低极性化合物的电喷雾质谱检测方法及几种中藏药材的活性成分研究

本学位论文首先报道了为解决低极性化合物的电喷雾质谱（ESI-MS）分析难题而建立的一种衍生化分析方法；然后从色谱-质谱联用分析、分离纯化和结构鉴定等方面分别报道了几种中藏药材的活性成分研究。论文由下述六章组成： 第一章报道了盐酸羟胺衍生化方法在电喷雾质谱 (ESI-MS) 分析中的应用。该方法利用盐酸羟胺和羰基成肟的快速反应，建立了针对三萜酮等含酮或醛羰基低极性化合物的ESI-MS 信号增强技术。此方法不仅可应用于增强羰基化合物的ESI-MS 质谱信号，还可检测化合物中羰基的个数以及辨别涉及羰基官能团的同分异构体。此外，通过简单的氧化反应，还可将该方法拓展到三萜醇、甾醇等含羟基的低极性化合物，增强它们的ESI-MS 信号。对比已报道的相关ESI-MS 增强质谱信号的衍生化方法，此方法有经济、实用、快速和简便的显著特点。 第二章是关于野生羌活及其栽培品种化学成分的色谱-质谱联用分析。对不同产地野生羌活生长过程中活性成分的动态变化、野生羌活不同形态部位和人工栽培羌活中的活性成分含量进行了HPLC 定量分析。结果表明主要活性成分羌活醇和异欧前胡素都随生长期存在规律性变化，羌活不同形态部位中的活性成分含量也有明显不同。这些实验结果有些较好地印证了传统中医的用药理论，有些也对羌活的传统使用方法提出了新的建议。 第三章介绍了几种传统中藏药材的色谱-质谱联用及串联质谱分析。通过GC-MS 方法，从藏药材长花党参挥发油中共分离鉴定出45 个化合物；利用HPLC方法测定了该藏药材中的主要化学成分——木犀草素的含量（0.7%）；利用串联质谱技术，对西番莲和射干中的主要成分进行了快速鉴定，从西番莲中鉴定了4个黄酮碳苷；从不同产地的射干和川射干中鉴定了8 个主要异黄酮成分，其中包括一个未见报道的化合物。 第四章的内容为藏药材石莲叶点地梅的活性成分研究。从植物石莲叶点地梅(Androsace integra (Maxim.) Hand.-Mazz.) 乙醇提取物的正丁醇萃取部分共分离和鉴定了6 个化合物，利用MS 和NMR 等现代波谱学技术阐明了它们的结构：其中包括4 个三萜类化合物：分别是androsacin (1)、 ardisiacrispin A (2) 、saxifragifolin A (3) 和20(29)-lupen-3-one (4)；一个神经酰胺：4-羟基-Δ8,9(Z)-鞘氨醇-2'-羟基正二十四碳酸酰胺(5)；一个甾体类化合物：胡萝卜苷(6)。化合物1为新的13,28-epoxy-oleanane 型三萜皂苷，在其结构表征的过程中，采用LC-MS 进行糖分析，获得了值得推广的好结果。通过活性筛选发现化合物1~3 对HepG2肝癌细胞表现出不同程度的抑制活性，其中化合物2 活性最好，其IG50 为1.65μg/mL。 第五章是关于一些传统中藏药材的农药活性筛选。利用Syngenta 公司的活性筛选平台对68 种传统中藏药材醇提物进行了抗菌和除草的生物源农药活性筛选。结果表明所筛选的68 种植物提取物中，共有14 种样品表现出明显的除草/杀虫活性，其中水母雪莲花、松萝和茯神木等植物提取物还具有多种生物活性。活性成分还有待进一步追踪分离、纯化和结构鉴定。 第六章为文献综述，概述了羌活药材的研究进展。对羌活属及药用羌活植物从分类学、本草学、品质评价、人工栽培、化学成分及药理作用等方面进行了文献归纳和总结。 In this dissertation, an electrospray ionization mass spectrometry (ESI-MS) signal enhancement method, as well as the work of bioactive components study, HPLC-MS/MS application, bioassay screening, chromatograph separation and structure identification of the metabolites in several medicinal herbs have been reported. First chapter expounded a rapid, simple ESI-MS sensitivity enhancement method for detecting carbonyl groups in natural products has been developed by using hydroxylamine hydrochloride (NH2OH·HCl) as a derivatization reagent. We use the oxime formed during the derivatization reactions and its Beckmann rearrangement intermediates as a means of detecting the carbonyl groups originally present in these triterpenoids. In comparison with other derivatization methods in the literature, this method is simple, specific and can be used to detect carbonyl groups in triterpenoids which have low polarity and are poorly or non-ionizable. Moreover, it can also be used to detect hydroxyl groups by using the Dess-Martin periodinane (DMP) to convert primary and secondary hydroxyls into carbonyl groups. Chapter 2 reported an HPLC-MS method for analyzing the main bioactive compounds in both wild and cultured Notopterygium incisum. The results indicated that the main bioactive compounds varied through different seasons regularly, and in different commercial parts of this herb the content of these compounds also differed from each other. The quantitative analysis results showed that in the traditional commercial parts, the content of main chemical constitutes in Silkworm Notopterygium, Bamboo Notopterygium and Irregular-nodal Notopterygium are higher than that in Striped