Kelp forest food webs as hot spots for the accumulation of microplastic and polybrominated diphenyl ether pollutants.

Kelp forests (KFs) are one of the most significant marine ecosystems in the planet. They serve as a refuge for a wide variety of marine species of ecological and economic importance. Additionally, they aid with carbon sequestration, safeguard the coastline, and maintain water quality. Microplastic (MP) and polybrominated diphenyl ethers (PBDEs) concentrations were analyzed across trophic levels in KFs around Todos Santos Bay. Spatial variation patterns were compared at three sites in 2021 and temporal change at Todos Santos Island (TSI) in 2021 and 2022. We analyzed these MPs and PBDEs in water, primary producers (Macrocystis pyrifera), grazers (Strongylocentrotus purpuratus), predators (Semicossyphus pulcher), and kelp detritus. MPs were identified in all samples (11 synthetic and 1 semisynthetic polymer) and confirmed using Fourier-transform infrared microspectroscopy-attenuated total reflectance (µ-FTIR-ATR). The most abundant type of MP is polyester fibers. Statistically significant variations in MP concentration were found only in kelps, with the greatest average concentrations in medium-depth kelps from TSI in 2022 (0.73 ± 0.58 MP g-1 ww) and in the kelp detritus from TSI in 2021 (0.96 ± 0.64 MP g-1 ww). Similarly, PBDEs were found in all samples, with the largest concentration found in sea urchins from Punta San Miguel (0.93 ± 0.24 ng g-1 ww). The similarity of the polymers can indicate a trophic transfer of MPs. This study shows the extensive presence of MP and PBDE subtropical trophic web of a KF, but correlating these compounds in environmental samples is highly complex, influenced by numerous factors that could affect their presence and behavior. However, this suggests that there is a potential risk to the systems and the services that KFs offer.

Dietary kelp meal improves serum antioxidants, intestinal immunity, and lipid metabolism in hybrid snakehead (Channa maculata ♀ × Channa argus ♂).

BACKGROUND: Dietary kelp possesses a variety of useful biological qualities but does not have a toxic effect on the host. In this study, we examine how kelp dietary supplementation enhances the serum biochemistry, intestinal immunity, and metabolism of hybrid snakehead. A total of 810 juvenile hybrid snakeheads (Channa maculata ♀ × Channa argus ♂), with an initial average weight of 11.4 ± 0.15 g, were allocated randomly to three treatment groups (three replicates per group). The fish were fed for 60 days with isonitrogenous and isolipidic diets. The groups were the control group (C) (20% high-gluten flour), the medium replacement group (MR) (10% high-gluten flour and 10% kelp meal), and the full replacement group (FR) (0% high-gluten flour and 15% kelp meal). RESULTS: The results showed that dietary kelp increased the activity of serum antioxidant enzymes significantly and decreased the content of serum malondialdehyde (MDA) in hybrid snakeheads, with significant changes in the FR group (P < 0.05). The intestinal morphology results showed that dietary kelp helped to increase the specific surface area of intestinal villi, which was beneficial for intestinal digestion and absorption. According to transcriptome and quantitative real-time polymerase chain reaction (qRT-PCR) analysis, dietary kelp can improve the expression of intestinal immunity and metabolism-related pathways. Among them, immune-related genes MHC1 and HSPA1 were significantly up-regulated, and IGH, MHC2, and IL-8 were significantly down-regulated (P < 0.05). Lipid metabolism-related genes DGAT2, FABP2, RXRα, and PLPP1 were all significantly up-regulated (P < 0.05). CONCLUSION: Dietary kelp can effectively improve the antioxidant function of hybrid snakeheads, improve intestinal morphology, reduce intestinal inflammation, and promote intestinal lipid synthesis and transportation, thereby improving intestinal immunity and metabolic functions. © 2024 Society of Chemical Industry.

Development of a kelp powder (Thallus laminariae) Standard Reference Material.

