Insights into the combined toxicity and mechanisms of BDE-47 and PFOA in marine blue mussel: An integrated study at the physiochemical and molecular levels.

The coexistence of multiple emerging contaminants imposes a substantial burden on the ecophysiological functions in organisms. The combined toxicity and underlying mechanism requires in-depth understanding. Here, marine blue mussel (Mytilus galloprovincialis L.) was selected and exposed to 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) and perfluorooctanoic acid (PFOA) individually and in combination at environmental related concentrations to elucidate differences in stress responses and potential toxicological mechanisms. Characterization and comparison of accumulation, biomarkers, histopathology, transcriptomics and metabolomics were performed. Co-exposure resulted in differential accumulation patterns, exacerbated histopathological alterations, and different responses in oxidative stress and biomarkers for xenobiotic transportation. Moreover, the identified differentially expressed genes (DEGs) and differential metabolites (DEMs) in mussels were found to be annotated to different metabolic pathways. Correlation analyses further indicated that DEGs and DEMs were significantly correlated with the above biomarkers. BDE-47 and PFOA altered the genes and metabolites related to amino acid metabolism, energy and purine metabolism, ABC transporters, and glutathione metabolism to varying degrees, subsequently inducing accumulation differences and combined toxicity. Furthermore, the present work highlighted the pivotal role of Nrf2-keap1 detoxification pathway in the acclimation of M. galloprovincialis to reactive oxygen species (ROS) stress induced by BDE-47 and PFOA. This study enabled more comprehensive understanding of combined toxic mechanism of multi emerging contaminants pollution.

Phenylhydrazone Alkaloids from the Deep-Sea Cold Seep Derived Fungus Talaromyces amestolkiae HDN21-0307.

Alkaloids with a phenylhydrazone architecture are rarely found in nature. Four unusual phenylhydrazone alkaloids named talarohydrazones A-D (1-4) were isolated from the deep-sea cold seep derived fungus Talaromyces amestolkiae HDN21-0307 using the one strain-many compounds (OSMAC) approach and MS/MS-based molecular networking (MN) combined with network annotation propagation (NAP) and the unsupervised substructure annotation method MS2LDA. Their structures were elucidated by spectroscopic data analysis, single-crystal X-ray diffraction, and quantum chemical calculations. Talarohydrazone A (1) possessed an unusual skeleton combining 2,4-pyridinedione and phenylhydrazone. Talarohydrazone B (2) represents the first natural phenylhydrazone-bearing azadophilone. Bioactivity evaluation revealed that compound 1 exhibited cytotoxic activity against NCI-H446 cells with an IC50 value of 4.1 µM. In addition, compound 1 displayed weak antibacterial activity toward Staphylococcus aureus with an MIC value of 32 µg/mL.

Metabolites and metabolic pathway analysis of sulfadimidine in carp (Cyprinus carpio) based on UHPLC-Q-orbitrap HRMS.

Sulfadimidine (SM2) is an N-substituted derivative of p-aminobenzenesulfonyl structure. This study aimed to analyze the metabolism of SM2 in carp (Cyprinus carpio). The carps were fed with SM2 at a dose of 200 mg/(kg · bw) and then killed. The blood, muscle, liver, kidney, gill, other guts, and carp aquaculture water samples were collected. The UHPLC-Q-Exactive Plus Orbitrap-MS was adopted for determining the metabolites of SM2 in the aforementioned samples. Twelve metabolites, which were divided into metabolites in vivo and metabolites in vitro, were identified using Compound Discoverer software. The metabolic pathways in vivo of SM2 in carp included acetylation, hydroxylation, glucoside conjugation, glycine conjugation, carboxylation, glucuronide conjugation, reduction, and methylation. The metabolic pathways in vitro included oxidation and acetylation. This study clarified the metabolites and metabolic pathways of SM2 in carp and provided a reference for further pharmacodynamic evaluation and use in aquaculture.

Integrative Inducer Intervention and Transcriptomic Analyses Reveal the Metabolism of Paralytic Shellfish Toxins in Azumapecten farreri.

