半枝莲多糖的提纯、结构解析及其抗氧化活性的研究
|
摘要
半枝莲为唇形科植物半枝莲Scutellaria barbata D.Don的干燥全草,分布在我国大部分地区。半枝莲作为传统的中药具有抗肿瘤、抑菌、解热等活性,其主要化学成分为黄酮类化合物、生物碱、挥发油及多糖等,多糖是半枝莲中的主要化学成分之一,有多种生物活性,但目前对半枝莲多糖的分离纯化及结构性质的研究报道甚少。本文研究了半枝莲多糖的提取、分离、纯化及硫酸化修饰,并且对半枝莲多糖SBP、SBP_1、SBP_2、SBPⅠ、SBP-L、SBP-L_1和SBP-L_2的体外抗氧化活性及其化学结构进行了研究,为今后半枝莲多糖更深入的研究与开发奠定了基础。研究结果如下:
1.在单因素实验基础上,设计正交实验优化了半枝莲粗多糖的提取工艺,得到热水提取半枝莲粗多糖的最佳工艺参数为:提取温度60℃,料液比1:20,提取时间2h,提取次数4次,半枝莲粗多糖的提取率为2.82%,并采用苯酚.硫酸法对多糖含量进行了测定。
2.进行了多糖脱色工艺的研究,对活性炭、中性氧化铝、大孔吸附树脂AB-8、LSA-21和NKA-9五种脱色剂的脱色性能进行了比较,从中筛选出了最佳的脱色剂为大孔吸附树脂AB-8。通过单因素实验确定了脱色的最佳条件是pH为7,温度为40℃,溶液中多糖含量为1046.5 mg/L。采用最佳条件脱色验证,测定多糖的保留率为88.46%,脱色率为90.65%。
3.脱色后的多糖(SBP)采用季铵盐沉淀法(Hexadecyltrimethylammoniumbromide,CTAB)进行分离,得到半枝莲多糖亚组分SBP_1和SBP_2。采用DE-52纤维素离子交换柱层析对多糖SBP进行了分级分离,得到水洗组分SBPⅠ用0.1 moL/LNaCl溶液洗脱得到SBPⅡ。利用Sephdex-200柱层析、高效液相色谱、紫外光谱和冻融法对SBPⅠ进行纯度鉴定,证明了SBPⅠ为组成均一的多糖组分。
4.对多糖进行了硫酸化修饰。采用氯磺酸-吡啶法对半枝莲多糖SBP进行硫酸酯化修饰,得到多糖SBP-L,其水溶性较原多糖明显提高,通过紫外分光光度计扫描在262 nm处有吸收峰,测得的SO_4~(2-)浓度为5.73 mg/mL,取代度为0.35。红外光谱显示在1258 cm~(-1)附近出现硫酸基(-O-SO~(3-))的特征吸收峰。
5.采用气相色谱法对半枝莲多糖的单糖组分进行了分析。比较了硅烷衍生化、糖腈衍生化法和糖醇衍生化三种衍生化方法,确定了糖腈乙酰化法为最佳的衍生化方法;通过单因素实验确定了硫酸水解半枝莲多糖的最佳条件为:硫酸浓度为0.5 mol/L、体积2 mL、水解时间12 h、温度100℃:气相色谱测定半枝莲多糖SBP由鼠李糖阿拉伯糖、木糖、甘露糖、葡萄糖和半乳糖组成,摩尔比为3.26:0.37:4.85:5.16:4.10:9.47,SBP-L由阿拉伯糖、甘露糖、葡萄糖和半乳糖组成,摩尔比为0.84:6.67:3.62:5.53,SBP1,SBP2,SBP-L1,SBP-L2和SBPⅠ都由鼠李糖木糖、甘露糖、葡萄糖和半乳糖组成,摩尔比依次为4.51:5.50:4.86:4.90:8.22,3.25:6.20:5.85:4.05:11.30,3.86:4.39:4.96:4.95:7.73,3.04:6.87:5.51:4.37:10.90,3.53:4.62:7.22:4.08:8.37。
6.利用高效液相色谱测定多糖的分子量,得到半枝莲多糖SBP1、SBP2、SBPⅠ、SBP-L_1和SBP-L_2的分子量依次为:290000Da、340000Da、300000Da、360000Da和390000Da;采用部分酸水解、高碘酸氧化、Smith降解结合气相色谱和红外光谱对半枝莲多糖进行结构分析,确定了多糖的结构:SBP_1和SBP_2糖链连接方式都是由β-(1→2)—吡喃葡聚糖及半乳糖构成主链,其中SBP_1由(1→3,4,6)—鼠李糖、阿拉伯糖、木糖、甘露糖和葡萄糖残基组成侧链分支,多糖中(1→6)键、(1→3)键、(1→2或1→4)键摩尔百分比约为9%:22%:69%,SBP_2由(1→3,4,6)—鼠李糖、阿拉伯糖、甘露糖和葡萄糖残基组成侧链分支,多糖中(1→6)键、(1→3)键、(1→2或1→4)键摩尔百分比约为12%:26%:62%:SBPⅠ以β—(1→2,3)—吡喃葡聚糖及半乳糖构成主链,由(1→4,6)—鼠李糖、阿拉伯糖、甘露糖和葡萄糖残基组成侧链分支,多糖中(1→6)键、(1→3)键、(1→2或1→4)键摩尔百分比约为10.6%:47%:42.4%:SBP-L_1以β—(1→2)—吡喃葡聚糖及半乳糖构成主链,由(1→3,4,6)—鼠李糖、阿拉伯糖、木糖、甘露糖和葡萄糖残基组成侧链分支,并存在少量α—糖苷键型吡喃糖基,多糖中(1→6)键、(1→3)键、(1→2或1→4)键摩尔百分比约为9.6%:38.6%:51.8%;SBP-L_2以β—(1→2)—吡喃葡聚糖及半乳糖、阿拉伯糖构成主链,由(1→3,4,6)—鼠李糖、甘露糖和葡萄糖残基组成侧链分支,多糖中(1→6)键、(1→3)键、(1→2或1→4)键摩尔百分比约为10.6%:24.2%:65.2%。
7.对七种半枝莲多糖(SBP、SBP1、SBP_2、SBPⅠ、SBP-L、SBP-L_1和SBP-L_2)的体外抗氧化活性进行了研究:采用邻苯三酚自氧化方法,测定半枝莲多糖对O~(2-)自由基的抑制作用;采用邻二氮菲-亚铁离子—H_2O_2体系测定了半枝莲多糖对羟基自由基的抑制作用;体外模拟胃液pH值条件下测定了半枝莲多糖对NO_2~-清除能力。结果表明,七种多糖均具有不同的抗氧化活性,呈剂量依赖关系。半枝莲多糖SBP的抗氧化性要强于分离后的多糖SBP_1和SBP_2,硫酸化修饰后的多糖SBP-L和SBP-L_2的抗氧化性较原多糖SBP和SBP_2的抗氧化性增强,多糖SBP-L_1较多糖SBP_1对羟基自由基和NO~(2-)的清除效果有所降低,对O~(2-)自由基清除效果有所提高。本文研究表明,几种半枝莲多糖都具有不同百分比的β—(1→3)葡聚糖苷键,因此,七种多糖出现不同程度的抗氧化性。
Scutellaria barbata D.Don,a herb belonging to the Lamiaceae family,is widely distributed in china.This herb is known in traditional Chinesse Medicine Ban-Zhi-Lian and has been used as an antitumor,anti-inflammatory,antifebrile agent.The main chemical compositions are flavonoids,alkaloid,essential oils and polysaccharide.Polysaccharide which is the one of major component of Scutellaria barbata D.Don have many kinds of bioactivities.But the study were seldom carried out on isolation,purification,structure and property of polysaccharides from Scutellaria barbata D.Don.This paper studied the extraction,separation,purification and sulfation of Scutellaria barbata D.Don polysaccharide,the antioxidant activity of SBP,SBP_1,SBP_2,SBPⅠ、SBP-L、SBP-L_1 and SBP-L_2 and their chemical structure,which gave a foundation for the further research..The main results and innovations were as follows:
1.Based on the single factor experiment,designing orthogonal experiments to optimize the extraction process of crude Scutellaria barbata polysaccharide,the optimal parameters of extracting Scutellaria barbata polysaccharide by hot water were:the extraction temperature was 60℃, solid-liquid ratio was 1:20,extraction time was 2 h and extraction times were 4,and the rate of polysaccharides extraction was 2.82%.The amount of carbohydrates was measured with phenol-sulfuric acid method.
