旱莲草多糖的提取及其抗氧化活性研究

采用水/甲醇混合溶剂浸泡提取旱莲草的多糖组分,利用硅胶柱层析和中压液相色谱,对多糖进行分离纯化,纯化后的多糖组分经薄层层析分析发现,这些多糖主要组成单糖均为葡萄糖和果糖。旱莲草多糖在总还原能力、总抗氧化能力和FRAP抗氧化能力测试中均显示较好的活性,能够有效清除羟自由基和DPPH自由基,在1.5 mg/m L的浓度下,对羟自由基的清除效率为(12.33~56.81)%,粗多糖组分在1 mg/m L的浓度下对DPPH自由基的清除效率为(17.88~84.47)%,精分多糖组分在5 mg/m L的浓度下,对DPPH自由基的清除效率为(18.64~91.35)%;极性小的多糖抗氧化活性显著优于极性大的多糖,抗氧化活性最高可达后者的11倍之多;粗多糖组分的抗氧化活性一般优于纯化后的相应精多糖组分,这说明旱莲草的不同多糖分子之间可能存在协同抗氧化作用。     

橘黄裸伞多糖的理化特性及体外抗氧化活性

采用水提醇沉、Sevage法去蛋白制备橘黄裸伞粗多糖,通过DEAE-52层析柱对粗多糖进行分离纯化,得到三种组分(GSPS-Ⅰ、GSPS-Ⅱ、GSPS-Ⅲ),主要对GSPS-Ⅱ、GSPS-Ⅲ进行研究。GSPS-Ⅱ、GSPS-Ⅲ的多糖含量分别为84.12%和80.35%,通过扫描电镜(SEM)观察发现GSPS-Ⅱ呈现六面体状结构,GSPS-Ⅲ呈现不规则的砖块状结构;通过红外光谱(IR)和氢核磁共振谱(1H NMR)对其检测,结果表明GSPS-Ⅱ、GSPS-Ⅲ均具有糖类物质的吸收峰且二者均含有α-和β-糖苷构型异头氢;采用清除DPPH、超氧阴离子、ABTS自由基实验评价其抗氧化活性,结果表明这两种多糖对本实验所测自由基均表现出一定的清除效果。综上所述,从橘黄裸伞分离纯化的多糖组分GSPS-Ⅱ、GSPS-Ⅲ具有良好的抗氧化活性。     

季也蒙假丝酵母胞外多糖的分离纯化及抗氧化活性研究

对一株源于海参肠道的季也蒙假丝酵母产胞外多糖进行分离纯化并对其抗氧化活性进行研究.采用乙醇沉淀、Sevag除蛋白等方法得到胞外粗多糖EPS.EPS经Sepharose强阴离子交换层析分离后,分别得到3个组分EPS1、EPS2和EPS3,对抗氧化活性较高的EPS2采用Sephacryal凝胶过滤层析进行纯化,得到1个单一组分EPS2-1,采用气相色谱法分析其单糖组成,并验证其抗氧化活性.结果表明:EPS2-1是由木糖、甘露糖、葡萄糖和半乳糖组成的;它具有较强的抗氧化活性,当其浓度为0.36 mg/mL时,对羟基自由基的清除率可达100％,明显高于同浓度下Vc对羟基自由基的清除率14.42％;当EPS2-1浓度为0.60 mg/mL时,对超氧阴离子自由基的清除率可达53.22％;同时EPS2-1表现出一定的还原力.研究证明该活性多糖具有很好的应用潜力,值得进一步研究开发.     

