Three plant species (Phylica paniculata Willd., Pergularia daemia Forssk. and Monsonia / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2006.

Aims. The objectives of this study were (i) to isolate and characterize by conventional column chromatography, structurally diverse triterpenes from G. lucidum to serve as chemical markers; (ii) to develop a high performance liquid chromatography (HPLC) method for the quality control and/or standardization of Lingzhi-containing products; (iii) to utilize and optimize operating conditions for the newer extraction technology: supercritical fluid extraction (SFE), in order to maximize yields of bioactive triterpenes, and to reduce time and costs. / Background. The dried fruiting body of Ganoderma lucidum, commonly known as Lingzhi, has been used extensively as a traditional Chinese medicine (TCM) for many centuries not only in China, but also in other countries such as Japan and Korea. In recent years, Lingzhi has also become a popular health supplement in many Western countries. The chemical composition of Lingzhi is complex, but it has been well documented that the lipophilic triterpenoid class of compounds possess a range of biological effects that include antitumor, immunomodulatory, cardiovascular, respiratory and antihepatotoxic activity. A major drawback in TCM research has been the lack of authentic chemical standards, and efficient methods for the extraction and analysis of bioactive fractions and/or single components. Conventional extraction methods for G. lucidum are time-consuming and laborious, and often result in low yields of useful chemical constituents. / Conclusion. This study enabled the development of a method for the simultaneous analysis of structurally diverse triterpenes with remarkably different chromatographic profiles. The isolated triterpenes, as chemical markers, and the HPLC method can readily be used for quality control and/or standardization purposes in evaluating Lingzhi-containing products. Optimization of operating conditions for SFE facilitated the rapid and selective extraction of acidic triterpenes from raw G. lucidum in significantly higher yields. / Methods. Raw material of G. lucidum was extracted with 80% ethanol; subjected to repeated column chromatography to purify triterpenes; and characterized by NMR (1H and 13C) and mass spectroscopy. Isolated lipophilic triterpenes were qualitatively and quantitatively analyzed by reversed-phase HPLC using an ODS column (150 x 4.6 mm) and PDA detection at 256 nm. The assay was validated over appropriate concentration ranges and benzophenone was used as an internal standard. Supercritical fluid extraction of G. lucidum was carried out using a commercial supercritical fluid extractor system Thar, SFE-1000M. Briefly, the raw powder of G. lucidum was soaked in ethanol containing 10% aqueous ammonia for 30 minutes prior to extraction. Extractions were performed at temperatures of 40, 50 and 60°C; and pressures of 200, 250, 300 and 350 bar; 5% ethanol was used as the co-solvent; and the flow rate of CO 2 was set at 20 g/min. / Results. Eight compounds were isolated and identified from G. lucidum: four triterpenes; namely, lucidenic acid N, ganoderic acid B, ganodermanontriol, and ganodermadiol; two steroids; ergosterol-7, 22-dien-3beta-ol and ergosterol peroxide; and two fatty acids, oleic acid and tetracosanoic acid. The four triterpenes were utilized as chemical markers, and the developed HPLC method was able to simultaneously analyze the structurally diverse components with good resolution. The total analysis run time was 110 min, and retention times (tR) were 14.88, 18.96, 63.88 and 90.73 min respectively, and the eluting system was a mixture of three solvents, methanol (A), acetonitrile (B) and 2% acetic acid solution (C): 0-22 min, 5% A, 25% B and 70% C; 22-85 min, gradient elution, the ratio changed gradually to 5% A, 80% B and 15% C; 85-110 min, gradient elution, the ratio changed gradually to 5% A, 85% B and 10% C. The validated HPLC method and the isolated chemical markers were effectively applied to determine the triterpenoid contents in a variety of commercial Lingzhi products. Supercritical fluid extraction conditions of: pressure 300 bar and temperature 50°C, gave the highest yields of triterpene-containing extracts. HPLC analysis of the SFE extracts showed predominantly acidic triterpenes such as lucidenic acid N and ganoderic acid B. / Hong Xin. / "December 2006." / Advisers: Ho Yee Ping; Albert H. L. Chow. / Source: Dissertation Abstracts International, Volume: 68-09, Section: B, page: 5968. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 149-175). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

