Marine bacterial surfactin CS30-2 induced necrosis-like cell death in Huh7.5 liver cancer cells

Marine bacterial strain Bacillus sp. CS30 exhibited high anticancer activity against Huh7.5 human liver cancer. We purified the corresponding anticancer agent by sequential acidic precipitation, methanol extraction, Sephadex LH-20 chromatography, and reversed phase high-performance liquid chromatography (RP-HPLC), then analyzed it in mass spectrometry. Based on the results of purification and mass spectrometry, we deduced that the anticancer agent was the same component as our previously purified antifungal agent surfactin CS30-2. However, to the best of our knowledge, this is the first report on the surfactin possessing both antifungal and anticancer activities. Surfactin CS30-2 was demonstrated to exhibit high anticancer activity in a dose-dependent manner against Huh7.5 liver cancer cells. Further investigation showed that surfactin CS30-2 induced the increased generation of reactive oxygen species (ROS) and severe disruption of cell membrane, thus leading to cell death. However, unlike previously reported surfactins, surfactin CS30-2 caused cancer cell death via necrosis instead of apoptosis.     

Extracts of the seaweed Bifurcaria bifurcata display antifungal activity against human dermatophyte fungi

Bifurcaria bifurcata is a seaweed of the Order Fucales(Ochrophyta, Phaeophyceae) that can be found all year round along the Portuguese Atlantic shore. Although it is considered edible in some countries,its biotechnological potential has not yet been assessed in detail and here we report its antimicrobial potential against human dermatophyte fungi. Three samples were harvested from Baleal Island(Peniche, Portugal)harvested in winter(BbPe); from Aguda Beach(Arcozelo, Vila Nova de Gaia, Portugal), harvested in autumn(BbAg) and a sample obtained by culturing the Aguda sample under laboratory conditions(BbLC).A broth macrodilution assay was applied to determine the MIC(minimum inhibitory concentration) and MLC(minimum lethal concentration) of the B. bifurcata extracts. Results show that the BbPe MeOH extracts were the most effective and had higher antifungal activity against all the tested dermatophyte strains than the BbLC and BaAg extracts [Epidermophyton floccosum FF9(100-200 μg/mL MIC; 200 μg/mL MLC), Microsporum canis FF1(400 μg/mL MIC; 400 μg/mL MLC), Trichophyton mentagrophytes FF7(100 μg/mL MIC; 800 μg/mL MLC), M. gypseum CECT 2908(800 μg/mL MIC; 800 μg/mL MLC), T.mentagrophytes var. interdigitale CECT 2958(800 μg/mL MIC; 800 μg/mL MLC), T. rubrum CECT 2794(200 μg/mL MIC; 400 μg/mL MLC)]. In fact, only E. floccosum FF9 was sensitive to BbAg and BbLC MeOH extracts. To our knowledge, this is the first report of antifungal activity of B. bifurcata against human dermatophyte fungi.     

Retreived bacteria from Noctiluca miliaris (green) bloom of the northeastern Arabian Sea

In recent years, seasonal blooms of the dinoflagellate Noctiluca miliaris have appeared in the open-waters of the northern Arabian Sea (NAS). This study provides the first characterization of bacteria from a seasonal bloom of green Noctiluca of NAS (20°N?17°N and 64°E–70°E), during the spring-inter-monsoon cruise of Sagar Sampada 253, in March 2007. Bacterial growth as assessed by most-probable number (MPN) and plate counts, revealed ‘variable-physiotypes’ over a wide range of salinities (0%–25% w/v NaCl), pH levels (5–8.5), and organic nutrient strengths, in comparison to non-bloom waters. MPN indices of bacteria in surface waters of bloom stations *DWK and *PRB, corresponded to (3.08–4.41)×10³ cells/mL at 3.5% NaCl (w/v), and (2.82–9.49)×10² cells/mL at 25% (w/v) NaCl in tryptone-yeast extract broth (TYE). Plate counts were (1.12–4)×10⁶ CFU/mL at 0% (w/v) NaCl, (1.28–3.9)×10⁶ CFU/mL at 3.5% (w/v) NaCl, and (0.4–7)×10⁴ CFU/mL at 25% NaCl (w/v) on TYE. One-tenth-strength Zobell’s gave (0.6–3.74)×10⁵ CFU/mL at pH 5 to (3.58–7.5)×10⁵ CFU/mL at pH 8.5. These bacteria were identified to the genera Bacillus, Cellulomonas, Staphylococcus, Planococcus, Dietzia, Virgibacillus, Micrococcus, Sporosarcinae, Leucobacter, and Halomonas. The identity of three strains (GUFBSS253N2, GUFBSS253N30, and GUFBSS253N84) was confirmed through 16S rDNA sequence homology as Bacillus cohnii, Bacillus flexus, and Bacillus cereus. The ～2–3-fold higher plate counts of culturable bacteria from the open-waters of the NAS indicate that these bacteria could critically determine the biogeochemical dynamics of the bloom and its milieu. The role of these bacteria in sustaining/terminating the bloom is under evaluation.     