苦荞多肽的纯化、结构与体外抗氧化活性研究CSCD

为了探究苦荞多肽粗提液抗氧化活性的主要来源,需要进一步对苦荞多肽进行分离纯化,通过结构鉴定来分析抗氧化活性与多肽结构之间的关系。该研究以苦荞功能提取物废渣为原料提取苦荞粗蛋白辅以植物乳杆菌发酵法制备苦荞多肽,采用大孔吸附树脂、葡聚糖凝胶、液相色谱对苦荞多肽粗提液进行分离纯化,并以抗氧化活性为指标,筛选抗氧化活性较强的组分,利用液相色谱-质谱联用技术鉴定抗氧化多肽结构。结果表明,纯度不同的苦荞多肽抗氧化能力也不同,经过Sephadex G-15葡聚糖凝胶分离纯化得到的T-3组分ABTS和DPPH自由基清除能力最优,清除率分别为96%、94%。纯化后苦荞多肽的抗氧化活性显著高于粗提液(P<0.05),但是抗氧化活性并没有完全与苦荞多肽纯度呈正相关,通过液相色谱进一步分离纯化T-3组分后得到的组分T-3-1,苦荞多肽纯度虽然达到了98%,但是对DPPH和ABTS抑制率不再增强,反而稍有下降,抑制率分别为89%和90%。由质谱鉴定结果得到,主要抗氧化肽的氨基酸序列为苯丙氨酸-脯氨酸-酪氨酸Phe-Pro-Tyr(FPY)和酪氨酸-亮氨酸-脯氨酸-苯丙氨酸Tyr-Leu-Pro-Phe(YLPF)。研究结果以期为苦荞多肽的进一步开发利用提供一定的理论基础。     

雪灵芝多糖的分离纯化及免疫活性评价

为分离纯化雪灵芝(Arenaria kansuensis)多糖,并对纯化组分进行分子量测定、单糖组分分析及免疫活性评价。实验采用水提醇沉法提取雪灵芝粗多糖(Arenaria kansuensis crude polysaccharide, AKCP);以DEAE-52纤维素柱对AKCP进行分离纯化,获得5个雪灵芝多糖组分AKP-1～AKP-5,进一步采用葡聚糖凝胶G-75柱对AKP-2进行分离纯化获得AKP-2a多糖组分。苯酚-硫酸法测定AKCP、AKP-2及AKP-2a的总糖含量分别为52%、70%和79%;凝胶渗透色谱-十八角度激光光散射(GPC-MALS)法检测AKP-2a的重均分子量Mw为2.07×10~5Da、数均分子量Mn为9.838×10~4Da;HPLC法检测AKP-2a是由半乳糖醛酸、甘露糖、核糖、鼠李糖、葡萄糖醛酸、葡萄糖、半乳糖、木糖、阿拉伯糖、岩藻糖10种单糖组成,其摩尔比为1∶0.25∶0.01∶0.20∶0.11∶0.25∶0.61∶0.07∶0.21∶0.12;以MTT法检测体外培养小鼠脾淋巴细胞增殖,AKCP、AKP-2及AKP-2a各浓度组SI水平,均明显高于对照组(P<0.05)。经NO释放实验及IFN-γELISA检测,AKP-2a各浓度组小鼠腹腔巨噬细胞培养上清中二者的水平,较对照组呈浓度依赖性增高(P<0.01)。综上结果,本研究通过分离纯化,获得了总糖含量较高的雪灵芝多糖AKP-2a组分,初步确定其分子量范围及单糖组成,并证实其具有激活淋巴细胞增殖、促进巨噬细胞功能的生物活性。     

阳春砂多糖的分离纯化及体外抗氧化作用研究

本实验研究了阳春砂多糖(AVP)及其纯化组分的抗氧化活性,首先采用DEAE-纤维素-52离子交换柱分级洗脱阳春砂粗多糖,再采用Sephadex G-100葡聚糖凝胶色谱柱进一步纯化得到纯化多糖;分别采用超氧阴离子自由基体系、羟基自由基体系、1,1-二苯基苦基苯肼自由基体系,对阳春砂多糖的抗氧化活性进行研究,并以维生素C为阳性对照物,实验结果表明阳春砂粗多糖及纯化组分对超氧阴离子自由基、羟基自由基、1,1-二苯基苦基苯肼自由基均有较强的清除能力,AVP-3体外抗氧化活性最强。     

