%), followed by the non-ferrous metals industry (30%), automobile industry (18%), and urban areas (6.3%). These results demonstrated the utility of stable isotopes in assessing the sources and contributions of PAHs in small-scale regions. However, there are still limitations in compound-specific isotope analysis of PAHs, including insufficient end-members for each source, difficulty in analysis, and the influence of non-point sources; thus, further study is needed to expand its application.

Monthly distribution of lipophilic marine biotoxins and associated microalgae in the South Sea Coast of Korea throughout 2021.

Aquaculture farms have been established along the South Sea Coast of Korea, supplying most of the seafood consumed domestically. However, annual harmful algal blooms pose a potential threat to seafood safety. This study aimed to determine the spatial and seasonal distributions of 12 lipophilic marine biotoxins (LMTs) in phytoplankton and mussels in the region in 2021. Solid-phase adsorption toxin tracking (SPATT) was used to monitor the cumulative compositions of LMTs in seawater. LMT concentrations were also determined in twelve commercially available species of domestic shellfish to evaluate the potential risks to human health. Gonyaulux spinifera and Dinophysis acuminata, causative microalgae of yessotoxins (YTXs) and pectenotoxins (PTXs), respectively, showed high densities in the region from May to July. This period corresponded to high LMT concentrations in phytoplankton and mussels. Phytoplankton mainly contained PTX-2 and homo-YTX, with a maximum concentration of 2300 ng g-1 wet weight (ww) in May. In contrast, mussels mainly contained homo-YTX and YTX, with a maximum concentration of 1300 ng g-1 ww in July. LMTs-producing microalgae showed low densities and concentrations after July, whereas mussels accumulated toxins until September. In the SPATT sampler, more diverse LMTs were detected than in seawater, phytoplankton, and mussels. For example, dinophysistoxin-1 and azaspiracid-2 were detected only in SPATT. YTXs were detected in domestic seafood samples, including mussels, red scallops, and pen shells, but the concentrations were below the European Food Safety Agency recommended standard of 3.75 mg YTX-eq. kg-1. Moreover, the hazard quotient was less than 100 in all scenarios, indicating that the human health risk was not significant. This study provides valuable data on monthly distribution patterns of LMTs in the South Sea Coast of Korea and can serve as baseline data for future management policies of marine biotoxins.

Characterization of microalgal toxicants in the sediments from an industrial area: Application of advanced effect-directed analysis with multiple endpoint bioassays.

Microalgal toxicants in sediments from an industrialized area (Ulsan Bay) in South Korea were identified using effect-directed analysis (EDA) with full-scan screening analysis (FSA) and microalgal bioassays with multiple endpoints. The growth rate and cell viability of three microalgae (Isochrysis galbana, Dunaliella tertiolecta, and Phaeodactylum tricornutum) were strongly inhibited following exposure to raw organic extracts of sediments from Site D5 (Woehang River). The polar fraction separated using a silica gel column significantly inhibited growth rate, esterase activity, cell membrane intensity, and chlorophyll a autofluorescence. In comparison, non- and mid-polar fractions induced non-toxic or esterase inhibition. Target toxicants, such as polycyclic aromatic hydrocarbons, styrene oligomers, and alkylphenols, were detected at low concentrations (450, 79, and 98 ng g-1 dw, respectively) in the sediment of D5, indicating the presence of unmonitored toxicants. FSA was performed for the polar fraction using LC-QTOFMS, and 31 candidates of toxicants were selected. Toxicological confirmation was conducted for 7 candidates for which standards are available. Out of these, 2-nitrophenol, 3-nitrophenol, and 4-nitrophenol showed significant microalgal toxicity; however, these compounds did not fully explain the induced toxicity. Overall, combining EDA and FSA with multiple endpoint bioassays demonstrated the benefits of characterizing the microalgal toxicants in the environments.

Molecular Characterization of Estrogen Receptor Agonists during Sewage Treatment Processes Using Effect-Directed Analysis Combined with High-Resolution Full-Scan Screening.

