|Applications of chromatographic, chemometric techniques and in-vitro bioassay to investigate the Chinese medicines, Radix Ginseng, Radix Panacis Quinquefolii, Ganoderma amboinense, Danggui Buxue Tang and RDLP
|Hong Kong Polytechnic University -- Dissertations.
|Department of Applied Biology and Chemical Technology
|xxvii, 184 leaves : ill. (some col.) ; 30 cm.
|Chinese medicine (CM) is now regarded as potential supplements and alterative medicine throughout the world in the past decades. However, quality, safety and efficacy of CMs are of great concern owing to limited scientific evidence available about its chemical composition, pharmacology and related properties. In this study, hyphenated chromatographic, chemometric data processing and biological screening techniques were applied to investigate the CMs, Radix Ginseng, Radix Panacis Quinquefolii and Ganoderma amboinense, and two CM formulations of RDLP and Danggui Buxue Tang (DBT) for both qualitative and quantitative analyses. Radix Ginseng and Radix Panacis Quinquefolii are two well-known and popular CMs and their dietary supplements can be found easily in health food stores. In this study, a fast extraction and a chromatographic procedure for high performance liquid chromatography-diode array detector-evaporative light scattering detector (HPLC-DAD-ELSD) were developed for introducing the basic concept of chemical analysis of natural products to tertiary students. The experiment designed was flexible for tertiary institute with different instruments. HPLC-DAD together with a chemometric method, evolving window orthogonal projections (EWOP) method was used to analyze the six nucleosides components, adenine, adenosine, inosine, guanosine, uracil and uridine, in the cap, the stipe and fruit body of Ganoderma amobinense (DH Lingzhi). Adenosine, guanosine, uridine and uracil, were identified in all the DH Lingzhi samples by using the experimental method. However, their accurate contents were difficult to determine by their peak areas as these their chromatograms were affected by the instrumental noise and other interferences. Hence, EWOP method was employed to eliminate the interferences in these data sets. As a result, both qualitative and quantitative analyses of the nucleosides components in DH Lingzhi samples were achieved. RDLP is a CM formulation developed by a Hong Kong CM practitioner and it is good for liver cancer. In this work, preparative HPLC-DAD and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell proliferation assay were applied respectively to analyze and fractionate the chemical constituents, and the safety and the anti-cancer activity of RDLP on liver cancer cell lines. The MTT cell proliferation assay was used as guidance for the fractionation of the chemical components of RDLP by PHPLC, so-called bioassay-guided fractionation approach. Four fractions of the RDLP extract were obtained, and their anti-cancer activities were studied. One of four fractions was found to contain potential bioactive components by us in RDLP. The RDLP extract was also less harmful to normal liver cell lines. DBT has been used for the improvement of the menopausal symptoms to woman since 1247 A. D. The formulation is composed by 10 qian Radix Astragali (Heungqi) and 2 qian Radix Angelicae Sinensis (Danggui). In this investigation, HPLC-DAD combined with Multicomponent spectral correlative chromatographic (MSCC) method Similarity index (SI) were utilized for investigating the chemical constitutions of DBT and its two herbs. Their extracts were prepared by the boiled and soaked methods. Then they were measured by HPLC-DAD. Moreover, the chromatographic data acquired were further treated with MSCC and SI to find out the chemical compositions of the samples. The variation of the chemical components found in the two DBT extracts was helpful to evaluate the bioactivities of DBT.
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