Identification of AOSC-binding proteins in neurons

Acidic oligosaccharide sugar chain （AOSC）, a D-mannuronic acid oligosaccharide, derived from brownalgaepolysaccharide, has been completed Phase I clinical trial in China as an anti-Alzheimer＇s Disease （AD） drug candidate. The identification of AOSC-binding protein（s） in neurons is very important for understanding its action mechanism. To determine the binding protein（s） of AOSC in neurons mediating its anti-AD activities, confocal microscopy, affinity chromatography, and liquid chromatography-tandem mass spectrometry （LC-MS/MS） analysis were used. Confocal microscopy analysis shows that AOSC binds to SH-SY5Y cells in concentration-, time-, and temperature-dependent fashions. The AOSC binding proteins were purified by affinity chromatography and identified by LC-MS/MS analysis. The results showed that there are 349 proteins binding AOSC, including clathrin, adaptor protein-2 （AP-2） and amyloid precursor protein （APP）. These results suggest that the binding/entrance of AOSC to neurons is probably responsible for anti-AD activities.     

化学诱导对2株海洋真菌次级代谢产物及其生物活性的影响

【目的】以2株海洋真菌为研究对象,探究不同种类及浓度的化学表观遗传修饰剂对其次级代谢产物化学多样性和生物活性的影响。【方法】基于化学表观遗传修饰策略,采用两种不同的化学表观遗传修饰剂5-氮杂胞嘧啶核苷(5-azaC)和丁酸钠,分别对Penicillium sp.019和Aspergillus terreus ZN4-5-4两株海洋来源真菌进行表观遗传修饰。通过观察真菌在培养基中的生长状态变化,并根据薄层层析(TLC)指纹图谱、高效液相色谱(HPLC)图谱、乙酰胆碱酯酶(AChE)抑制、1,1-二苯基-2-三硝基苯肼(1,1-Diphenyl-2-picryl-hydrazyl,DPPH)自由基清除和卤虫致死活性筛选模型等技术手段,从中筛选出代谢产物丰富、AChE抑制活性与抗氧化活性产物丰富且毒性小的发酵培养条件。【结果】添加1 mmol/mL的丁酸钠诱导剂可使菌株019和ZN4-5-4的次生代谢产物的种类增加,且同时能提高该菌株次级代谢产物的AChE的抑制活性和DPPH自由基清除活性。因此,确定菌株019和ZN4-5-4的最优发酵条件均是添加浓度为1 mmol/mL丁酸钠的PDB培养基。【结论】化学表观遗传修饰策略指导下,添加诱导剂5-azaC或者丁酸钠对2株海洋真菌次生代谢产物的产生有影响。     

Chemical constituents and antioxidant activities of waste liquid extract from Apostichopus japonicus Selenka processing

