Fucoxanthin was extracted from the intact rhizoid of Laminariajaponica Aresch with dimethyl sulfoxide （DMSO）, and then recovered from the DMSO extract by partitioning into ethyl acetate and subsequent evaporation. S...Fucoxanthin was extracted from the intact rhizoid of Laminariajaponica Aresch with dimethyl sulfoxide （DMSO）, and then recovered from the DMSO extract by partitioning into ethyl acetate and subsequent evaporation. Some isolation conditions such as solvent volume and extraction time were screened. The quantity and quality of the extracted fucoxanthin were determined by spectral analysis （absorption spectra and fluorescence emission spectra）. The results indicated that： （1） the average total content of fucoxanthin was 122.1μg in 1 g of fresh L. japonica rhizoid; （2） in comparison with the widely used organic solvent, acetone, DMSO was much more effective for the extraction of fucoxanthin; （3） both DMSO volume and extraction time influenced extraction efficiency such as the recovery rate and purity of fucoxanthin （1 g of fresh L. japonica rhizoid treated with 4 mL DMSO for 60 min, yielded 〉 88% of the total fucoxanthin with purity 0.63）; （4） when （NH4）2SO4 concentration was in the range of 0.5-1.0 mol/L, the pigments rapidly and entirely moved from DMSO into the ethyl acetate phase; （5） the ethyl acetate and DMSO were recycled using a rotary evaporator.展开更多
Effects of various concentrations of two heavy metals, namely Cd and Cu, on gametophytes of Laminariajaponica Aresch were determined by recording morphological changes of gametophytes, determining pH values and the he...Effects of various concentrations of two heavy metals, namely Cd and Cu, on gametophytes of Laminariajaponica Aresch were determined by recording morphological changes of gametophytes, determining pH values and the heavy metal content of the culture solution, calculating the germination rate of sporophytes, and observing heavy metal （Cd） distribution using a fluorescence microscope. The results showed that heavy metals damaged the gametophytes, and were even lethal, and that the higher the concentration of heavy metal ions, the greater the injury to gametophytes. Gametophytes could not survive in culture solutions containing more than 100 mg/L Cd and 50 mg/L Cu and were only able to survive in culture solution containing a mixture of Cd and Cu up to a concentration of 10 mg/L, which indicates that gametophytes have a higher tolerance to Cd than Cu and that multiple heavy metal ions in solution markedly aggravate the damage to gametophytes compared with individual heavy metal ions. With increases in the concentration of the heavy metal, the burgeoning rate of sporophytes decreased acutely, and solutions containing multiple heavy metal ions caused even more marked harm to sporophytes than solutions containing a single heavy metal ion, because most sporophytes died in mixed solutions. The pH value of the culture medium dropped immediately at the beginning （the first day） of treatment, increased over the following days, and then decreased again. The pH of culture media containing multiple heavy metal ions showed greater variation than media containing a single heavy metal ion, with the extent of the decrease in pH of culture media containing multiple ions being greatest during the last period of the experiment. With increases in the concentration of heavy metals, the capacity of gametophytes to accumulate these ions increased. The blue fluorescent light emitted by the Cd-and Cd-binding protein complex existing in gametophytes in media containing different concentrations of Cd showed clearly the distribution o展开更多