Notopterygium. This result is tally with the traditionally concept that the quality of Notopterygium, Bamboo Notopterygium and Irregular-nodal Notopterygium are better than that of Striped Notopterygium, which means that the quality of rhizomes is better than main roots. The chemical constituents of cultured N. incisum is reported for the first time in this dissertation and the analysis results showed some growth curves of chemical constituents in this plant, but still left some questions unanswered. Chapter 3 discussed the GC/LC-MS analysis of the traditional Chinese medicines Codonopsis thalictrifolis, Passiflora incarnate, Belamcanda chinensis and Passiflora incarnate. The main constituent, luteolin was isolated and identified from the traditional Tibet medicine of C. thalictrifolis. The quantitative analysis by HPLC has revealed that the content of luteolin in this herb is 0.7%. GC-MS was employed to analyzed chemical constituents of the essential oil from the flower of C. thalictrifolis. More than 60 peaks were detected and 45 of them were identified by comparing their spectra with that of the standards in the database and literatures. ESI-MS/MS was used to analyze the n-butanol extract of Passiflora incarnate. Based on the information of pseudo molecular ions and fragment ions of the glycosides, four major flavone-C-glycosides have been detected and identified as 7-methoxyluteolin-6-C-β-D-glucopyranoside, vitexin, swertisin and orientin. The isoflavone compounds in theextracts of three samples of B. chinensis collected in Gansu, Sichuan and Hunan, and the extract of Iris tectorum collected in Sichuan were analyzed by using TOF-HRMS and IT-MS. From the extracts of these herbs, a new isoflavone, identified as 5’,5,6,7-tetrahydroxy-3’4’-dimethoxyl isoflavon, and 7 known ones have been identified by analyzing the fragmentation patterns and their molecular formulas given by HRMS and the tandem mass spectrometry acquired by IT-MS. Chapter 4 elucidated the isolation and identification of a new triterpene saponin, androsacin (1), along with five known compounds (2-6) were isolated from the whole plants of Androsace integra (Maxim.) Hand.-Mazz., an herb used in traditional Chinese and Tibetan medicine. The chemical structure of the new compound was established as 3β-O-{β-D-glucopyranosyl-(1→4)-O-β-D-xylopyranosyl-(1→2)-O-β-D-glucopyranosyl-(1→4)-[O-β-D-glucopyranosyl-(1→2)]-α-L-arabinopyranosyl}-16α-hydroxy-13β,28-epoxy-olean-30-al by analyzing its MS, 1D- and 2D-NMR spectra. Compound 2 was cytotoxic toward HepG2 cancer cell with the GI50 value of 1.65 μg/mL. Chapter 5 described the biogenic pesticide activity screening of 68 traditional Chinese and Tibetan medicine extractions. The intention of this study is to explore bioactive natural compounds from these traditional medicinal herbs for biogenic insecticides use. Based on Syngenta’s bioassay, 14 extractions of these traditional medicines showed pesticide activities, and some of them had multi-activities on antibacterial and insecticidal. Chapter 6 is a review on the chemical and bioactivity research progress of Notopterygium incisum and N. forbesii.