Endocrine-disrupting potential was evaluated during the sewage treatment process using in vitro bioassays. Aryl hydrocarbon receptor (AhR)-, androgen receptor (AR)-, glucocorticoid receptor (GR)-, and estrogen receptor (ER)-mediated activities were assessed over five steps of the treatment process. Bioassays of organic extracts showed that AhR, AR, and GR potencies tended to decrease through the sewage treatment process, whereas ER potencies did not significantly decrease. Bioassays on reverse-phase high-performance liquid chromatography fractions showed that F5 (log KOW 2.5-3.0) had great ER potencies. Full-scan screening of these fractions detected two novel ER agonists, arenobufagin and loratadine, which are used pharmaceuticals. These compounds accounted for 3.3-25% of the total ER potencies and 4% of the ER potencies in the final effluent. The well-known ER agonists, estrone and 17ß-estradiol, accounted for 60 and 17% of the ER potencies in F5 of the influent and primary treatment, respectively. Fourier transform ion cyclotron resonance mass spectrometry analysis showed that various molecules were generated during the treatment process, especially CHO and CHOS (C: carbon, H: hydrogen, O: oxygen, and S: sulfur). This study documented that widely used pharmaceuticals are introduced into the aquatic environments without being removed during the sewage treatment process.

Spatiotemporal distribution characteristics of yessotoxins and pectenotoxins in phytoplankton and shellfish collected from the southern coast of South Korea.

The distribution characteristics of lipophilic marine biotoxins (LMTs), such as yessotoxins (YTXs) and pectenotoxins (PTXs) in phytoplankton, mussels, and commercial seafood were determined for the southern coast of South Korea. Gonyaulax spinifera and Dinophysis acuminata, which are the causative microalgae of YTXs and PTXs, were recorded during summer. Homo-YTX and PTX-2 were predominantly detected in phytoplankton (max: 5.7 µg g-1 ww), whereas only YTXs were detected in mussels (max: 1.1 µg g-1 ww). LMT concentrations in mussels were positively correlated with those in phytoplankton. However, there was a 1-month time gap in maximum LMT concentrations between mussels and phytoplankton. Homo-YTX was detected in commercial seafood, including red scallop and comb pen shell. However, homo-YTX concentrations in shellfish were below the recommended value of the European Food Safety Authority (3.75 mg YTX equivalents kg-1); thus, the consumption of this seafood was not considered to be a significant risk for human health.

Identification of novel polar aryl hydrocarbon receptor agonists accumulated in liver of black-tailed gulls in Korea using advanced effect-directed analysis.

Although bioaccumulation of persistent organic pollutants in seabirds has been examined, few studies have been conducted to identify previously unidentified substances. Here, aryl hydrocarbon receptor (AhR) agonists were identified in livers of black-tailed gulls from South Korea using effect-directed analysis combined with full-scan screening analysis. Significant AhR-mediated potencies were observed in the polar fractions of liver extracts using H4IIE-luc bioassay. Eight known polar AhR agonists accounted for 11-20% of the total AhR-mediated potencies in the polar fractions; hydrocortisone and rutaecarpine were the major contributors. Twenty-two AhR agonist candidates in the polar fractions were identified using liquid chromatography-quadrupole time-of-flight mass spectrometry during a six-step selection process. Of these, [10]-gingerol, angelicin, corticosterone, eupatilin, etofenprox, oxadixyl, and tretinoin were identified as novel AhR agonists. The contribution to potencies increased with inclusion of novel AhR agonists (27-52%); corticosterone and [10]-gingerol contributed significantly. Quantitative structure-activity relationship suggested that the novel AhR agonists have other potential toxic effects, including carcinogenicity and mutagenicity. Polar AhR agonists have been used for pharmaceuticals and pesticides. Some novel AhR agonists have log KOW > 2 and log KOA ≥ 6, which indicates that these compounds can be biomagnified in air-breathing organisms, such as seabirds.