A Standard Reference Material (SRM) of seaweed, SRM 3232 Kelp Powder (Thallus laminariae) has been developed to support food and dietary supplement measurements in compliance with the Food Safety Modernization Act (FSMA) and the Dietary Supplement Health and Education Act of 1994 (DSHEA). The material was characterized for nutritional minerals, arsenic species, isomers of vitamin K1, proximates, and toxic elements. Kelp is a rich source of vitamins and minerals, and it is an excellent source of dietary iodine. Kelp also contains a large amount of arsenic, which is toxic as inorganic species but much less so as organic species. To capture the dietary profile of kelp, certified values were issued for As, Ca, Cd, Cr, Cu, Fe, Hg, I, K, Mg, Mn, Mo, Na, Pb, and Zn. Reference values for proximates were assigned. For the first time, a certified value for iodine, reference values for isomers of vitamin K1, and reference values for arsenic species including arsenosugars were assigned in a seaweed. SRM 3232 fills a gap in Certified Reference Materials (CRMs) needed for quality assurance and method validation in the compositional measurements of kelp and similar seaweeds used as food and as dietary supplements. Graphical Absract Arsenic species and isomers of vitamin K1 were determined in the development of SRM 3232 Kelp Powder (Thallus laminariae).

Copper nanoclusters as fluorescence-quenching probes for the quantitative analysis of total iodine.

Tannic acid-coated copper nanoclusters (CuNCs@TA) were synthesized and used quantitatively to analyze iodine in kelp. Compared with other methods for iodine detection, the proposed method showed excellent performance. The iodine-induced linear decrease in the fluorescence intensity of CuNCs@TA allowed the quantitative detection of iodine in the range 20-100 µM, and the limit of detection for iodine was 18 nM. The probe can be used for the determination of iodine in real samples with reliable and accurate results. Modified Stern-Volmer equation and thermodynamic calculation studies were used to discuss the quenching mechanism.

An approach for identification and determination of arsenic species in the extract of kelp.

The National Institute of Standards and Technology is developing a kelp powder standard reference material (SRM) in support of dietary supplement measurements. Edible seaweeds such as kelp and laver consumed as diet or dietary supplement contain tens of mg/kg arsenic. The speciation information of arsenic in the seaweed should be provided because the total arsenic alone does not fully address the safety issue of the dietary supplement as the value assignment is originally intended. The inability to avail all arsenic species for value assignment measurements prevented the certification of arsenic species in the candidate SRM; however, approximately 70 % of total arsenic extracted with a 1:1 volume fraction of methanol:water mixture allowed arsenic speciation values to be assigned to a procedure-defined extract, which may be used for method validation in research to improve upon current extraction and measurement practices. Arsenic species in kelp and laver were identified using electrospray ionization ion trap time of flight mass spectrometry (ESI-IT-TOF). Arsenosugars As(328), As(482), and As(392) were found in the kelp candidate SRM while As(328) and As(482) were found in GBW 08521, a certified reference material (CRM) of laver produced by the National Institute of Metrology of China (NIM). A discovery that the digests of kelp and laver contained only dimethylarsinic acid led to the conclusion that the seaweeds did not contain detectible levels of arsenobetaine, arsenocholine or trimethylarsine oxide that could overlap with the peaks of arsenosugars in the separation. The mean ± s of (5.68 ± 0.28) mg/kg and (13.43 ± 0.31) mg/kg found for As(482) and As(392) in kelp, respectively, using instrumental neutron activation analysis (INAA) demonstrated that value assignment measurement of arsenosugars was possible without arsenosugar calibration standards.

[Metabolomic approach to evaluating the effect of the mixed decoction of kelp and licorice on system metabolism of SD rats].