A number of recent research studies have been done on different species of the plants of the genus Phyllanthus. The plants are widely distributed in most tropical and subtropical countries and have long been used for the treatment of liver diseases in China and India. / Hepatitis B virus (HBV) is a major pathogen of human viral hepatitis. It has been estimated that 350 million people are chronic carriers of HBV throughout the world. Increasing evidence indicates that persistent viral infection of the liver is associated with cirrhosis and hepatocellular carcinoma. Hepatitis B virus belongs to a family of DNA viruses called hepadnaviruses. The current treatments of HBV infection with interferon or lamivudine have several disadvantages, and there appears to be much room for improvement in terms of medical treatment. / My project research focuses on two poorly characterized Indian Phyllanthus species called Phyllanthus nanus ("PN") and Phyllanthus niruri ("PI"). In my studies, random amplified polymorphic DNA ("RAPD") technique and high performance liquid chromatography ("HPLC") fingerprinting were used to authenticate different species of Phyllanthus. Both aqueous and ethanolic extracts of PN and PI were prepared to study their cytotoxicity in hepatoma cell lines. The effect of these extracts on hepatitis B virus was also examined in the HBV-genome integrated cell lines - PLC/PRF/5 (Alexander) and HepG2 2.2.15. Microparticle enzyme immunoassay ("MEIA") and enzyme-linked immunosorbent assay were used to measure the amount of hepatitis B surface antigen ("HBsAg") and hepatitis B e antigen ("HBeAg") secretion from the cell lines. RT-PCR was used to detect the change in HBsAg mRNA's expression level in the drug-treated cell lines. Real-time PCR was also employed to examine the effect of drug treatment on the level of HBV DNA replication and the amount of virions secreted into the medium. The experimental results showed that both the aqueous and ethanolic extracts of PN and PI exerted suppressive effect on HBsAg secretion and HBsAg mRNA level. The PN and PI ethanolic extracts also showed mild suppression of viral replication in vitro. The ethanolic extract of PN seemed to be more potent in suppressing HBV than the other extracts tested. (Abstract shortened by UMI.) / Lam Wai Yip. / "June 2005." / Advisers: Mary Waye; Vincent Ooi. / Source: Dissertation Abstracts International, Volume: 67-07, Section: B, page: 3594. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 217-234). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract in English and Chinese. / School code: 1307.

Ganoderma lucidum and Cordyceps sinensis are medicinal fungi commonly used in Chinese medicine. The allied species of G. lucidum, G. sinense and G. tsugae as well as the allied species of C. sinensis, C. militaris are also commercially available as health supplements. The present study aimed at evaluating and comparing the biological activities of the different parts of fruiting bodies of Ganoderma species (whole fruiting body, pileus, stipe, spores and spore oil) and Cordyceps species (whole fruiting body, stroma and larva/pupa). / The extracts of C. sinensis and C. mililaris could stimulate secretion in human airway epithelial cell Calu-3, implying the potentials of these extracts to modulate the mucociliary clearance. The potencies of the stimulatory effects of the stroma of fruiting bodies showed stronger effects than the larva/pupa. All parts of C. sinensis and the stroma of C. mililaris could modulate the proliferation of human peripheral blood mononuclear cells and the different potencies of immunomodulatory effects were also observed. / The results demonstrated that the hot water extracts of G. lucidum, G. sinense and G. tsugae possessed antiproliferative effects on human breast cancer cell lines MCF-7 and MDA-MB-231. The extracts from the stipes showed stronger inhibitory activities on cancer cell proliferation than those from pilei. Furthermore, the extracts from the whole fruiting body and stipe of G. lucidum possessed strong antitumor effects on the nude mice bearing MCF-7 xenografts and the BALB/c mice bearing sarcoma S-180 allografts as well as strong immunomodulatory effects in terms of stimulating splenic lymphocyte proliferative responses. The oral administrations of spores and spore oil did not show significant inhibition on MCF-7 xenografts growth but they inhibited sarcoma allografts growth effectively. / The strong biological effects of the stipe of Ganoderma and the stroma of Cordyceps were showed for the first-time. This study sheds light on how the fruiting bodies of Ganoderma or Cordyceps should be used to achieve the most out of their pharmacological properties. This study also demonstrated the novel application of Cordyceps in promoting secretion in human airway submucosal glands, which reinforces the rationale of using this fungus for treating respiratory diseases. / Yue Gar Lee Grace. / "August 2006." / Advisers: Leung Ping Chung; Kwok Pui Fung; Bik San Clara Lau. / Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1584. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 318-340). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Ingestion of extracts prepared from various medicinal plants to induce or augment labour is common amongst Black South African women during the late stages of pregnancy. This applies particularly to the rural areas where modern health care facilities are often lacking. Many of these plants have not been investigated scientifically and one needs to substantiate claims of quality, safety and efficacy. Furthermore, it is believed that the consumption of these plant extracts can result in foetal meconium staining at delivery. An investigation into the uterotonic properties of three plants viz. Ekebergia capensis Sparrm. Clivia miniata (Lindl.) Regel. and Grewia occidentalis L. were carried out using guinea pig uterine smooth muscle in vitro. Supercritical fluid extraction was performed with water modified supercritical carbon dioxide to extract the uterotonic components. An attempt was also made to couple supercritical fluid extraction directly on-line to the bioassay so that on line screening of crude plant extracts could be performed within short periods of time. The effects of supercritical CO2 decompression on temperature and pH of the muscle bathing solution were considered since these factors affect muscle contractility. The direct effects of excess CO2 on intracellular mechanisms were eliminated by constructing a CO2 reduction interface together with passage of carbogen which aided in the rapid displacement of excess CO2, As samples of these extracts were found to induce muscle contraction, supercritical fluid fractionation (SFF) was performed by sequentially increasing the fluid density. Extracted fractions were obtained by sequentially increasing the pressure at constant temperature and modifier concentration in an attempt to identify the active fractions. Extractions were performed at 200 atm, 300 atm and 400 atm respectively. Subsequent testing of these fractions enabled the detection of active and inactive fractions as well as a fraction that had a spasmolytic effect on uterine muscle. The 400 atm extracts of E. capensis and C. miniata displayed maximum activity while only the 300 atm extract of G. occidentalis induced uterine muscle contraction. Subsequent analysis of the sequentially extracted fractions, by high performance liquid chromatography and micellar electrokinetic capillary chromatography revealed that certain compounds present in the fractions that stimulated muscle contraction, were sensitive to the extraction pressure hence making it possible to determine the compounds that were likely to be active. Column chromatography followed by various spectroscopic techniques were performed in an attempt to isolate and elucidate the structures of the compounds that were present in the plant extracts. The extract of Ekebergia capensis yielded five known compounds (B-sitosterol, oleanonic acid, 3-epioleanolic acid, 2,3,22,23-tetrahydroxy-2,6,1 0, 15,19 ,23-hexamethyl-6, 10, 14, 18- tetracosatetrene and 7-hydroxy-6-methoxy coumarin. The extract of Clivia miniata yieded linoleic acid and 5-hydroxymethyl-2-furancarboxaldehyde while the extract of Grewia occidentalis yielded 3-(4-hydroxy-3-methoxyphenyl)-2-propenal, a novel compound 2,2' ,6,6'-tetramethoxy-4'-al-4-(w-oxo-E-propenyl)-biphenyl and oleanonic acid. The pure compounds were further evaluated pharmacologically to identify the active components and assess the physiological mode of action by the use of various receptor blockers. Oleanonic acid, 3-epioleanolic acid, linoleic acid and 5- hydroxymethyl-2-furancarboxaldehyde and 3-(4-hydroxy-3-methoxyphenyl)-2-propenal were found to induce an agonistic muscle response. All these compounds were observed to mediate their effects through the cholinergic receptors. The results obtained in this study supports the claim of these plants possessing uterotonic properties. / Thesis (Ph.D.)-University of Natal, Durban, 1997.