2.This paper studied on de-coloration of Scutellaria barbata polysaccharide.which compared the de-coloration capacity of five different kinds of de-coloration agents:active carbon,neutral alumina,macroporous adsorption resins AB-8,LSA-21 and NKA-9.The better de-coloration agents was AB-8.Decoloring experiments were carried out by single factor experiment.The best conditions for de-coloration were determined as follows:the pH is 7,the temperature is 40℃,the content of polysaccharides is 1046.5 mg/L.Adopt the best condition,holding rate of polysaccharides is 88.46%,de-coloration rate is 90.65%.
3.Polysaccharides(SBP) were treated after de-coloring by quaternary ammonium salt precipitation method.SBP_1 and SBP_2 were obtained.SBP was isolated using DE-52 cellulose chromatography in series processes.SBPⅠand SBPⅡwas obtained respectively eluting the DE-52 cellulose chromatography with distilled water and the solution of 0.1 moL/L NaCI.SBPⅠwas identified as homogeneous polysaccharide component by SephadexG-200 gel chromatography,HPLC,UV spectrum and freeze-thawing.
4.Chemical modification of sulfation reaction about SBP,Scutellaria barbata polysaccharide Sulfate(SBP-L) was obtained and its water-sohibility was increased.By scanning it with UV spectrum,the results showed that it had absorption peaks at 262 nm.The content of sulphate groups was 5.73 mg/mL,the proportion of sulfate substitution was 2.89.IR showed that it has a-O-SO~(3-) absorption peaks at 1258 cm~(-1).
5.The monosaccharide components of Scutellaria barbata polysaccharide were measured by GC.Three methods of TMS,acety.lated aldononitriles and sugar alcohol acetates were compared to confirm acetylated aldononitriles which is one of the best methods.Hydrolyzing experiment were carried out by single factor experiment.The best conditions were determined as follows:the concentration of sulphuric acid was 0.5mol/L,the volume was 2 mL,the time was 12 h and the temperature was 100℃.GC result showed that SBP was mainly composed of rhamnose,arabinose, xylose,mannose,glucose and galactose,and the mole proportion of monosaccharide was 3.26:0.37:4.85:5.16:4.10:9.47.SBP-L was mainly composed of arabinose,mannose,glucose and galactose,and the mole proportion of monosaccharide was 0.84:6.67:3.62:5.53.SBP_1,SBP_2,SBP-L_1, SBP-L_2 and SBP I were mainly composed of rhamnosexylose,mannose,glucose,galactose and the mole proportion of monosaccharide respectively were 4.51:5.50:4.86:4.90:8.22,3.25:6.20:5.85: 4.05:11.30,3.86:4.39:4.96:4.95:7.73,3.04:6.87:5.51:4.37:10.90,3.53:4.62:7.22:4.08:8.37.
6.The average molecular weights of SBP_1,SBP_2,SBPⅠ,SBP-L_1 and SBP-L_2 were approximately 290000,340000,300000,360000 and 390000Da.,determined respectively by HPLC. Partial hydrolysis with sulphuric acid,periodate oxidation,Smith degradation,GC and IR was used to determine their structure.The results showed that:The main chain of SBP_1 and SBP_2 was made up ofβ—(1→2)—pyranose glucosan and galactose,and the side chain of SBP_1 was composed of(1→3, 4,6)—rhamnose,arabinose,xylose,mannose and glucose.The mole proportion of bond(1→6): (1→3):(1→2 or 1→4)was 9%:22%:69%.The side chain of SBP_2 was composed of(1→3, 4,6)—rhamnose,arabinose,mannose and glucose.The mole proportion of bond(1→6):(1→3): (1→2 or 1→4) was 12%:26%:62%.The main chain of SBPⅠwas made up ofβ—(1→2,3)—pyranose glucosan and galactose,and the side chain of SBPⅠwas composed of(1→4,6)—rhamnose,arabinose,mannose and glucose.The mole proportion of bond(1→6):(1→3):(1→2 or 1→4)was 10.6%:47%:42.4%.The main chain of SBP-L_1 was made up ofβ—(1→2)—pyranose glucosan and galactose,and the side chain of SBP-L_1 was composed of(1→3,4,6)—rhamnose, arabinose,xylose,mannose and glucose and had a fewα—pyranose glucesidal bond.The mole proportion of bond(1→6):(1→3):(1→2 or 1→4) was 9.6%:38.6%:51.8%.The main chain of SBP-L_2 was made up ofβ—(1→2)—pyranose glucosan,galactose and arabinose,and the side chain of SBP-L_2 was composed of(1→3,4,6)—rhamnose,mannose and glucose.The mole proportion of bond(1→6):(1→3):(1→2 or 1→4) was10.6%:24.2%:65.2%.
7.Antioxidative activities of seven kinds of Scutellaria barbata polysaccharides(SBP,SBP_1, SBP_2,SBPⅠ,SBP-L,SBP-L_1 and SBP-L_2) in vitro were studied:the capability of the polysaccharides to eliminate O~(2-) was determined through pyrogallol auto oxidation;the ability of the polysaccharides to scavenge hydroxyl radicals was determined by the system of ferrosin-iron-H_2O_2 and the ability of the polysaccharides to scavenge NO_2~- in the condition of pH for simulation gastric juice in vitro.The result showed that the seven kinds of Scutellaria barbata polysaccharides had different antioxidation activities in vitro,and their activities depended on their amounts.The antioxidation activities of SBP was better than separate polysaccharides SBP_1 and SBP_2.The antioxidant activities of sulfation of Scutellaria barbata polysaccharide SBP-L and SBP-L_2 were increased than SBP and SBP_2,through comparing with SBP_1 and SBP-L_1 the ability of the SBP_1 to scavenge hydroxyl radicals and NO_2~- is better,but the capability of the polysaccharides to eliminate O~(2-) was reverse.This paper studied that seven kinds of Scutellaria barbata polysaccharides had different antioxidation activities in vitro because all kinds of them had different percent ofβ-(1→3) glucesidal bond.
