反相高效液相色谱法测定紫草素衍生物

紫草(gromwell)为紫草科多年生草本植物,是常用中药,因其根呈紫色而得名。药理学研究发现紫草提取物具有抑菌、抗炎、抗肿瘤以及抗生育等作用。对紫草素衍生物的测定方法主要有比色法、重量法、紫外-可见分光光度法、薄层扫描法、薄层-分光光度法、单扫描极谱法以及正相高效液相色谱法。所有这些方法主要用于测定紫草提取物中的萘醌类化合物总量或主要单体含量。本文建立了一种分离快、柱效高、准确度高、重现性好的反相高效液相色谱(RP—HPLC)同时测定紫草素衍生物的新方法。     

毛细管电泳法测定紫草中的紫草素

建立了毛细管电泳高频电导法测定紫草药材中左旋紫草素含量的分析方法。以融硅毛细管(150μm×70 cm)为分离柱,研究了缓冲液的种类、浓度、添加剂种类与添加量、分离电压和进样量等因素对分离和检测的影响,优化选择1.0 mmol/L H3BO3 3.0 mmol/L三乙胺缓冲液为电泳介质,分离电压18.0 kV,可实现分离检测。在优化条件下左旋紫草素线性范围为10.0~250 mg/L;线性相关系数为0.9962;检出限为5.0 mg/L(S/N=3)。2批样品不同浓度添加水平的日内和日间RSD均小于4%,两批药材的加标回收率分别为93.9%~97.4%和93.1%~101%。该方法简便、快速、灵敏度高,可以用于紫草药材的质量控制。     

薄层色谱—化学发光联用技术测定紫草素含量

１引言紫草为紫草科紫草属植物，新疆紫草和紫草属植物紫草的根，为常用中药，紫草素为其有效成份。药理实验证实紫草素具有抗肿瘤、抗炎、抗菌，促伤口愈合等作用。紫草素测定方法一般采用分光光度法、重量法、薄层扫描法。本文根据化学发光原理，以鲁米诺－过氧化氢－钻离子（Ｃｏ２＋）为发光体系，利用过氧化氢定量氧化紫草素苯环上的羟基，从而消耗发光体系中的Ｈ２Ｏ２来降低发光强度，实现发光值动态猝灭。在一定范围内猝灭强度与紫草素含量呈线性关系。本法具有灵敏度高、仪器简单、线性范围宽等优点，是测定萘酯类药物的又一新方法…     

紫草油有效成分的高效液相色谱测定法及其在超临界流体萃取制备紫草油中的应用

紫草提取制备成的紫草油能够预防及治疗婴儿尿布疹、皮肤溃烂、湿疹等多种皮肤疾患,临床应用非常广泛,超临界流体萃取是紫草有效成分提取的优选方法.该文建立了紫草油有效成分的高效液相色谱(HPLC)测定方法,并以紫草油所含的有效成分含量为评价指标,采用三因素三水平正交试验法对紫草超临界流体萃取制备过程中的几个重要因素(萃取压力...     

大孔吸附树脂对紫草宁的吸附性的研究及从细胞培养液中提取紫草宁的 …

以配制的紫草宁水溶液为对象，研究了大孔吸附树脂提取水溶液中紫草宁的方法，包括树脂的筛选，紫草宁的浓度，溶液ｐＨ及盐离子浓度对吸附的影响；解吸剂的选择，确定了适宜的吸附和解吸条件。最后，采用真实的细胞培养体系进行间歇的边培养边吸附分离研究，紫草宁产率提高了７６．４％，为今后实现细胞培养与分离相耦合的连续化操作提供充分的准备工作。     

紫草素的电化学伏安行为研究

紫草素在0.2 mol/L HAc-NaAc缓冲溶液(pH4.3)中,产生灵敏的线性扫描极谱峰,峰电位EP=-0.186 V(vs.Ag/AgCl)。峰电流与紫草素浓度在5×10-7~1×10-5mol/L范围内呈良好的线性关系。可用于紫草中紫草素含量的测定。用线性扫描和循环伏安法测试表明,该体系属具有反应物吸附性的可逆过程体系。     

紫草素的电化学研究

建立了测定紫草素的单扫描极谱分析方法;在0.03mol/LNa2CO3+0.2mol/LNaOH(pH=12.63)底液中,紫草素于-0.60V(SCE)处有一灵敏的二次导数还原峰,电流峰高与浓度在7.0×10-8～5.6×10-7mol/L和6.0×10-7～4.2×10-5mol/L范围内呈线性关系,检出限为3.5×10-8mol/L;用于中药紫草中紫草素的测定,结果良好;同时还研究了紫草素的电极反应机理.     