Apos tichopus japonicus Selenka is an ideal tonic food that is used traditionally in many Asian countries, and it contains many bioactive substances, such as antioxidant, antimicrobial, and anticancer materials. To convert waste liquid generated during production into a useful resource, extract from waste liquid was isolated by column chromatography and studied by the pyrogallol autoxidation and 1,10-phenanthroline-Fe²⁺ oxidation methods. Results show that the extract scavenged about 91% of the superoxide anion radical at a concentration of 1.4 mg/mL and 24% of the hydroxyl radical at 3.3 mg/mL. Four compounds were isolated and identified from the extract: 2,4-dihydroxy-5-methyl-1,3-azine; 2,4-dihydroxy-1,3-diazine; 3-O-[β-D-quinovopranosyl-(1→2)-4-O-sodium sulfate-β-D-xylopranosyl]-holosta-9(11)-ene-3β,12α,17α-triol; and 24-ethyl-5α-cholesta-7-ene-3β-O-β-D-xylopyranoside. All of these compounds are known in A. japonicus, and were found in the waste liquid for the first time.     

Metabolomic Fingerprint of the Model Ciliate,Tetrahymena thermophila Determined by Untargeted Profiling Using Gas Chromatography-Mass Spectrometry

The ciliate Tetrahymena is a valuable model organism in the studies of ecotoxicology. Changes in intracellular metabolism are caused by exogenous chemicals in the environment. Intracellular metabolite changes signify toxic effects and can be monitored by metabolomics analysis. In this work, a protocol for the GC-MS-based metabolomic analysis of Tetrahymena was established. Different extraction solvents showed divergent effects on the metabolomic analysis of Tetrahymena thermophila. The peak intensity of metabolites detected in the samples of extraction solvent Formula 1(F1) was the strongest and stable, while 61 metabolites were identified. Formula 1 showed an excellent extraction performance for carbohydrates. In the samples of extraction solvent Formula 2(F2), 66 metabolites were characterized, and fatty acid metabolites were extracted. Meanwhile, 57 and 58 metabolites were characterized in the extraction with Formula 3(F3) and Formula 4(F4), respectively. However, the peak intensity of the metabolites was low, and the metabolites were unstable. These results indicated that different extraction solvents substantially affected the detected coverage and peak intensity of intracellular metabolites. A total of 74 metabolites(19 amino acids, 11 organic acids, 2 inorganic acids, 11 fatty acids, 11 carbohydrates, 3 glycosides, 4 alcohols, 6 amines, and 7 other compounds) were identified in all experimental groups. Among these metabolites, amino acids, glycerol, myoinositol, and unsaturated fatty acids may become potential biomarkers of metabolite set enrichment analysis for detecting the ability of T. thermophila against environmental stresses.     

The effect of salinity on Fucus ceranoides (Ochrophyta,Phaeophyceae) in the Mondego River (Portugal)

Algae(and their extracts) are increasingly important for pharmaceutical applications due to the diversity of useful compounds they contain. The genus Fucus contains one of the most studied species,Fucus vesiculosus. The species F. ceranoides differs from the others of the genus by presenting longitudinal air-vesicles and a capacity to survive at low salinities. It is an alga that inhabits the Mondego River estuary(Portugal), at the southern limit of its distribution, and can serve as a role model to understand the effect of a salt gradient on the production of bioactive compounds. We assessed the phenolic content and antioxidant activity of different F. ceranoides extracts(e.g. methanolic, aqueous and polysaccharide) prepared from samples harvested from two different zones to evaluate if the adaptation of F. ceranoides to different salinity levels influenced its chemical composition. The antioxidant activity of the extracts was determined using 1,2-diphenyl-picrylhydrazyl(DPPH) and 2.2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS)radicals. These assays demonstrated that the methanolic extract of lyophilized F. ceranoides that grew at low salinities was the most bioactive, i.e. DPPH(IC_(50)=50.39 μg/mL) and ABTS(TEAC=2.42). The total phenolic content(Folin-Ciocalteu method) and the methanolic extract of the lyophilized F. ceranoides collected from a low salinity habitat exhibited the highest phenolic content(PGE=49.48 μg/mg of lyophilized extract)amongst those sampled. Thin layer chromatography(TLC) and Fourier transform infrared spectroscopy(FTIR) were used for the identification of compounds in the extracts. This characterization allowed confirmation that the various extracts contained almost the same compounds but with notable quantitative differences. Based on these results, we conclude that there were differences in the quantity of the compounds due to the effect of salinity. The drying methods used were also found to have influenced the quality of the extracted compounds.     