Multimedia distributions and the fate of microcystins from freshwater discharge in the Geum River Estuary, South Korea: Applicability of POCIS for monitoring of microalgal biotoxins.

Here, we investigated the characteristics of the environmental multimedia distribution of microcystins (MCs) introduced from freshwater discharge through the estuary dam of the Geum River. In addition, the applicability of a passive sampling device (polar organic chemical integrative sampler, POCIS) for monitoring MCs was evaluated. Surface water, suspended solids (SS), sediments, and oysters were collected from the inner and outer estuary dam. Seven MC variants were analyzed using HPLC-MS/MS. POCIS was deployed at three sites over one week, and MCs were monitored for four weeks from August to September 2019. Before POCIS was deployed in the field, compounds-specific sampling rates of MCs were determined as functions of water temperature (10, 20, and 30 °C), flow rate (0, 0.38, and 0.76 m s-1), and salinity (0, 15, and 30 psu) in the laboratory. The sampling rates of MCs in POCIS increased significantly with increasing water temperature and flow rate, whereas salinity did not significantly affect the sampling rates between freshwater and saltwater. The MCs in the Geum River Estuary mainly existed as particulate forms (mean: 78%), with relatively low proportions of dissolved forms (mean: 22%), indicating that MCs were mainly contained in cyanobacterial cells. There was no significant correlation among the concentrations of MCs in water, SS, sediments, and oysters. Time-weighted average concentrations of MCs from POCIS were not significantly correlated with the concentrations of MCs in water and oysters. The metabolites of MCs, including MC-LR-GSH, MC-LR-Cys, MC-RR-GSH, and MC-RR-Cys, were detected in oysters (no metabolites were detected in POCIS). Overall, POCIS can be useful for monitoring dissolved MCs in the aquatic ecosystem, particularly in calculating time-weighted average concentrations, but it seems to have limitations in evaluating the contamination status of total MCs, mainly in particulate form.

Novel polar AhR-active chemicals detected in sediments of an industrial area using effect-directed analysis based on in vitro bioassays with full-scan high resolution mass spectrometric screening.

Studies investigating aryl hydrocarbon receptor (AhR)-active compounds in the environment typically focus on non- and mid-polar substances, such as PAHs; while, information on polar AhR agonists remains limited. Here, we identified polar AhR agonists in sediments collected from the inland creeks of an industrialized area (Lake Sihwa, Korea) using effect-directed analysis combined with full-scan screening analysis (FSA; using LC-QTOFMS). Strong AhR-mediated potencies were observed for the polar and latter fractions of RP-HPLC (F3.5-F3.8) from sediment organic extracts in the H4IIE-luc in vitro bioassays. FSA was performed on the corresponding fractions. Twenty-eight tentative AhR agonists were chosen using a five-step process. Toxicological confirmation using bioassay revealed that canrenone, rutaecarpine, ciprofloxacin, mepanipyrim, genistein, protopine, hydrocortisone, and medroxyprogesterone were significantly active. The relative potencies of these AhR-active compounds compared to that of benzo[a]pyrene ranged from 0.00002 to 2.0. Potency balance analysis showed that polar AhR agonists explained, on average, ~6% of total AhR-mediated potencies in samples. Some novel polar AhR agonists also exhibited endocrine-disrupting potentials capable of binding to estrogen and glucocorticoid receptors, as identified by QSAR modeling. In conclusion, the focused studies on distributions, sources, fate, and ecotoxicological effects of novel polar AhR agonists in the environment are necessary.

Distribution of microcystins in environmental multimedia and their bioaccumulation characteristics in marine benthic organisms in the Geum River Estuary, South Korea.