Paralytic shellfish toxins (PSTs) are widely distributed neurotoxins, and the PST metabolic detoxification mechanism in bivalves has received increasing attention. To reveal the effect of phase I (cytochrome P450)-II (GST)-III (ABC transport) metabolic systems on the PST metabolism in Azumapecten farreri, this study amplified stress on the target systems using rifampicin, dl-α-tocopherol, and colchicine; measured PST levels; and conducted transcriptomic analyses. The highest toxin content reached 1623.48 µg STX eq/kg in the hepatopancreas and only 8.8% of that in the gills. Inducer intervention significantly decreased hepatopancreatic PST accumulation. The proportional reductions in the rifampicin-, dl-α-tocopherol-, and colchicine-induced groups were 55.3%, 50.4%, and 36.1%, respectively. Transcriptome analysis showed that 11 modules were significantly correlated with PST metabolism (six positive/five negative), with phase I CYP450 and phase II glutathione metabolism significantly enriched in negatively correlated pathways. Twenty-three phase I-II-III core genes were further validated using qRT-PCR and correlated with PST metabolism, revealing that CYP46A1, CYP4F6, GSTM1, and ABCF2 were significantly correlated, while CYP4F11 and ABCB1 were indirectly correlated. In conclusion, phase I-II-III detoxification enzyme systems jointly participate in the metabolic detoxification of PSTs in A. farreri. This study provides key data support to profoundly elucidate the PST metabolic detoxification mechanism in bivalves.

A novel ZIF-L/PEI thin film nanocomposite membrane for removing perfluoroalkyl substances (PFASs) from water: Enhanced retention and high flux.

Membrane separation technology is widely recognized as an effective method for removing perfluoroalkyl substances (PFASs) in water treatment. ZIF-L, a metal-organic framework (MOF) family characterized by its mat-like cavities and leaf-like morphology, has garnered considerable interest and has been extensively employed in fabricating thin-film nanocomposite (TFN) membranes. In this study, a robust, high-performance TFN membrane to remove PFASs in a nanofiltration (NF) process was created through an interfacial polymerization approach on the surface of polysulfone (PSF), incorporating ZIF-L within the selective layer. The TFN membrane modified by adding 5 wt% ZIF-L (relative to the weight of ethylene imine polymer (PEI)) exhibits 2.3 times higher water flux (up to 47.56 L·m-2·h-1·bar-1) than the pristine thin film composite membrane (20.46 L·m-2·h-1·bar-1), and the rejection for typical PFASs were above 95 % (98.47 % for perfluorooctanesulfonic acid (PFOS) and 95.85 % for perfluorooctanoic acid (PFOA)). The effectiveness of the ZIF-L/PEI TFN membrane in retaining representative PFASs was examined under various conditions, including different pressures, feed concentrations, aqueous environments, and salt ions. Notably, the experiments demonstrated that even after contamination with humic acid (HA), >88 % of the water flux could be restored by washing. Additionally, density functional theory (DFT) calculations were employed to predict the distinct intermolecular interactions between PFASs and ZIF-L as well as PEI. These calculations provide additional insights into the interception mechanism of TFN membranes towards PFASs. Based on this study, TFN membranes incorporating MOF as nanofillers show great potential as an effective method for purifying PFASs from aqueous environments and possess superior environmental sustainability and cost-effectiveness.

Contamination Status and Risk Assessment of Paralytic Shellfish Toxins in Shellfish along the Coastal Areas of China.

Paralytic shellfish toxins (PSTs) are widely distributed in shellfish along the coast of China, causing a serious threat to consumer health; however, there is still a lack of large-scale systematic investigations and risk assessments. Herein, 641 shellfish samples were collected from March to November 2020, and the PSTs' toxicity was detected via liquid chromatography-tandem mass spectrometry. Furthermore, the contamination status and potential dietary risks of PSTs were discussed. PSTs were detected in 241 shellfish samples with a detection rate of 37.60%. The average PST toxicities in mussels and ark shells were considerably higher than those in other shellfish. The PSTs mainly included N-sulfonylcarbamoyl toxins (class C) and carbamoyl toxins (class GTX), and the highest PST toxicity was 546.09 µg STX eq. kg-1. The PST toxicity in spring was significantly higher than those in summer and autumn (p < 0.05). Hebei Province had the highest average PST toxicity in spring. An acute exposure assessment showed that consumers in Hebei Province had a higher dietary risk, with mussels posing a significantly higher dietary risk to consumers. This research provides reference for the green and sustainable development of the shellfish industry and the establishment of a shellfish toxin prevention and control system.

Risk characteristics of shellfish toxins in Mytilus unguiculatus around the Zhoushan Islands, East China Sea.