辣椒叶多糖抗氧化作用研究

目的:从辣椒叶中分离纯化活性多糖并考察其抗氧化活性。方法:采用水提醇沉法提取多糖,Sevag法脱蛋白,得辣椒叶粗多糖;分别使用超纯水、0.06 mol/L NaCl溶液、0.18 mol/L NaCl溶液作为洗脱液,通过DEAE-52离子交换柱色谱纯化得到三种辣椒叶多糖LD-0、LD-0.06、LD-0.18,测定多糖含量。DPPH、ABTS法检测多糖体外抗氧化作用。以小鼠血清、肝组织中总超氧化物歧化酶活性、过氧化氢酶活性及丙二醛含量为指标,考察小鼠灌胃LD-0.06多糖对脂质过氧化模型的影响。结果:辣椒叶粗多糖、LD-0、LD-0.06、LD-0.18多糖含量分别为9.92%、43.14%、82.97%、37.63%,其中LD-0.06多糖含量最高。体外抗氧化实验结果显示,三种结果均具备较好的清除能力,其中LD-0.06对ABTS+、DPPH·的清除效果最好,IC50值分别为0.58 mg/m L和0.60 mg/m L,结果与对照组在0.05水平具有显著性差异,说明辣椒叶多糖提取物是一种有效的天然抗氧化剂。在脂质过氧化模型小鼠体内,与模型组比较,LD-0.06多糖能显著增强小鼠血清和肝组织中的T-SOD与CAT活性,降低MDA的含量,且剂量越高,体内抗氧化能力越强。结论:辣椒叶多糖提取物具有一定的抗氧化作用,为进一步开发利用辣椒资源提供了理论依据。     

不同分子量菊芋多糖的生物活性研究

研究不同醇沉组份的菊芋多糖分子量和其抑制α-葡萄糖苷酶活性与抗氧化活性差异。菊芋水提物分级醇沉获得粗多糖,过Sephadex G-50凝胶柱进行纯化,得到重均分子量为1959、2180、2746、2011 Da的多糖组分:JAP-1、JAP-2、JAP-3和JAP-4。测定这些组分体外抑制α-葡萄糖苷酶活性和对DPPH与羟基自由基清除能力。结果表明,JAP-4具有较明显的抑制α-葡萄糖苷酶活性,在5 mg/mL时抑制率为20.32%。JAP-1、JAP-2、JAP-3和JAP-4均有显著的抗氧化活性,其中JAP-2对DPPH自由基和羟基自由基的清除活性优于其他组分。JAP-2在2 mg/mL时对DPPH自由基和羟基自由基清除率分别达到84.50%和89.74%,接近于Vc的抗氧化活性。不同分子量菊芋多糖的活性存在差异,可能是由于分子量的不同所导致。     

羧甲基茯苓多糖体外抗氧化活性研究

目的：比较不同浓度洗脱液洗脱得到的羧甲基茯苓多糖的抗氧化活性。方法：以茯苓为原料提取茯苓多糖,进行羧甲基取代反应,分离和纯化得到了均一性羧甲基茯苓多糖CMP-1、CMP-2、CMP-3、CMP-4,通过测定还原能力、DPPH自由基清除率、羟基自由基清除率、超氧阴离子自由基清除率比较其体外抗氧化活性。结果：羧甲基茯苓多糖均表现出与浓度正相关的体外抗氧化活性,其中CMP-4具有相对更强的体外抗氧化活性。结论：所得羧甲基茯苓多糖样品具有不同的体外抗氧化能力,随着洗脱液浓度增加,抗氧化活性增强。     

鸡枞菌粉不同组分的体外抗氧化活性研究

目的:研究鸡枞(Termitomyces albuminosus)菌粉不同组分的体外抗氧化活性.方法:通过分离纯化,鸡枞菌粉中皂甙和多糖被富集于两个不同的组分.以Vc,没食子酸和TSHQ为对照,测定不同组分的还原力、对二苯代苦味酰肼自由基(DPPH)和超氧阴离子自由基O-2的清除能力,及其对亚油酸过氧化的抑制作用.结果:鸡枞菌粉不同组分均具有一定的抗氧化活性.结论:鸡枞具有较好的抗氧化活性,为开发鸡枞产品提供了理论依据.     