In the present study, ultrasonic extraction technique （UET） is used to improve the yield of polysaccharides from Lami- naria japonica （LJPs）. And their antioxidative as well as glycosidase inhibitory activities ar...In the present study, ultrasonic extraction technique （UET） is used to improve the yield of polysaccharides from Lami- naria japonica （LJPs）. And their antioxidative as well as glycosidase inhibitory activities are investigated. Box-Behnken design （BBD） combined with response surface methodology （RSM） is applied to optimize ultrasonic extraction for polysaccharides. The optimized conditions are obtained as extraction time at 54 min, ultrasonic power at 1050 W, extraction temperature at 80℃ and ratio of material to solvent at 1：50 （g mL-1）. Under these optimal ultrasonic extraction conditions, an actual experimental yield （5.75% ＋ 0.3%） is close to the predicted result （5.67%） with no significant difference （P〉0.05）. Vitro antioxidative and glycosidase inhibitory activities tests indicate that the crude polysaccharides （LJP） and two major ethanol precipitated fractions （LJP1 and LJP2） are in a concentration-dependent manner. LJP2 （30%-60% ethanol precipitated polysaccharides） possesses the strongest α-glucosidase in- hibitory activity and moderate scavenging activity against hydroxyl radicals （66.09% ±2.19%, 3.0 mg mL-l）. Also, the inhibitory activity against a-glucosidase （59.08% ± 3.79%, 5.0 mg mL-1） is close to that of acarbose （63.99% ± 3.27%, 5.0 mg mL-l）. LJP 1 （30% ethanol precipitated polysaccharides） exhibits the strongest scavenging activity against hydroxyl radicals （99.80%q-0.00%, 3.0mg mL-1） and moderate a-glucosidase inhibitory activity （47.76%± 1.92%, 5.0 mgmL-1）. LJP shows the most remarkable DPPH scav- enging activity （66.20%±0.11%, 5.0mgmL-1） but weakest a-glucosidase inhibitory activity （37.77%±1.30%, 5.0mgmL-1）. How- ever, all these LJPs exert weak inhibitory effects against a-amylase. These results show that UET is an effective method for extract- ing bioactive polysaccharides from seaweed materials. LJP 1 and LJP2 can be developed as a potential ingredient in hypoglycemic agents or functional food for t展开更多
Fucoidan, a group of sulfated heteropolysaccharides, was extracted from Laminaria japonica, an important economic alga species in China. The anticoagulant activity of fucoidan and its derivatives (including sulfated, ...Fucoidan, a group of sulfated heteropolysaccharides, was extracted from Laminaria japonica, an important economic alga species in China. The anticoagulant activity of fucoidan and its derivatives (including sulfated, phosphorylated, and aminated fucoidan) was examined using in-vitro anticoagulant systems. The correlation between chemical variations within the fucoidan group and anticoagulant activity was determined. The in-vitro anticoagulant properties of fucoidan and its derivatives were determined by measuring activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT). The results indicate anticoagulant activity in all samples using APTT and TT assays; however, only the fucoidan derivatives affected the PT assay. Thus, the fucoidan derivatives were able to inhibit both intrinsic and extrinsic blood coagulants. Fucoidan (FPS) and its derivatives presented better anticoagulant activity than low molecular weight fucoidan (DFPS) and its derivatives, suggesting that molecular weight and proper conformation are contributing factors for anticoagulant activity of polysaccharides. Amino groups have a positive charge and can thus change the charge density of fucoidan. Accordingly, among the tested samples, aminated fucoidan (NF) was the most active reflecting the importance of charge density for anticoagulant activity. Available data obtained using in-vitro models suggest that the sulfate content, sulfate/total-sugar ratio, molecular weight, and the substituted group of fucoidan are important factors for anticoagulant activity but that the influence of sulfate, phosphate and amino groups on anticoagulant activity was different.展开更多
