Comparative characterization of polysaccharide fractions in grapes]

Different methods of isolating polysaccharides from grapes were evaluated with respect to the three indictrial grape varieties Traminer pink, Rkatsiteli, Cabernet-Sovinjon. Hot water extration was shown to be the best method in the isolation of water-soluble polysaccharides in primary wine-making. The qualitative and quantitative composition of the isolated polysaccharides was found to depend on the method of isolating polysaccharide fractions. The polysaccharides of grapes were heterogeneous in their molecular weight. The results of periodate oxidation of high molecular weight carbohydrates suggest a branched structure of polymers of water-soluble polysaccharides of grapes.     

The quantitative immunochemical determination of pneumococcal and meningococcal capsular polysaccharides by light scattering rate nephelometry

A quantitative nephelometric method was used for the measurement of the individual pneumococcal, as well as meningococcal, polysaccharides in the polyvalent vaccine final containers. This method is simple, rapid, inexpensive, and provides both qualitative and quantitative analyses of the polyvalent polysaccharide vaccines. By this method the individual pneumococcal types, 1, 2, 3, 4, 6A, 7F, 8, 9N, 12F, 14, 18C, 19F, 23F and 25 polysaccharides, were found to be present at 90-114% of the manufacturer's indicated concentrations; meningococcal group A, C, Y and W135 polysaccharides were at 90-108% of the manufacturer's listed concentrations. This nephelometric method coupled with gel filtration can also be used for measurement of the molecular sizes or stability of individual polysaccharides in the final container. Pneumococcal polysaccharide types 3, 6A, 9N and 19F, used as representative types, were treated with 0.5 N hydrochloric acid. The molecular sizes for types 3 and 9 N polysaccharides were stable to acid treatment. In contrast, types 6A and 19F polysaccharides were degraded. Heating meningococcal groups A, C, Y and W135 polysaccharides at 37 degrees C for 48 h did not affect their molecular size in the polyvalent vaccine.     

乌梅丸水提液中多糖理化性质分析及预防结肠炎的作用

目的:考察乌梅丸水提液中多糖含量,建立HPLC法分析乌梅丸多糖分子量及单糖组成方法,观察其对右旋葡聚糖硫酸钠(DSS)所致小鼠溃疡性结肠炎(UC)的预防作用,为乌梅丸临床用药及药理活性成分的研究提供依据。方法:采用水提醇沉法提取多糖;硫酸-苯酚法测定乌梅丸多糖含量;凝胶色谱法-龙智达分子量工作站测定多糖分子量;PMP柱前衍生化法HPLC分析单糖组成;DSS(3%)法建立UC小鼠模型。ICR小鼠50只随机分为5组:正常组,模型组,乌梅丸多糖2.5%,5%,10%剂量组,每组10只。造模前7天,在乌梅丸多糖治疗各组的小鼠饮水中添加乌梅丸多糖,一直给药维持至实验结束(第14天)。结果:葡萄糖在0~0.08 mg·m L-1浓度范围内呈良好的线性关系(r=0.998);右旋葡聚糖酐标准品分子量在2500~2000000 Da范围内呈良好线性关系(r=0.998);乌梅丸水提液中多糖含量为40.3%,纯化的乌梅丸多糖糖含量为91.6%;分子量范围在171343~525009 Da之间,分布系数D=1.18;其多糖的单糖组成主要为甘露糖、葡萄糖醛酸、葡萄糖、半乳糖和木糖组成,其相对含量比为1.8:1.0:19.3:32.8:4.2;乌梅丸多糖能降低结肠组织损伤程度。结论:多糖可能是乌梅丸汤剂中有效的活性成分,对小鼠UC有一定的防治作用,其理化性质分析方法简便快速,结果准确,重复性好,可作为乌梅丸多糖的质量控制方法。     

Influence of laminarin polysaccahrides on oxidative damage

Laminarin is a tropical plant traditionally used in Chinese medicine. In this experiment, Laminarin polysaccharides were analysed using GC-MS method and result showed that the polysaccharides contained mannose (3.27%), arabinose (8.61%), glucose (4.23%), galactose (12.12%), fucose (46.93%). Laminarin polysaccharides were tested to evaluate their effect on lung oxidative stress and lipid peroxidation in rats. The animal were divided into model and polysaccharides-treated animal. Laminarin polysaccharides were administered by gavage over a 14-day period. The results indicated that Laminarin polysaccharides significantly normalized catalase (CAT) activity (P<0.01), increased glutathione peroxidase (GPx) activity (P<0.05), superoxide dismutase (SOD) activity and reduced malondialdehyde (MDA) concentrations in animal. It could be concluded that Laminarin polysaccharides appeared to be more effective in reducing sepsis-induced oxidative stress, lipid peroxidation in animal.     