The Zhoushan Islands, are an important area for Mytilus unguiculatus aquaculture, and are threatened by potentially harmful algal blooms. However, a full understanding of the risks posed by their toxin residues is still lacking. M. unguiculatus samples were collected from the area between 2020 and 2021 and analyzed for their toxin profiles to assess the contamination status of shellfish toxins. The main toxins detected were the paralytic shellfish toxins (PSTs), gymnodimine (GYM), and domoic acid (DA). Nine PSTs components were detected, the dominant ones being C1, C2, and GTX5, with an overall detection rate of 85.7 %. The detection rate of DA was 55.05 %, and GYM was detected in all samples. The toxin levels in the samples were significantly lower than the European Union regulatory limits, but toxin contamination was generally universal.

Adaptive responses of geographically distinct strains of the benthic dinoflagellate, Prorocentrum lima (Dinophyceae), to varying light intensity and photoperiod.

The toxic Prorocentrum lima complex can potentially cause serious harm to the benthos and entire food chain. Studies have revealed physiological differences in strains from different regions related to local environment, while differences in the adaptive responses of P. lima complex should be urgently assessed. Hence, this study explored the adaptive responses to varying light intensities and photoperiods of two P. lima complex strains SHG101 and 3XS34, isolated from the Bohai Sea and the South China Sea, respectively. We found the highest cell density of 7.49 × 104 cells mL-1 recorded in the 3XS strain in the stationary phase with high light intensity exposure. No significant difference was observed in growth rate among SHG groups, however, significant differences were found among 3XS groups ranging from 0.176 to 0.311 d-1. Three key pigments Chl a, Peri, and Fuco accounted for up to 60% of the total pigments. Production and concentrations of pigments and Fv/Fm values exhibit a significant negative correlation with high light intensity and growth. Conversely, total diarrhetic shellfish toxin content and the proportion of diol esters increased to varying degrees after high intensity light exposure, with 3XS strain under high light intensity and a photoperiod of light and darkness (12L:12D) consistently exhibiting the highest levels, finally reaching a maximum (21.6 pg cell-1) at day 28. A shortened photoperiod of high light intensity (8L:16D) resulted in impaired recovery compared with 12L:12D. Furthermore, 3XS showed more delayed and intense adaptive responses, indicating a stronger tolerance compared to SHG. Collectively, these results directly characterized variation in the adaptive responses of geographically distinct strains of P. lima complex, highlighting the previously ignored potential risk diversity of this species.

Variation and characterization of prometryn in oysters (Crassostrea gigas) after seawater exposure.

Prometryn (PRO) is frequently detected in shellfish of international trade among triazine herbicides because of its wide application in agriculture and aquaculture worldwide. Nevertheless, the variations of PRO remain unclear in aquatic organisms, which affect the accuracy of their food safety risk assessment. In the present study, the tissue-specific accumulation, biotransformation, and potential metabolic pathway of PRO were reported in oyster species Crassostrea gigas for the first time. The experiments were conducted through semi-static seawater exposure with low and high concentrations of PRO (at nominal concentrations of 10 and 100 µg/L) via daily renewal over 22 days, followed by 16 days of depuration in clean seawater. The characterization of prometryn in oysters was then evaluated through the bioaccumulation behavior, elimination pathway and metabolic transformation, comparing with other organisms. The digestive gland and gonad were found to be the main target organs during uptake. In addition, the highest bioconcentration factor of 67.4 ± 4.1 was observed when exposed to low concentration. The level of PRO in oyster tissues rapidly decreased within 1 day during depuration, with an elimination rate of >90 % for the gill. Moreover, four metabolites of PRO were identified in oyster samples of exposed groups, including HP, DDIHP, DIP, and DIHP, in which HP was the major metabolite. Considering the mass percentage of hydroxylated metabolites higher than 90 % in oyster samples, PRO poses a larger threat to aquatic organisms than rat. Finally, the biotransformation pathway of PRO in C. gigas was proposed, the major metabolic process of which was hydroxylation along with N-dealkylation. Meanwhile, the newly discovered biotransformation of PRO in oyster indicates the importance of monitoring environmental levels of PRO in cultured shellfish, to prevent possible ecotoxicological effects as well as to ensure the safety of aquatic products.

Perfluoroalkyl substances (PFASs) in aquatic products from the Yellow-Bohai Sea coasts, China: Concentrations and profiles across species and regions.