The aim of the study is to evaluate the effects of the single and mixed decoction of Thallus laminariae (kelp) and Glycyrrhiza glabra (licorice) on the metabolism and their difference. The mixed decoction of kelp and licorice and the single decoction were made and intragastrically administered to the SD rats. The effect on system metabolism, the toxicity of liver and kidney were assessed by GC-MS profiling of the endogenous molecules in serum, routine biochemical assays and histographic inspection of tissues from SD rats, separately. The mixed decoction of kelp and licorice induced more obvious pathological abnormalities in SD rats than a single decoction of kelp, while the extracts of licorice did not show any pathological change. Neither the mixed, nor the single decoction showed abnormal histopathology. After intragastric administration of extracts for 5 days, the mixed decoction induced a decrease of ALT (no significant change in the groups of single decoction) and an increase of BUN (so did the single decoction of kelp). Metabolomic profile of the molecules in serum revealed that the metabolic patterns were all obviously affected for the three groups, i.e., the mixed and single decoction of kelp and licorice. The rats given with the single decoction of kelp showed a similar pattern to that of the mixed decoction, indicating that the kelp primarily contributed the perturbation of metabolism for the mixed decoction. All three groups induced a decrease of branched chain amino acids, TCA cycle intermediates and glycolysis intermediates (e.g., pyruvic acid and lactic acid) and an increase of 3-hydroxybutyric acid. Kelp decoction showed stronger potential in reducing TCA cycle intermediates and glycolysis intermediates than the other two groups, while the levels of branched chain amino acids were the lowest after licorice extracts were given. These results suggested that the effect of the mixed decoction on metabolism was closely associated with both kelp and licorice. The continuous administration of single decoction of kelp and the mixed decoction of licorice and kelp resulted in pathological abnormalities in kidney of SD rats. The mixed decoction of kelp and licorice distinctly perturbed sera molecules and hence system metabolism, which showed associated with those of kelp and licorice. Although the metabolic effect was associated with both kelp and licorice, the results suggested kelp contributed to it primarily.

Ocean acidification significantly alters the trace element content of the kelp, Saccharina latissima.

Seaweeds are ecosystem engineers that can serve as habitat, sequester carbon, buffer ecosystems against acidification, and, in an aquaculture setting, represent an important food source. One health issue regarding the consumption of seaweeds and specifically, kelp, is the accumulation of some trace elements of concern within tissues. As atmospheric CO2 concentrations rise, and global oceans acidify, the concentrations of elements in seawater and kelp may change. Here, we cultivated the sugar kelp, Saccharina latissima under ambient (~400 µatm) and elevated pCO2 (600-2400 µatm) conditions and examined the accumulation of trace elements using x-ray powder diffraction, sub-micron resolution x-ray imaging, and inductively coupled plasma mass spectrometry. Exposure of S. latissima to higher concentrations of pCO2 and lower pH caused a significant increase (p < 0.05) in the iodine and arsenic content of kelp along with increased subcellular heterogeneity of these two elements as well as bromine. The iodine-to­calcium and bromine-to­calcium ratios of kelp also increased significantly under high CO2/low pH (p < 0.05). In contrast, high CO2/low pH significantly reduced levels of copper and cadmium in kelp tissue (p < 0.05) and there were significant inverse correlations between concentrations of pCO2 and concentrations of cadmium and copper in kelp (p < 0.05). Changes in copper and cadmium levels in kelp were counter to expected changes in their free ionic concentrations in seawater, suggesting that the influence of low pH on algal physiology was an important control on the elemental content of kelp. Collectively, these findings reveal the complex effects of ocean acidification on the elemental composition of seaweeds and indicate that the elemental content of seaweeds used as food must be carefully monitored as climate change accelerates this century.

Seaweeds as a major source of dietary microplastics exposure in East Asia.

Microplastics (MPs) occurrence in marine ecosystems is well known, but their accumulation in seaweeds and subsequent human exposure remain understudied. This research quantifies MPs presence in two commonly consumed seaweeds, kelp (Saccharina japonica) and nori (Pyropia yezoensis), in East Asia, revealing widespread contamination dominated by microfibers (<500 µm). Based on dietary patterns, human uptake through seaweed consumption was estimated and quantified. Notably, Chinese people consume an estimated 17,034 MPs/person/year through seaweed consumption, representing 13.1% of their total annual MPs intake. This seaweeds-derived exposure surpasses all other dietary sources, contributing up to 45.5% of overall MPs intake. The highest intake was in South Korea, followed by North Korea, China, and Japan. This research identifies seaweeds as a major, previously overlooked route of dietary MPs exposure. These findings are crucial for comprehensive risk assessments of seaweed consumption and the development of mitigation strategies, particularly for populations in East Asian countries.

Ecklonia cava Polyphenols Have a Preventive Effect on Parkinson's Disease through the Activation of the Nrf2-ARE Pathway.