紫草素及去氧紫草素的ESI离子阱质谱裂解分析

应用电喷雾离子阱多级质谱技术对紫草素及其活性衍生物去氧紫草素的结构和质谱裂解规律进行比较研究,并在负离子模式下解析了紫草素的主要特征碎片离子及其裂解规律。紫草素负离子模式下的主要碎片为m/z 287,269,259,218,190及173;其中,碎片m/z 218是其特征峰,碎片m/z 269和m/z 259均可进一步裂解为m/z 241,碎片m/z 190为m/z 259和m/z 218的共同产物离子;此外,碎片m/z 190和m/z 173均可进一步裂解为m/z 162。去氧紫草素负离子模式下主要裂解为碎片m/z 271、253、228和203;其中,碎片m/z253是其特征峰。紫草素和去氧紫草素均能发生过渡态氢重排β-裂解和连续的CO中性丢失。     

大学化学综合实验教学实践——紫草药材提取实验

常见中药材紫草的主要活性成分为羟基萘醌类化合物，其具有多种药理作用。将紫草中羟基萘醌类化合物的提取引入有机化学基础实验课程，对于可变参数——提取时间和加醇倍量进行研究，学生自行设计并实施实验。该实验综合使用有机化学及分析化学实验基础操作的同时，提升学生学习兴趣，提高学生实验设计、数据处理及分析的能力。     

紫草素在玻碳电极上的电化学行为及应用

采用电化学测试方法测定了紫草素在玻碳电极上的电化学行为。结果表明,紫草素在玻碳电极产生一对氧化还原波,在pH为3.98及HAc-NaAc(20%乙醇)缓冲溶液中。证实该反应是以吸附控制为主的电极过程。同时,以氧化峰为对象的循环伏安法研究了峰电流与药物浓度的关系。峰值电流与紫草素浓度在2.08×10~(-8)～1.82×10~(-6)mol·L~(-1)范围内具有良好的线性关系。该方法可用于中草药紫草中紫草素及其衍生物总含量的测定,样品无需预处理即可直接测定。通过方波伏安法测出新疆特有紫草素的平均含量为9.063m·mg~(-1),相对标准偏差3.08%,证明采用此方法测定紫草素含量是可行的。     

天山花楸果实中微量元素及氨基酸的检测分析

为探讨天山花楸果实中微量元素与药理学的相互关系,采用原子吸收分光光度仪和氨基酸自动分析仪分别测定了花楸果实中8种微量元素及17种氨基酸。结果表明,天山花楸果实中含有比较丰富的人体必需氨基酸及微量元素,它与天山花楸的临床保健作用有一定的相关性,具有很高的开发价值。     

叶菜型甘薯茎尖中微量元素和氨基酸的含量分析

采用等离子体发射光谱法测定了叶菜型甘薯茎尖中K、Ca、Mg、Fe、Zn、Cu、Se等7种微量元素的含量,氨基酸分析仪测定了18种氨基酸的含量。结果表明,与其它12种叶菜类蔬菜相比较,叶菜型甘薯茎尖中K、Ca、Se含量较高;含有18种氨基酸,人体必需的氨基酸种类齐全,含量较高的氨基酸是天冬氨酸、谷氨酸、亮氨酸;高Se是叶菜型甘薯茎尖的显著特点。为探讨叶菜型甘薯茎尖中微量元素和氨基酸与保健功能之间的关系提供了有用的数据。     