Perfluoroalkyl substances (PFASs) are emerging contaminants capable of harming human health, primarily via ingesting aquatic products. The current study monitored a survey of 23 PFASs in 1049 aquatic products from the coasts of the Yellow-Bohai Sea in China to comprehensively investigate the concentrations and distributions of PFASs. PFOA, PFOS, PFNA, PFOSA, and PFUdA were more predominantly and frequently detected than other PFASs in all samples, dominating PFAS patterns in aquatic products. The mean levels of ∑PFASs in different species followed the order: marine shellfish > marine crustaceans > fish > cephalopods > sea cucumber. Profiles of PFASs differ between species, suggesting species-specific accumulation plays a role. Various aquatic species are potential environmental bioindicators that signal individual PFAS contamination. For instance, clams can act as a potential PFOA bioindicator. High ∑PFAS levels in some sites (such as Binzhou, Dongying, Cangzhou, and Weifang) could be attributed to industrial activities involving fluoropolymer manufacture. The differences between PFAS concentrations and profiles in aquatic products across the study regions have been proposed as PFAS fingerprints of the Yellow-Bohai Sea coasts. Analyses of principal components and Spearman correlations indicated that the precursor biodegradation possibly contribute to C8-C10 PFCAs in the study samples. This study reported a wide presence of PFASs in different species of aquatic products across the Yellow-Bohai Sea coasts. The potential health risks that PFASs pose in certain species (such as marine shellfish and marine crustaceans) should not be neglected.

Seasonal variations of heavy metals in seawater and integrated poly-cultured scallop Chlamys farreri in Ailian Bay, northern China.

Seasonal variations of heavy metals in integrated poly-cultured scallops and seawater from Ailian Bay, northern China were analyzed to reveal the potential factor in bioaccumulation of metals in scallop Chlamys farreri. Results showed that heavy metals (Cu, Zn, As, Cd, Cr, Pb and Hg) in seawater were much below the maximum permissible limits and showed no seasonal changes, but were consistent with the growing period of the poly-cultivated kelp. The content of Zn in scallop tissues was highest with an average value of 88.35 ± 11.50 mg/kg, and Hg content was lowest (0.046 ± 0.025 mg/kg). The accumulation of Cu, As, Cd and Hg in scallops presented a significant seasonal change, and they were closely correlated with the physicochemical quality instead of heavy metals in seawater. Cadmium provided 88.9 % of the total hazard index for adults and 72.2 % for children. Arsenic should also be paid more attention in the risk assessment of human health.

Characteristic Profile of the Hazardous, Nutritional, and Taste-Contributing Compounds during the Growth of Argopecten irradians with Different Shell Colors.

Bay scallops (Argopecten irradians; A. irradians) are shellfish with high nutritional and economic value. However, nutritional studies on A. irradians with different shell colors are limited. This study examines the hazardous, nutritional, and taste-contributing compounds during the growth of A. irradians with different shell colors. During the growth of A. irradians, the hazardous contents were below the standard limit. Changes in the nutritional and taste-contributing compounds between months were more significant than shell color. Bay scallops had more fats, total fatty acids, and taste-contributing compounds in August and more proteins, essential fatty acids, vitamin D, vitamin B12, Cu, and Zn in September and October. In October, the golden shell color strain had more proteins, essential fatty acids, vitamin D, vitamin B12, Cu, and Zn, while the purple shell color strain had more taste-contributing compounds. A. irradians had better taste in August and higher nutritional value in September and October. In October, the golden shell color strain has higher nutritional value, and the purple shell color strain has better commercial value and taste. The correlation analysis indicates that the nutritional quality of bay scallops is affected by age (months), shell color, and seawater environment.

Residue Analysis and Assessment of the Risk of Dietary Exposure to Domoic Acid in Shellfish from the Coastal Areas of China.

Harmful algal blooms in Chinese waters have caused serious domoic acid (DA) contamination in shellfish. Although shellfish are at particular risk of dietary exposure to DA, there have been no systematic DA risk assessments in Chinese coastal waters. A total of 451 shellfish samples were collected from March to November 2020. The presence of DA and four of its isomers were detected using liquid chromatography-tandem mass spectrometry. The spatial-temporal distribution of DA occurrence and its potential health risks were examined. DA was detected in 198 shellfish samples (43.90%), with a maximum level of 942.86 µg/kg. DA was recorded in all 14 shellfish species tested and Pacific oysters (Crassostrea gigas) showed the highest average DA concentration (82.36 µg/kg). The DA concentrations in shellfish showed distinct spatial-temporal variations, with significantly higher levels of occurrence in autumn than in summer and spring (p < 0.01), and particularly high occurrence in Guangdong and Fujian Provinces. The detection rates and maximum concentrations of the four DA isomers were low. While C. gigas from Guangdong Province in September showed the highest levels of DA contamination, the risk to human consumers was low. This study improves our understanding of the potential risk of shellfish exposure to DA-residues.