Parkinson's disease (PD) is a degenerative neurological disorder defined by the deterioration and loss of dopamine-producing neurons in the substantia nigra, leading to a range of motor impairments and non-motor symptoms. The underlying mechanism of this neurodegeneration remains unclear. This research examined the neuroprotective properties of Ecklonia cava polyphenols (ECPs) in mitigating neuronal damage induced by rotenone via the activation of the nuclear factor erythroid 2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway. Using human neuroblastoma SH-SY5Y cells and PD model mice, we found that ECP, rich in the antioxidant polyphenol phlorotannin, boosted the gene expression and functionality of the antioxidant enzyme NAD(P)H quinone oxidoreductase-1. ECP also promoted Nrf2 nuclear translocation and increased p62 expression, suggesting that p62 helps sustain Nrf2 activation via a positive feedback loop. The neuroprotective effect of ECP was significantly reduced by Compound C (CC), an AMP-activated protein kinase (AMPK) inhibitor, which also suppressed Nrf2 nuclear translocation. In PD model mice, ECPs improved motor functions impaired by rotenone, as assessed by the pole test and wire-hanging test, and restored intestinal motor function and colon tissue morphology. Additionally, ECPs increased tyrosine hydroxylase expression in the substantia nigra, indicating a protective effect on dopaminergic neurons. These findings suggest that ECP has a preventative effect on PD.

Structural characterization and immunomodulating assessment of ultra-purified water extracted fucoidans from Saccharina latissima, Alaria esculenta and Laminaria hyperborea.

Fucoidans, a group of high molecular weight polysaccharides derived mainly from brown algae, are characterized by their high fucose content, degree of sulfation (DS), and intra- and interspecific structural variation. Fucoidans are increasingly recognized due to various reported bioactivities, potentially beneficial for human health. To unlock their potential use within biomedical applications, a better understanding of their structure-functional relationship is needed. To achieve this, systematic bioactivity studies based on well-defined, pure fucoidans, and the establishment of standardized, satisfactory purification protocols are required. We performed a comprehensive compositional and structural characterization of crude and ultra-purified fucoidans from three kelps: Saccharina latissima (SL), Alaria esculenta (AE) and Laminaria hyperborea (LH). Further, the complement-inhibiting activity of the purified fucoidans was assessed in a human whole blood model. The purification process led to fucoidans with higher DS and fucose and lower concentrations of other monosaccharides. Fucoidans from SL and LH resembles homofucans, while AE is a heterofucan rich in galactose with comparably lower DS. Fucoidans from SL and LH showed complement-inhibiting activity in blood and blood plasma, while no inhibition was observed for AE under the same conditions. The results emphasize the importance of high DS and possibly fucose content for fucoidans' bioactive properties.

In vivo speciation studies and antioxidant properties of bromine in Laminaria digitata reinforce the significance of iodine accumulation for kelps.

The metabolism of bromine in marine brown algae remains poorly understood. This contrasts with the recent finding that the accumulation of iodide in the brown alga Laminaria serves the provision of an inorganic antioxidant - the first case documented from a living system. The aim of this study was to use an interdisciplinary array of techniques to study the chemical speciation, transformation, and function of bromine in Laminaria and to investigate the link between bromine and iodine metabolism, in particular in the antioxidant context. First, bromine and iodine levels in different Laminaria tissues were compared by inductively coupled plasma MS. Using in vivo X-ray absorption spectroscopy, it was found that, similarly to iodine, bromine is predominantly present in this alga in the form of bromide, albeit at lower concentrations, and that it shows similar behaviour upon oxidative stress. However, from a thermodynamic and kinetic standpoint, supported by in vitro and reconstituted in vivo assays, bromide is less suitable than iodide as an antioxidant against most reactive oxygen species except superoxide, possibly explaining why kelps prefer to accumulate iodide. This constitutes the first-ever study exploring the potential antioxidant function of bromide in a living system and other potential physiological roles. Given the tissue-specific differences observed in the content and speciation of bromine, it is concluded that the bromide uptake mechanism is different from the vanadium iodoperoxidase-mediated uptake of iodide in L. digitata and that its function is likely to be complementary to the iodide antioxidant system for detoxifying superoxide.