食物类型和浓度对双齿许水蚤摄食的影响

在不同饵料条件下,对双齿许水蚤(Schmackeria dubin)的清滤率(mL.只-1.h-1)、滤食率(细胞.只-1.h-1)和摄食率(mg.只-1.h-1)进行了测定。结果表明:以湛江等鞭金藻(Isochrysis zhanjiangensis)为饵料,其浓度为15×104～120×104 mL-1时,双齿许水蚤的摄食率达到较大;分别饲以同生物量的牟氏角毛藻(Chaetoceros müelleri)、亚心形扁藻(Platymonas subcordiformis)、湛江等鞭金藻、小球藻(Chlorella sp.)时,饵料为角毛藻和扁藻时双齿许水蚤的摄食率达到较大;饲以同生物量的牟氏角毛藻、亚心形扁藻、湛江等鞭金藻、小球藻、颤藻(Oscillatoria sp.)、小环藻(Cyclotella sp.)六种藻的混合饵料时,双齿许水蚤对湛江等鞭金藻的摄食率和选食率均为最大,分别为0.0458 mg.只-1.h-1和17.1%。可见,饵料浓度、饵料种类、混合饵料对双齿许水蚤的清滤率和摄食率均有显著影响(P<0.05)。     

Antimicrobial Activity of Ulva reticulata and Its Endophytes

Seaweeds are known to exhibit various antimicrobial properties,since it harbours an enormous range of indigenous bioactive compounds.The emergence of drug resistant strains has directed to the identification of prospective metabolites from seaweed and its endophytes,thereby exploiting the properties in resisting bacterial diseases.The current study was aimed to assess the antimicrobial activity of extracts obtained from Ulva reticulate,for which metabolites of Ulva reticulata and its endophytes were extracted and assessed against human pathogens like Staphylococcus aureus,Escherichia coli,Pseudomonas aeruginosa,Salmonella typhi,and Bacillus subtilis.It was observed that the hexane extract of isolate VITDSJ2 was effective against all the tested pathogens but a significant inhibition was observed for Staphylococcus aureus and Escherichia coli.Further,Gas chromatography coupled with Mass spectroscopy(GC-MS) revealed the existence of phenol,3,5-bis(1,1-dimethylethyl) in the crude hexane extract which is well-known to possess antibacterial activity.The effective isolate VITDSJ2 was identified to be the closest neighbour of Pseudomonas stutzeri by phenotypic and genotypic methods.The crude extracts of the seaweed Ulva reticulata was also screened for antibacterial activity and the hexane extract was effective in showing inhibition against all the tested pathogens.The compound in the crude extract of Ulva reticulata was identified as hentriacontane using GC-MS.The extracts obtained from dichloromethane did not show significant activity in comparison with the hexane extracts.Hence the metabolites of Ulva reticulata and the bacterial secondary metabolites of the endophytes could be used in the treatment of bacterial infections.     

Lipid oxidation and fatty acid composition in salt-dried yellow croaker (<Emphasis Type="Italic">Pseudosciaena polyactis</Emphasis>) during processing