1.在单因素实验基础上,设计正交实验优化了半枝莲粗多糖的提取工艺,得到热水提取半枝莲粗多糖的最佳工艺参数为:提取温度60℃,料液比1:20,提取时间2h,提取次数4次,半枝莲粗多糖的提取率为2.82%,并采用苯酚.硫酸法对多糖含量进行了测定。
2.进行了多糖脱色工艺的研究,对活性炭、中性氧化铝、大孔吸附树脂AB-8、LSA-21和NKA-9五种脱色剂的脱色性能进行了比较,从中筛选出了最佳的脱色剂为大孔吸附树脂AB-8。通过单因素实验确定了脱色的最佳条件是pH为7,温度为40℃,溶液中多糖含量为1046.5 mg/L。采用最佳条件脱色验证,测定多糖的保留率为88.46%,脱色率为90.65%。
3.脱色后的多糖(SBP)采用季铵盐沉淀法(Hexadecyltrimethylammoniumbromide,CTAB)进行分离,得到半枝莲多糖亚组分SBP_1和SBP_2。采用DE-52纤维素离子交换柱层析对多糖SBP进行了分级分离,得到水洗组分SBPⅠ用0.1 moL/LNaCl溶液洗脱得到SBPⅡ。利用Sephdex-200柱层析、高效液相色谱、紫外光谱和冻融法对SBPⅠ进行纯度鉴定,证明了SBPⅠ为组成均一的多糖组分。
4.对多糖进行了硫酸化修饰。采用氯磺酸-吡啶法对半枝莲多糖SBP进行硫酸酯化修饰,得到多糖SBP-L,其水溶性较原多糖明显提高,通过紫外分光光度计扫描在262 nm处有吸收峰,测得的SO_4~(2-)浓度为5.73 mg/mL,取代度为0.35。红外光谱显示在1258 cm~(-1)附近出现硫酸基(-O-SO~(3-))的特征吸收峰。
5.采用气相色谱法对半枝莲多糖的单糖组分进行了分析。比较了硅烷衍生化、糖腈衍生化法和糖醇衍生化三种衍生化方法,确定了糖腈乙酰化法为最佳的衍生化方法;通过单因素实验确定了硫酸水解半枝莲多糖的最佳条件为:硫酸浓度为0.5 mol/L、体积2 mL、水解时间12 h、温度100℃:气相色谱测定半枝莲多糖SBP由鼠李糖阿拉伯糖、木糖、甘露糖、葡萄糖和半乳糖组成,摩尔比为3.26:0.37:4.85:5.16:4.10:9.47,SBP-L由阿拉伯糖、甘露糖、葡萄糖和半乳糖组成,摩尔比为0.84:6.67:3.62:5.53,SBP1,SBP2,SBP-L1,SBP-L2和SBPⅠ都由鼠李糖木糖、甘露糖、葡萄糖和半乳糖组成,摩尔比依次为4.51:5.50:4.86:4.90:8.22,3.25:6.20:5.85:4.05:11.30,3.86:4.39:4.96:4.95:7.73,3.04:6.87:5.51:4.37:10.90,3.53:4.62:7.22:4.08:8.37。
6.利用高效液相色谱测定多糖的分子量,得到半枝莲多糖SBP1、SBP2、SBPⅠ、SBP-L_1和SBP-L_2的分子量依次为:290000Da、340000Da、300000Da、360000Da和390000Da;采用部分酸水解、高碘酸氧化、Smith降解结合气相色谱和红外光谱对半枝莲多糖进行结构分析,确定了多糖的结构:SBP_1和SBP_2糖链连接方式都是由β-(1→2)—吡喃葡聚糖及半乳糖构成主链,其中SBP_1由(1→3,4,6)—鼠李糖、阿拉伯糖、木糖、甘露糖和葡萄糖残基组成侧链分支,多糖中(1→6)键、(1→3)键、(1→2或1→4)键摩尔百分比约为9%:22%:69%,SBP_2由(1→3,4,6)—鼠李糖、阿拉伯糖、甘露糖和葡萄糖残基组成侧链分支,多糖中(1→6)键、(1→3)键、(1→2或1→4)键摩尔百分比约为12%:26%:62%:SBPⅠ以β—(1→2,3)—吡喃葡聚糖及半乳糖构成主链,由(1→4,6)—鼠李糖、阿拉伯糖、甘露糖和葡萄糖残基组成侧链分支,多糖中(1→6)键、(1→3)键、(1→2或1→4)键摩尔百分比约为10.6%:47%:42.4%:SBP-L_1以β—(1→2)—吡喃葡聚糖及半乳糖构成主链,由(1→3,4,6)—鼠李糖、阿拉伯糖、木糖、甘露糖和葡萄糖残基组成侧链分支,并存在少量α—糖苷键型吡喃糖基,多糖中(1→6)键、(1→3)键、(1→2或1→4)键摩尔百分比约为9.6%:38.6%:51.8%;SBP-L_2以β—(1→2)—吡喃葡聚糖及半乳糖、阿拉伯糖构成主链,由(1→3,4,6)—鼠李糖、甘露糖和葡萄糖残基组成侧链分支,多糖中(1→6)键、(1→3)键、(1→2或1→4)键摩尔百分比约为10.6%:24.2%:65.2%。
7.对七种半枝莲多糖(SBP、SBP1、SBP_2、SBPⅠ、SBP-L、SBP-L_1和SBP-L_2)的体外抗氧化活性进行了研究:采用邻苯三酚自氧化方法,测定半枝莲多糖对O~(2-)自由基的抑制作用;采用邻二氮菲-亚铁离子—H_2O_2体系测定了半枝莲多糖对羟基自由基的抑制作用;体外模拟胃液pH值条件下测定了半枝莲多糖对NO_2~-清除能力。结果表明,七种多糖均具有不同的抗氧化活性,呈剂量依赖关系。半枝莲多糖SBP的抗氧化性要强于分离后的多糖SBP_1和SBP_2,硫酸化修饰后的多糖SBP-L和SBP-L_2的抗氧化性较原多糖SBP和SBP_2的抗氧化性增强,多糖SBP-L_1较多糖SBP_1对羟基自由基和NO~(2-)的清除效果有所降低,对O~(2-)自由基清除效果有所提高。本文研究表明,几种半枝莲多糖都具有不同百分比的β—(1→3)葡聚糖苷键,因此,七种多糖出现不同程度的抗氧化性。
Scutellaria barbata D.Don,a herb belonging to the Lamiaceae family,is widely distributed in china.This herb is known in traditional Chinesse Medicine Ban-Zhi-Lian and has been used as an antitumor,anti-inflammatory,antifebrile agent.The main chemical compositions are flavonoids,alkaloid,essential oils and polysaccharide.Polysaccharide which is the one of major component of Scutellaria barbata D.Don have many kinds of bioactivities.But the study were seldom carried out on isolation,purification,structure and property of polysaccharides from Scutellaria barbata D.Don.This paper studied the extraction,separation,purification and sulfation of Scutellaria barbata D.Don polysaccharide,the antioxidant activity of SBP,SBP_1,SBP_2,SBPⅠ、SBP-L、SBP-L_1 and SBP-L_2 and their chemical structure,which gave a foundation for the further research..The main results and innovations were as follows:
1.Based on the single factor experiment,designing orthogonal experiments to optimize the extraction process of crude Scutellaria barbata polysaccharide,the optimal parameters of extracting Scutellaria barbata polysaccharide by hot water were:the extraction temperature was 60℃, solid-liquid ratio was 1:20,extraction time was 2 h and extraction times were 4,and the rate of polysaccharides extraction was 2.82%.The amount of carbohydrates was measured with phenol-sulfuric acid method.