This study was undertaken to redress the scant knowledge regarding the chemistry and mode of action of pregnancy-related traditional medicines, or isihlambezo (Zulu), which are used by 60 to 80% of women in South Africa. The three selected plants are among the six most frequently cited species from the approximately 90 used by traditional healers. The purpose of the study was to identify components which could cause uterine contractions, those with nutritional value for the foetus and mother, and those with any toxic effects. Plant root extracts were purified via silica gel column chromatography and bioassays were carried out on the fractions, using isolated rat uterine tissue. Purified compounds were identified via spectral techniques, and some were characterised by comparison to authentic standards using HPLC, and others by matching their GC-MS spectra to library standards. Thirty-eight compounds were identified in total, the majority of these being novel to the species concerned. Those isolated from Combretum kraussii were 1 sitosterol, 2 combretastatin, 3 3',4-tri-O-methylellagic acid, 4 combretastatin B-1, 5 combretastatin A-1, 6 3,3'-di-O-ellagic acid lactone, 7a ellagic acid lactone, 7b ellagic acid, 8 and 9 a mixture of combretastatin B-1 and A-1 glucosides, 10 and 11 partly characterised glucosides of ellagic acid. Those isolated from Gunnera perpensa were 12 3',4-tri-methylellagic acid, 13 ellagic acid lactone, 14 1,1'-biphenyl-4,4'-diacetic acid, 15 p-hydroxybenzaldehyde, 16 Z-methyl lespedezate, 17 and 18 partly characterized higher glucosides of Z-methyllespedezate. Those isolated rom Rhoicissus tridentata were 19 (-)-epigallocatechin, 20 (+)-gallocatechin, 21 procyanidin B3, 22 procyanidin B4, 23 (+)-catechin hydrate, 24 (+)-mollisacacidin, 25 (+)-epicatechin, 26 fisetinidol-(4a-8) catechin, 27 (-)-fisetinidol, 28 fisetinidol-(4b-8)catechin, 29 gallic acid, 30 epicatechin-3-0-gallate, 31 partly characterized hydrogel of glucose, 32 sitosterol, 33 sitosterolin, 34 y-sitosterol, 35 oleanolic acid, 36 lupen-3-one, 37 20-epi-y-taraxastananol and 38 triacontanol. The compounds with the greatest in vitro uteroactivity were predominantly proanthocyanidins or phenolic glucosides. It is proposed that effects of phenolic glucosides could be due to the interaction of the sugar moiety as well as the phenolic moiety with the receptor site in muscle tissue. The corresponding phenolic aglycones isolated were only moderately uterotonic, or unreactive by comparison. Non-polar compounds such as sitosterol and sitosterolin showed minimal enhancement of the uterine response at low concentrations, and inhibition at higher concentrations. Aqueous root extracts of the plants were all found to be non-toxic according to cell-viability tests using monkey vero cells and human fibroblasts. Extracts are therefore considered safe for human consumption, although it is recommended that Rhoicissus tridentata be used with caution because it showed the lowest cell viability of the three species, and uterine hyperstimulation has been attributed to this species, as well as CNS depression and respiratory arrest. Ions which could be nutritionally beneficial in pregnancy, calcium, iron, and phospate, were present in low in aqueous extracts. Levels of calcium and potassium ions were considered to be too low to directly stimulate uterine muscle. Proanthocyanidins, combretastatins, ellagic acid derivatives and phytosterols, with health-promoting properties, were also identified. / Thesis (Ph.D.)-University of KwaZulu-Natal, Durban, 2004.