三裂叶蟛蜞菊中氨基酸成分和矿物元素的分析

为更好地开发利用蟛蜞菊这一中草药资源,采用氨基酸分析仪和原子吸收光谱仪测定了游离氨基酸的含量和矿物元素的含量。结果表明,蟛蜞菊中富含多种成分的氨基酸,其中谷氨酸、天门冬氨酸、亮氨酸含量较高,且含有7种人体必需的氨基酸;还含有丰富的钙、镁、铁、锰、锌、铜和硒等矿物元素;蟛蜞菊中高含量的氨基酸成分及矿物元素与其多种生理、药理活性作用密切相关。     

Origins of opposite syn-anti diastereoselectivities in primary and secondary amino acid-catalyzed intermolecular aldol reactions involving unmodified alpha-hydroxyketones

The effects of different amino acid catalysts on the stereoselectivity of the direct intermolecular aldol reactions between alpha-hydroxyketones and isobutyraldehyde or 4-nitrobenzaldehyde have been studied with the aid of density functional theory methods. The transition states of the crucial C-C bond-forming step with the enamine intermediate addition to the aldehyde for the proline and threonine-catalyzed asymmetric aldol reactions are reported. B3LYP/6-31+G** calculations provide a good explanation for the opposite syn vs anti diastereoselectivity of these two kinds of amino acid catalysts (anti-selectivity for the secondary cyclic amino acids proline, syn-selectivity for the acyclic primary amino acids like threonine). Calculated and observed diastereomeric ratio and enantiomeric excess values are in good agreement.     

Common prevalence of alanine and glycine in mobile reactive centre loops of serpins and viral fusion peptides: Do prions possess a fusion peptide?

Summary Serpin reactive centre loops and fusion peptides released by proteolytic cleavage are particularly mobile. Their amino acid compositions reveal a common and unusual abundance of alanine, accompanied by high levels of glycine. These two small residues, which are not simultaneously abundant in stable helices (standard or transmembrane), probably play an important role in mobility. Threonine and valine (also relatively small amino acids) are also abundant in these two kinds of peptides. Moreover, the known 3D structures of an uncleaved serpin reactive centre and a fusion peptide are strikingly similar. Such sequences possess many small residues and are found in several signal peptides and in PrP, a protein associated with spongi-form encephalopathies and resembling virus envelope proteins. These properties may be related to the infection mechanisms of these diseases.     

Low-Temperature Heat Capacity and Thermal Decomposition of Crystalline[Ho（Thr）（H2O）5]Cl3

Rare earth elements have been widely used in many areas. Rare earth complex bearing an amino acid was synthesized to study the influence and the long-term effect of rare earth elements on environment and human beings,because amino acid is the basic unit of the living things. Previous work on these kinds of comidex is focused on synthesis and characterization of them. But thermodynamic data have seldom been reported. Here we present the thermod~nRmle study of [ Ho (Thr)(H20 )5]Cl3. The heat capecity of Holmium complex with threonine,[Ho(Thr)(H2O)5]Cl3,was measured with an automatic adiabatic calorimeter in the temperature range from 79K to 330K and no thermal anomaly was found in this range,Thermodynamic functions relative to standard state 298.15K were derived from the heat capactiy data.Thermal decomposition behavior of the complex in nitrogen atmosphere in the range from 300K to 900K was studied by thermogravimetric(TG) technique and a possible decompostion mechanism was proposed according to the TG-DTG results.     

Dual substrate recognition of aminotransferases

Pyridoxal 5'-phosphate-dependent aminotransferases reversibly catalyzes the transamination reaction in which the alpha-amino group of amino acid 1 is transferred to the 2-oxo acid of amino acid 2 (usually 2-oxoglutarate) to produce the 2-oxo acid of amino acid 1 and amino acid 2 (glutamate). An aminotransferase must thus be able to recognize and bind two kinds of amino acids (amino acids 1 and 2), the side chains of which are different in shape and properties, from among many other small molecules. The dual substrate recognition mechanism has been discovered based on three-dimensional structures of aromatic amino acids, histidinol phosphate, glutamine:phenylpyruvate, acetylornithine, and branched-chain amino acid aminotransferases. There are two representative strategies for dual substrate recognition. An aromatic amino acid aminotransferase prepares charged and neutral pockets for acidic and aromatic side chains, respectively, at the same place by a large-scale rearrangement of the hydrogen-bond network caused by the induced fit. In a branched-chain aminotransferase, the same hydrophobic cavity implanted with hydrophilic sites accommodates both hydrophobic and acidic side chains without side-chain rearrangements of the active-site residues, which is reminiscent of the lock and key mechanism. Dual substrate recognition in other aminotransferases is attained by combining the two representative methods.     