Effect of steaming process on the structural characteristics and antioxidant activities of polysaccharides from Polygonatum sibiricum rhizomes

Polygonatum sibiricum (P. sibiricum) rhizomes are widely used as a tonic and functional food, and are often processed to enhance their tonic function by repeated steaming and drying. As the most important constituent, the polysaccharide from P. sibiricum rhizomes (PSP) has demonstrated various activities, but the alteration of structural characteristics and activities of the purified PSPs during steaming process was rarely investigated. To well understand the effect of steaming process on the polysaccharides of P. sibiricum, neutral polysaccharides from P. sibiricum rhizomes (PSP0?~?PSP9) after steaming were first isolated and purified, and then the chemical properties and antioxidant activities were determined. The results showed that the molar ratios of monosaccharides in PSPs were different. The molecular weights of PSPs were increased significantly after the fourth steaming. Morphological studies showed that the surface of PSPs became much tighter during the steaming process. Fourier transform infrared spectroscopy spectra displayed the polysaccharides had similar backbones and chemical groups. Furthermore, the antioxidant activity of PSPs was measured through radical scavenging tests. It was found that the radical scavenging activity of PSPs was elevated strikingly after steaming, and increased gradually with numbers of steaming process. The biological and chemical variance of PSPs revealed considerable segregation of PSP0, PSP1?~?PSP4 and PSP5?~?PSP9. In conclusion, our results proposed the fourth time as the optimal number of steaming to extract functional polysaccharide from P. sibiricum rhizomes.

     

Optimization of the Extraction Process and Antioxidant Activity of Polysaccharide Extracted from Centipeda minima

The purpose of this study is to optimize the extraction process and study antioxidant activity of Polysaccharide extracted from Centipeda minima. The Box-Behnken design-response surface methodology was adopted to optimize the extraction process of polysaccharides from Centipeda minima. We purified the crude polysaccharides from Centipeda minima, as well as determined the purity, monosaccharide composition, and molecular weight of the purified fraction. Fourier transform infrared spectrometer (FT-IR) and scanning electron microscopy (SEM) were used to analyze the structural features of the polysaccharides. Further, we investigated the antioxidant activities of different fractions of polysaccharides. Consequently, the results showed that the optimum extraction conditions for polysaccharides were: a liquid-solid ratio of 26 mL/g, extraction temperature of 85.5 °C, and extraction time of 2.4 h. Moreover, the yield of polysaccharides measured under these conditions was close to the predicted value. After purification, we obtained four components of Centipeda minima polysaccharides (CMP). The purity, monosaccharide composition, molecular weight, and structural characteristics of CMP were different, but with similar infrared absorption spectra. CMP exhibited a typical infrared absorption characteristic of a polysaccharide. Besides, CMP displayed good antioxidant activity, with potential to scavenge DPPH radical, hydroxyl radical, and superoxide radical. Therefore, this study provides a reference for future research on the structure and biological activity of CMP, and lays a theoretical foundation for food processing and medicinal development of CMP.     

绵马贯众多糖超声提取工艺优化及抗氧化活性分析

绵马贯众是中国传统常用中药,本研究以温度、时间、超声功率、液料比为影响因子,多糖得率为评价指标,通过响应面法优化超声辅助提取绵马贯众多糖的工艺条件,同时测定其基本理化性质及抗氧化活性。研究结果表明,绵马贯众多糖的最佳提取工艺条件为:温度64℃、时间60 min、超声功率210 W、液料比27 mL/g。此时多糖得率为9.57%,与预测值接近。理化性质分析表明绵马贯众多糖为含少量蛋白的酸性多糖。体外抗氧化研究表明绵马贯众多糖具有很强的DPPH自由基清除活性,IC50值为0.29 mg/mL;较好的羟基自由基清除活性,其IC50值为1.10 mg/mL;对DNA的氧化损伤有显著的保护作用。绵马贯众多糖可以作为一种潜在的抗氧化剂应用于食品和化妆品等领域。     

