库拉索芦荟多糖的分离纯化和含量测定

目的　分离纯化库拉索芦荟多糖并建立含量测定方法。方法　醇沉法提取库拉索芦荟多糖,Sephadex G-100凝胶柱色谱纯化。以甘露糖为标准,苯酚硫酸法测定库拉索芦荟多糖含量。结果　经分离纯化得到的主要组分库拉索芦荟多糖I含量为81.11%,测定平均回收率为100.3%。结论　该方法可从库拉索芦荟中提取分离纯度较高的多糖,含量测定方法简便、快速、重现性好。     

库拉索芦荟多糖的分离纯化和含量测定

目的分离纯化库拉索芦荟多糖并建立含量测定方法.方法醇沉法提取库拉索芦荟多糖,Sephadex G-100凝胶柱色谱纯化.以甘露糖为标准,苯酚-硫酸法测定库拉索芦荟多糖含量.结果经分离纯化得到的主要组分库拉索芦荟多糖Ⅰ含量为81.11%,测定平均回收率为100.3%.结论该方法可从库拉索芦荟中提取分离纯度较高的多糖,含量测定方法简便、快速、重现性好.     

芦荟多糖的分离纯化及其相对分子质量和含量测定

目的 研究库拉索芦荟多糖的分离纯化,测定分离得到的纯化多糖PSA2的相对分子质量及其多糖和蛋白质的含量. 方法采用水提醇沉法提取粗多糖,反复冻融法除蛋白质,DEAE-cellulose 52和Sephadex G-100凝胶色谱柱纯化多糖,得到纯化组分(PSA2),高效凝胶渗透色谱(HPGPC)法测定PSA2的相对分子质量,苯酚 硫酸法测定其多糖含量,考马斯亮蓝G-250法测定其蛋白质含量. 结果 PSA2的相对分子质量为8 457.4,其多糖含量为 61.0%,蛋白质含量为0.7%. 结论 多糖的提取工艺简单可行,HPGPC法,苯酚 硫酸法和考马斯亮蓝G-250法准确快捷.     

库拉索芦荟粗多糖提取工艺优化

目的优化库拉索芦荟粗多糖提取工艺。方法采用正交设计方法对库拉索芦荟粗多糖的提取工艺进行优化，采用苯酚-硫酸法测定多糖含量。结果库拉索芦荟多糖提取的最佳工艺条件为液料比1：1，温度80℃，醇沉浓度80％，提取4h。结论优化工艺可为库拉索芦荟多糖的开发利用提供参考。     

红芪多糖的提取分离纯化及组成分析

目的获得红芪多糖纯品并且得到其单糖组成。方法通过分步醇沉法和凝胶柱色谱获得红芪多糖纯品,用气相色谱法测其组成。结果红芪多糖1和2经Sephadex G-200纯化后分别得到两个组分,红芪多糖3通过Sephadex G-25得到两个组分;红芪多糖1,2,3均由鼠李糖、阿拉伯糖、木糖、葡萄糖、半乳糖五种单糖组成。结论分步醇沉法获得的三种多糖1,2,3经Sephadex G-200和Sephadex G-25纯化可获得红芪多糖纯品,GC法测其组成方法简便可行。     

红芪多糖的提取分离纯化及组成分析

目的 获得红芪多糖纯品并且得到其单糖组成.方法 通过分步醇沉法和凝胶柱色谱获得红芪多糖纯品,用气相色谱法测其组成.结果 红芪多糖1和2经Sephadex G-200纯化后分别得到两个组分,红芪多糖3通过Sephadex G-25得到两个组分;红芪多糖1,2,3均由鼠李糖、阿拉伯糖、木糖、葡萄糖、半乳糖五种单糖组成.结论 分步醇沉法获得的三种多糖1,2,3 Sephadex G-200 和Sephadex G-25纯化可获得红芪多糖纯品,GC法测其组成方法简便可行.     