The present study was designed to investigate the effects of Laminaria japonica(Laminaria) on pharmacokinetics of glycyrrhetinic acid(GA) following oral administration of Liquorice extract in rats.Following oral admin...The present study was designed to investigate the effects of Laminaria japonica(Laminaria) on pharmacokinetics of glycyrrhetinic acid(GA) following oral administration of Liquorice extract in rats.Following oral administrations of single-dose and multi-dose Liquorice extract and Liquorice-Laminaria extract,respectively,plasma samples were obtained at various times and the concentrations of GA,liquiritigenin,and isoliquiritigenin were measured by LC-MS.The effects of Laminaria extract on pharmacokinetics of GA were also investigated,following single-dose and multidose of glycyrrhizic acid(GL).The effects of Laminaria extract on intestinal absorption of GA and GL were studied using the in situ single-pass intestinal perfusion model.The metabolism of GL to GA in the contents of small and large intestines was also studied.The results showed Liquorice-Laminaria extract markedly increased the plasma concentration of GA,accompanied by a shorter Tmax.Similar alteration was observed following multidose administration.However,pharmacokinetics of neither liquiritigenin nor isoliquiritigenin was affected by Laminaria.Similarly,Laminaria markedly increased concentration and decreased Tmax of GA following oral GL were observed.The data from the intestinal perfusion model showed that Laminaria markedly increased GL absorption in duodenum and jejunum,but did not affect the intestinal absorption of GA.It was found that Laminaria enhanced the metabolism of GL to GA in large intestine.In conclusion,Laminaria increased plasma exposures of GA following oral administration of liquorice or GL,which partly resulted from increased intestinal absorption of GL and metabolism of GL to GA in large intestine.展开更多
为了纯化海带褐藻糖胶及研究褐藻糖胶结构,利用DEAE-Sepharose Fast Flow离子交换柱分离海带褐藻糖胶粗品,得到LP-1、LP-2、LP-3和LP-4 4个组分,通过Sephadex G-150凝胶柱对硫酸根含量较高组分LP-2、LP-3和LP-4进行进一步分离纯化及分...为了纯化海带褐藻糖胶及研究褐藻糖胶结构,利用DEAE-Sepharose Fast Flow离子交换柱分离海带褐藻糖胶粗品,得到LP-1、LP-2、LP-3和LP-4 4个组分,通过Sephadex G-150凝胶柱对硫酸根含量较高组分LP-2、LP-3和LP-4进行进一步分离纯化及分子量测定,最终LP-2获得两种均一多糖LP-21和LP-22,其分子量分别为123和105 k Da,LP-3和LP-4为均一多糖,分子量分别为74和32 k Da。通过高效液相色谱法分析得率较高的组分(LP-21、LP-22和LP-3)单糖组成,用红外色谱法分析其结构。高效液相色谱结果表明,分级纯化后的组分(LP-21、LP-22和LP-3)主要由L-岩藻糖、D-半乳糖组成,还有少量D-葡萄糖、D-甘露糖、D-氨基葡萄糖、L-鼠李糖、D-木糖和D-葡萄糖醛酸。红外光谱分析表明,LP-21和LP-22的硫酸根主要结合在岩藻糖C_4位,LP-3硫酸根主要结合在岩藻糖C_2或C_3位上。展开更多
In this study, we evaluated the anti-proliferative activity of phlorotannins derived from brown algae Laminariajaponica Aresch extracts on the human hepatocellular carcinoma cell （BEL-7402） and on routine leukemic c...In this study, we evaluated the anti-proliferative activity of phlorotannins derived from brown algae Laminariajaponica Aresch extracts on the human hepatocellular carcinoma cell （BEL-7402） and on routine leukemic cells （P388） by MTT assay. Cells were incubated with 100 μg/mL of the phlorotannin extract （PE） for 48 h. The inhibitory rate of PE on BEL-7402 and P388 cells was 30.20±1.16% and 43.44±1.86%, respectively, and the half-inhibitory concentration of PE （IC50） on P388 and BEL-7402 cells was 120 μg/mL and 〉200 μg/mL, respectively. Microscopic observation shows that the morphologic features of tumor cells treated with PE and 5-fluorouracil are markedly different from the normal control group. The inhibitory rate of fraction A2 isolated from PE by sephadex LH-20 for BEL-7402 and P388 cells at the sample concentration of 70.42 μg/mL was 61.96±7.02% and 40.47±8.70%, respectively. The apoptosis peak for fraction A2 was the most profound of all fractions used in the flow cytometry assay. The results indicate that the anti-proliferative of this algal extract is associated with the total phlorotannin content.展开更多