黄精种子营养成分的测定与分析

采用化学分析的方法,对黄精(Polygonatum sibiricum Red.)种子主要营养成分--粗淀粉、粗脂肪、粗蛋白的含量进行了测定,并采用气相色谱法、氨基酸自动分析仪分别对其脂肪酸和氨基酸的组分作了分析,以明确黄精种子的营养成分及其利用价值.结果显示:(1)黄精种子的营养成分以淀粉(73.70%)为主,脂肪含量为(10.74%),蛋白质含量为(8.21%);(2)脂肪酸中以不饱和脂肪酸(89.52%)为主,其中人体必需的多不饱和脂肪酸亚油酸占49.77%;(3)氨基酸种类齐全,其中人体必需氨基酸蛋氨酸、赖氨酸、异亮氨酸的含量比小麦、玉米等作物的含量高.结果说明,黄精种子具有较高的营养价值,在食品、医疗保健方面具有一定开发前景.     

Identification of structure and antioxidant activity of a fraction of polysaccharide purified from Dioscorea nipponica Makino

A large number of polysaccharides are present in boiling-water extraction of Dioscorea nipponica Makino. A DEAE-Sepharose CL-6B column chromatography was used to isolate the major polysaccharides from D. nipponica Makino. The largest amount of fraction of polysaccharide was subjected to further purification by gel-filtration on Sephadex G-100. The purified fraction was a neutral polysaccharide and a single peak in HPLC with Sugar KS-804 column, with a molecular weight of 38,000, and comprised mainly of glucose and fructose (45:1). Analysis by Periodate oxidation–Smith degradation indicated that there were 5.9%(1→)-glycosidic linkages, 4.94% (1 → 2)-glycosidic linkages, 61.16% (1 → 4)-glycosidic linkages, and 28% (1 → 3)-glycosidic linkages. On the basis of superoxide radical assay, hydroxyl radical assay, and self-oxidation of 1,2,3-phentriol assay, its antioxidant activity was investigated. This purified fraction of polysaccharide exhibited equivalent inhibiting power for self-oxidation of 1,2,3-phentriol to Vc, a little higher scavenging activity of superoxide radical and hydroxyl radical than Vc, and should be explored as a novel potential antioxidant.     

响应面优化塔拉种子多糖的超声波提取及其抗氧化性研究

为获得塔拉多糖超声波提取的最佳工艺,利用中心组合实验设计原理,采用四因素三水平的响应面分析法,获得多元二次线性回归方程,以多糖提取率为响应值做响应面。并考察塔拉多糖的体外抗氧化活性。结果表明：塔拉多糖的最佳提取工艺条件为,超声辅助提取超声功率363 W,超声温度56℃,超声时间23 min,料液比1：17（g·mL^-1）,塔拉粗多糖的最大提取率（25.56%）与理论推测值（25.71%）相差较小。经纯化后塔拉多糖提取率为17.82%。以VC为阳性对照,对不同纯度的塔拉多糖进行抗氧化性的研究,发现其纯度越高,对ABTS自由基的清除能力越强。     

Antioxidant acitivity in vitro and in vivo of the capsule polysaccharides from Streptococcus equi subsp. zooepidemicus

Crude capsule polysaccharides (CCP) were prepared from the culture of Streptococcus equi subsp. zooepidemicus C55129 and were partially purified through an anion-exchange column chromatography to afford partially purified capsule polysaccharides (PCP). The main component of CCP and PCP was hyaluronic acid. In vitro antioxidant assay, the capsule polysaccharides showed strong inhibition of lipid peroxidation and hydroxyl radical scavenging activity and moderate 1,1-diphenyl-2-picryldydrazyl radical scavenging activity. In addition, CCP exhibited much stronger reductive power than PCP. For antioxidant testing in vivo, CCP and PCP were orally administrated over a period of 15 days in a d-galactose induced aged mice model. As results, administration of capsule polysaccharides inhibited significantly the formation of malondialdehyde in mice livers and serums and raised the activities of antioxidant enzymes and total antioxidant capacity in a dose-dependent manner. However, the antioxidant activity of CCP was lower than that of PCP. The results suggest that the capsule polysaccharides from Streptococcus equi subsp. zooepidemicus C55129 have direct and potent antioxidant activities.     

