The Isolation, Purification, Structure Identification and Bio-activity Assay of the Effective Components in Licorice and the Study of the Fingerprinting based on Activity

中药有效成分是其发挥药效作用的物质基础，对评价中药材的质量、制定中药质量控制标准控制、优化制剂工艺、揭示中药作用机制、实现中药现代化具有重要意义。甘草酸和甘草黄酮是中药甘草中主要的有效成分，目前的分离纯化方法存在着繁琐、工作量大的缺点，用做质量控制指标成分的标准品也由于制备困难无法满足市场需求，在活性研究方面也存在某些空白，本论文在甘草有效成分的分离纯化,结构鉴定,生物活性测试及指纹图谱技术方面进行了大量有益的研究探索。建立了一种采用大孔树脂吸附解吸甘草提取液制备具有抗溃疡活性而无假醛固酮样副作用的无酸甘草黄酮提取物DGL的简便有效的方法。采用优选出的极性大孔树脂XDA-1，在最佳分离条件下，...Active constituents in Traditional Chinese Medicine (TCM) are the basis of their pharmacology. They are very important in the evaluation of the quality of the herbs, the establishment of the standard for quality control, the optimization of the preparative technology, the research of the action mechanism and the modernization of TCM. Glycyrrhizic acid (GA) and licorice flavonoids (LF) are the mai...学位：理学博士院系专业：化学化工学院化学系_分析化学学号：B20022500

水体沉积物中有机碳和有机分子碳稳定同位素研究进展

有机碳是指存在于沉积物总有机质(TOC)中的碳。由于同位素之间的物理、化学性质的不同,生物在生存活动过程利用环境中碳的同时改变了其同位素的比值,即产生同位素效应(isotopic effect)。通过检测来自于生物有机质中的碳同位素比值,就有可能对沉积物所在地的环境变化信息进行重建,有机碳碳同位素的研究成果已在全球碳循环、古气候变化、生物演化、地层对比研究等领域得到了应用。水体沉积物能够较好地保存环境中的有机质信息,以及良好的时间次序[1]。水体沉积物中有机碳和有机分子碳的碳同位素研究为研究近百万年来全球及局部地区古气候变化,追踪沉积物中有机质的生物先质来源,了解过去环境中生态系统状况等环境提供了良好方法[2~6]。通常研究的水体沉积物主要分为海洋沉积物(marine sediments)和湖泊沉积物(lacustrine sedi-ments)。对于自全新世以来的气候环境变化,最初大多是通过海洋沉积物中有机物的同位素分析得到的,但是海洋沉积物不能反映内陆的气候、环境变化。而通过研究湖泊沉积物则可以获得相关地区过去诸如气温、湿度、植被覆盖等环境信息[7]。并且当沉积物中存在有大量有机物质时,其同位素分馏特征受..

Determination of Carthamin in Safflower(Carthamus tinctorius L)by RP-HPLC

经从红花 (CarthamustinctoriusL)中分离纯化、HPLC检测、质谱鉴定得红花红色素纯品 ,并用其作为标准品 ,采用反相高效液相色谱法 ,测定了不同产地红花和商业红花产品中红花红色素的含量。该方法准确性高、重现性好 ,平均回收率为98.1%。The carthamin,extracted from the safflower(Carthamus tinctorius L),was purified and identified byMS.Itwas then used as a standard for the reversed phase high performance liquid chromatographic determiˉnation of carthamin in safflower collected fromdifferentareas and in commercial safflower products.HPLC sepˉaration was carried out on an Alltima C 18 column(5μm,150×4.6mm)with a mixture of acetonitrile,methanol and water(30∶10∶60by volume)at pH=3as mobile phase.The linear range was0.05～0.2g/Lwith a correlation coefficient of0.999.The average recovery and RSD were98.1%and1.9%,respecˉtively

Study on the free radical scavenging activity of polyphenolic compounds by on-line high-flux screening method