Lactic acid fermentation in the development of a seaweed sauerkraut-style product: Microbiological, physicochemical, and sensory evaluation.

Consumption of nutrient-rich seaweeds and fermented nondairy foods represent fast growing trends among health-minded consumers. Assessment of lacto-fermented seaweed was performed to address these trends, and to offer shelf-life extension and product diversification for fresh kelps. The objectives were to evaluate the effects of kelp species and inclusion level on fermentation kinetics, physicochemical quality, safety, shelf-life, and consumer acceptability of a seaweed sauerkraut-style product. Six formulations with different inclusion levels (25, 50, and 75%) of shredded kelp (sugar kelp, SK or winged kelp, WK) were mixed with cabbage, 2% salt, and inoculated with Lactobacillus plantarum (approximately 106 CFU/g) and Leuconostoc mesenteroides (approximately 101 CFU/g). Products were processed in triplicate, fermented until a target pH of ≤4.6 was achieved, and sampled periodically for 60 days. Kelp species and inclusion level significantly affected most variables tested. The most rapid fermentation (3 days), as evidenced by pH decrease, lactic acid bacteria counts, and lactic acid levels, was noted in WK formulations. Some SK formulations took up to 14 days to achieve the target pH, and coliforms persisted to a greater extent in the SK formulations. Higher levels of kelp decreased the fermentation rate and concentration of fermentable sugars in the brine, but increased the total phenolic content and antioxidant activity of the sauerkrauts. Despite differences in instrumental color (L* a* b* ) and texture (shear force) among formulations, overall liking as rated by a consumer panel was not significantly affected by species or inclusion level. Results support the use of lacto-fermentation in the production of refrigeration-stable seaweed sauerkraut-style product. PRACTICAL APPLICATION: Health-conscious consumers are becoming increasingly interested in plant-based diets and fermented foods, and the development of novel seaweed sauerkraut-style products can help to meet these needs. This study demonstrated the successful production of a sauerkraut-style product formulated with up to 50% farm-raised kelp. Physical, chemical, microbiological, and consumer acceptability testing established lactic acid fermentation as a viable method for shelf life extension and value addition of fresh kelps. These results provide science-based information on an alternative processing method for cultivated seaweeds and can assist the industry in product diversification efforts.

Characterization of nutritionally important lipophilic constituents from brown kelp Ecklonia radiata (C. Ag.) J. Agardh.

This research study presents information for the first time on the nutritionally relevant lipophilic compounds obtained from Ecklonia radiata, a poorly studied brown kelp. The major lipophilic compounds were analyzed utilizing liquid chromatography (LC)-tandem mass spectrometry (MS/MS) and gas chromatography (GC)-mass spectrometry (MS). The LC-MS/MS results revealed the presence of eight major lipophilic compounds, including sterols, carotenoids, vitamin E, and phylloquinone (vitamin K1). Quantitative analysis showed that fucosterol was the most predominant phytosterol in the fronds and stipes of E. radiata. The carotenoids (all-E)-fucoxanthin and (all-E)-ß-carotene were present in higher yield. In terms of vitamin E, α-tocopherol was identified as the main tocol. The coenzyme, phylloquinone, important for protein synthesis, was also identified in E. radiata. GC-MS identified 13 fatty acids with palmitic (C16:0) and oleic acid (C18:1n9c) present in the highest quantities. To our knowledge, this is the first report on E. radiata, and the valuable data presented herein can be used as a baseline for developing novel nutraceuticals.

The dual effects of riboflavin and kelp polyphenol extracts on the gel properties of myofibrillar protein from Scomberomorus Niphonius under UVA irradiation.

In the present study, effects of riboflavin (RF) and kelp polyphenol extracts (KPE) on mackerel (Scomberomorus Niphonius) myofibrillar protein (MP) gel were studied with or without ultraviolet A (UVA) irradiation treatment. The gel strength was increased with the addition of RF and KPE under UVA irradiation. Analysis of the proteins in the gel indicated that the carbonyl content increased, while the contents of total sulfhydryl and amino groups decreased. The proteins appeared to have no α-helix structures, and the endogenous tryptophan content appeared to decrease. The results of SDS-PAGE indicated that the RF and KPE treated samples under UVA irradiation showed massive MP cross-linking by covalent bonds. Electron spin resonance (ESR) results indicated that UVA irradiation generated free radicals in RF and KPE, which ultimately led to an improvement in MP gel properties. It also indicated that KPE could prevent the occurrence of peroxidation to improve the gel properties.