An effective and accurate method was developed for the extraction of astaxanthin from Laminariajapo- nica with subsequent separation by ionic liquid-based monolithic cartridge. The optimized extraction conditions in- ...An effective and accurate method was developed for the extraction of astaxanthin from Laminariajapo- nica with subsequent separation by ionic liquid-based monolithic cartridge. The optimized extraction conditions in- cluding extraction solvent（ethanol）, extraction time（90 min） and ultrasonic power（75 W） were obtained by systema- tical investigation. Chromatographic analysis was performed on a C18 column with ultraviolet（UV） detection at 476 nm, and a solution consisting of methanol/acetonitrile/H2O/dichloromethane（83：6：6：5, volume ratio） was used as the mobile phase at a flow rate of 0.7 mL/min. After ionic liquid-based monolithic solid phase extraction, 17.82 μg/g as- taxanthin was obtained from Laminaria japonica. This ionic liquid-based monolithic cartridge exhibits high affinity and selectivity for astaxanthin, and it can be potentially used as the stationary phase of high performance liquid chromatography（HPLC）.展开更多
The amplified fragment length polymorphic DNA (AFLP) technique was adopted to estimate the population genetic polymorphism among 30 sporophytes of Laminaria japonica collected from a cultivating farm in Rongcheng,Chin...The amplified fragment length polymorphic DNA (AFLP) technique was adopted to estimate the population genetic polymorphism among 30 sporophytes of Laminaria japonica collected from a cultivating farm in Rongcheng,China.Three methods were used for genomic DNA extraction from Laminaria japonica sporophyte and only the products obtained using the improved genomic DNA extraction kit method proved qualified for AFLP analysis.The parameters of the method were optimized.Samples of forty milligrams and the cell lysis time of 120 min were suggested to replace the parameters recommended by the manufacturer.Thirty individuals of Laminaria japonica from the same cultivating site were investigated using one pair of selective primers.A total of 21 loci were obtained and 17 of them were polymorphic.The mean percent age of polymorphic loci of this population was 80.95%.The Nei's gene diversity (H) within this population was 0.3028 and the average Shannon's Information index (I) was 0.4498.A genetic distance matrix among different individuals was constructed as well.Through this study,an applicable AFLP genetic analysis working system for Laminaria japonica sporophyte was established.The results of this research also revealed a high level of genetic diversity within the studied population.展开更多
文摘Fucoxanthin was extracted from the intact rhizoid of Laminariajaponica Aresch with dimethyl sulfoxide （DMSO）, and then recovered from the DMSO extract by partitioning into ethyl acetate and subsequent evaporation. Some isolation conditions such as solvent volume and extraction time were screened. The quantity and quality of the extracted fucoxanthin were determined by spectral analysis （absorption spectra and fluorescence emission spectra）. The results indicated that： （1） the average total content of fucoxanthin was 122.1μg in 1 g of fresh L. japonica rhizoid; （2） in comparison with the widely used organic solvent, acetone, DMSO was much more effective for the extraction of fucoxanthin; （3） both DMSO volume and extraction time influenced extraction efficiency such as the recovery rate and purity of fucoxanthin （1 g of fresh L. japonica rhizoid treated with 4 mL DMSO for 60 min, yielded 〉 88% of the total fucoxanthin with purity 0.63）; （4） when （NH4）2SO4 concentration was in the range of 0.5-1.0 mol/L, the pigments rapidly and entirely moved from DMSO into the ethyl acetate phase; （5） the ethyl acetate and DMSO were recycled using a rotary evaporator.