Transcriptome Analysis Reveals MAPK/AMPK as a Key Regulator of the Inflammatory Response in PST Detoxification in Mytilus galloprovincialis and Argopecten irradians.

Paralytic shellfish toxins (PSTs) are an increasingly important source of pollution. Bivalves, as the main transmission medium, accumulate and metabolize PSTs while protecting themselves from damage. At present, the resistance mechanism of bivalves to PSTs is unclear. In this study, Mytilus galloprovincialis and Argopecten irradians were used as experimental shellfish species for in situ monitoring. We compared the inflammatory-related gene responses of the two shellfish during PSTs exposure by using transcriptomes. The results showed that the accumulation and metabolism rate of PSTs in M. galloprovincialis was five-fold higher than that in A. irradians. The inflammatory balance mechanism of M. galloprovincialis involved the co-regulation of the MAPK-based and AMPK-based anti-inflammatory pathways. A. irradians bore a higher risk of death because it did not have the balance system, and the regulation of apoptosis-related pathways such as the PI3K-AKT signaling pathway were upregulated. Taken together, the regulation of the inflammatory balance coincides with the ability of bivalves to cope with PSTs. Inflammation is an important factor that affects the metabolic pattern of PSTs in bivalves. This study provides new evidence to support the studies on the resistance mechanism of bivalves to PSTs.

Dimeric Tetracenomycin Derivatives from a Taklamakan Desert-Derived Streptomyces sp. HDN154193.

Bitetracenomycin A (1) and its diastereomers [(±)-bitetracenomycin B, (±)-2] were discovered from the cultures of Streptomyces sp. HDN154193. Compounds 1 and (±)-2 were the first tetracenomycin dimers obtained from a natural source with sp3 methine protons at the bridge positions (C-12/12'), which also exhibited broad-spectrum antibacterial activity. The racemate (±)-2 was semisynthesized and separated into enantiomers (+)-2 and (-)-2, and the absolute configurations were determined by specific rotation and ECD data. These metabolites exhibited potent antibacterial activity especially against drug-resistant strains (MRSA and MRCNS) with MIC values ranging from 1.0 to 1.9 µg/mL.

Effects of single and combined exposure to BDE-47 and PFOA on distribution, bioaccumulation, and toxicity in blue mussel (Mytilus galloprovincialis).

The contamination of 2,2',4,4'-Tetrabrmodiphenyl ether (BDE-47) and perfluorooctanoic acid (PFOA) has drawn a worldwide attention over the risks in ecological and food safety. In this work, blue mussel (Mytilus galloprpvincialis) was employed to investigate the combined effects of BDE-47 (10 ng mL-1) and PFOA (100 ng mL-1) on tissue distribution, accumulation, elimination, and toxicity. Results suggested that BDE-47 and PFOA accumulated mostly in digestive gland, followed by gills and gonad, and M. galloprovincialis displayed higher accumulation capacity to BDE-47 than PFOA. Co-exposure treatment reduced the accumulation of BDE-47, and enhanced the accumulation of PFOA. Furthermore, biochemical and histopathological tests revealed that the aggravated toxicity in co-exposure groups was mainly attributed to the oxidative stress and damage of tissue structure. This work could be helpful to get a better understanding of the combined behaviors and cumulative risks of BDE-47 and PFOA in marine ecosystem.

Simultaneous determination of triazine herbicides and their metabolites in shellfish by HPLC-MS/MS combined with Q/E-Orbitrap HRMS.