Identification of polysaccharides from pericarp tissues of litchi (Litchi chinensis Sonn.) fruit in relation to their antioxidant activities

A large number of polysaccharides are present in the pericarp tissues of harvested litchi fruits. A DEAE Sepharose fast-flow anion-exchange column and a Sephadex G-50 gel-permeation column were used to isolate and purify the major polysaccharides from litchi fruit pericarp tissues. Antioxidant activities of these major polysaccharide components were also evaluated. An aqueous extract of the polysaccharides from litchi fruit pericarp tissues was chromatographed on a DEAE anion-exchange column to yield two fractions. The largest amount of the polysaccharide fraction was subjected to further purification by gel filtration on Sephadex G-50. The purified product was a neutral polysaccharide, with a molecular weight of 14 kDa, comprised mainly of 65.6% mannose, 33.0% galactose and 1.4% arabinose. Analysis by Smith degradation indicated that there were 8.7% of (1-->2)-glycosidic linkages, 83.3% of (1-->3)-glycosidic linkages and 8.0% of (1-->6)-glycosidic linkages in the polysaccharide. Furthermore, different polysaccharide fractions extracted and purified from litchi fruit pericarp tissues exhibited strong antioxidant activities. Among these fractions, the purified polysaccharide had the highest antioxidant activity and should be explored as a novel potential antioxidant.     

Extraction of polysaccharides and the antioxidant activity from the seeds of Plantago asiatica L

The extraction conditions of polysaccharides from Plantago asiatica L. seeds were investigated. Four parameters affecting the polysaccharides extraction, extraction times, water to sample, extraction temperature and single extraction time, were determined by orthogonal experiments. Under the optimized conditions, the polysaccharides yield of P. asiatica L. seeds was 2.467%. The antioxidant activities of the polysaccharides were investigated. The reducing power of the polysaccharides was dose dependent, and the reducing capacity of the polysaccharides was inferior to butylated hydroxytoluene, which is known to be a strong reducing agent. The scavenging rates of the polysaccharides on superoxide and 1,1-diphenyl-2-picrylhydrazyl radicals were79.7% and 81.4%, at polysaccharides concentration of 0.75 mg/mL, respectively, a scavenging rates approximately similar to that of 0.75 mg/mL ascorbic acid (83.5% and 85.1%, respectively). Furthermore, it exhibited a moderate concentration-dependent ABTS radical scavenging activity, ferrous ion chelating potency and H₂O₂ scavenging activity. The data obtained in the in vitro models clearly establish the antioxidant potency of the polysaccharides extracted from Semen Plantaginis.     

响应面法优化黄绿卷毛菇子实体多糖的提取工艺及其体外抗氧化活性研究

以采自四川石渠的黄绿卷毛菇Floccularia luteovirens为研究对象,采用水提醇沉的经典方法以及单因素试验,并利用Box-Benhnken Design（BBD）中心组合试验设计原理,以提取温度、时间以及料液比3个影响因素作为自变量,多糖提取率为考察指标,设计3因素3水平的响应面试验,优化黄绿卷毛菇子实体多糖（FLPs）的提取工艺,并通过?OH和O₂ ^-?自由基清除能力考察FLPs的抗氧化能力。最终确定最优水提工艺为提取温度89.31℃,提取时间5.08h,料液比1:48.54（g/mL）,且FLPs具有良好的?OH和O₂ ^-?自由基清除能力,具有较强的体外抗氧化活性。同时,验证试验证明了该设计方法和模型的准确性和可行性,且在该工艺条件下,多糖提取率有所提高,可为其多糖功能性食品和药品的开发提供理论依据。