目的:构建植物化学物在线清除自由基活性高通量筛选技术,创建植物化学物抗氧化活性研究新方法。方法:以附带2个检测器的高效液相色谱(HPlC)装置和流动注射技术为手段,引入二苯基三硝基苯肼(dPPH)有机自由基分别与经HPlC分离的植物化学物发生在线清除反应,优化流速、反应管尺寸等条件,比较反应前后dPPH吸收信号的变化。结果:确定和验证了两类植物多酚化学物茶多酚和丹酚酸在线清除自由基活性HPlC-dPPH表征方法。反应环聚醚醚酮(PEEk)盘管规格为长10M、内径0.254MM,dPPH·浓度为1.0x10-5MOl/l,流速为0.7~0.8Ml/MIn。结论:HPlC-dPPH流动注射在线表征技术能够实现对多组分植物化学物抗氧化能力的高通量筛选。Objective:To construct a high-flux screening procedure by on-line free radical scavenging activity method of phyto- chemicals, and create a new evaluation system of antioxidant activity of natural plant materials.Methods:A HPLC system with two detectors and flow injection techniques was employed.An organic free radicals DPPH was induced on-line HPLC-DPPH and reaction with the single phytochemical components isolated by HPLC.To optimize reactive conditions including flow rate and the size of reaction coil, and detect the DPPH signals before and after reaction.Results:A new on-line HPLC-DPPH method was established for a high-flux screening of several free radical scavenging components exampled as Tea polyphenols and Salvianolic acids.The reaction coil of PEEK was 10 m length and inner diameter 0.254 mm, the concentration and flow velocity of DPPH was 1.0 × 10-5mol/L and 0.7～0.8 ml/ min, respectively.Conclusion:The PHLC-DPPH flow injection hyphenated technique could realize high-flux screening analysis for antioxidant of phytochemicals.江苏省高校自然科学研究计划资助项目(05KJB330083

西洋参多糖及总皂苷中无机元素的ICP/MS法测定

采用电感耦合等离子发射光谱—质谱联用法(ICP/MS),对西洋参样品、西洋参多糖、西洋参总皂苷中无机元素Mg、Al、P、Ca、V、Cr、Mn、Fe、Co、Ni、Cu、Zn、As、Se、Sr、Mo、Cd、Pb的含量进行测定分析。结果表明,西洋参多糖中各种元素的含量均明显高于西洋参样品,而不同方法提取的多糖中元素含量有一定差异;西洋参总皂苷中Cr、Ni、Cu、Zn、Pb元素的含量明显高于西洋参样品,而其它元素含量较低。这就为西洋参多糖、西洋参皂苷的综合开发利用及其药效提供理论依据

Spectroscopic Characterization on Interaction of Gold (Au ~(3+)) Biosorption by Bacillus megaterium D01

对休眠的巨大芽孢杆菌 (Bacillusmegatherium)D0 1菌体吸附Au3 + 的作用过程进行了谱学表征 .运用AAS考察了pH、时间和温度对D0 1菌体吸附Au3 + 过程的化学动力学和热力学相关参数的影响 .D0 1菌粉中硫元素含量的EDX分析说明该菌体中对Au3 + 具有还原作用的L 半胱氨酸和蛋氨酸的含量极少 ;D0 1菌体水解后葡萄糖含量的UV vis测定说明该菌体水解产物中含有一定量的还原糖 .空白的和吸附Au3 + 的D0 1菌体的FTIR检测表明该菌体细胞壁肽聚糖层糖类化合物的羟基和肽链侧链氨基酸残基离子化羧基为吸附Au3 + 的活性基团 ;肽聚糖层部分多糖的水解产物低聚糖、二糖及单糖等还原糖的半缩醛羟基游离态醛基为电子供体 ,将Au3 + 原位还原成Au0 .葡萄糖和Au3 + 相互作用的XRD和FTIR表征证明Au3 + 是在还原糖的醛基上直接被还原成Au0Biosorptive interaction of gold with resting cell of Bacillus megatherium D01 biomass has been characterized by spectroscopic techniques. The effects of pH, time and temperature on the correlation parameters of chemical kinetics and thermodynamics of the binding reaction has been investigated through the determination of the gold ion binding to the biomass using atomic absorption spectrophotometry (AAS). The analysis for sulfur content in dry powder of the D01 biomass by energy dispersive X-ray (EDX) shows that cysteine and methionine being capable of reducing Au 3+ to Au0 is very small, whereas the glucose content in hydrolysates of the biomass analyzed by ultraviolet-visible spectrophotometry (UV-vis) indicates that the amount of the reducing sugars in the biomass is much larger than 3.33%. The Fourier transform infrared (FTIR) spectroscopy on blank and gold-loaded biomass demonstrates that the active groups such as the hydroxyl group of saccharides and the ionized carboxyl group of amino acid residues on the cell wall seem to be the sites for Au 3+ binding, and the free aldehyde group of the hemiacetalic hydroxyl group from reducing sugars, i.e. the hydrolysates of the polysaccharides, serving as the electron donor, in situ reduces the Au 3+ to Au0. X-ray powder diffractometry (XRD) and FTIR spectroscopic characterizations of the interaction of Au 3+ with glucose confirm that the reduction of Au 3+ to Au0 can directly occur in the aldehyde group of the reducing sugars