Lipid oxidation in salt-dried yellow croaker (Pseudosciaena polyactis) was evaluated during processing with commonly used analytical indices, such as the peroxide value (POV), the thiobarbituric acid reactive substances (TBARS) value, and oxidative-relative lipoxygenase (LOX) activity. Additionally, fatty acids were analyzed using gas chromatography-mass spectrometry. Both POV and TBARS increased significantly (P < 0.05) at the rinsing stage. POV reached its peak value of 3.63 meq O₂ per kg sample at the drying stage, whereas TBARS constantly increased from 0.05 to 0.20 mg MDA per kg sample. Processing of salt-dried yellow croaker had an extremely significant (P < 0.01) effect on LOX activity. Twenty-six fatty acids were identified. Combined eicosapentaenoic acid (EPA; C20:5n3) and docosahexaenoic acid (DHA; C22:6n3) content varied between (19.20 ± 0.37) mg g^?1 and (23.45 ± 1.05) mg g^?1. The polyunsaturated fatty acid/saturated fatty acid (PUFA/SFA) ratio in yellow croaker was 0.73–1.10, and the n-6/n-3 PUFA ratio was approximately 0.13–0.20. The contents of most fatty acids varied significantly (P < 0.05) during the different processing stages, and these differences were caused by lipid oxidation. C18:0, C16:1n7, C19:0, and C22:6n3 showed clear changes in principle component one of a principle components analysis. These fatty acids are potential markers for evaluating lipid oxidation in fish muscle because there was a significant correlation between these markers and TBARS and LOX activity (P < 0.05) with Pearson’s coefficients > 0.931.     

A Bioassay for the Cytotoxicity of Gemcitabine Using the Marine Ciliate Euplotes vannus

This study investigated the cytotoxicity of gemcitabine using the marine ciliate Euplotes vannus as the test organism.The median lethal concentrations(LC50 values)were determined using acute toxicity tests within an exposure time of 30 min with 0,6,12,24,and 48 mg mL^-1 gemcitabine.The median inhibition effect(IC50 value)on the growth of the ciliate cells was examined using chronic toxicity tests within 5 days(120 h)after exposure for 30 min with 0,0.7,3.5,7,and 14 mg mL^-1 gemcitabine.The 30-min LC50value was 10.66-mg mL 1.The LC50 values decreased with increasing exposure times and well fitted to the toxicity curve equation LC50=10.93+28.4e^-0.19t(R2=0.93;P<0.05,t=exposure time).The IC50 value for growth rates was 7.05 mg mL^-1,and the inhibition effect on growth rates well fitted to the model equation r%=0.8681e^-0.0782Cgem(r%means growth rate with inhibition by gemcitabine,Cgem means concentrations of gemcitabine,R^2=0.99 and P<0.05).The LC50 values of a wide range of gemcitabine concentrations could therefore be predicted for any given exposure time.These results suggest that the clinical dose of gemcitabine(20 mg mL^-1)was higher than the 30-min LC50 value,which was almost the same as the 6-min LC50 value(19.88 mg mL^-1)for E.vannus cells.The results also demonstrate that E.vannus can be used as a robust test organism for bioassays of chemotherapeutic drugs during short exposure periods.     

双棘黄姑鱼血清性类固醇激素的周年变化

采用放射免疫法(RIA)测定双棘黄姑鱼(Nibea diacanthus)血清性类固醇激素的周年变化。结果表明:雌鱼血清雌二醇(Estadiol-17β)浓度一年出现两次峰值,均在成熟产卵前一个月,即4月[(338±59)pg/mL]和9月[(296±48)pg/mL];血清睾酮(T)浓度在卵黄发生期急剧上升,并在卵巢成熟期达到最高值,分别是5月(2.506±0.262 ng/mL)和10月份(2.268±0.243 ng/mL),分别在血清雌二醇浓度两次峰值出现1个月之后,因此,雌鱼的血清雌二醇浓度和血清睾酮浓度均与性腺发育相关。雄鱼血清睾酮浓度的两次峰值均出现在性腺已发育成熟的5月份(2.307±0.792 ng/mL)和10月份(2.286±0.607 ng/mL),与性腺发育有明显的相关性;而血清雌二醇浓度全年维持在一个相对稳定的水平,与雄性性腺发育相关性不明显。     