基金中国科学院资助项目，the Project of ScientificInnovation of the Chinese Academy of Sciences，国家科技攻关项目
文摘Effects of various concentrations of two heavy metals, namely Cd and Cu, on gametophytes of Laminariajaponica Aresch were determined by recording morphological changes of gametophytes, determining pH values and the heavy metal content of the culture solution, calculating the germination rate of sporophytes, and observing heavy metal （Cd） distribution using a fluorescence microscope. The results showed that heavy metals damaged the gametophytes, and were even lethal, and that the higher the concentration of heavy metal ions, the greater the injury to gametophytes. Gametophytes could not survive in culture solutions containing more than 100 mg/L Cd and 50 mg/L Cu and were only able to survive in culture solution containing a mixture of Cd and Cu up to a concentration of 10 mg/L, which indicates that gametophytes have a higher tolerance to Cd than Cu and that multiple heavy metal ions in solution markedly aggravate the damage to gametophytes compared with individual heavy metal ions. With increases in the concentration of the heavy metal, the burgeoning rate of sporophytes decreased acutely, and solutions containing multiple heavy metal ions caused even more marked harm to sporophytes than solutions containing a single heavy metal ion, because most sporophytes died in mixed solutions. The pH value of the culture medium dropped immediately at the beginning （the first day） of treatment, increased over the following days, and then decreased again. The pH of culture media containing multiple heavy metal ions showed greater variation than media containing a single heavy metal ion, with the extent of the decrease in pH of culture media containing multiple ions being greatest during the last period of the experiment. With increases in the concentration of heavy metals, the capacity of gametophytes to accumulate these ions increased. The blue fluorescent light emitted by the Cd-and Cd-binding protein complex existing in gametophytes in media containing different concentrations of Cd showed clearly the distribution o
基金supported by the Project of National Key Technology Research and Development Program for the 12th Five-year Plan(No.2012BAD33B10)Public Science and Technology Research Funds Projects of Ocean(No.201305018-2)+4 种基金the Innovative Development of Marine Economy Regional Demonstration Projects(Nos.SZHY2012-B01-004,GD2013-B03-001)the National Scien-ce Foundation for Young Scientists of China(No.31101271)the Natural Science Foundation of Guangdong Province(Nos.2014A030310338,2014A030310351)the Comprehensive Strategic Cooperation Programs between the Guangdong Province and Chinese Academy of Sciences(No.2011B090300057)the Frontier Science Program for Young Scientists of South China Sea Institute of Oceanology,Chinese Academy of Science(No.SQ 201017)
文摘In the present study, ultrasonic extraction technique （UET） is used to improve the yield of polysaccharides from Lami- naria japonica （LJPs）. And their antioxidative as well as glycosidase inhibitory activities are investigated. Box-Behnken design （BBD） combined with response surface methodology （RSM） is applied to optimize ultrasonic extraction for polysaccharides. The optimized conditions are obtained as extraction time at 54 min, ultrasonic power at 1050 W, extraction temperature at 80℃ and ratio of material to solvent at 1：50 （g mL-1）. Under these optimal ultrasonic extraction conditions, an actual experimental yield （5.75% ＋ 0.3%） is close to the predicted result （5.67%） with no significant difference （P〉0.05）. Vitro antioxidative and glycosidase inhibitory activities tests indicate that the crude polysaccharides （LJP） and two major ethanol precipitated fractions （LJP1 and LJP2） are in a concentration-dependent manner. LJP2 （30%-60% ethanol precipitated polysaccharides） possesses the strongest α-glucosidase in- hibitory activity and moderate scavenging activity against hydroxyl radicals （66.09% ±2.19%, 3.0 mg mL-l）. Also, the inhibitory activity against a-glucosidase （59.08% ± 3.79%, 5.0 mg mL-1） is close to that of acarbose （63.99% ± 3.27%, 5.0 mg mL-l）. LJP 1 （30% ethanol precipitated polysaccharides） exhibits the strongest scavenging activity against hydroxyl radicals （99.80%q-0.00%, 3.0mg mL-1） and moderate a-glucosidase inhibitory activity （47.76%± 1.92%, 5.0 mgmL-1）. LJP shows the most remarkable DPPH scav- enging activity （66.20%±0.11%, 5.0mgmL-1） but weakest a-glucosidase inhibitory activity （37.77%±1.30%, 5.0mgmL-1）. How- ever, all these LJPs exert weak inhibitory effects against a-amylase. These results show that UET is an effective method for extract- ing bioactive polysaccharides from seaweed materials. LJP 1 and LJP2 can be developed as a potential ingredient in hypoglycemic agents or functional food for t