2.This paper studied on de-coloration of Scutellaria barbata polysaccharide.which compared the de-coloration capacity of five different kinds of de-coloration agents:active carbon,neutral alumina,macroporous adsorption resins AB-8,LSA-21 and NKA-9.The better de-coloration agents was AB-8.Decoloring experiments were carried out by single factor experiment.The best conditions for de-coloration were determined as follows:the pH is 7,the temperature is 40℃,the content of polysaccharides is 1046.5 mg/L.Adopt the best condition,holding rate of polysaccharides is 88.46%,de-coloration rate is 90.65%.
3.Polysaccharides(SBP) were treated after de-coloring by quaternary ammonium salt precipitation method.SBP_1 and SBP_2 were obtained.SBP was isolated using DE-52 cellulose chromatography in series processes.SBPⅠand SBPⅡwas obtained respectively eluting the DE-52 cellulose chromatography with distilled water and the solution of 0.1 moL/L NaCI.SBPⅠwas identified as homogeneous polysaccharide component by SephadexG-200 gel chromatography,HPLC,UV spectrum and freeze-thawing.
4.Chemical modification of sulfation reaction about SBP,Scutellaria barbata polysaccharide Sulfate(SBP-L) was obtained and its water-sohibility was increased.By scanning it with UV spectrum,the results showed that it had absorption peaks at 262 nm.The content of sulphate groups was 5.73 mg/mL,the proportion of sulfate substitution was 2.89.IR showed that it has a-O-SO~(3-) absorption peaks at 1258 cm~(-1).
5.The monosaccharide components of Scutellaria barbata polysaccharide were measured by GC.Three methods of TMS,acety.lated aldononitriles and sugar alcohol acetates were compared to confirm acetylated aldononitriles which is one of the best methods.Hydrolyzing experiment were carried out by single factor experiment.The best conditions were determined as follows:the concentration of sulphuric acid was 0.5mol/L,the volume was 2 mL,the time was 12 h and the temperature was 100℃.GC result showed that SBP was mainly composed of rhamnose,arabinose, xylose,mannose,glucose and galactose,and the mole proportion of monosaccharide was 3.26:0.37:4.85:5.16:4.10:9.47.SBP-L was mainly composed of arabinose,mannose,glucose and galactose,and the mole proportion of monosaccharide was 0.84:6.67:3.62:5.53.SBP_1,SBP_2,SBP-L_1, SBP-L_2 and SBP I were mainly composed of rhamnosexylose,mannose,glucose,galactose and the mole proportion of monosaccharide respectively were 4.51:5.50:4.86:4.90:8.22,3.25:6.20:5.85: 4.05:11.30,3.86:4.39:4.96:4.95:7.73,3.04:6.87:5.51:4.37:10.90,3.53:4.62:7.22:4.08:8.37.
6.The average molecular weights of SBP_1,SBP_2,SBPⅠ,SBP-L_1 and SBP-L_2 were approximately 290000,340000,300000,360000 and 390000Da.,determined respectively by HPLC. Partial hydrolysis with sulphuric acid,periodate oxidation,Smith degradation,GC and IR was used to determine their structure.The results showed that:The main chain of SBP_1 and SBP_2 was made up ofβ—(1→2)—pyranose glucosan and galactose,and the side chain of SBP_1 was composed of(1→3, 4,6)—rhamnose,arabinose,xylose,mannose and glucose.The mole proportion of bond(1→6): (1→3):(1→2 or 1→4)was 9%:22%:69%.The side chain of SBP_2 was composed of(1→3, 4,6)—rhamnose,arabinose,mannose and glucose.The mole proportion of bond(1→6):(1→3): (1→2 or 1→4) was 12%:26%:62%.The main chain of SBPⅠwas made up ofβ—(1→2,3)—pyranose glucosan and galactose,and the side chain of SBPⅠwas composed of(1→4,6)—rhamnose,arabinose,mannose and glucose.The mole proportion of bond(1→6):(1→3):(1→2 or 1→4)was 10.6%:47%:42.4%.The main chain of SBP-L_1 was made up ofβ—(1→2)—pyranose glucosan and galactose,and the side chain of SBP-L_1 was composed of(1→3,4,6)—rhamnose, arabinose,xylose,mannose and glucose and had a fewα—pyranose glucesidal bond.The mole proportion of bond(1→6):(1→3):(1→2 or 1→4) was 9.6%:38.6%:51.8%.The main chain of SBP-L_2 was made up ofβ—(1→2)—pyranose glucosan,galactose and arabinose,and the side chain of SBP-L_2 was composed of(1→3,4,6)—rhamnose,mannose and glucose.The mole proportion of bond(1→6):(1→3):(1→2 or 1→4) was10.6%:24.2%:65.2%.
7.Antioxidative activities of seven kinds of Scutellaria barbata polysaccharides(SBP,SBP_1, SBP_2,SBPⅠ,SBP-L,SBP-L_1 and SBP-L_2) in vitro were studied:the capability of the polysaccharides to eliminate O~(2-) was determined through pyrogallol auto oxidation;the ability of the polysaccharides to scavenge hydroxyl radicals was determined by the system of ferrosin-iron-H_2O_2 and the ability of the polysaccharides to scavenge NO_2~- in the condition of pH for simulation gastric juice in vitro.The result showed that the seven kinds of Scutellaria barbata polysaccharides had different antioxidation activities in vitro,and their activities depended on their amounts.The antioxidation activities of SBP was better than separate polysaccharides SBP_1 and SBP_2.The antioxidant activities of sulfation of Scutellaria barbata polysaccharide SBP-L and SBP-L_2 were increased than SBP and SBP_2,through comparing with SBP_1 and SBP-L_1 the ability of the SBP_1 to scavenge hydroxyl radicals and NO_2~- is better,but the capability of the polysaccharides to eliminate O~(2-) was reverse.This paper studied that seven kinds of Scutellaria barbata polysaccharides had different antioxidation activities in vitro because all kinds of them had different percent ofβ-(1→3) glucesidal bond.
引文
[1]肖海涛,李铣.半枝莲的化学成分[J].沈阳药科大学学报,2006,23(10):637-640.
[2]张素华,辛春兰.半枝莲的药理作用及应用[J].中医杂志,2007,6(48):572.
[3]邹箴蕾,吴启南.半枝莲的化学成分及药理作用研究进展[J].时珍国医国药,2005,16(2):149-150.
[4]杨蓓芬,李钧敏,邵红.半枝莲的次生代谢产物含量测定与体外抑菌活性的研究[J].四川中医,2005,23(11):35-36.
[5]袁崇均,王笳,杨红,黄卫平.正交试验法优选半枝莲提取工艺的研究[J].华西药学杂志,2002,17(2):112-113.
[6]邸多隆,刘丽春,刘晔玮,刘霞,蒋生祥.半枝莲提取工艺的研究[J].中成药,2003,7(25):530-532.
[7]张海方,许化溪.半枝莲化学成分及生物学活性的研究进展[J].江苏大学学报(医学版),2006,1(16):74.
[8]Sato Yoichi,et al.Phytochemical flavones isolated from Scutellaria barbara and antibacterial activity against methicillin-resistant Staphylococcus aureus[J].Journal of Ethnopharmacology,2000(72):483-488.
[9]Yu JQ,Lei JC,Yu HD,et al.Chemical composition and antimicrobial of the essential oil of Scutellaria barbata[J].Phytochemistry,2004,65:881-884.
[10]贺莉娟,梁逸曾,赵晨曦.唇形科植物挥发油化学成分的GC/MS研究[J].化学学报,2007,3(65):227-232.