2-C-支链-1,6-脱水吡喃葡萄糖的开环反应及吡喃烯糖的合成研究

在糖化学合成中，1,6-脱水吡喃糖不仅是合成具有生物活性低聚糖、糖共体、抗原、抗体以及天然产物等化合物重要原料，而且还是许多具有生物活性的天然产物的结构单元。同时，它还具有[3,2,1]的双环缩醛结构，使其在糖化学合成中具有高的立体选择性和区域选择性，同时减少了C-1 和C-6 位的保护和去保护的优点。此外，环内的缩醛开环后，又可以相应地在C-1 和C-6 位进行官能团转化以及糖苷化反应。 本文报道了一种新的1,6-脱水吡喃糖的合成方法，并设计合成了2-C-支链-1,6-脱水吡喃葡萄糖1-195、1-197、1-198 以及2-C-支链-6-硫代1,6-脱水吡喃葡萄糖1-225。到目前为止，1,6-脱水糖开环并进行糖苷化反应，存在选择性较差、产率低的缺点。我们发现，在乙腈做溶剂的条件下，NiCl5 能高立体选择性高产率地催化化合物1-195、1-197、1-198 开环并与ROH、RSH 发生糖苷化反应。在NiCl5-乙腈条件下，合成了一系列2-C-支链-α-糖苷和2-C-支链-β-硫代糖苷，并对2-C-支链1,6-脱水吡喃葡萄糖的生成机理以及开环机理进行了探讨。 烯糖在糖化学合成中是重要的起始原料，从Fischer 首次合成烯糖至今，一直不断地有新的合成方法出现。但目前文献报道的方法存在所用试剂有毒、价格贵和操作繁琐等缺点。我们对Fischer-Zach 方法进行了改进， 发现Zn-NaH2PO4-H2O 和Zn-PEG600-H2O 体系都能很好地合成烯糖。该方法具有条件温和、绿色环保、操作简单的优点。在Zn-NaH2PO4 溶液或Zn-PEG600 条件下，以溴代糖为原料，高产率地合成一系列的烯糖。 The 1,6-anhydrohexopyranoses are crucial subunits of myriad bioactive nature products, as well as important syntons of carbohydrate chemistry which have been extensively used to prepare the biologically potential oligosaccharides, glycoconjugates, antibiotics, and structurally varied nature products. Their particular [3.2.1] bicyclic skeleton makes them have high regio- and stereo-control in a variety of reactions, and such structure avoids protecting hydroxyl groups at C1 and C6.Additionally, the cleavage of the internal acetal under acidic conditions could be beneficial for further transformations of functional group and glycosylation of the corresponding pyranosyl sugar at the C6 or C1 site. Herein we developed a novel approach to prepare the 1,6-anhydrohexopyranose, and synthesized the 2-C-branched-1,6-anhydrohexopyranose 1-195, 1-197, 1-198 and 2-C-branched-6-thio-1,6-anhydrohexopyranose 1-225. Until now, glycosylation of 1,6-anhydrohexopyranoses has been limited because of the low yields and low stereoselectivity. In this paper, we found that NiCl5-MeCN system could selectively cleave the ring of 1,6-anhydrohexopyranoses with alcohols and thiols at room temperature in high yields. A series of 2-C-branched-α-glycosides and 2-C-branched-β-thioglycosides have been synthesized via NiCl5-catalyzed. Furthermore, we investigated the formation and ring-opening mechanism of 2-C-acetylmethyl-1,6-anhydrohexopyranose. Glycals are significant starting material in carbohydrate chemistry. After the Fischer-Zach method for forming glucal was reported for the first time, the numerous synthetic methods for glycals have been explored. However, there are several drawbacks in the existing methods, such as the usage of very expensive and toxic reagents, intricate operation, and the influence of acid-sensitive and base-sensitive functional group. We improved the Fischer-Zach method and developed a facile, mild and environmentally benign methodology towards the synthesis of the glycals in Zn-NaH2PO4-H2O or Zn-PEG600-H2O system. Our method involves the treatment of glycosyl bromides with Zn in NaH2PO4 aqueous solution or PEG600-H2O at room temperature, affording various glycals in excellent yields.

