不同方法对寡聚甘露糖醛酸铬配合物分子量测定的影响

目的探讨不同方法对于测定具有抗2型糖尿病活性的海洋寡聚甘露糖醛酸铬配合物(HS203)分子量测定的影响。方法采用聚丙烯酰胺凝胶电泳(PAGE),高效凝胶渗透色谱(HPGPC)及高效凝胶渗透色谱与多角度激光光散射器联用(HPGPC-MALLS)法分别测定HS203的分子量,并考察了不同色谱柱以及不同分子量对照品的选择对于HS203分子量测定结果的影响。结果 PAGE法表明HS203的聚合度为5~20之间,通过计算得出HS203的重均分子量范围在2 400~2 600Da。在HPGPC分析过程中,不同色谱柱对于HS203分子量测定结果影响不大,采用右旋糖酐作为对照品测得分子量为6 000~7 10...     

锦灯笼中酸浆苦素类化学成分的研究

目的 研究锦灯笼95％乙醇提取物的石油醚、氯仿、醋酸乙酯萃取部位中化学成分.方法 利用硅胶柱色谱、反相ODS柱色谱、制备液相色谱等方法进行分离、纯化,根据化合物的理化性质和光谱数据鉴定其结构.结果 分离得到了7个化合物,经结构鉴定分别为酸浆苦素A(1)、酸浆苦素D(2)、酸浆苦素L(3)、酸浆苦素O(4)、木犀草素(5...     

美花铁线莲地上部分的皂苷类成分研究

目的研究美花铁线莲地上部分的皂苷类化学成分。方法将干燥的样品用70%乙醇回流提取,所得浸膏分步萃取得到总皂苷,再利用正相硅胶柱色谱、凝胶柱色谱、反相硅胶柱色谱、制备高效液相色谱等方法分离得到单体化合物,根据其理化性质与波谱数据鉴定化学结构。结果分离得到5个单体皂苷,分别鉴定为clematiganoside A(1)、huzhangoside D(2)、huzhangoside B(3)、cauloside G(4)和loniceroside C(5)。结论所有化合物均为首次从该种植物中分离得到,其中皂苷5是从本属植物中首次发现。     

抗生素质控分析中HPLC色谱柱的选择

依据各类抗生素不同的理化性质和各类色谱柱不同填料的选择性。结合实践经验并参考国内外相关文献．较全面地综述抗生素质控分析中HPLC色谱柱的选择。     

板蓝根中的一个新异香豆素

目的对板蓝根的化学成分进行研究。方法采用硅胶柱色谱、凝胶柱色谱进行分离,通过理化性质和波谱分析方法鉴定化合物结构。结果从正丁醇层中分离得到化合物Ⅰ(indigotiiso coumarinA)。结论化合物Ⅰ为新化合物,命名为板蓝根异香豆素A(indigotiisocoumarinA ,3 ( 3 pyrrole) 6 hydroxy 7 hydroxymethyl isocoumarin)     

穿山龙总皂苷的HPLC指纹图谱研究

目的:建立穿山龙总皂苷的高效液相色谱(HPLC)指纹图谱,为其质量评价提供依据。方法:测定9批不同来源的穿山龙样品。色谱柱为Kromasil C8(250mm×4.6mm,5μm),流动相为水-乙腈(梯度洗脱),流速为1.0mL·min-1,检测波长为203nm,柱温为30℃。结果:共标示出穿山龙皂苷类化合物17个特征指纹峰;不同来源的穿山龙总皂苷色谱峰峰面积比值有一定的差异。结论:HPLC指纹图谱分析法能简便、快速地区分不同来源的穿山龙,为全面控制穿山龙药材的质量提供依据。     

中药连翘的化学成分研究

目的:对中药连翘的化学成分进行研究。方法:利用大孔吸附树脂、硅胶柱色谱、Sephadex LH-20柱色谱、ODS柱色谱和制备液相等色谱手段进行分离纯化,根据化合物的理化性质和波谱学等方法鉴定其结构。结果:从木犀科连翘属植物连翘的95%乙醇提取物中分离得到6个化合物,分别鉴定为对羟基苯甲酸(1),对乙醇基苯甲醛(2),对羟基苯乙酸(3),对香豆酸(4),对羟基苯基乙酸乙酯(5),1-(4-羟基苯基)-2,3-二羟基丙酮(6)。结论:化合物2、5、6为首次从连翘属(Forsythia)中分离得到。     

一枝蒿的化学成分分离与鉴定

目的 研究菊科蓍属药用植物一枝蒿(Achillea alpina L.)的化学成分.方法 对一枝蒿体积分数95％乙醇提取物的二氯甲烷和乙酸乙酯部位采用硅胶柱色谱、Sephadex LH-20、聚酰胺柱色谱和高效液相(HPLC)色谱等技术分离纯化,根据理化性质并应用波谱数据鉴定各化合物结构.结果 从一枝蒿中分离鉴定了10...     