[11]许益民,郭立伟,陈建伟.半枝莲多糖的分离、纯化及其理化性质[J].天然产物研究与开发,1992,1(4):1-5.
[12]孟延发,李志孝,张立.半枝莲多糖组分的纯化性质及分析[J].生物化学杂志,1993,2(9):224-228.
[13]《全国中草药汇编》编写组.全国中草药汇编(上册)[M].北京:人民卫生出版社,1990.224.
[14]Wang ZQ,Xu FM,YanXZ,et al.Scutebarbatine A,a new neoclerodane-type diterpenoid alkaloid from Scutellaria barbata[J].Chinese Chemical Letters,1996,7(4):333-334.
[15]陈柏君,高山林,余国奠.黄芩及其同属药用植物研究进展[J].中国野生植物 资源,1999,18(3):22-24.
[16]李洁.半枝莲对人肺巨细胞癌细胞株PG细胞bcl-2基因表达的影响[J].中药药理与临床,2007,23(1):46-47.
[17]宋寄春,谭诗云,陈明锴.半枝莲对人大肠癌细胞增殖抑制及诱导调亡的作用[J].咸宁学院学报(医学版),2007,2(21):106-108.
[18]Lee TK,Lee DK,Kim DI,et al.Inhibitory effects of Scutellaria barbata D.Don on human uterine leiomyomal smooth muscle cell proliferation through cell cycle analysis[J].Int Immunopharmacol,2004,4(3):447-454.
[19]Lee TK,Kim DI,Song YL,et al.Differential inhibition of Scutellaria barbata D.Don(Lamiaceae) on HCG-promoted proliferation of cultured uterine leiomyomal and myometrial smooth muscle cells[J].Immunopharmacol Immunotoxicol,2004,26(3):329-342.
[20]王刚,董玫,刘秀书,汤建华.半枝莲醇提物抗肿瘤活性的研究[J].现代中西医结合杂志,2004,13(9):1141-1142.
[21]李洁,孙兆贵.半枝莲诱导人肺巨细胞癌细胞株PG细胞凋亡的初步研究[J].中国中医药科技,2007,6(14):441.
[22]张春玲,胡峻峰等.Ames试验检测几种中草药及绿茶的抗诱变作用[J].卫生毒理学杂志,2002,1(16):66.
[23]张春玲,胡峻峰等.几种中草药及绿茶对B(a)P和NNK的抗诱变作用[J].癌变·畸变·突变,2003,2(15):101-102.
[24]Wong BY,Lau BH,Tadi PP,et al.Chinese medicinal herbs modulate mutagenesis,DNA binding and metabolism of aflatoxin B1[J].Mutat Res,1992,279(3):209-216.
[25]Wong BY,Lau BH,Teel RW.Chinese medicinal herbs modulate mutagenesis,DNA binding and metabolismofbenzo[α]pyrene7,8-dihydrodiol and benzo[α]pyrene7,8-dihydrodiol-9,10-epoxide[J].Cancer Lett,1992,62(2):123-131.
[26]Wong BY,Lau BH,Yamasaki T,et al.Modulation of cytochrome P-450IA1-mediated mutagenicity,DNA binding and metabolism of benzo[α]pyrene by Chinese medicinal herbs[J].Cancer Lett,1993,68(1):75-82.
[27]Wong BY,Lau BH,Yamasaki T,et al.Inhibition of dexamethasone-induced cytochrome P450-mediated mutagenicity and metabolism of aflatoxin B1 by Chinese medicinal herbs[J].Eur J Cancer Prev,1993,2(4):351-356.
[28]祝勇军,张春容,曹伦.半枝莲抗肿瘤作用研究进展[J].中国中医急症,2006, 5(15):533.
[29]蔡璇,彭松等.五种中药对139株微生物体外抗菌活性的实验研究[J].药学服务与研究,2005,5(3):267-270.
[30]佐藤阳一.从生药成分中开发抗MRSA感染的药物:关于半枝莲的抗菌活性成分[J].国外医学(中医中药分册),1998,20(2):46.
[31]陆平成,许益民.半枝莲多糖对细胞免疫的调节作用[J].南京中医学院学报,1989,2(1):32-33.
[32]马维坤,仰榴青等.半枝莲多糖提取工艺及其免疫活性初步研究[J].江苏大学学报(医学版),2007,4(17):316-317.
[33]余建清,柳惠斌等.半枝莲总黄酮对红细胞膜脂质过氧化损伤的保护作用[J].中国药师,2005,11(8):897-899.
[34]龙盛京.17种清热中药抗活性氧作用的研究[J].中草药,1999,30(10):40.
[35]杨容,王志等.中药半枝莲中黄酮类化合物的抗氧化活性[J].河北大学学报(自然科学版),2007,5(27):490-492.
[36]王转子,支德娟,关红梅.半枝莲多糖和白花蛇舌草多糖抗衰老作用的研究[J].中国兽医医药杂志,1999,4:5-7.
[37]佟继铭,陈光晖等.半枝莲的解热作用实验研究[J].中国民族民间医药杂志,1999,38:166-167.
[38]佟继铭,刘玉玲等.野黄芩甙的解热作用研究[J].承德医学院学报,1999,2(16):101-103.
[39]刘天龙,许剑琴.多糖现代研究及应用进展[J].中国兽医杂志,2004,3(40):24.
[40]张兴桃,高贵珍.多糖研究进展[J].宿州学院学报,2005,5(20):85.
[41]丁一芳,李连闯,赵玺.多糖的研究进展[J].黑龙江医药,2006,2(19):123.
[42]王红英.中药多糖研究进展[J].实用医技杂志,2006,6(13):1021.
[43]赛德艾合买提.浅谈多糖的研究进展[J].伊犁师范学院学报,2006,3:77.
[44]陈靠山.功能多糖的研究进展[J].皖南医学院学报,2005,3(24):161.
[45]卢艳花.中药有效成分提取分离技术[M].北京:化学工业出版社,2005.298-304.
[46]刘训红,阚毓铭,王玉玺.太子参多糖的研究[J].中草药,1993,24(3):119.
[47]Awal A,Haq QN,Quader A,et al.Structural study of a polusaccharide from the seeds of borassus flabellifer linn.Carbohyrate Research,1995,227:189.
[48]伊艳,高文宏,于淑娟.多糖提取技术的研究进展[J].食品工业科技,2007,2(28):248-249.
[49]廉宜君,谷新利.中草药多糖的提取分离及其免疫药理作用研究[J].四川畜牧 兽医,2007,3:33.
[50]马淑淘,张天民,刘立.硫酸软骨素生产工艺探讨[J].中国药学杂志,1993,28(12):741.
[51]沈渤江,张天民,吴柯.不同工艺制备的硫酸软骨素的理化性质差异[J].中国医药工业杂志,1990,21(5):208.
[52]林元藻.生化制药[M].北京:人民卫生出版社,1998.
[53]吴敏春,陈琼华等.毛木耳多糖的分离、分析及免疫药理活性的研究[J].中国药科大学学报,1991,22(2):97.
[54]徐惠,于志浩,苏富强等.螺旋藻粘多糖的分离及其免疫学作用[J].中国生化药物杂志,1997,18(2):72.
[55]田庚元,王晨,冯宇澄.枸杞子糖蛋白的分离纯化、物化性质及糖肽键特征[J].生物化学与生物物理学报,1995,27(2):201.