Purification of Mycobacterial Deoxyribonucleic Acid

Impurities believed to be polysaccharides have been found in mycobacterial deoxyribonucleic acid (DNA) preparations. Agar-gel diffusion of the DNA preparations against concanavalin A indicated the presence of three polysaccharides and was used to follow the purification procedures. The polysaccharides appeared to be the same for all strains studied. Precipitation of DNA with cetyltrimethylammonium bromide was used to separate impurities from some DNA preparations. The presence of the contaminants was found to affect markedly the determination of the guanine plus cytosine content according to a method dependent on the ratio of absorbancies at 260 and 280 nm; the impurities did not affect the determination by the method of thermal denaturation. The presence of a DNA-polysaccharide complex is suggested.     

Fluorescent derivatization of polysaccharides and carbohydrate-containing biopolymers for measurement of enzyme activities in complex media

Fluorescence derivatization provides a means of tracing the dynamics of polysaccharides even in the presence of high concentrations of other organic compounds or salts. A method of labeling polysaccharides with fluoresceinamine was extended to polysaccharides of a wide range of chemical composition, and alternative means of preparation were established for polysaccharides not initially amenable to column chromatography. The polysaccharides were activated with cyanogen bromide, coupled to fluoresceinamine, and separated from unreacted fluorophore via gel filtration chromatography or dialysis. Since the resulting derivatized polysaccharides proved to be stable to further physical and chemical manipulation, methods were also developed for re-activation and labeling with a second fluorophore, as well as for tethering the labeled polysaccharides to agarose beads. As an example of the application of this approach, five distinct fluorescently-labeled polysaccharides (pullulan, laminarin, xylan, chondroitin sulfate, and alginic acid) were used to investigate the activities and structural specificities of extracellular enzymes produced in situ by marine microbial communities, providing a means of measuring specifically the activities of endo-acting extracellular enzymes and avoiding use of low molecular mass substrate proxies. These labeled polysaccharides could be used to explore the dynamics of polysaccharides in other types of complex media, as well as to investigate the activities and specificities of endo-acting enzymes in other systems.     

草果多糖的含量测定

目的：为了充分利用草果资源,减少资源浪费,研究草果多糖含量的测定.方法：利用热水浸泡充分提取草果多糖,并用Sevag法脱蛋白法将其提纯,然后进行草果多糖含量的测定.结果：样品多糖的含量测得为0.312％,回收率为97.14％,RSD =0.827％ （n =5）.结论：广西贵港草果中含有一定的多糖,且该方法操作简便,准确,可靠,可作为检验草果多糖含量的较好方法.     

近红外光谱测定灵芝子实体多糖含量

灵芝具有多种药理活性,多糖是其主要活性成分之一.目前灵芝多糖的定量常采用的比色法,使用比较繁琐,也缺乏一定的安全环保性.应用近红外光谱对灵芝进行多糖定量分析,发现对子实体直接分析存在定量不准的问题.为了快速准确地评估灵芝子实体多糖含量,本研究采用对灵芝子实体水提物进行近红外光谱检测与分析,由此建立了较好的定量模型.该模...     

检测四价脑膜炎球菌多糖疫苗中A群多糖含量ELISA法的建立

目的建立双抗体夹心ELISA法,对A群流脑多糖抗原进行特异性定量测定。方法制备抗A群多糖的特异性多克隆抗体,所得抗血清经辛酸-硫酸铵沉淀法纯化后,用过碘酸钠法制备辣根过氧化物酶标记多克隆抗体。分别以抗A群多糖多克隆抗体作为包被抗体及酶标二抗,建立双抗体夹心ELISA法,优化反应条件,对A群多糖抗原进行特异性定量测定。结果一系列验证试验表明,该法特异性较好,未检出与C、Y、W135群多糖的交叉反应;1.25～20 ng/mL多糖浓度范围的剂量反应曲线线性最佳,相关系数大于0.98,经实验内10次及不同试验间以16、84、ng/mL测定3次A群多糖中的含量,变异系数在6.3%～11.5%间,回收率在91.8%～105.9%之间,符合常规质控要求,检测限量为4 ng/mL。采用该法测定3批ACYW135群四价脑膜炎球菌多糖疫苗中A群多糖含量、分子大小及回收率的结果均符合规程草案质量标准。结论建立的双抗体夹心ELISA法可尝试用于ACYW135群脑膜炎球菌多糖疫苗中A群多糖的关键质量指标的检测。     

Useful applications of DOSY experiments for the study of mushroom polysaccharides

DOSY analysis has been performed on different mushroom extracts and fractions with the aim to describe a general method for the study of high molecular weight polysaccharides. These NMR experiments can be exploited to monitor the fractionation pathways performed on crude extracts in order to isolate polysaccharides. DOSY can also rapidly verify the purity of the isolated compounds, as well as evaluate their molecular size. In spite of the complexity of DOSY spectra of mixtures, this NMR technique seems to be a valid analytical tool that could be adopted as a routine method for the study of polysaccharides from different sources.     