温莪术中的愈创木烷型倍半萜

目的对温莪术(Curcuma wenyujin)的化学成分进行研究。方法采用硅胶柱色谱、ODS柱色谱、Sephadex LH-20柱色谱以及HPLC进行分离和纯化,并依据理化性质和谱学分析鉴定其化学结构。结果从温莪术的体积分数为50%的乙醇溶液提取物中分离得到7个愈创木烷型倍半萜,分别鉴定为zedoarondiol(1)i、sozedoarondiol(2)a、erugidiol(3)z、edoalactone A(4)z、edoalactone B(5)、zedoalactone C(6)和zedoarolide B(7)。结论化合物1-7均为首次从该植物中分离得到。     

海洋来源真菌草酸青霉(Penicillium oxalicum)次级代谢产物的分离与鉴定

目的研究海洋来源真菌草酸青霉(Penicillium oxalicum)的次级代谢产物。方法采用重结晶、硅胶柱色谱、Sephadex LH-20柱色谱、开放ODS柱色谱等方法进行分离纯化;根据理化性质及光谱数据确定化合物的结构。结果分离得到10个化合物,分别鉴定为N-acetyl-hydrazinobenzoic acid(1)、N-[2-cis-(4-hydroxyphenyl)ethenyl]formamide(2)、p-hydroxyphenylacetamide(3)、penipanoid A(4)、penipanoid C(5)、2-(4-hydroxybenzyl)quinazolin-4(3H)-one(6)、oxaline(7)、meleagrin(8)、(3R,4R)-3,4,8-trihydroxy-3,4-dihydro-1(2H)-naphthalenone(9)和(3R,4R)-3,4,6,8-tetrahydroxy-3,4-dihydro-1(2H)-naphthalenone(10)。结论化合物1为新天然产物,化合物9、10为首次从青霉属真菌中分离得到。     

短蛸章鱼多糖的提取分离和基本性质分析

目的从短蛸(Octopus ocellatus)的胴体和腕部提取多糖,对短蛸不同部位提取的粗多糖的基本理化性质进行分析和比较,并对粗多糖进行分离和纯化。方法依次采用冷水,木瓜蛋白酶和胰蛋白酶联合酶解的方法提取多糖,采用Sevage法和等电点法去除粗多糖中的蛋白。对短蛸不同部位提取粗多糖的总糖、蛋白、糖醛酸、硫酸根含量和重均相对分子质量、单糖组成等性质进行分析和比较,并采用Sephacryl S-300凝胶柱层析和QSepharose4 Fast Flow离子交换层析方法对粗多糖进行分离和纯化。结果从短蛸胴体中提取的粗多糖中总糖和糖醛酸的含量明显高于腕部粗多糖,且蛋白含量较腕部多糖低;水提多糖中总糖和糖醛酸的含量明显低于酶提多糖,且蛋白含量高于酶提多糖;短蛸胴体粗多糖的单糖组成相对简单,主要含有Glc和少量Man、GlcNAc、GlcUA,而腕部粗多糖的单糖组成较为复杂,除主要含有Man、GlcNAc、GlcUA和GlcN外,还含有少量Gal和Fuc等;各多糖的重均相对分子质量在127.9～266.4kD之间;水提多糖和酶提多糖在SephacrylS-300凝胶柱层析上的洗脱曲线也明显不同。结论采用不同方法、从...     