A method is presented for the simultaneous identification of nine compounds in samples of A. membranaceus and A. membranaceus var. mongholicus. Compounds identified in the extracts of the two plants included glycosides, saponins and flavonoids. They are identified as Calycosin-7-O-beta-D-glucoside (C1), Ononin (C2), (6aR,11aR)-3-hydroxy- 9,10-dimethoxypterocarpan-3-O-beta-D-glucoside (C3), (3R)-7,2'-dihydr- oxy-3',4'-dimethoxyisoflavan-7-O-beta-D-glucoside (C4), Calycosin (C6), Astragaloside IV (C5 ), Formononetin (C7), (6aR,11a R)-3-hydroxy-9,10-di-methoxypterocarpan (C8), and Isomucronulatol (C9). / An HPLC-DAD-MS method is proposed for the differentiation of Rhizoma Coptidis and Cortex Phellodendri samples. This method can also be used to identify two common species of Rhizome Coptidis, i.e., C. chinensis and C. deltoidea, and two species of Cortex Phellodendri, i.e., P. chinensis and P. amurense. From the experiment results, there are thirteen, twelve and seven common components found in samples Rhizoma Coptidis, P. amurense and P. chinensis, respectively. Nine compounds in Rhizoma Coptidis were identified to be alkaloids. The common components in Cortex Phellodendri included four alkaloids and two lactones, i.e., obaculacotone and obacunone, present in all samples of P. amurense. / High Performance Liquid Chromatography-Atmospheric Pressure Chemical Ionization Mass-Spectrometry has been applied to the analysis and standardization of Chinese Herbal Medicines. The applications included quantitative study of Astragaloside in Radix Astragali, investigation on the chromatographic fingerprint of Radix Astragali, differentiation of Cortex Phellodendri and Rhizoma Coptidis, and identification of constituents in Sanhuang Xiexin Decoction. / In the quantitative study of Astragaloside, an Multiple Reaction Monitoring scan mode was used. The linearity between 2 and 500 mg/L is 0.9996. The precision of injection and reproducibility of method is 1.72% and 3.27% respectively. A total of 20 samples from local market and mainland China were analyzed and the results are comparable to those obtained from HPLC-ELSD analysis. / The present study also proposed a HPLC separation and online identification for the 15 constituents in a composite Chinese herbal formula, the Sanhuang Xiexin Decoction. It provided a possible starting point to evaluate related herbal preparations containing Rhizoma Coptidis, Radix Scutellariae and Rhizoma Rhei. Thirteen constituents in the decoction were identified, including five major alkaloids from Rhizoma Coptidis, five anthraquinones from Rhizoma Rhei and two favonoids and one glycoside from Radix Scutellariae. / "November 2007." / Adviser: Chi Chun Tao. / Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4726. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 177-200). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Shum Ka Chiu. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 174-182). / Abstracts in English and Chinese. / Acknowledgement --- p.ii / Abstract --- p.iii / 摘要 --- p.vi / Table of content --- p.viii / List of figures --- p.xvi / List of tables --- p.xxii / Abbreviations --- p.xxv / Chapter Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Importance of authentication of Traditional Chinese Medicines --- p.1 / Chapter 1.1.1 --- Confusing nomenclatures --- p.1 / Chapter 1.1.2 --- Similar morphologies of different medicinal materials --- p.2 / Chapter 1.1.3 --- Toxicities of medicinal materials --- p.2 / Chapter 1.1.4 --- Conservation of natural products --- p.2 / Chapter 1.2 --- TCM listed in the Pharmacopoeia of People's Republic of China --- p.3 / Chapter 1.3 --- Overview of mis-use and intoxication of TCM --- p.4 / Chapter 1.4 --- Ordinances regulating