| Objective:To establish the new quality control and evaluation system of Kansui Radix and to promote the internationalization of Kansui Radix quality standard, systematic study of Kansui Radix were conducted in this thesis under the guidance of theories of Traditional Chinese Medicine and using international guidelines and quality standards for reference combining with modern scientific and technological methods, including herbal textual research, literature review, pharmacognostic studies, qualitative, and quantitative analysis, HPLC fingerprint construction, safety evaluation, pharmacodynamic study and other aspects.Main Research Contents and Methods:1.Herbal textual research and literature review--- Herbal textual research on Kansui Radix in the aspects of name, habitat, process, flavor of herbal, formulation and clinical application was done by consulting material medica of the past dynasties and modern literatures. Meanwhile, literatures of Kansui Radix at home and abroad in the aspects of chemical constituents, detection method, pharmacological action and clinical application were also reviewed herein.2.Pharmacognostic study--- Character analysis was conducted involving external morphology, size and color as well. Moreover, microscopic and TLC were used in pharmacognosy identification of Kansui Radix.3.Examination items determination--- Water content, total ash and extract of Kansui Radix from different habitats were determined following the guidance in the 2015 edition of Chinese Pharmacopoeia4.Safety evaluation of Kansui Radix--- The quantity of aflatoxins, organochloride pesticides (OCPCs), heavy metal and deleterious elements (As, Cd, Hg and Pd) in Kansui Radix was respectively determinated by HPLC-FLD after immunoaffinity column with post-column photochemical derivatization, gas chromatography-mass spectrometry (GC-MS) and inductively coupled plasma-mass spectrometry (ICP-MS).5.Fingerprint study---, the HPLC fingerprint of Kansui Radix was established by optimizing the extraction techniques and HPLC parameters.6. Quantitative analysis--- Euphol in Kansui Radix was extracted completely and was determined with standard curve method under the same HPLC conditions in Kansui Radix fingerprint study.7.Pharmacodynamic study--- The inhibitory effects of the different polar extracts of Kansui Radix and index component on the proliferation of MKN45 cell were investigated by MTT colorimetric assay.8.Statistical analysis--- The measurement uncertainty for the methods of quantitative analysis of Euphol, determinations of water content, total ash and extract of Kansui Radix was investigated according to Evaluation and Expression of Uncertainty in Measurement, using GUM (ISO) and QUAM (EURACHEM) for reference.Results:1.Ten batches of sample are all identified as Kansui Radix. Their morphological and microscopical characters are similar, and the difference of external morphology maybe due to different living environment.2.The TLC identification method of Kansui Radix was developed.3.HPLC fingerprint of Kansui Radix was established, of which nine common characteristic peaks were marked with euphol as internal reference.4.The euphol was completely extracted in Kansui Radix and was determined with standard curve method. The evaluation of measurement uncertainty for this method was investigated with the statistical limit also calculated for the first time.5.The quantity of aflatoxins in Kansui Radix was determined for the first time by HPLC-FLD after immunoaffinity column with post-column photochemical derivatization. The upper limit of aflatoxins in Kansui Radix was recommended.6.The quantity of 20 OCPCs in Kansui Radix was determined for the first time by gas chromatography-mass spectrometry (GC-MS), the utmost limits of 20 OCPCs in Kansui Radix was recommended.7.The quantity of 4 heavy metals and deleterious elements in Kansui Radix was determined for the first time by inductively coupled plasma-mass spectrometry (ICP-MS), the upper limits of 4 heavy metals and deleterious elements were recommended.8.The results of pharmacodynamic study showed that all different polar extracts and index component of Kansui Radix, especially euphol, displayed inhibitory effect on the proliferation of MKN45 cell.9.The contents of water, total ash and extract of Kansui Radix were determined, and the evaluation of measurement uncertainty for these methods were also investigated for the first time.10.The quality standard (draft) and instructions for drafting were formulated.Moreover the advanced evaluation system of quality and safety of Kansui Radix were established herein.Conclusion:Kansui Radix has been clinically used for the treatment of gastric and esophagus cancer because of its high anticancer activity. However, study on the quality control and evaluation of Kansui Radix are not enough.For instance, Kansui Radix has not been embodied in international standards, such as USP, BP, EP, JP.Although Kansui Radix has been list in Chinese Pharmacopoeia, the overall quality control and safety evaluation of Kansui Radix are still insufficient.In this thesis, the key techniques of quality control and safety evaluation index of Kansui Radix were systematically studied exemplified by herbal textual research, literature review, pharmacognostic studies, qualitative and quantitative analysis, HPLC fingerprint, pharmacodynamic study. The safety of Kansui Radix was evaluated by determining the contents of Aflatoxins, OCPCs, heavy metals and deleterious elements, the utmost limits of which were finally recommended using USP and other international standards for references. Furthermore, the measurement uncertainty for the quantity analysis of euphol, water, total ash and extract was also evaluated using GUM and QUAM for reference.In conclusion, all the achievement herein provided scientific and systematic methods and experimental basis for the quality control and evaluation of Kansui Radix, which showed a groundbreaking significance to promote the internationalization of Kansui Radix. |
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