[56]王元凤,金征宇.茶多糖脱色的研究[J].食品与发酵工业[J],2004,30(12):60-65.
[57]王松柏,秦雪梅,郭小青,王光丽,张丽增.树脂对防风粗多糖脱色效果[J].应用化学,2005,22(10):1308-1311.
[58]向东,王国政,赖凤英,陈健.活性炭对南瓜粗多糖液的脱色研究[J].河南科学,2004,22(6):780-782.
[59]朱越雄,石亮,曹广力.野生树舌多糖的脱色和脱蛋白[J].氨基酸和生物资源,2005,27(3):1-3.
[60]梁忠岩,张翼伸等.长白山松杉灵芝子实体水溶多糖的分离鉴定与结构研究[J].生物化学与生物物理学报,1993,25(1):59.
[61]张维杰.糖复合物生化研究技术[M].浙江:浙江大学出版社,1999:8.
[62]Methods in Enzymology,1990,28-193.
[63]张翼伸.有关复合物的分级纯化、结构确定、生物活性的几个问题[J].生命的化学,1994,14(6):42.
[64]Jann B,Alexander S,Shashkov,Kochanowski H,et al.Structure of the O16polysaccharide from Escherichia coli O16:KI:an NMR investigation[J].Carbohydrate Research,1994,264:305.
[65]Sandeep S,Glushka J,Halbeek H,et al.Structure of the capsular polysaccharide of clostridium perfringers hobbs 5 as determined by NMR apectroscopy[J].Carbohydrate Research,1997,299:119.
[66]Hanson GC,Bouhours JF,Karlsson H.Analysis of sialic acid-containing mucin oligosaccharides from porcine small intestine by high-temperature gas chromatography-mass spectrometry their dimethylamides[J].Carbohydrate Research,1991,221:179.
[67]Anello JD,Gray GR.An improved procedure for the analysis of linkage positions in 2-acetamido-D-glucopyranosyl residues by the reductive cleavage method[J].Carbohydrate Research,1994,251:115.
[68]Wang N,Gray GR,et al.Reductive cleavage of permthylated polysaccharides with borane-methylsulfide comples and butyltin trichloride[J].Carbohydrate Research,1997,298:131.
[69]Christine RR,Gray GR,et al.Reductive cleavage of the positional in omers of benzoylated and methylated methyl α-D-mannopyranoside[J].Carbohydrate Research,1997,298:243.
[70]Kobata A,Fukuda M,et al.Structural characterization of oligosaccharides from glycoproteins.4A.Glycoidase treatment and other methods,including methylation[J].Glycobiology IRL press,1993:165.
[71]Monteuil J,Chaplin MF,et al.Glycoproteins[J].Carbohydrate analysis,1994:181.
[72]Jacob GS,Lennarz WJ,et al.Glycosidases in stuctrual analysis[J].Methods in Rnzymol,1994,230:280.
[73]Dwek RA.Analysis of flycoprotein associated oligosaccharides[J].Ann Rev Biochem,1993,62:65.
[74]王希.多糖的药理作用研究[J].本溪冶金高等专科学校学报,2003,5(2):6.
[75]王健,龚兴国.多糖的抗肿瘤及免疫调节研究进展[J].中国生化药物杂志,2001,22(1):52.
[76]张志浩.多糖的免疫调节作用及研究进展[J].山西食品工业,2003,2:10.
[77]何朝勇,王立为.多糖的免疫调节作用综述[J].安徽中医学院学报,2002,21(4):62.
[78]卢杏通,戴镜红等.多糖类免疫调节剂研究概况[J].动物医学进展,2003,24(1):10-12.
[79]白日霞.多糖免疫药物研究进展[J].大连民族学院学报,2003,5(1):38.
[80]邓小云,丁登峰等.植物多糖药理作用研究进展[J].中医药导报,2006,9(12):86.
[81]石磊,陈靠山.植物多糖药理作用研究进展[J].曲阜师范大学学报,2005,3(31):100-101.
[82]Kiho T,Morimoto H,Kobayashi T,et al.Effect of a polysaccharide(TAP)from the bodies of Tremella aurantia on glucose metabolism in mouse liver[J].Biosci,B iotechnol,Biochem.2000,64:417-419.
[83]熊学敏,叶士玲,许春波等.南瓜多糖对四氧嘧啶糖尿病大鼠的降糖作用[J].江西中医学院学报,1998,10(4):174.
[84]陈传平,张庭廷等.柚肉多糖的提取及其降血脂作用研究[J].中国实验方剂学杂志,2007,1(13):29-31.
[85]孔庆胜等.南瓜多糖的分离、纯化及其降血脂作用[J].中国生化药物杂志,2000,21(3):130-132.
[86]罗祖友,吴季勤,吴谋成.植物多糖的抗氧化与抗病毒活性[J].湖北民族学院学报(自然科学版),2007,1(25):77.
[87]Hu XL,You JY,BC,et al.Determination of total flavonoids in Scutellaria barbata D.Don by dynamic ultrasonic extraction coupled with on-line spectrophotometry[J].Elsevier ScienceDirect,2008,610:217-223.
[88]Kim DI,Lee TK,Lim IS,et al.Regulation of IGF-I production and proliferation of human leiomyomal smooth muscle cells by Scutellaria barbata D.Don in vitro:isolation of flavonoids of apigenin and luteolon as acting compounds[J].Elsevier ScienceDirect,2005,205:213-224.
[89]Lee TK,Lee DK,Kim DI,et al.Inhibitory effects of Scutellaria barbata D.Don on human uterine leiomyomal smooth muscle cell proliferation through cell cycle analysis[J].International Immunopharmacology,2004,4:447-454.
[90]Yin XL,Zhou JB,et al.Anticancer activity and mechanism of Scutellaria barbata extract on human lung cancer cell line A549[J].Life Sciences,2004,75:2233-2244.
[91]Dai SJ,Tao JY,et al.Neo-Clerodane diterpenoids from Scutellaria barbata with cytotoxic activities[J].Phytochemisty,2006,67:1326-1330.
[92]张赛丹,方岩雄,陈敏敏等.几种传统中药中多糖的提取及抗肿瘤活性研究[J].中药材,2007,2(30):179-182.
[93]李洁.不同产地半枝莲中多糖含量的比较研究[J].中国中医药信息杂志,2006,10(13).
[94]邢文善,纪耀华等.半枝莲粗多糖的提取及总糖的含量测定[J].吉林中医药,2005,7(25):56.
[95]孟延发,李志孝等.半枝莲多糖SPS4组分的纯化性质及分析[J].生物化学杂 志,1993,2(9):224-228.
[96]薛梅,王自军,周静.半枝莲中总黄酮和多糖的微波提取与含量测定[J].时珍国医国药,2005,16(10):965-966.
[2]张素华,辛春兰.半枝莲的药理作用及应用[J].中医杂志,2007,6(48):572.
[3]邹箴蕾,吴启南.半枝莲的化学成分及药理作用研究进展[J].时珍国医国药,2005,16(2):149-150.
[4]杨蓓芬,李钧敏,邵红.半枝莲的次生代谢产物含量测定与体外抑菌活性的研究[J].四川中医,2005,23(11):35-36.
[5]袁崇均,王笳,杨红,黄卫平.正交试验法优选半枝莲提取工艺的研究[J].华西药学杂志,2002,17(2):112-113.
[6]邸多隆,刘丽春,刘晔玮,刘霞,蒋生祥.半枝莲提取工艺的研究[J].中成药,2003,7(25):530-532.