Acid-free preparation and characterization of kelp (Laminaria japonica) nanocelluloses and their application in Pickering emulsions.

Cellulose nanofibers (CNFs) from kelp were prepared by cellulase treatment with lengths greater than 3 µm. CNFs were further oxidized by TEMPO-oxidized system, and the lengths of the oxidized CNFs (TEMPO-CNFs) were 0.6-1 µm. AFM and TEM images showed that intertwined CNFs fibers were divided into individual nanofibrils. The crystallinity of TEMPO-CNFs increased to 66.5 %. TGA analysis indicated that TEMPO-CNFs were more sensitive to temperature than cellulose and CNFs. FT-IR spectra revealed no changes in the basic cellulose structures of CNFs and TEMPO-CNFs. In the sunflower oil/water (20/80, v/v) model emulsions, the oil droplet sizes were less than 20 µm in CNFs emulsions, which became smaller in TEMPO-CNFs emulsions. Delamination was found in CNFs emulsions after three days of storage. Addition of NaCl increased the volumes of TEMPO-CNFs emulsions but enlarged the oil droplets sizes. TEMPO-CNFs emulsions had the largest volume with smallest and most homogeneous oil droplets at pH 3. TEMPO-CNFs emulsions showed good stability after storage for 30 days. Further, TEMPO-CNFs could also emulsify 50 % (v/v) of sunflower oil. All these results indicated that TEMPO-CNFs can be used in preparing Pickering emulsions.

Facile Removal of Phytochromes and Efficient Recovery of Pesticides Using Heteropore Covalent Organic Framework-Based Magnetic Nanospheres and Electrospun Films.

Nontargeted analysis of food safety requires selective removal of interference matrices and highly efficient recovery of chemical hazards. Porous materials such as covalent organic frameworks (COFs) show great promise in selective adsorption of matrix molecules via size selectivity. Considering the complexity of interference matrices, we prepared crystalline heteropore COFs whose two kinds of pores have comparable sizes to those of several common phytochromes, main interference matrices in vegetable sample analysis. By controlling the growth of COFs on the surface of Fe3O4 nanoparticles or by utilizing a facile co-electrospinning method, heteropore COF-based magnetic nanospheres or electrospun nanofiber films were prepared, respectively. Both the nanospheres and the films maintain the dual-pore structures of COFs and show good stability and excellent reusability. Via simple magnetic separation or immersion operation, respectively, they were successfully used for the complete removal of phytochromes and highly efficient recovery of 15 pesticides from the extracts of four vegetable samples, and the recoveries are in the range of 83.10-114.00 and 60.52-107.35%, respectively. Film-based immersion operation gives better sample pretreatment performance than the film-based filtration one. This work highlights the great application potentials of heteropore COFs in sample pretreatment for nontargeted analysis, thus opening up a new way to achieve high-performance sample preparation in many fields such as food safety analysis, environment monitoring, and so on.

Microbiological evaluation of the efficacy of two new biodetergents on multidrug-resistant nosocomial pathogens.

BACKGROUND: In the last few years, several outbreaks of nosocomial infections caused by multidrug-resistant pathogenic agents have been observed, and various biocides products were developed in order to control this phenomenon. We investigated the efficacy of two natural biodetergents composed of plants and kelps extracts, BATT1 and BATT2, against multidrug-resistant strains. METHODS: In-vitro antibacterial efficacy of BATT1 and BATT2 against nosocomial multidrug-resistant isolates was assessed using a suspension-inhibition test, with and without bovine serum albumin (BSA). The test was also carried out on glass surfaces with and without BSA. RESULTS: In vitro tests with both biocidal disinfectants at 25% concentration demonstrated an overall drop in bacterial, mould and yeast counts after 10 min of contact with or without organic substances. For Pseudomonas aeruginosa, it was necessary to use undiluted disinfectants with and without an organic substance. The same results were obtained in tests carried out on glass surfaces for all strains. CONCLUSIONS: The natural products BATT1 and BATT2 behave like good biocides even in presence of organic substances. The use of both disinfectants may be beneficial for reducing hospital-acquired pathogens that are not susceptible to disinfectants.However, it has to be stressed that all these experiments were carried out in vitro and they still require validation from use in clinical practice.