基金Supported by the Ocean Public Welfare Scientific Research Project, State Oceanic Administration of the People's Republic of China (No. 201005024)
文摘Fucoidan, a group of sulfated heteropolysaccharides, was extracted from Laminaria japonica, an important economic alga species in China. The anticoagulant activity of fucoidan and its derivatives (including sulfated, phosphorylated, and aminated fucoidan) was examined using in-vitro anticoagulant systems. The correlation between chemical variations within the fucoidan group and anticoagulant activity was determined. The in-vitro anticoagulant properties of fucoidan and its derivatives were determined by measuring activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT). The results indicate anticoagulant activity in all samples using APTT and TT assays; however, only the fucoidan derivatives affected the PT assay. Thus, the fucoidan derivatives were able to inhibit both intrinsic and extrinsic blood coagulants. Fucoidan (FPS) and its derivatives presented better anticoagulant activity than low molecular weight fucoidan (DFPS) and its derivatives, suggesting that molecular weight and proper conformation are contributing factors for anticoagulant activity of polysaccharides. Amino groups have a positive charge and can thus change the charge density of fucoidan. Accordingly, among the tested samples, aminated fucoidan (NF) was the most active reflecting the importance of charge density for anticoagulant activity. Available data obtained using in-vitro models suggest that the sulfate content, sulfate/total-sugar ratio, molecular weight, and the substituted group of fucoidan are important factors for anticoagulant activity but that the influence of sulfate, phosphate and amino groups on anticoagulant activity was different.
基金supported by funding from the National Basic Research Program of China(973 Program)(Nos.2011CB505300,2011CB505303)
文摘The present study was designed to investigate the effects of Laminaria japonica(Laminaria) on pharmacokinetics of glycyrrhetinic acid(GA) following oral administration of Liquorice extract in rats.Following oral administrations of single-dose and multi-dose Liquorice extract and Liquorice-Laminaria extract,respectively,plasma samples were obtained at various times and the concentrations of GA,liquiritigenin,and isoliquiritigenin were measured by LC-MS.The effects of Laminaria extract on pharmacokinetics of GA were also investigated,following single-dose and multidose of glycyrrhizic acid(GL).The effects of Laminaria extract on intestinal absorption of GA and GL were studied using the in situ single-pass intestinal perfusion model.The metabolism of GL to GA in the contents of small and large intestines was also studied.The results showed Liquorice-Laminaria extract markedly increased the plasma concentration of GA,accompanied by a shorter Tmax.Similar alteration was observed following multidose administration.However,pharmacokinetics of neither liquiritigenin nor isoliquiritigenin was affected by Laminaria.Similarly,Laminaria markedly increased concentration and decreased Tmax of GA following oral GL were observed.The data from the intestinal perfusion model showed that Laminaria markedly increased GL absorption in duodenum and jejunum,but did not affect the intestinal absorption of GA.It was found that Laminaria enhanced the metabolism of GL to GA in large intestine.In conclusion,Laminaria increased plasma exposures of GA following oral administration of liquorice or GL,which partly resulted from increased intestinal absorption of GL and metabolism of GL to GA in large intestine.