Identification of Phenylethanoid Glycosides in Plant Extract of Plantago asiatica L. by Liquid Chromatography-Electrospray Ionization Mass Spectrometry

The present work describes a liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) method for rapid identification of phenylethanoid glycosides in plant extract from Plantago asiatica L. By using a binary mobile phase system consisting of 0.2% acetic acid and acetonitrile under gradient conditions, a good separation was achieved on a reversed-phase C-18 column. The [M-H](-) ions, the molecular weights, and the fragment ions of phenylethanoid glycosides were obtained in the negative ion mode using LC-ESI-MS. The identification of the phenylethanoid glycosides (peaks 1-3) in the extract of P. asiatica L. was based on matching their retention time, the detection of molecular ions, and the fragment ions obtained by collision-induced dissociation (CID) experiments with those of the authentic standards and data reported in the literature.

Separation and determination of flavone and xanthone glycosides in Tibetan folk medicinal species Swertia mussotii and S-franchetiana by capillary electrophoresis

The contents of five pharmacologically active flavone and xanthone glycosides, namely, swertianolin, swertisin, isoorientin, mangiferin, and 7-O-[alpha-L-rhamnopyranosyl-(1 -> 2)-beta-D-xylopyranosyl]-1,8-dihydroxy-3-methoxyxanthone, extracted from Tibetan folk medicinal species Swertia mussotii and S. franchetiana were determined by capillary electrophoresis with diode-array detection. The separation of five components has been optimized with a capillary column with a total length of 48.5 cm and effective length of 40 cm (50 mu m i.d). The influence of the running buffer, the sodium dodecyl sulfonate (SDS) concentration, organic modifier, etc. on the resolution was evaluated. The background electrolyte contained 30 mM borate buffer, 28 mM SDS, 1.0% (v/v) acetonitrile, and was adjusted to pH 9.0 with 0.1 M NaOH. A good baseline resolution was obtained for the separation of five components within 5 min with the working voltage of 24 kV and a column temperature of 25 degrees C. The established method was rapid and reproducible for the separation and determination of five flavone and xanthone glycosides from the extracts of S. mussotii and franchetiana plant samples.

Nonequilibrium microstructures and their evolution in a Fe-Cr-W-Ni-C laser clad coating

The laser-solidified microstructural and compositional characterization and phase evolution during tempering at 963 K were investigated using an analytical transmission electron microscope with energy dispersive X-ray analysis. The cladded alloy, a powder mixture of Fe, Cr, W, Ni, and C with a weight ratio of 10:5:1:1:1, was processed with a 3 kW continuous wave CO2 laser. The processing parameters were 16 mm/s beam scanning speed, 3 mm beam diameter. 2 kW laser power, and 0.3 g/s feed rate. The coating was metallurgically bonded to the substrate, with a maximum thickness of 730 mu m, a microhardness of about 860 Hv and a volumetric dilution ratio of about 6%. Microanalyses revealed that the cladded coating possessed the hypoeutectic microstructure comprising the primary dendritic gamma-austenite and interdendritic eutectic consisted of gamma-austenite and M7C3 carbide. The gamma-austenite was a non-equilibrium phase with extended solid solution of alloying elements and a great deal of defect structures, i.e. a high density of dislocations, twins, and stacking faults existed in gamma phase. During high temperature aging, in situ carbide transformation occurred of M7C3 to M23C6 and M6C. The precipitation of M23C6, MC and M2C carbides from austenite was also observed.