贵州松河煤矿晚二叠世煤的有机地球化学特征

以贵州松河煤矿晚二叠世煤层为研究对象,进行了详细的有机地球化学研究。利用偏光显微镜观察煤岩组分和测定镜质体反射率,利用气相色谱（GC）和色谱-质谱联用（GC-MS）分别对煤样中饱和烃和芳香烃进行测定,并计算相关的有机地球化学参数。结果显示,松河煤样镜质组含量最高,镜质体平均反射率（R_o）为1.21%。正烷烃碳数分布范围为C₁₂-C₂₉,主峰碳为C₁₄或C₁₅,∑C_21-/∑C₂₂₊值较高,奇偶优势指数OEP值接近于1,姥植比（Pr/Ph）在1.40~2.65之间;检测到的芳烃种类较多,以来源于树脂类有机质的萘系和菲系化合物为主,甲基取代同系物中热力学稳定异构体的含量相对较高。分析表明,松河煤的成煤母质主要为陆相高等植物,但也伴随海相低等生物的贡献;煤样成熟度较高,较高的芴系化合物和苯并萘并噻吩含量反映煤层主要形成于还原环境之中。      

Purification and characterization of cold-active endo-1,4-β-glucanase produced by Pseudoalteromonas sp. AN545 from Antarctica

A bacterium hydrolyzing carboxymethylcellulose, isolated from Antarctic sea ice, was identified as Pseudoalteromonas sp. based on 16S rDNA gene sequences and named as Pseudoalteromonas sp. AN545. The extracellular endo-1,4-β-glucanase AN-1 was purified successively by ammonium sulfate precipitation, DEAE-Sepharose ion exchange chromatography and Sephadex G-75 gel filtration chromatography. The molecular mass of AN-1 was estimated to be 47.5 kDa utilizing SDS-PAGE and gel chromatography analysis. AN-1 could hydrolyze caboxymethylcellulose, avicel and β-glucan, but not cellobiose, xylan and p-Nitrophenyl-β-D-glucopyranoside. The optimal temperature and pH for the β-glucanase activity of AN-1 were determined to be at 30°C and pH 6.0, respectively. AN-1 was stable at acidic solutions of pH 5.0-6.5 and temperatures below 30°C for 1 h. Moreover, the specific activity was enhanced by Ca2+ and Mg2+, and inhibited by Cu2+. The kinetic parameters Michaelis constant (Km) and maximum velocity (Vmax) of AN-1 were 3.96 mg/mL and 6.06×10-2 mg/(min·mL), respectively.     

Chemical constituents of marine medicinal mangrove plant Sonneratia caseolaris

Twenty-four compounds including eight steroids (1-8), nine triterpenoids (9-16, 24), three flavonoids (20-22), and four benzenecarboxylic derivatives (17-19, 23) were isolated and identified from stems and twigs of medicinal mangrove plant Sonneratia caseolaris. The structures of the isolated compounds were determined by extensive analysis of their spectroscopic data. Among these metabolites, compounds 1, 4-20 and 22-24 were isolated and identified for the f'urst time from S. caseolaris. In the in vitro cytotoxic assay against SMMC-7721 human hepatoma cells, compound 21 (3',4',5,7-tetrahydroxyflavone)exhibited significant activity with IC50 2.8 μg/mL, while oleanolic acid (14), 3,3'-di-O-methyl ether ellagic acid (18), and 3,3',4-O-tri-O-methyl ether ellagic acid (19) showed weak activity. None of these compounds displayed significant antibacterial activites.     

扁舵鲣抗氧化肽分离纯化及结构鉴定

【目的】从扁舵鲣(Auxis thazard)的木瓜蛋白酶酶解产物中分离鉴定具有较高1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除率的抗氧化肽。【方法】用木瓜蛋白酶水解扁舵鲣,以DPPH自由基清除能力为检测指标,通过超滤、凝胶过滤层析和反向高效液相色谱分离抗氧化肽,再经过超高效液相/三重四级杆飞行时间质谱(UPLC/Xevo G2-XS QTOF)进行结构鉴定。【结果】从酶解产物中获得3种抗氧化肽,其氨基酸序列分别为β-丙氨酸-1-甲基-L-组氨酸(241 u)、Gly-Ala-Gly-Gly-Pro(357 u)和Val-Glu(246 u)。【结论】扁舵鲣的木瓜蛋白酶酶解产物含有抗氧化活性的肽类,可为其抗氧化肽的开发提供理论依据。     

Mercury (Hg2+) effect on enzyme activities and hepatopancreas histostructures of juvenile Chinese mitten crab Eriocheir sinensis