Direct speciation analysis of organic mercury in fish and kelp by on-line complexation and stacking using capillary electrophoresis.

To determine organic mercury (Hg) species that could not be detected by ultraviolet (UV), a highly automated on-line complexation method was established, which combined with normal stacking by capillary electrophoresis-diode array detector. The approach was based on the fact that the compounds and complex reagent interacted to form hydrophilic chelates under the effect of the separation voltage, which was effectively separated and detected by UV. Key parameters, such as the type and concentration of complex reagent, separation voltage and so on were systematically investigated. Under the optimized conditions, the precision and repeatability were in the range of 0.16-3.31% and 0.17-1.21%, respectively. Furthermore, PhHg, EtHg and MeHg were effectively separated and determined in fresh fish (Silver carp) muscle and kelp (Kombu) with the recoveries of 84.63-111.39% and 75.68-114.76%, respectively. The proposed method had the advantages of easy-operating, cost-efficient, stable and reliable compared to off-line complexation method.

Combination of curcuma zedoary and kelp inhibits growth and metastasis of liver cancer in vivo and in vitro via reducing endogenous H2S levels.

The combination of traditional Chinese medicines can improve the efficacy of cancer treatment. Furthermore, the combination of the traditional Chinese medicine curcuma zedoary and kelp was used to enhance the effect of the dissipation of blood stasis in pediatrics during the Song Dynasty. Curcumenol and laminarin, the main components of curcuma zedoary and kelp, are also reported to have a wide range of biological activities. Therefore, we hypothesize that the combination of curcuma zedoary and kelp may prevent the development of cancer. The aim of this research was to confirm whether a combination of curcuma zedoary and kelp could inhibit the proliferation and metastasis of hepatoma cells and consequently improve prognosis. In this study, we firstly found in H22-bearing mice that the combination of curcuma zedoary and kelp inhibited tumor growth and the expression of metastasis-related proteins (MMPs, VEGF, pAkt, pERK1/2). Meanwhile, the decreased cystathionine beta synthase (CBS, an endogenous hydrogen sulfide (H2S) synthetase) level was also observed in H22-bearing mice admistrated with the combination of curcuma zedoary and kelp. It was also observed that the combination of curcumenol and laminarin inhibited the proliferation, migration and invasion in human hepatoma HepG2 cells. Furthermore, we investigated the potential inhibiting mechanism of the combination of curcumenol and laminarin on HepG2 cell proliferation and metastasis. Our previous research showed that a CBS/H2S system was vital for maintaining the proliferation in hepatoma cells. Here, we found that the levels of pSTAT3 and BCL-2 were decreased in CBS knockdown HepG2 cells and the combination of curcumenol and laminarin significantly decreased the H2S level in a dose-dependent manner and down-regulated the levels of pSTAT3 and BCL-2 in HepG2 cells. Angiogenesis, positively regulated by the vascular endothelial growth factor (VEGF), is essential for human cancer metastasis. In the present study, we found that the combination of curcumenol and laminarin could significantly down-regulate the expression levels of VEGF and its downstream key genes pAkt and pERK1/2. Furthermore, previous research showed that hydrogen sulfide could stimulate angiogenesis. Here, we also observed the reduction of the VEGF, Akt, pAkt, ERK1/2 and pERK1/2 proteins levels and the inhibition of proliferation and metastasis in CBS knockdown HepG2 cells. Moreover, exogenous H2S rescued the cytological results caused by the combination of curcumenol and laminarin. Taken together, the combination of curcuma zedoary and kelp could inhibit the proliferation and metastasis of liver cancer cells in vivo and in vitro by inhibiting endogenous H2S production and down-regulating the pSTAT3/BCL-2 and VEGF pathway, which provides strong evidence for the application of curcuma zedoary and kelp in treatments of liver cancer.