铁基抗高温磨损激光熔覆涂层强韧设计和研究II:Fe-Cr-C-W-Ni激光熔覆层的微观组织及性能

本文以对热工模具进行失效分析的基础上，利用激光熔覆技术，在５ＣｒＭｎＭｏ基础上设计并制备强韧兼备的抗高温磨损涂层。

裂纹顶端位错的运动和小范围屈服的K_(Ⅱc)

本文从裂纹顶端位错运动的简化模型估计形成塑性区所消耗的功;从而计算临界裂纹扩展力。由于Ⅰ型加载和Ⅱ型加载的应力状态有所不同,裂纹顶端塑性功耗也会不一样。基于上述思想,考虑了小范围屈服的K_(Ⅰc)和K_(Ⅱc)的关系,得出K(Ⅱc)>K(Ⅰc)的结论,符合实验结果;并就K(Ⅱc)/K(Ⅰc)的比值与材料性能和温度的关系作了讨论。

铁基抗高温磨损激光熔覆涂层强韧设计和研究 Ⅱ:Fe-Cr-C-W-Ni激光熔覆层的微观组织及性能

利用ＯＭ、ＳＥＭ、ＴＥＭ研究了Ｆｅ－Ｃｒ－Ｃ－Ｗ－Ｎｉ激光熔覆涂层熔覆态及其高温时效态的微观组织结构。结果表明激光熔覆层组织细小，具有强韧两相组成（奥氏体和Ｍ_７Ｃ_３碳化物）的微观结构特征，高温时效处理组织中有Ｍ_（２３）Ｃ_６、Ｍ_６Ｃ、Ｍ_２Ｃ等新碳化物形成。显微硬度和冲击磨损实验证实了激光熔覆态和峰值时效态熔覆层均具有良好的力学性能。

钾离子通道Kir2.1/C-末端功能区的分子动力学模拟

<正>内向整流型钾离子通道(Kir,inwardlyrectifyingpotassium)在细胞激活、细胞内外钾离子K~+的动态平衡、胰岛素分泌等细胞生理过程中起重要作用。而细胞内各种不同因素和第二信使对Kir的调控则是实现其不同生理功能的途径。已有实验结果表明,4,5二磷酸磷脂酰肌醇(PIP2,phosphatidylinositol4,5-bisphosphate)与Kir相互作用的强弱决定了Kir对各种调控因素的响应程度。根据Kir2.1与Kir3.1胞内C-末端X-ray

Visual C++ 6.0学习教程

MicrosoftVisualC十十6.0作为Microsoft Visual Studio的重要组成部分，包含了迄今为止功能最为强大的基于Windows的应用框架，在同类产品中处于领先地位。VisualC十十6.0是Microsoft迄今为止最全面、最完善的程序开发工具，为了适应各种编程风格，该软件提供了各种各样的辅助工具，在发挥编程能力和提高灵活性方面达到了空前的水平。与以往VisualC十十的各种版本相比较，VisualC十十6.0在编程环境、程序语言技术等方面做了许多改进，从而使VisualC十十更加适合专业程序员快速进行应用程序的开发。