Chinese medicines as natural products --- p.7 / Chapter 1.4.1 --- Laws governing Chinese medicine --- p.7 / Chapter 1.4.2 --- Laws governing endangered species --- p.8 / Chapter 1.5 --- Current technologies in the authentication of Traditional Chinese Medicines and their limitations --- p.9 / Chapter 1.6 --- Historical applications of Radix Aconiti --- p.12 / Chapter 1.7 --- Modern applications of Radix Aconiti --- p.16 / Chapter 1.8 --- Research on Radix Aconiti and its chemical components --- p.17 / Chapter 1.8.1 --- Chemistry --- p.17 / Chapter 1.8.2 --- Pharmacology --- p.19 / Chapter 1.8.3 --- Molecular interaction --- p.22 / Chapter 1.9 --- Brief review on the systematics and phylogeny of Aconitum --- p.23 / Chapter 1.10 --- Historical applications of Radix Aucklandiae and related materials --- p.25 / Chapter 1.11 --- Modern applications of Radix Aucklandiae and related material --- p.27 / Chapter 1.12 --- Research on Aucklandiae and related material and their chemical components --- p.28 / Chapter 1.12.1 --- Chemistry --- p.28 / Chapter 1.12.2 --- Pharmacology --- p.29 / Chapter 1.13 --- Brief review on the systematics and phylogeny of Aucklandia and related medicinal species --- p.31 / Chapter 1.14 --- Authentication by DNA sequencing --- p.33 / Chapter 1.14.1 --- Introduction --- p.33 / Chapter 1.14.2 --- Criteria of sequence markers --- p.36 / Chapter 1.14.3 --- Model used to process polymorphism in DNA sequences --- p.37 / Chapter 1.15 --- Screening for novel markers --- p.38 / Chapter 1.15.1 --- Reason for screening novel markers --- p.38 / Chapter 1.15.2 --- Basic principle --- p.39 / Chapter 1.16 --- Introduction to gas chromatography- mass spectrometry --- p.40 / Chapter 1.16.1 --- Basic principles and components of GC-MS --- p.41 / Chapter 1.16.2 --- Advantages and limitations of GC-MS --- p.42 / Chapter 1.16.3 --- Usage of GC-MS on natural product analysis --- p.43 / Chapter 1.16.4 --- Chemometric analysis --- p.44 / Chapter 1.17 --- Objectives --- p.46 / Chapter Chapter 2. --- Materials and Methods --- p.47 / Chapter 2.1 --- Plant samples --- p.47 / Chapter 2.1.1 --- Samples of Aconitum --- p.47 / Chapter 2.1.2 --- Samples of Aucklandia and related species --- p.51 / Chapter 2.2 --- DNA extraction method --- p.58 / Chapter 2.2.1 --- Reagents --- p.58 / Chapter 2.2.2 --- Methods --- p.59 / Chapter 2.3 --- Chemical extraction methods --- p.61 / Chapter 2.4 --- Chemical standard extraction and purification method --- p.62 / Chapter 2.5 --- DNA sequencing --- p.63 / Chapter 2.5.1 --- Reagents --- p.63 / Chapter 2.5.2 --- Methods --- p.65 / Chapter 2.6 --- Genomic subtraction --- p.70 / Chapter 2.7 --- Search for species-specific markers from the subtraction library --- p.74 / Chapter 2.8 --- Gas chromatography- mass spectrometry --- p.74 / Chapter 2.9 --- GC-MS chemometric analysis --- p.75 / Chapter Chapter 3. --- Authentication of Aconitum by DNA Sequencing --- p.76 / Chapter 3.1 --- Introduction --- p.76 / Chapter 3.2 --- Methods --- p.77 / Chapter 3.3 --- Results - 5S spacer --- p.77 / Chapter 3.3.1 --- Sequence information --- p.77 / Chapter 3.3.2 --- Sequence similarity --- p.78 / Chapter 3.3.3 --- Phylogram study --- p.81 / Chapter 3.4 --- Results -psbA-trnH --- p.85 / Chapter 3.4.1 --- Sequence information --- p.85 / Chapter 3.4.2 --- Sequence similarity --- p.85 / Chapter 3.4.3 --- Phylogram study --- p.87 / Chapter 3.5 --- Discussion --- p.91 / Chapter 3.5.1 --- Overview of nuclear ribosomal 5S spacer --- p.91 / Chapter 3.5.2 --- Extensive polymorphism of 5S spacer --- p.91 / Chapter 3.5.3 --- Distribution of samples in the phylograms constructed by 5S spacer --- p.93 / Chapter 3.5.4 --- Utility of 5S spacer for authentication --- p.94 / Chapter 3.5.5 --- Overview of psbA-trnH