Spatio-temporal distributions and bioaccumulation characteristics of freshwater cyanobacterial toxins, such as microcystins (MCs) in the Geum River Estuary, South Korea, were investigated during summer. Environmental multimedia samples (water, suspended particulate matter (SPM), and sediments) and tidal flat organisms (polychaetes, decapods, amphipods, and bivalves) were collected from regions inside and outside of the estuary dam for MCs analysis. Phytoplankton communities in the Geum River (freshwater) and estuarine area (brackish water) were also analyzed in order to understand the relationship with MCs concentrations. Seasonal variation in the structure of phytoplankton communities was detected in the Geum River, with a relatively high density of Cyanophyta in summer. MC concentrations were strongly correlated to water temperature, chlorophyll a, and cyanobacterial density. MC-LR was the most abundant MC variants in environmental samples. Dissolved MCs remained for longer periods and were more widely distributed in the coastal environments compared to particulate MCs. The distribution coefficients between water and SPM (Kd-SPM) and between water and sediments (Kd-sediment) of MCs showed that the phase shift of MCs in the environmental samples occurred in the estuary. Kd-SPM declined from the inside to outside regions of the estuary dam, and was mainly attributed to differences in the half-lives of MCs in dissolved (4.7 d for MC-LR) and particulate phases (0.44-0.52 d for MC-LR). Species-specific bioaccumulation of MCs occurred in tidal flat organisms, with relatively high bioaccumulation factors of MCs being detected in polychaetes and decapods compared to amphipods and bivalves. Overall, this study advances our understanding on the distribution, transport, fate, and bioaccumulation of MCs in estuarine and coastal environments.

Trophic transfer of persistent toxic substances through a coastal food web in Ulsan Bay, South Korea: Application of compound-specific isotope analysis of nitrogen in amino acids.

Trophic magnification factor (TMF) of persistent toxic substances (PTSs: Hg, PCBs, PAHs, and styrene oligomers (SOs)) in a coastal food web (12 fish and four invertebrates) was determined in Ulsan Bay, South Korea. The nitrogen stable isotope ratios (δ15N) of amino acids [δ15NGlu-Phe based on glutamic acid (δ15NGlu) and phenylalanine (δ15NPhe)] were used to estimate the trophic position (TPGlu-Phe) of organisms. The TPGlu-Phe of organisms ranged from 1.64 to 3.69, which was lower than TP estimated by δ15N of bulk particulate organic matter (TPBulk: 2.46-4.21). Mercury and CB 138, 153, 187, and 180 were biomagnified through the whole food web (TMF > 1), while other PTSs, such as PAHs and SOs were not (biodilution of SOs firstly reported). In particular, the trophic transfer of PTSs was pronounced in the resident fish (e.g., rock bream, sea perch, Korean rockfish). Of note, CB 99, 101, 118, and 183 were additionally found to be biomagnifying PTSs in these species. Thus, fish residency appears to represent an important factor in determining the TMF of PTSs in the coastal environment. Overall, δ15NGlu-Phe provided accurate TPs of organisms and could be applied to determine the trophic transfer of PTSs in coastal food webs.

Study on synthesis and application of tetrabasic lead sulfate as the positive active material additive for lead-acid batteries.

Tetrabasic lead sulfate (4BS) was used as a positive active material additive for lead-acid batteries, which affirmatively affected the performance of the battery. Herein, tetrabasic lead sulfate was synthesized from scrap lead paste that was formed through the production process of the lead-acid batteries. This solves the disposing problem of the scrap lead paste that is challenging in the production of the lead-acid batteries. Scrap lead paste was first pre-treated and the 4BS with high purity and crystalline was synthesized by sintering at the temperature of 450°C and hold time of 7 h. As demonstrated by X-ray diffraction and scanning electron microscopy test and Material Studio software calculation, the purity of synthesized 4BS is higher than 98 wt%, small particles have pillar forms and are evenly distributed. Moreover, the synthesized 4BS of 1 wt% was added to the positive lead paste and then valve-regulated lead-acid battery was made after the pasting, curing and formation processes. The effectiveness of the lead-acid batteries after adding 4BS as crystal seeds was evaluated, and the 100% charge-discharge cycle life of the new battery (523 times) was about 1.4 times higher than that of general lead-acid batteries (365 times).