[7]张海方,许化溪.半枝莲化学成分及生物学活性的研究进展[J].江苏大学学报(医学版),2006,1(16):74.
[8]Sato Yoichi,et al.Phytochemical flavones isolated from Scutellaria barbara and antibacterial activity against methicillin-resistant Staphylococcus aureus[J].Journal of Ethnopharmacology,2000(72):483-488.
[9]Yu JQ,Lei JC,Yu HD,et al.Chemical composition and antimicrobial of the essential oil of Scutellaria barbata[J].Phytochemistry,2004,65:881-884.
[10]贺莉娟,梁逸曾,赵晨曦.唇形科植物挥发油化学成分的GC/MS研究[J].化学学报,2007,3(65):227-232.
[11]许益民,郭立伟,陈建伟.半枝莲多糖的分离、纯化及其理化性质[J].天然产物研究与开发,1992,1(4):1-5.
[12]孟延发,李志孝,张立.半枝莲多糖组分的纯化性质及分析[J].生物化学杂志,1993,2(9):224-228.
[13]《全国中草药汇编》编写组.全国中草药汇编(上册)[M].北京:人民卫生出版社,1990.224.
[14]Wang ZQ,Xu FM,YanXZ,et al.Scutebarbatine A,a new neoclerodane-type diterpenoid alkaloid from Scutellaria barbata[J].Chinese Chemical Letters,1996,7(4):333-334.
[15]陈柏君,高山林,余国奠.黄芩及其同属药用植物研究进展[J].中国野生植物 资源,1999,18(3):22-24.
[16]李洁.半枝莲对人肺巨细胞癌细胞株PG细胞bcl-2基因表达的影响[J].中药药理与临床,2007,23(1):46-47.
[17]宋寄春,谭诗云,陈明锴.半枝莲对人大肠癌细胞增殖抑制及诱导调亡的作用[J].咸宁学院学报(医学版),2007,2(21):106-108.
[18]Lee TK,Lee DK,Kim DI,et al.Inhibitory effects of Scutellaria barbata D.Don on human uterine leiomyomal smooth muscle cell proliferation through cell cycle analysis[J].Int Immunopharmacol,2004,4(3):447-454.
[19]Lee TK,Kim DI,Song YL,et al.Differential inhibition of Scutellaria barbata D.Don(Lamiaceae) on HCG-promoted proliferation of cultured uterine leiomyomal and myometrial smooth muscle cells[J].Immunopharmacol Immunotoxicol,2004,26(3):329-342.
[20]王刚,董玫,刘秀书,汤建华.半枝莲醇提物抗肿瘤活性的研究[J].现代中西医结合杂志,2004,13(9):1141-1142.
[21]李洁,孙兆贵.半枝莲诱导人肺巨细胞癌细胞株PG细胞凋亡的初步研究[J].中国中医药科技,2007,6(14):441.
[22]张春玲,胡峻峰等.Ames试验检测几种中草药及绿茶的抗诱变作用[J].卫生毒理学杂志,2002,1(16):66.
[23]张春玲,胡峻峰等.几种中草药及绿茶对B(a)P和NNK的抗诱变作用[J].癌变·畸变·突变,2003,2(15):101-102.
[24]Wong BY,Lau BH,Tadi PP,et al.Chinese medicinal herbs modulate mutagenesis,DNA binding and metabolism of aflatoxin B1[J].Mutat Res,1992,279(3):209-216.
[25]Wong BY,Lau BH,Teel RW.Chinese medicinal herbs modulate mutagenesis,DNA binding and metabolismofbenzo[α]pyrene7,8-dihydrodiol and benzo[α]pyrene7,8-dihydrodiol-9,10-epoxide[J].Cancer Lett,1992,62(2):123-131.
[26]Wong BY,Lau BH,Yamasaki T,et al.Modulation of cytochrome P-450IA1-mediated mutagenicity,DNA binding and metabolism of benzo[α]pyrene by Chinese medicinal herbs[J].Cancer Lett,1993,68(1):75-82.
[27]Wong BY,Lau BH,Yamasaki T,et al.Inhibition of dexamethasone-induced cytochrome P450-mediated mutagenicity and metabolism of aflatoxin B1 by Chinese medicinal herbs[J].Eur J Cancer Prev,1993,2(4):351-356.
[28]祝勇军,张春容,曹伦.半枝莲抗肿瘤作用研究进展[J].中国中医急症,2006, 5(15):533.
[29]蔡璇,彭松等.五种中药对139株微生物体外抗菌活性的实验研究[J].药学服务与研究,2005,5(3):267-270.
[30]佐藤阳一.从生药成分中开发抗MRSA感染的药物:关于半枝莲的抗菌活性成分[J].国外医学(中医中药分册),1998,20(2):46.
[31]陆平成,许益民.半枝莲多糖对细胞免疫的调节作用[J].南京中医学院学报,1989,2(1):32-33.
[32]马维坤,仰榴青等.半枝莲多糖提取工艺及其免疫活性初步研究[J].江苏大学学报(医学版),2007,4(17):316-317.
[33]余建清,柳惠斌等.半枝莲总黄酮对红细胞膜脂质过氧化损伤的保护作用[J].中国药师,2005,11(8):897-899.
[34]龙盛京.17种清热中药抗活性氧作用的研究[J].中草药,1999,30(10):40.
[35]杨容,王志等.中药半枝莲中黄酮类化合物的抗氧化活性[J].河北大学学报(自然科学版),2007,5(27):490-492.
[36]王转子,支德娟,关红梅.半枝莲多糖和白花蛇舌草多糖抗衰老作用的研究[J].中国兽医医药杂志,1999,4:5-7.
[37]佟继铭,陈光晖等.半枝莲的解热作用实验研究[J].中国民族民间医药杂志,1999,38:166-167.
[38]佟继铭,刘玉玲等.野黄芩甙的解热作用研究[J].承德医学院学报,1999,2(16):101-103.
[39]刘天龙,许剑琴.多糖现代研究及应用进展[J].中国兽医杂志,2004,3(40):24.
[40]张兴桃,高贵珍.多糖研究进展[J].宿州学院学报,2005,5(20):85.
[41]丁一芳,李连闯,赵玺.多糖的研究进展[J].黑龙江医药,2006,2(19):123.
[42]王红英.中药多糖研究进展[J].实用医技杂志,2006,6(13):1021.
[43]赛德艾合买提.浅谈多糖的研究进展[J].伊犁师范学院学报,2006,3:77.
[44]陈靠山.功能多糖的研究进展[J].皖南医学院学报,2005,3(24):161.
[45]卢艳花.中药有效成分提取分离技术[M].北京:化学工业出版社,2005.298-304.
[46]刘训红,阚毓铭,王玉玺.太子参多糖的研究[J].中草药,1993,24(3):119.
[47]Awal A,Haq QN,Quader A,et al.Structural study of a polusaccharide from the seeds of borassus flabellifer linn.Carbohyrate Research,1995,227:189.
[48]伊艳,高文宏,于淑娟.多糖提取技术的研究进展[J].食品工业科技,2007,2(28):248-249.
[49]廉宜君,谷新利.中草药多糖的提取分离及其免疫药理作用研究[J].四川畜牧 兽医,2007,3:33.
[50]马淑淘,张天民,刘立.硫酸软骨素生产工艺探讨[J].中国药学杂志,1993,28(12):741.