spacer --- p.94 / Chapter 3.5.6 --- Distribution of samples in the phylograms constructed by psbA-trnH spacer --- p.95 / Chapter 3.5.7 --- A distinctive region of inversion --- p.96 / Chapter 3.5.8 --- Utility of psbA-trnH for authentication --- p.97 / Chapter Chapter 4. --- Screening for Novel Markers for Authentication of Aconitum --- p.98 / Chapter 4.1 --- Introduction --- p.98 / Chapter 4.2 --- Methods --- p.99 / Chapter 4.3 --- Results - subtracted clones --- p.99 / Chapter 4.4 --- Results - SSH6 --- p.104 / Chapter 4.4.1 --- Sequence information --- p.104 / Chapter 4.4.2 --- Sequence similarity --- p.105 / Chapter 4.5 --- Results-SSH15 --- p.107 / Chapter 4.5.1 --- Sequence information --- p.107 / Chapter 4.5.2 --- Sequence similarity --- p.107 / Chapter 4.5.3 --- Phylogram study --- p.109 / Chapter 4.6 --- Results-SSH45 --- p.113 / Chapter 4.6.1 --- Sequence information --- p.113 / Chapter 4.6.2 --- Sequence similarity --- p.113 / Chapter 4.6.3 --- Phylogram study --- p.115 / Chapter 4.7 --- Discussion --- p.119 / Chapter 4.7.1 --- Utility of subtraction in screening markers --- p.119 / Chapter 4.7.2 --- SSH6 --- p.121 / Chapter 4.7.3 --- SSH15 --- p.122 / Chapter 4.7.4 --- SSH45 --- p.123 / Chapter 4.7.5 --- Hybridization in Aconitum --- p.124 / Chapter 4.7.6 --- Inferring species identities of samples from the market --- p.126 / Chapter 4.8 --- Conclusion --- p.128 / Chapter Chapter 5. --- Assessment of Aucklandia lappa and Related Species by GC-MS --- p.129 / Chapter 5.1 --- Introduction --- p.129 / Chapter 5.2 --- Methods --- p.130 / Chapter 5.3 --- Results --- p.130 / Chapter 5.3.1 --- Extraction of essential oil --- p.130 / Chapter 5.3.2 --- GC-MS analysis --- p.131 / Chapter 5.3.3 --- Peak alignment and hierarchical cluster analysis --- p.133 / Chapter 5.3.4 --- Purification of chemical markers from Aucklandia lappa --- p.148 / Chapter 5.3.5 --- Standardization of the purified chemical markers --- p.148 / Chapter 5.3.6 --- Quantitative analysis of chemical markers --- p.152 / Chapter 5.4 --- Discussion --- p.154 / Chapter 5.4.1 --- Analysis of chemical composition --- p.154 / Chapter 5.4.2 --- A comparison on chemometric methods --- p.154 / Chapter 5.4.3 --- Similarity of chemical profiles --- p.156 / Chapter 5.4.4 --- Dendrogram analysis --- p.157 / Chapter 5.4.5 --- Utility of GC-MS in authentication of A. lappa and related species --- p.159 / Chapter 5.4.6 --- Limitations --- p.159 / Chapter 5.4.7 --- Comparison with molecular data --- p.161 / Chapter 5.4.8 --- Contents of dehydrocostuslactone and costunolide --- p.163 / Chapter 5.4.9 --- Locality study --- p.164 / Chapter 5.5 --- Conclusion --- p.165 / Chapter Chapter 6. --- General Discussion --- p.167 / Chapter 6.1 --- DNA sequencing --- p.168 / Chapter 6.2 --- Genomic subtraction --- p.169 / Chapter 6.3 --- Future work on molecular authentication --- p.170 / Chapter 6.4 --- Future work on authentication of Aconitum --- p.170 / Chapter 6.5 --- Gas chromatography- mass spectrometry --- p.171 / Chapter 6.6 --- Future work on authentication by GC-MS --- p.172 / Chapter 6.7 --- Future work on authentication of Aucklandia lappa and related species … --- p.173 / References --- p.174 / Appendix A. Sequence Alignment of 5S Spacer from Aconitum Species --- p.183 / Appendix B. Sequence Alignment of psbA- trnH Spacer from Aconitum Species --- p.188 / Appendix C. Sequences of Subtracted Clones from Aconitum --- p.191 / Appendix D. Sequence Alignment of SSH6 from Aconitum Species --- p.194 / Appendix E. Sequence Alignment of SSH15 from Aconitum Species --- p.195 / Appendix F. Sequence Alignment of SSH45 from Aconitum Species --- p.200 / Appendix G. Gas Chromatograms of Essential Oil Extracts of Aucklandia lappa and Related Species --- p.202