Reduction in vegetative growth of the water mold Saprolegnia parasitica (Coker) by humic substance of different qualities

Humic substances (HS) account for 50-80% of the dissolved organic matter in non-eutrophicated freshwater ecosystems. HS are not inert, but are taken up by and may interact with aquatic organisms. However, at present no information is available on the interaction of HS with fungi, for instance, the fish-pathogenous species Saprolegnia parasitica. To fill this gap, we tested effects of HS on S. parasitica growth in-vitro using 25-500mgL(-1) carbon of HS on GY-agar. We investigated 20 HS including natural organic matter (NOM) samples, two lignite derived HS, and one synthetic HS. The aim was (1) to find out, if there are inhibiting effects and (2) if potential effects can be explained by humic matter properties by structure activity relationships. The growth of S. parasitica was related to the growth on HS-free agar controls. Characterization of HS and NOMs included elemental analysis, high-pressure size exclusion chromatography (HPSEC), UV/VIS, FTIR-, and EPR-spectroscopy in order to obtain information on elemental and structural composition including various metals, molecular weights of the HS fraction, aromaticity, free organic radicals, and functional groups. NOMs with high moieties of high-molecular carbohydrates supported the growth of S. parasitica, all other HS and NOMs reduced it. However, no inhibition of the development of the sporangia and primary zoospores was found. Therefore, the impact of the HS on S. parasitica has to be classified as fungistatic, rather than fungicidal. Synthetic and lignite-derived HS were among the most efficient HS sources. Growth inhibition was correlated (p<0.05) with the molecular weights of the HS-fraction, sUVa, COOH groups, C and H. Our results suggest that especially HS with higher molecular weights and aromaticity which contain a high number of organic radicals are the most efficient in reducing fungal growth. Furthermore, highly functionalized HS seem to be important for the observed effect. The development of internal oxidative stress could be a mechanism explaining the observed growth inhibition of S. parasitica.     

Caffeic acid methyl and ethyl esters exert potential antidiabetic effects on glucose and lipid metabolism in cultured murine insulin-sensitive cells through mechanisms implicating activation of AMPK

Context: Caffeic acid methyl (CAME) and ethyl (CAEE) esters stimulate glucose uptake and AMP-activated protein kinase (AMPK) in C2C12 myocytes (ATCC^® CRL-1772^TM).

Objective: Effects of CAME and CAEE were now assessed on myocyte glucose transporter GLUT4 activity and expression, on hepatic gluconeogenesis and on adipogenesis as well as major underlying signaling pathways.

Materials and methods: GLUT4 protein translocation was studied in L6 GLUT4myc cells, glucose-6-phospatase (G6Pase) in H4IIE hepatocytes and adipogenesis in 3T3-L1 adipocytes. Key modulators were measured using western immunoblot. Cells were treated for 18?h with either CAME or CAEE at various concentrations (12.5–100?μM).

Results: Myocyte glucose uptake rose from 10.1?±?0.5 to 18.7?±?0.8 and 21.9?±?1.0?pmol/min/mg protein in DMSO-, CAME- and CAEE-stimulated cells, respectively, similar to insulin (17.7?±?1.2?pmol/min/mg protein), while GLUT4myc translocation increased significantly by 1.70?±?0.18, by 1.73?±?0.18- and by 1.95?±?0.30-fold (relative to DMSO), following insulin, CAME and CAEE stimulation, respectively. CAME and CAEE suppressed hepatocyte G6Pase by 62.0?±?6.9% and 62.7?±?6.0% with IC₅₀ of 45.93 and 22.64?μM, respectively, comparable to insulin (70.7?±?2.3% inhibition). Finally, CAME and CAEE almost abrogated adipogenesis (83.3?±?7.2% and 97.3?±?3.0% at 100?μM; IC₅₀ of 13.8 and 12.9?μM, respectively). The compounds inhibited adipogenic factors C/EBP-β and PPAR-γ and stimulated AMPK activity in the three cell-lines.

Discussion and conclusions: CAME and CAEE exerted antidiabetic activities in insulin-responsive cells through insulin-independent mechanisms involving AMPK and adipogenic factors.     

硫酸多糖电泳方法的研究

目的研究几种海洋硫酸多糖的电泳行为。方法采用醋酸纤维膜电泳法和聚丙烯酰胺凝胶电泳(PAGE)法 ,染色剂选用爱茜蓝。结果几种海洋硫酸多糖的醋酸纤维膜电泳均呈单一斑点 ,与标准肝素相比 ,几种海洋硫酸多糖的迁移值不同 ;在建立的PAGE电泳条件下肝素获得了很好的分离 ,几种海洋硫酸多糖的PAGE电泳区带均呈连续分布 ,并利用电泳图象扫描软件进行了相对分子量分布分析。结论醋酸纤维膜电泳法可用于海洋硫酸多糖的定性鉴别 ,PAGE法可用于海洋硫酸多糖相对分子量分布的测定     