[51]沈渤江,张天民,吴柯.不同工艺制备的硫酸软骨素的理化性质差异[J].中国医药工业杂志,1990,21(5):208.
[52]林元藻.生化制药[M].北京:人民卫生出版社,1998.
[53]吴敏春,陈琼华等.毛木耳多糖的分离、分析及免疫药理活性的研究[J].中国药科大学学报,1991,22(2):97.
[54]徐惠,于志浩,苏富强等.螺旋藻粘多糖的分离及其免疫学作用[J].中国生化药物杂志,1997,18(2):72.
[55]田庚元,王晨,冯宇澄.枸杞子糖蛋白的分离纯化、物化性质及糖肽键特征[J].生物化学与生物物理学报,1995,27(2):201.
[56]王元凤,金征宇.茶多糖脱色的研究[J].食品与发酵工业[J],2004,30(12):60-65.
[57]王松柏,秦雪梅,郭小青,王光丽,张丽增.树脂对防风粗多糖脱色效果[J].应用化学,2005,22(10):1308-1311.
[58]向东,王国政,赖凤英,陈健.活性炭对南瓜粗多糖液的脱色研究[J].河南科学,2004,22(6):780-782.
[59]朱越雄,石亮,曹广力.野生树舌多糖的脱色和脱蛋白[J].氨基酸和生物资源,2005,27(3):1-3.
[60]梁忠岩,张翼伸等.长白山松杉灵芝子实体水溶多糖的分离鉴定与结构研究[J].生物化学与生物物理学报,1993,25(1):59.
[61]张维杰.糖复合物生化研究技术[M].浙江:浙江大学出版社,1999:8.
[62]Methods in Enzymology,1990,28-193.
[63]张翼伸.有关复合物的分级纯化、结构确定、生物活性的几个问题[J].生命的化学,1994,14(6):42.
[64]Jann B,Alexander S,Shashkov,Kochanowski H,et al.Structure of the O16polysaccharide from Escherichia coli O16:KI:an NMR investigation[J].Carbohydrate Research,1994,264:305.
[65]Sandeep S,Glushka J,Halbeek H,et al.Structure of the capsular polysaccharide of clostridium perfringers hobbs 5 as determined by NMR apectroscopy[J].Carbohydrate Research,1997,299:119.
[66]Hanson GC,Bouhours JF,Karlsson H.Analysis of sialic acid-containing mucin oligosaccharides from porcine small intestine by high-temperature gas chromatography-mass spectrometry their dimethylamides[J].Carbohydrate Research,1991,221:179.
[67]Anello JD,Gray GR.An improved procedure for the analysis of linkage positions in 2-acetamido-D-glucopyranosyl residues by the reductive cleavage method[J].Carbohydrate Research,1994,251:115.
[68]Wang N,Gray GR,et al.Reductive cleavage of permthylated polysaccharides with borane-methylsulfide comples and butyltin trichloride[J].Carbohydrate Research,1997,298:131.
[69]Christine RR,Gray GR,et al.Reductive cleavage of the positional in omers of benzoylated and methylated methyl α-D-mannopyranoside[J].Carbohydrate Research,1997,298:243.
[70]Kobata A,Fukuda M,et al.Structural characterization of oligosaccharides from glycoproteins.4A.Glycoidase treatment and other methods,including methylation[J].Glycobiology IRL press,1993:165.
[71]Monteuil J,Chaplin MF,et al.Glycoproteins[J].Carbohydrate analysis,1994:181.
[72]Jacob GS,Lennarz WJ,et al.Glycosidases in stuctrual analysis[J].Methods in Rnzymol,1994,230:280.
[73]Dwek RA.Analysis of flycoprotein associated oligosaccharides[J].Ann Rev Biochem,1993,62:65.
[74]王希.多糖的药理作用研究[J].本溪冶金高等专科学校学报,2003,5(2):6.
[75]王健,龚兴国.多糖的抗肿瘤及免疫调节研究进展[J].中国生化药物杂志,2001,22(1):52.
[76]张志浩.多糖的免疫调节作用及研究进展[J].山西食品工业,2003,2:10.
[77]何朝勇,王立为.多糖的免疫调节作用综述[J].安徽中医学院学报,2002,21(4):62.
[78]卢杏通,戴镜红等.多糖类免疫调节剂研究概况[J].动物医学进展,2003,24(1):10-12.
[79]白日霞.多糖免疫药物研究进展[J].大连民族学院学报,2003,5(1):38.
[80]邓小云,丁登峰等.植物多糖药理作用研究进展[J].中医药导报,2006,9(12):86.
[81]石磊,陈靠山.植物多糖药理作用研究进展[J].曲阜师范大学学报,2005,3(31):100-101.
[82]Kiho T,Morimoto H,Kobayashi T,et al.Effect of a polysaccharide(TAP)from the bodies of Tremella aurantia on glucose metabolism in mouse liver[J].Biosci,B iotechnol,Biochem.2000,64:417-419.
[83]熊学敏,叶士玲,许春波等.南瓜多糖对四氧嘧啶糖尿病大鼠的降糖作用[J].江西中医学院学报,1998,10(4):174.
[84]陈传平,张庭廷等.柚肉多糖的提取及其降血脂作用研究[J].中国实验方剂学杂志,2007,1(13):29-31.
[85]孔庆胜等.南瓜多糖的分离、纯化及其降血脂作用[J].中国生化药物杂志,2000,21(3):130-132.
[86]罗祖友,吴季勤,吴谋成.植物多糖的抗氧化与抗病毒活性[J].湖北民族学院学报(自然科学版),2007,1(25):77.
[87]Hu XL,You JY,BC,et al.Determination of total flavonoids in Scutellaria barbata D.Don by dynamic ultrasonic extraction coupled with on-line spectrophotometry[J].Elsevier ScienceDirect,2008,610:217-223.
[88]Kim DI,Lee TK,Lim IS,et al.Regulation of IGF-I production and proliferation of human leiomyomal smooth muscle cells by Scutellaria barbata D.Don in vitro:isolation of flavonoids of apigenin and luteolon as acting compounds[J].Elsevier ScienceDirect,2005,205:213-224.
[89]Lee TK,Lee DK,Kim DI,et al.Inhibitory effects of Scutellaria barbata D.Don on human uterine leiomyomal smooth muscle cell proliferation through cell cycle analysis[J].International Immunopharmacology,2004,4:447-454.
[90]Yin XL,Zhou JB,et al.Anticancer activity and mechanism of Scutellaria barbata extract on human lung cancer cell line A549[J].Life Sciences,2004,75:2233-2244.
[91]Dai SJ,Tao JY,et al.Neo-Clerodane diterpenoids from Scutellaria barbata with cytotoxic activities[J].Phytochemisty,2006,67:1326-1330.
[92]张赛丹,方岩雄,陈敏敏等.几种传统中药中多糖的提取及抗肿瘤活性研究[J].中药材,2007,2(30):179-182.
[93]李洁.不同产地半枝莲中多糖含量的比较研究[J].中国中医药信息杂志,2006,10(13).
[94]邢文善,纪耀华等.半枝莲粗多糖的提取及总糖的含量测定[J].吉林中医药,2005,7(25):56.
[95]孟延发,李志孝等.半枝莲多糖SPS4组分的纯化性质及分析[J].生物化学杂 志,1993,2(9):224-228.
[96]薛梅,王自军,周静.半枝莲中总黄酮和多糖的微波提取与含量测定[J].时珍国医国药,2005,16(10):965-966.