Biosynthetic phage display: a novel protein engineering tool combining chemical and genetic diversity

BACKGROUND: Molecular diversity in nature is developed through a combination of genetic and chemical elements. We have developed a method that permits selective manipulation of both these elements in one protein engineering tool. It combines the ability to introduce non-natural amino acids into a protein using native chemical ligation with exhaustive targeted mutagenesis of the protein via phage-display mutagenesis. RESULTS: A fully functional biosynthetic version of the protease inhibitor eglin c was constructed. The amino-terminal fragment (residues 8-40) was chemically synthesized with a non-natural amino acid at position 25. The remaining carboxy-terminal fragment was expressed as a 30-residue peptide extension of gIIIp or gVIIIp on filamentous phage in a phage-display mutagenesis format. Native chemical ligation was used to couple the two fragments and produced a protein that refolded to its active form. To facilitate the packing of the introduced non-natural amino acid, residues 52 and 54 in the carboxy-terminal fragment were fully randomized by phage-display mutagenesis. Although the majority of the observed solutions for residues 52 and 54 were hydrophobic - complementing the stereochemistry of the introduced non-natural amino acid - a significant number of residues (unexpected because of stereochemical and charge criteria) were observed in these positions. CONCLUSIONS: Peptide synthesis and phage-display mutagenesis can be combined to produce a very powerful protein engineering tool. The physical properties of the environment surrounding the introduced non-natural residue can be selected for by evaluating all possible combinations of amino acid types at a targeted set of sites using phage-display mutagenesis.     

氨基酸卟啉锌配合物对氨基酸酯的分子识别研究(英文)

用紫外可见吸收光谱滴定方法研究了一种新型的苏氨酸卟啉锌配合物(主体)对氨基酸酯(客体)的分子识别.这种锌卟啉可以与氨基酸酯形成 1:1和 1:2的两种加合物.氨基酸酯的氨基首先与氨基酸残基的羧基作用形成 1:1的加合物,然后与锌卟啉的中央锌原子配位形成 1:2的加合物,客体分子与主体分子上的氨基酸残基之间的排斥作用以及主客体之间的色散力作用是主体分子能识别客体分子的另外两种作用。     

伊敏褐煤中微量元素的地球化学特征及其无机-有机亲和性分析

采用脱矿物质、提取腐殖酸等方法结合ICP-MS对伊敏15_上、16_下煤中的32种微量元素的含量及其赋存特征进行了分析。与地壳克拉克值及中国侏罗-白垩纪煤和世界褐煤中微量元素含量相比,伊敏褐煤中微量元素没有明显富集。脱矿物质处理后,Ni明显富集,As略比原煤高,其他微量元素都低于原煤。原煤提取腐殖酸和脱矿物煤提取腐殖酸中Ni、Mo、Cd、Sn、W明显富集,这表明Ni、Mo、Cd、Sn、W和腐殖酸形成了稳定的有机态化合物。原煤提取腐殖酸残煤和脱矿物煤提取腐殖酸残煤中V、Ni、As富集,表明V、Ni、As与煤的大分子结构形成了稳定的有机态化合物。根据微量元素和灰分的相关性系数,把微量元素分为以下几类:无机富集元素Cr、U;亲无机元素Cu、Cd、In、Sn、Ga、Y、Zr、Hf、Bi、Th;偏无机元素Be、Sc、Rb、Sr、Nb、Cs、Ta、Pb;偏有机元素V、As、Tl、Ba、Se;亲有机元素Li、Co、W;有机富集元素Ni、Zn、Mo、Sb。