白花蛇舌草多糖的分离提取及含量测定

采用热水浸提法提取白花蛇舌草水溶性多糖,薄层层析法鉴定其多糖的单糖组成。通过正交试验优选浸提显色条件,以苯酚-硫酸法制得有色糖醛衍生物,用分光光度法在490nm波长处测定吸光度,其曲线方程为Y=0．01361x-0．08161,相关系数r=0．9997。结果表明,白花蛇舌草多糖由鼠李糖、葡萄糖、半乳糖及甘露糖等组成,含量为15．10％,回收率达95．68％。     

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。     

Studies on Sophora alopecuroides Gum

The seeds of Sophora alopecuroides L, contained 12.8 per cent of endosperm, from which the water-soluble polysaccharide gum was isolated and purified by fractionation with alcohol three times. Hydrolysis of the polysacchatides with sulphuric acid produced galactose and mannose in the molar ratio of 1:2.37, as determined by the phenolsulphuric acid method and by paper chromatographic method. The total sugar content reached 67-70%. The polysaccharides, after complete hydrolysis, reduction with NaBH4 and acetylation, ridded acetates. An analysis on them by gas-chromatography showed that they contained galactose and mannose in the molar ratio of 1:1.83, and the total sugar content reached 56%. In order to identify the linkages of the polysaccharides isolated from the seeds of Sophora alopecuroides, they were methylated by using the methods of Hokomori, Conval and Albersheim. The fully methylated polysaccharides were hydrolyzed and then reduced with NaBH4, followed by acetylation. Finally they yielded acetates. An analysis on IR data, the results of acid hydrolysis, periodate oxidation and Smith type degradation, as well as the acetates by GC, GC-MS and 1HNMR showed that the linkages of Sophora alopecuroides. gum may be, The purified polysaccharides seem to be homogenous after ultracentrifugation analysis. The sedimentation coefficient of the gum obtained from repeated purification with alcohol was 5.31, its intrinsic viscosity was 6.25 dl/g and its molecular weight was estimated to be 635000. By the light-scattering method, however, its molecular weight was estimated to be 715000. The crude polysacchafides contained 44 per cent of the water-soluble polysaccharides.     

A sensitive staining method for detecting acidic polysaccharides in cellulose acetate and agarose gels

A sensitive staining method has been developed for the detection of acidic polysaccharides in cellulose acetate and agarose gels. The method is based on the precipitation of bovine serum albumin by acidic polysaccharides at acidic pH values and the subsequent staining of precipitated protein with amido black or Coomassie brilliant blue R-250 stains. The detection limit of acidic polysaccharides is 15-40 ng on cellulose acetate strips and 50-150 ng on agarose plates. The sensitivity of the described staining technique is of the same order for a wide range of acidic polysaccharides of different origin in contrast to Alcian blue and toluidine blue stains, which detect only mucopolysaccharides of animal origin at comparable levels. The method was also applied to the colorimetric quantitative determination of acidic polysaccharides after electrophoretic separation.     

灰树花多糖的提取及抗氧化活性

采用单因素和正交试验法对热水浸提、超声波辅助和微波辅助提取灰树花多糖的工艺进行研究,分别获得了3种方法的最佳条件,并发现微波辅助法最佳,其最优条件为:微波功率800 W、微波时间3 min,浸提2h,多糖得率为13.05％,比热水浸提法提高了51.22％.利用提取的灰树花多糖进行抗氧化性研究,发现灰树花多糖对O2-·和·OH都具有一定的清除作用,且多糖浓度与其对O2-·的清除作用存在量效关系,但不如·OH明显.     

A novel high-performance anion-exchange chromatographic method for the analysis of carrageenans and agars containing 3,6-anhydrogalactose

A novel method has been developed to determine the sugar composition of 3,6-anhydrogalactose-containing polysaccharides, such as carrageenan and agar. The method is based on reductive hydrolysis with a methylmorpholine-borane complex in the presence of acid and subsequent high-performance anion-exchange chromatography analysis of the alditols without any derivatization. The method was validated by 13C NMR analysis of six carrageenans and three agars and by a previously used method based on derivatization to alditol acetates and gas-liquid chromatography analysis. The new method was found to be superior to the gas-liquid chromatography method as the analysis time was less than half. Also it was found to be more accurate and reproducible and no derivatization was required. The analysis of the six different carrageenan samples revealed that homogeneous mu- and nu-carrageenan, theoretically without 3,6-anhydrogalactose residues, cannot be isolated from red seaweeds. Consequently, the question arose if mu- and nu-carrageenans at all are present in seaweeds and if the current hypotheses regarding biosynthesis of carrageenans in the seaweeds are correct. The data demonstrated that carrageenans are highly complex natural polysaccharides, which are more irregular than assumed hitherto. The new analytical technique will permit elucidation of the detailed structure of seaweed polysaccharides and determination of their structure-property relationships.     