金线莲多糖的分离纯化与含量测定

目的：从金线莲中分离纯化多糖，并建立金线莲多糖含量测定方法。方法：用水提醇沉法提取金线莲多糖，用SephadexG-100凝胶柱色谱纯化多糖；并用改良的苯酚-硫酸法测定多糖含量。结果：测得金线莲粗多糖生药量为37．45％，精制金线莲多糖（AFPS）含量为72．84％，AFPS生药量为27．14％，测定平均回收率为100．40％。结论：金线莲中多糖含量较高，具有开发利用价值。     

天麻多糖的分离纯化与抗氧化活性研究北大核心CSCD

目的从天麻中分离纯化出活性多糖并考查其抗氧化活性。方法用水提醇沉法提取天麻粗多糖,Sevage法脱去其中的蛋白质,依次通过D101大孔吸附树脂、DEAE-52离子交换色谱及Sephadex G-100凝胶色谱纯化,得到2个天麻多糖(GEP):GEP1-G和GEP2-G,测定其清除1,1-二苯基-2-三硝基苯肼(DPPH)自由基、超氧阴离子自由基和羟基自由基的能力,以评价其抗氧化作用。结果天麻中粗多糖提取率为5.6%,经D101大孔吸附树脂纯化后,粗多糖中多糖含量为65.7%;DEAE-52纯化后,有6个洗脱组分GEP 1~6;组分GEP1和GEP2再经Sephadex G-100纯化得到纯品多糖GEP1-G和GEP2-G,其多糖含量分别是97.3%和98.1%。在抗氧化活性实验中:当GEP1-G和GEP2-G浓度为1 mg·mL^(-1)时,两者对DPPH的清除率分别为44.50%和25.60%,对超氧阴离子抑制率分别为33.32%和21.55%,对羟自由基抑制率分别为39.50%和22.80%。结论通过大孔吸附树脂、离子交换色谱和凝胶过滤色谱得到了纯品天麻多糖,其具有一定的抗氧化作用。     

蒽酮-硫酸法测定库拉索芦荟多糖含量

目的:建立测定库拉索芦荟多糖含量的方法.方法:采用先醇沉后水提的方法分离提取芦荟多糖,以甘露糖标准液绘制标准曲线,用蒽酮-硫酸分光光度法测定多糖含量,检测波长为610 nm.结果:芦荟各部位多糖含量不同,凝胶、叶皮和全叶的多糖含量分别为0.952,1.673,0.992 mg·g-1,平均回收率为99.57%.结论:该测定方法简便、快速、重现性好,对库拉索芦荟多糖含量的测定可获得满意结果.     

气相色谱法测定吐鲁番无核白葡萄树伤流液组成多糖的单糖

目的对吐鲁番葡萄树伤流液多糖进行分离纯化,并分析其单糖组成。方法采用醇沉法提取多糖,Sevage法脱蛋白,经DEAE-52纤维素柱及Sephadex G-150凝胶柱分离纯化,采用气相色谱分析单糖组成。结果用体积分数80%乙醇沉淀、Sevage法除蛋白,得葡萄树伤流液粗多糖,粗多糖经过DEAE-52纤维素柱分离得到8个多糖组分,编号分别为:组分-1,组分-2,组分-3,组分-4,组分-5,组分-6,组分-7,组分-8。其中4个多糖组分经过Sephadex G-150纯化后,通过GC分析组分-1和组分2含有鼠李糖、阿拉伯糖、木糖、甘露糖、葡萄糖、半乳糖;组分-3含有鼠李糖、阿拉伯糖、甘露糖、葡萄糖、半乳糖;组分-5含有鼠李糖、甘露糖、葡萄糖、半乳糖。结论该方法可较好地分离纯化葡萄树伤流液多糖。采用GC分析多糖的单糖组成,研究结果适用于多糖中的单糖组成分析。     

制何首乌多糖的纯度鉴定和理化性质研究

目的对制何首乌多糖进行纯度鉴定和理化性质研究。方法采用水提醇沉法提取制何首乌多糖,经过纤维素和凝胶柱层析纯化后,纯化后的多糖PRPM-1、PRPM-2和PRPM-3分别用Sepha-dexG-100柱层析和紫外吸收光谱检测其纯度;通过物理和化学方法分析多糖理化性质。结果制何首乌多糖为均一多糖。结论该测定方法准确,重复性好。     

Purification of two thrombin-like enzymes from the venom of Agkistrodon caliginosus (kankoku-mamushi)

Two thrombin-like enzymes were purified from the venom of Agkistrodon caliginosus. Both were homogeneous on polyacrylamide gel electrophoresis, hydrolyzed $\text{N-α-}\text{tosyl}\text{-}\text{l}\text{-}\text{arginine}$ methylester and did not show any caseinolytic activity. The molecular weights of the enzymes were estimated to be about 34,000–37,000 and 42,000–43,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by gel-filtration on Sephadex G-100 column. One enzyme was purified from the venom by gel-filtration on Sephadex G-100 column, CM-Sephadex C-50 and DEAE-Sephadex A-50 chromatographies, affinity chromatography on p-aminobenzamidine-ε-aminocaproic acid-Sepharose 4B and gel-filtration on Sephadex G-100 column. The second enzyme was separated from the first enzyme by the DEAE-Sephadex A-50 chromatography mentioned in the above procedure and was isolated by affinity chromatography on arginine-Sepharose 4B and gel-filtration on Sephadex G-100 column. From 4 g of the venom, 0.88 mg of the first enzyme and 1.18 mg of the second enzyme were obtained, with specific activities of 118 and 139 NIH units per mg of protein, respectively.     