We studied the effects of mercury (Hg²⁺) on antioxidant and digestive enzyme activities in terms of LC₅₀ value and on hepatopancreas histostructures of juvenile Chinese mitten crabs Eriocheir sinensis in 40-day exposure to various concentrations of Hg²⁺ (0, 0.01, 0.05, 0.10, 0.20, and 0.30 mg/L). The results show that the activities of superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase (CAT) significantly increased in the concentrations of 0.01 and 0.05 mg/L, while that of enzyme decreased in 0.10, 0.20 and 0.30 mg/L treatments. Meanwhile, Hg²⁺ disrupted the histostructures of the hepatopancreas, causing decreases in activities of pepsin, tryptase, amylase, and cellulose, which are synthesized in the hepatopancreas. Moreover, as the Hg²⁺ concentration increased, the survival rate of the crabs decreased, worst at 56.57% in 0.30 mg/L. Therefore, although crabs are able to tolerate low levels of mercury pollution, high levels lead to cellular injury and tissue damage in hepatopancreas, which then loses some of its vital physiological functions such as absorption, storage, and secretion.     

Spatial Distributions and Potential Sources of Long Chain(C_(30),C_(32) 1,15-) Alkyl Diols in Surface Sediments from Eastern China Marginal Seas

Long chain alkyl diols have shown important potential for the reconstruction of sea surface temperature,productivities and upwelling conditions in marine or lacustrine environments.However,little is known about the distribution and sources of the diols in eastern China marginal seas(CMS),which are areas of important organic carbon sink.Here the contents of C_(30) and C_(32)1,15-diols were analyzed in 181 surface sediments from eastern CMS.The similar distribution pattern and strong linear correlation between C_(30) and C_(32) diols indicated that they had similar biological source,with a dominance of C_(30) diol.Their contents ranged from 7-2726 ng g~(-1) for C_(30) diol and 5-669 ng g~(-1) for C_(32) diol,and both showed higher values mainly in the mud area of the Yellow Sea,while the TOC normalized contents showed a more obvious seaward increasing trend.The similar distribution pattern and significant positive correlation between diols and the other marine biomarkers(brassicasterol,dinosterol,C_(37) alkenones) indicated C_(30) and C_(32) diols in eastern CMS were mainly from marine algae.This conclusion was also supported by principal component analysis(PCA).Our results also showed that sediment diol contents were generally related to marine productivity,suggesting that diols could be applied for marine productivity reconstruction in eastern CMS.     

Isolation,Identification, and Quantitative Determination of Saponin in Apostichopus japonicus by HPLC-DAD

Apostichopus japonicus is an important invertebrate that is widely used as a tonic food in Asian countries.The purpose of this study is to purify and identify a class of compound,the saponins,from the body wall of A.japonicus,and to establish a new me-thod to determine the quantity of saponins in the sea cucumber.In this study,the saponins of A.japonicus,cladoloside A(CA),were ob-tained from 80%ethanol extract by column chromatography for the first time and were characterized using the spectral method.The resulting purified saponins were then profiled using ¹HNMR,¹³CNMR,and ESI-MS,which revealed the CA molecular formula of C₅₃H₈₂O₂ and contained a triterpenoid backbone,a methylglucopyranosyl moiety,a quinovopyranosyl,and two xylopyranosyls.A me-thod for the quantitative determination of CA,comprising microwave-assisted extraction,high-performance liquid chromatography,and diode array detector method,was established.Extraction efficiency was optimized by changing microwave power,extraction sol-vent,volume,time,and temperature.Results showed that under the optimum conditions(extraction time of 10 min,temperature of 45℃,and solvent of 25 mL 70%ethanol under 400 W),the detection limit of CA was 0.0015 mg mL^?1,and the recoveries of CA from samples at spiking levels of 10,20,and 50μg g^?1 ranged from 90.1%-104%.The proposed method was successfully applied to ana-lyze the saponins in different tissues of A.japonicus collected in different seasons.The method developed in this study can provide quantitative technical support for the quality control of A.japonicus.