An evaluation of tritium and fluorescence labelling combined with multi-detector SEC for the detection of carbonyl groups in polysaccharides

The carbonyl content of a pectic polysaccharide from Sphagnum papillosum (sphagnan) and periodate oxidised alginates was investigated using three different carbonyl labelling strategies combined with size-exclusion chromatography (SEC) with multi-angle laser light scattering (MALLS) and on-line fluorescence or off-line tritium detection. The labelling strategies were tritium incorporation via NaB³H₄ reduction, and fluorescent labelling with carbazole carbonyl oxyamine (CCOA), or 2-aminobenzamide (2-AB), respectively. Carbonyl quantification was based on labelled pullulan, dextran and alginate standards possessing only the reducing end carbonyl group. As a result the carbonyl distribution in the polysaccharides could be determined. In sphagnan it was found that the carbonyl content increased with increasing molecular weight, whereas in periodate oxidised alginate the carbonyl content was as expected independent of the molecular weight. The methods proved useful for carbonyl detection in water soluble polysaccharides in general. The tritium incorporation method was preferred for alkali stable polysaccharides, while the CCOA method was most suitable for acid stable polysaccharides with low carbonyl content. The 2-AB method is applicable for all polysaccharides tested with varying carbonyl content; however, it lacks the ability to detect ketone functionalities.     

Cytochemistry and distribution of polysaccharides in an electroreceptor: the tuberous organ of Gnathonemus petersii (Mormyrids).

The polysaccharides were studied in an electroreceptor organ, the tuberous organ of Gnathonemus petersii (Mormyridae). Histochemical methods (P.A.S., alcian blue, toluidine blue and iron colloidal reactions) allowed us to demonstrate the existence of glycogen in the sensory cytoplasm, and P.A.S. positive polysaccharides in the sensory cavity. The polysaccharides were shown to be amylase proof; they display an acidity due to the existence of sulphated radicals. The histochemical study was completed by a cytochemical analysis: a treatment with thiocarbohydrazide (TCH) according to the Thiery's method. This method allowed us to estimate the glycogen concentration, its localization, and relationship with cellular organites within the sensory cytoplasm, as well as to differentiate the highly glycogenous type II cells of the platform from the other accessory cells (Derbin and Szabo, 1968). After a treatment for 20 hours with TCH, silver stained grains were visible on the polysaccharide filaments of the sensory chamber, between the microvilli and the vacuoles of the epidermal cells lining to the sensory cavity. Silver grains coated the outer surface of the microvilli. Such polysaccharides were not identical to the filamentous polysaccharides of the cavity. In order to determine the cytochemical localization of the polysaccharide acid groups, sections were stained with iron salts. The colloidal iron constitutes a deposit opaque to electrons and located both on the filamentous polysaccharides of the sensory cavity and in the vacuoles of the epidermal cells, indicating that only these filamentous polysaccharides display acid radicals.     

Polysaccharide analysis using carbohydrate gel electrophoresis: a method to study plant cell wall polysaccharides and polysaccharide hydrolases.

A method to characterize plant cell wall polysaccharides is presented. The complexity of the polymer structures and the large number of different charged and uncharged monosaccharides that make up plant polysaccharides have previously made analysis technically demanding and laborious. Polysaccharide analysis using carbohydrate gel electrophoresis (PACE) relies on derivatization of reducing ends of sugars and oligosaccharides with a fluorophore, followed by electrophoresis under optimized conditions in polyacrylamide gels. We show that PACE is a sensitive and simple tool for studying the monosaccharide composition of polysaccharides and of cell wall preparations. In combination with specific hydrolases, it can be used to analyze the structure of polysaccharides. Moreover, the specificity and kinetics of the plant polysaccharide hydrolases themselves can be quickly and effectively studied. PACE can detect as little as 500 fmol of monosaccharides and 100 fmol of oligosaccharides, and it is fast and quantitative.     

聚酰胺树脂分离纯化芦荟多糖的研究

文章采用聚酰胺吸附树脂,采用水、0．2M／LNaCl溶液、5％NaOH水溶液为洗脱剂梯度洗脱分离纯化,得到了三个芦荟多糖组分,通过IR、UV、GC等手段对其进行了分析。以甘露糖为标准,采用硫酸．苯酚法,测得水洗和NaOH水溶液洗脱所得多糖含量分别为90．10％和93．28％;以木糖、甘露糖、葡萄糖、半乳糖为单糖标准的GC分析结果显示：水洗所得多糖主要以甘露糖为主,同时含有少量的葡萄糖。