文摘为了纯化海带褐藻糖胶及研究褐藻糖胶结构,利用DEAE-Sepharose Fast Flow离子交换柱分离海带褐藻糖胶粗品,得到LP-1、LP-2、LP-3和LP-4 4个组分,通过Sephadex G-150凝胶柱对硫酸根含量较高组分LP-2、LP-3和LP-4进行进一步分离纯化及分子量测定,最终LP-2获得两种均一多糖LP-21和LP-22,其分子量分别为123和105 k Da,LP-3和LP-4为均一多糖,分子量分别为74和32 k Da。通过高效液相色谱法分析得率较高的组分(LP-21、LP-22和LP-3)单糖组成,用红外色谱法分析其结构。高效液相色谱结果表明,分级纯化后的组分(LP-21、LP-22和LP-3)主要由L-岩藻糖、D-半乳糖组成,还有少量D-葡萄糖、D-甘露糖、D-氨基葡萄糖、L-鼠李糖、D-木糖和D-葡萄糖醛酸。红外光谱分析表明,LP-21和LP-22的硫酸根主要结合在岩藻糖C_4位,LP-3硫酸根主要结合在岩藻糖C_2或C_3位上。
基金Supported by the National Key Technology Research & Development Program of the 11th Five Year Plan of China (No. 2006BAD30B01)the National Natural Science Foundation of China (No. 30871945)
文摘In this study, we evaluated the anti-proliferative activity of phlorotannins derived from brown algae Laminariajaponica Aresch extracts on the human hepatocellular carcinoma cell （BEL-7402） and on routine leukemic cells （P388） by MTT assay. Cells were incubated with 100 μg/mL of the phlorotannin extract （PE） for 48 h. The inhibitory rate of PE on BEL-7402 and P388 cells was 30.20±1.16% and 43.44±1.86%, respectively, and the half-inhibitory concentration of PE （IC50） on P388 and BEL-7402 cells was 120 μg/mL and 〉200 μg/mL, respectively. Microscopic observation shows that the morphologic features of tumor cells treated with PE and 5-fluorouracil are markedly different from the normal control group. The inhibitory rate of fraction A2 isolated from PE by sephadex LH-20 for BEL-7402 and P388 cells at the sample concentration of 70.42 μg/mL was 61.96±7.02% and 40.47±8.70%, respectively. The apoptosis peak for fraction A2 was the most profound of all fractions used in the flow cytometry assay. The results indicate that the anti-proliferative of this algal extract is associated with the total phlorotannin content.
基金Supported by the Korea Sea Grant Program(Gyeong-gi Sea Grant) Funded by the Ministry of Land,Transport and Maritime Affairs,Korea
文摘An effective and accurate method was developed for the extraction of astaxanthin from Laminariajapo- nica with subsequent separation by ionic liquid-based monolithic cartridge. The optimized extraction conditions in- cluding extraction solvent（ethanol）, extraction time（90 min） and ultrasonic power（75 W） were obtained by systema- tical investigation. Chromatographic analysis was performed on a C18 column with ultraviolet（UV） detection at 476 nm, and a solution consisting of methanol/acetonitrile/H2O/dichloromethane（83：6：6：5, volume ratio） was used as the mobile phase at a flow rate of 0.7 mL/min. After ionic liquid-based monolithic solid phase extraction, 17.82 μg/g as- taxanthin was obtained from Laminaria japonica. This ionic liquid-based monolithic cartridge exhibits high affinity and selectivity for astaxanthin, and it can be potentially used as the stationary phase of high performance liquid chromatography（HPLC）.
基金funded by the ‘908’ Marine Survey Project of Shandong Province (SD-908-01-01-05.06)
文摘The amplified fragment length polymorphic DNA (AFLP) technique was adopted to estimate the population genetic polymorphism among 30 sporophytes of Laminaria japonica collected from a cultivating farm in Rongcheng,China.Three methods were used for genomic DNA extraction from Laminaria japonica sporophyte and only the products obtained using the improved genomic DNA extraction kit method proved qualified for AFLP analysis.The parameters of the method were optimized.Samples of forty milligrams and the cell lysis time of 120 min were suggested to replace the parameters recommended by the manufacturer.Thirty individuals of Laminaria japonica from the same cultivating site were investigated using one pair of selective primers.A total of 21 loci were obtained and 17 of them were polymorphic.The mean percent age of polymorphic loci of this population was 80.95%.The Nei's gene diversity (H) within this population was 0.3028 and the average Shannon's Information index (I) was 0.4498.A genetic distance matrix among different individuals was constructed as well.Through this study,an applicable AFLP genetic analysis working system for Laminaria japonica sporophyte was established.The results of this research also revealed a high level of genetic diversity within the studied population.