Triazine herbicides are used extensively in agriculture and aquaculture worldwide because of their broad effectiveness in weed control. However, after they are discharged into the sea, they seriously contaminate aquatic ecosystems and threaten aquatic organisms, especially shellfish. Currently, there are no established methods for the detection and confirmation of triazine herbicides and their metabolites in biological matrixes. Hence, the food safety of aquatic products cannot be accurately evaluated, which creates a technical barrier against international aquatic product trade. In this study, for the first time, a method was developed for the analysis and confirmation of seven triazine herbicides and 13 metabolites in shellfish, based on alkaline acetonitrile extraction and neutral Al2O3 cartridge purification coupled with internal standard calibration. Specifically, quantitative and qualitative analysis was conducted using high-performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-MS/MS), and accurate identification was carried out by quadrupole orbitrap high-resolution mass spectrometry (Q/E Orbitrap HRMS). The results showed that target analytes demonstrated good linearity within the corresponding range (R2 > 0.995). The limit of detection and limit of quantitation of the proposed method were 0.1 and 0.3 µg/kg, respectively. The average recoveries of analytes were between 70.0% and 120% when spiked at three levels with blank oyster (Crassostrea gigas) as the matrix, and the relative standard deviations (RSDs) were all less than 12% (n=6). The proposed method was successfully applied for the detection of triazine herbicide residues in oyster samples during actual breeding, and the presence of DIP, HP, DEHA, and other metabolites in positive samples was confirmed by Q/E Orbitrap HRMS. This method exhibits high accuracy, high sensitivity, and good reproducibility. It has promising application prospects in the field of hazard analysis and the positive identification of aquatic products.

Semicarbazide Accumulation, Distribution and Chemical Forms in Scallop (Chlamys farreri) after Seawater Exposure.

Semicarbazide is a newly recognized marine pollutant and has the potential to threaten marine shellfish, the ecological equilibrium and human health. In this study, we examined the accumulation, distribution, and chemical forms of semicarbazide in scallop tissues after exposure to 10, 100, and 1000 µg/L for 30 d at 10 °C. We found a positive correlation between semicarbazide residues in the scallops and the exposure concentration (p < 0.01). Semicarbazide existed primarily in free form in all tissues while bound semicarbazide ranged from 12.1 to 32.7% and was tissue-dependent. The time for semicarbazide to reach steady-state enrichment was 25 days and the highest levels were found in the disgestive gland, followed by gills while levels in gonads and mantle were similar and were lowest in adductor muscle. The bioconcentration factor (BCF) of semicarbazide at low exposure concentrations was higher than that at high exposure concentrations. These results indicated that the scallop can uptake semicarbazide from seawater and this affects the quality and safety of these types of products when used as a food source.

Antibacterial p-Terphenyl with a Rare 2,2'-Bithiazole Substructure and Related Compounds Isolated from the Marine-Derived Actinomycete Nocardiopsis sp. HDN154086.

Assisted by MS/MS-based molecular networking and X-ray diffraction analysis, five new p-terphenyl derivatives, namely, nocarterphenyls D-H (1-5), were obtained and characterized from the cultures of the marine sediment-derived actinomycete Nocardiopsis sp. HDN154086. The skeleton of nocarterphenyl D (1) was defined to possess a rare 2,2'-bithiazole scaffold, naturally occurring for the first time, and nocarterphenyls E-H (2-5) are p-terphenylquinones with unusual thioether linked fatty acid methyl ester substitutions. Compound 1 showed promising activity against multiple bacteria with MIC values ranging from 1.5 to 6.2 µM, and 2 exhibited notable antibacterial activity against MRSA which surpassed the positive control ciprofloxacin.

Nontarget Screening and Toxicity Evaluation of Diol Esters of Okadaic Acid and Dinophysistoxins Reveal Intraspecies Difference of Prorocentrum lima.

High-resolution mass spectrometry (HRMS) analysis with the assistance of molecular networking was used to investigate intracellular toxin profiles of five Prorocentrum lima (P. lima) strains sampled from the north Yellow Sea and South China Sea. Mice were used as a model species for testing the acute toxicity of intracellular okadaic acid (OA) and dinophysistoxins (DTXs) in free and esterified states. Results showed that OA and DTX1 esterified derivatives were detected in all P. lima samples, accounting for 55%-96% of total toxins in five strains. A total of 24 esters and 1 stereoisomer of DTX1 (35S DTX1) were identified based on molecular networking and MS data analysis, 15 esters of which have been reported first. All P. lima strains displayed specific toxin profiles, and preliminary analysis suggested that toxin profiles of the five P. lima strains might be region-related. Moreover, acute toxicity in mice suggested higher toxicity of esters compared with free toxins, which highlights the importance and urgency of attention to esterified toxins in P. lima.