牛膝多糖工艺研究

牛膝根经热水提取、乙醇分级沉淀，鞣酸去除蛋白质，再经ＤＥＡＥ－Ｓｅｐｈａｒｎｓｅｇｆａｔ－ｆｌｏｗ柱和ＳｅｐｈａｄｅｘＧ－２００凝胶过滤，洗液对蒸馏水稼析、冻干。所得的经纯化的牛膝多糖分子量为１．４４ｋｄ，纯度９８．６％。     

天麻多糖提取工艺及纯化研究

目的:比较天麻多糖(GEP)超声波提取和水提工艺的多糖提取率,探讨超声技术在多糖提取方面的优势。方法:利用超声波提取工艺从中药天麻中提取天麻多糖,并用sevag法脱蛋白,经透析、冷冻干燥得天麻粗多糖。然后通过Sephadex G-75凝胶色谱柱将其进行分离纯化,得到两个天麻多糖分级组分GEPⅠ和GEPⅡ。再利用紫外可见分光光度法对GEPⅠ和GEPⅡ的纯度进行鉴定。结果:在提取温度60℃、料水比1:40、提取时间0.5h的条件下,用超声波回流提取3次,是天麻多糖超声提取的最佳方法。在相同因素的条件下,得到的提取率较传统的水提取法提高了11.2%,同时缩短了提取时间。结论:用超声波提取,相比传统水提醇沉法,在提取率方面得到了显著的提高。     

Hydrolysis of ester-type drugs by the purified esterase from human intestinal mucosa.

Esterase from human intestinal mucosa was purified 210 fold by solubilization with Triton X-100, chromatography on DEAE-cellulose, Sephadex G-100 and hydroxylapatite, and isoelectric focusing. The purified esterase showed a single band by polyacrylamide gel electrophoresis. The molecular weight of the purified esterase was estimated to be about 55,000 by gel filtration on Sephadex G-150, and the isoelectric point was 5.02. The purified esterase was strongly inhibited by diethyl p-nitrophenyl phosphate (E-600) and diisopropyl fluorophosphate (DFP), and was not inhibited by eserine sulfate and p-chloromercuribenzoate. The purified esterase from human intestinal mucosa was found to be one of the carboxylesterases. The purified esterase hydrolyzed ester-type drugs, i.e., aspirin, clofibrate, indanyl carbenicillin and procaine, but did not hydrolyze amide-type drugs and choline-type drugs.     

巴西蘑菇Agaricus blazei Murill水溶性多糖的分离纯化

从巴西蘑菇子实体中提取的水溶性粗多糖经过脱蛋白、脱色等步骤获得半纯品多糖。依次采用DE—AB纤维素和Sepharose CL-4B柱层析进一步纯化后，获得5种水溶性多糖组分。其中主要组分AB-I、AB-Ⅱ-b和AB-Ⅲ-b得率分别为3．3％、1．52％、0．85％。Sephadex G-l00凝胶层析、旋光度和HPLC鉴定各组分已达到层析纯。     

Properties of NAD glycohydrolase purified from five-pace snake (Agkistrodon acutus) venom

NAD glycohydrolase (NADase) (E.C. 3.2.2.5) from five-pace snake (Agkistrodon acutus) venom was purified to electrophoretic homogeneity through a 4-step isolation procedure, including column chromatography using DEAE-Sephadex A-50, Sephadex G-75, CM Sephadex C-50 and Sephadex G-100. The final product was 11.8-fold purified with a 3.9% yield. The pure enzyme showed maximal activity at about 40 degrees C with optimal pH at 7.5. It was a glycoprotein with a pI of 7.6. Its mol. wt was respectively 98,000 as measured by gel filtration and 50,000, by SDS-PAGE. There was only one N-terminal residue, proline. NADase is thus composed of two identical subunits in each molecule. The enzyme contained copper ions. NADase activity was lost when the copper enzyme complex was treated with EDTA. The Km of the enzyme for beta-NAD, NADP and beta-NGP were 0.50 mM, 0.13 mM and 0.16 mM respectively.