本书内容丰富、图文并茂，是一本适合各种读者学习VisualC十十6.0的优秀参考书。

目 录
第一章 VisualC十十6.0简介及安装
1.1VisualC十十6.0新特性
1.2viSualC十十6.0开发环境简介
1.3如何学习使用VisualC十十6.0
1.4VisualC十十6.0的安装
第二章 走进C十十的世界
2.1类和对象的简介
2.2继承和多态性――一个具体的例子
2.3内嵌对象
2.4在栈中申请对象
2.5全程对象的申请
2.6对象之间的相互关系――指针数据成员
2.7this指针的使用
2.8对指针的引用
2.9友元类和友元函数
2.10静态类成员
2.11重载运算符
2.12从代码中分离出类定义
2.13匈牙利表示法
第三章 VisualC十十6.0的编程环境
3.1VisualC十十6.0主窗口
3.2VisualC十十6.0工具栏
3.3VisualC十十6.0菜单栏
3.4项目与项目工作区
3.5资源与资源编辑器
第四章 编一个最简单的VC十十程序
4.1什么是AppWizard？
4.2迎接你的第一个AppWizard程序
4.3“Iamaprogrammer.”在哪儿？
第五章 程序框架入门
5.1一个简化过的程序框架
5.2WinMain（）：第一个动作
5.3登记窗口类
5.4创建一个窗口
5.5显示窗口
5.6显示出那条消息
5.7窗口类与窗口对象
第六章 消息循环
6.1在消息循环中兜圈子
6.2对事件做出响应：WindowFun（）
6.3响应不同的消息
6.4现在你还跟得上吗？
6.5设备界面进行交互
第七章 精通程序框架
7.1WinMain（）函数在哪儿？
7.2应用程序框架和源文件
7.3工具条、状态条和打印等选项
7.4程序的控制流程
第八章 使用classWizard编程
8.1使用ClassWizard添加消息处理函数
8.2classWizard功能介绍
8.3传送鼠标消息
8.4保存鼠标绘图的信息
第九章 视图与文档
9.1Document－View模式
9.2从视图中分离出文档
9.3保存文档
9.4再访MyProg2.cpp
第十章 对象连接与嵌入（OLE）及其自动化
10.1公共对象模式（COM）
10.2类厂（classfactory）
10.3OLE自动化
10.4IDispatch接口
第十一章 动态连接库（DLLs）
11.1为什么使用DLL
11.2传统的DLL
11.3MFC库DLL
11.4MyProg4A――编写自己的类库扩展DLL
11.5MyProg4B――使用MFC库扩展DLL
11.6资源访问
第十二章 图形设备接口
12.1设备环境类
12.2GDI对象
12.3Windows的颜色映射
12.4映射方式
12.5字体
12.6MyProg3例程序
12.7MyProg3B程序
12.8MyPr0g3C例程序――使用CScrollView
第十三章 对话框
13.1在状态条上显示对话控件的帮助信息
13.2利用Fi1eOpen通用对话框打开多个文件
13.3定制通用文件对话框
13.4扩展和缩减一个对话框
13.5显示一个模式或无模式对话框
13.6编写定制的DDX/DDV例程
第十四章 剖析工具Spy十＋
14.1窗体
14.2消息
14.3进程与线程
第十五章 代码调试
15.1TRACE
15.2调试框架
15.3自我诊断
15.4调试代码的作用
15.5用Dump（）显示对象的信息
15.6检查内存

Development And Characterization Of Composite Ni-Cr-C-Caf2 Laser Cladding On Gamma-Tial Intermetallic Alloy

A process of laser cladding Ni-CF-C-CaF2 mixed powders to form a multifunctional composite coatingd on gamma-TiAl substrate was carried out. The microstructure of the coating was examined using XRD, SEM and EDS. The coating has a unique microstructure consisting of primary dendrite or short-stick TiC and block Al4C3 carbides reinforcement as well as fine isolated spherical CaF2 solid lubrication particles uniformly dispersed in the NiCrAlTi (gamma) matrix. The average microhardness of the composite coatings is approximately HV 650 and it is 2-factor greater than that of the TiAl substrate. (C) 2008 Elsevier B.V. All rights reserved.

Improved fluorescence from Tm3+/Er3+/Ce3+ triply doped bismuth-silicate glasses for S+C-bands amplifiers

Fluorescence of Tm3+/Er3+ codoped bismuth-silica (BS) glasses and the sensitization of Ce3+ are investigated. It shows that Ce3+ codoping with Tm3+/Er3+ in BS glasses results in a quenching of Tm3+ ion emission from F-3(4) to the H-3(6) level. Consequently, the 1.47 mu m emission occurs after the population inversion between the H-3(4) and F-3(4) levels. Furthermore, the codoped glasses show the broad emission spectra over the whole S and C bands with full-width at half-maximum (FWHM) up to about 119nm, as it combines 1.55 mu m emission band of Er3+ with 1.47 mu m emission band of Tm3+ under 800nm excitation.