中国超大城市空气污染和人体健康 (APHH-Beijing) 中英联合研究计划 最终报告

Chinese versio

Application of Macroporous Resins in Separating Licorice Flavonoids and Glycyrrhizic Acid

本文在比较了四种大孔树脂对甘草酸和甘草总黄酮静态分离性能的基础上,筛选出大孔树脂XDA-1,对它的动态吸附分离条件如上样液浓度、洗脱溶剂及洗脱流速进行了进一步的研究。结果表明,树脂XDA-1对甘草总黄酮和甘草酸的吸附量大,易于洗脱,分离效果好,最终的黄酮收集液中不含甘草酸。本实验的优化条件为:上样液pH=5,甘草酸浓度为1.5 mg／mL,总黄酮浓度为0.75 mg／mL,上样流速为3 BV／h;洗脱液采用1.5 BV／h 45％的乙醇为最佳。再用含1％NaOH的80％乙醇以1.5 BV／h流速洗脱,得到甘草酸。总黄酮的收率为69.7％,甘草酸收率为52％,纯度为65.5％。Glycyrrhizic acid (GA) and licorice flavonoids (LF) are main bioactive components in licorice. However, long-term excessive intake of glycyrrhizic acid could cause side effect such as sodium retention and hypertension. In order to separate these components individually, and to produce licorice extracts aiming at different applications, a preparative scale separation technique is needed. Against this background, the sorption and desorption properties of licorice flavonoids (LF) and glycyrrhizic acid (GA) on macroporous resins including XDA-1, LSA-10, D101 and LSA-20 have been studied to gain better understandings on the separation characteristics of these resins. The adsorption capability of the resins is found to be strongly dependent on the pH value of the feed solution. Adsorption isotherms were obtained by batch experiments and the data are fitted to the Freundlich equation to evaluate the adsorption properties of these resins. XDA-1 is found to provide the best fit to the Freundlich isotherm. It has much higher adsorption capacity for GA and LF than other resins. XDA-1 shows a higher adsorption affinity towards LF than that of GA based on the calculated adsorption isotherms. Dynamic adsorption and desorption on the selected XDA-1 resin were carried out in a laboratory packed column (0. 5 cm×30 cm) to obtain optimal parameters for LC separation. A simple preparative separation procedure for LF and GA from licorice root was established. An LF enriched extract containing up to 60% LF but without GA contamination; and a GA extract with 65. 6% chromatographic purity have been prepared by the method

Determination of Yellow Pigments in Safflower (Carthamus tinctorius L) by RP-HPLC

本文分离纯化了红花(CarthamustinctoriusL)中SaffloweryellowA,Safflo-minA,SaffloweryellowB,SafflominC四种红花黄色素,并用所纯化的红花黄色素做标准品,用反相高效液相色谱法,梯度洗脱,同时测定了不同产地红花中四种红花黄色素的含量。该方的法准确度高,重现性好。A HPLC method for the determination of yellow pigments in safflowers collected in different regions was investigated.Safflower yellow A,safflomin A,safflower yellow B and safflomin C were isolated as standards.Separation of yellow pigments was carried out on a reversed-phase column by a gradient elution.The method developed was rapid and accurate

Inhibitory Effects of Several Licorice Flavonoids on the Monophenolase Activity of Tyrosinase

研究了甘草中四种黄酮类化合物甘草甙、异甘草素葡萄糖芹菜甙、异甘草甙和甘草查耳酮甲对酪氨酸酶单酚酶活性的抑制。结果表明异甘草素葡萄糖芹菜甙、异甘草甙和甘草查耳酮甲的IC50分别为0.072mM,0.038mM,0.0258mM,它们都属于竞争性抑制剂,甘草甙没有抑制活性。研究表明,异甘草素葡萄糖芹菜甙、异甘草甙和甘草查耳酮甲可以作为潜在的美白化妆品添加剂。The inhibitory effects of several licorice flavonoids including liquiritin, licuraside, isoliquiritin and licochalcone A on the monophenolase activity of tyrosinase were studied. The results indicated that the IC_50 values of licuraside, isoliquiritin and licochalcone A were 0.072 mM,0.038 mM,0.0258 mM, respectively, and they are all competitive inhibitors. These flavonoids have the potential to be further developed into effective skin whitening agents. Different from the above flavonoids, no inhibitory activity was observed for liquiritin.国家自然科学基金重点项目(20235020