子囊霉素的分离与鉴别

应用大孔吸附树脂柱层析的方法从他克莫司产生菌的发酵提取液中分离得到化合物FW99802,经重结晶后得到纯度大于98%的结晶性粉末。对其理化性质的研究及光谱分析结果表明它与子囊霉素同质。     

猪肝组织中硫酸乙酰肝素的分离纯化及二糖组成分析

目的从少量猪肝组织中分离纯化硫酸乙酰肝素(HS)并进行二糖组成分析。方法分别采用胰蛋白酶及中性蛋白酶对猪肝组织进行酶解,再分别采用AG 50W-X2强阳离子交换色谱、Sephadex G-10凝胶过滤色谱、Q-Sepharose Fast Flow强阴离子交换色谱法纯化。将纯化的HS分别采用肝素酶Ⅰ和Ⅲ降解,所得到的二糖经2-氨基苯甲酰胺标记后,采用PA03色谱柱分析其二糖组成。结果经色谱和电泳分析,所得到的HS不含蛋白质及其它糖胺聚糖,其二糖组成以非硫酸化和低硫酸化二糖为主。结论该方法可以有效分离、纯化猪肝组织中的HS,并适用于少量组织中HS二糖组成分析。     

凉粉草中抗缺氧化学成分

目的研究凉粉草(Mesona chinensisBenth)中的抗缺氧化学成分。方法用硅胶柱色谱、Sephadex LH 20柱色谱、ODS柱色谱及制备HPLC色谱分离纯化化合物;通过化合物的理化常数、1H NMR和13C NMR波谱数据,鉴定化合物结构;通过大鼠肾上腺嗜铬细胞瘤克隆化细胞株(PC12)实验,对各单体化合物进行活性评价。结果从凉粉草体积分数为65%的乙醇提取物的氯仿层和乙酸乙酯层分离得到8个化合物:咖啡酸(1)、3(4乙氧基3羟基苯基)烯丙酸(2)、咖啡酸乙酯(3)、山奈酚(4)、高山黄芩素(5)、2十六烷基-十八烷酸(6)、熊果酸(7)和豆甾醇(8);应用PC12细胞缺氧模型在DMSO溶液中测得化合物1、4、5、6、7、8在不同质量浓度下具有不同的抗缺氧活性。结论首次从凉粉草属中分离得到化合物2、3、5、6;化合物1、4、5、6、7、8有一定的抗缺氧活性,化合物4、5、7、8有较好的抗缺氧活性。     

卵磷脂质量对甲氨喋呤(MTX)脂质体包裹率的影响

本文研究了卵磷脂的理化性质与MTX脂质体包裹率的关系。分别测定了两种卵磷脂(大豆卵磷脂、蛋黄卵磷脂)的氧化指数、碘价、紫外吸收光谱及薄层层析,并用具有不同氧化指数的卵磷脂用二次乳化蒸发法制备MTX脂质体,测定它们包裹MTX的百分率。初步表明:卵磷脂对药物包裹率随其氧化指数的升高而降低,同时薄层层析的杂质点亦增多。实验结果提示,用氧化指数在0.2以下,碘价在60左右,以及薄层层析一个点的卵磷脂制备MTX脂质体可以得到较高的包裹率。     

Study of physico-chemical properties of novel highly sulfated,aromatic, mimetics of heparin and heparan sulfate

Heparin (H) and heparan sulfate (HS) play major roles in a number of biological processes. Yet, H/HS-based pharmaceutical agents are also associated with multiple adverse effects. This has led to the concept of designing noncarbohydrate, aromatic mimetics that modulate H/HS function. In this work, we study a library of synthetic, aromatic H/HS mimetics for their capillary electrophoretic profiles, the acid and base stability, and aqueous–organic partitioning property. The nonsugar H/HS mimetics exhibit electrophoretic properties similar to sulfated oligosaccharides suggesting that the mimetics can be rapidly and quantitatively analyzed. Stability studies show that the mimetics are essentially stable under neutral and basic conditions in a manner similar to the heparins, but are considerably unstable under acidic conditions in contrast to heparins. The measurement of partition coefficients show major differences within the sulfated mimetics as well as between the measured and calculated log P values. Understanding these physico-chemical properties is expected to have significant implications in the pharmaceutical development of this growing class of molecules. © 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 99: 1207–1216, 2010