| ObjectiveUnder the guidance of theories of Traditional Chinese Medicine and combining with modern scientific and technological methods, we take Chinese medicine Gekko swinhonis as the research object to establish the key techniques of quality control and evaluation system of animal drug Gekko swinhonis by systematic study of Gekko swinhonis from different habitats from the aspects of herbal textual research, literature review, pharmacognostic studies, qualitative and quantitative analysis, fingerprinting construction and pharmacodynamic study.Main Research Contents and Methods1. Herbal textual research and literature review at home and abroad---Herbal textual research on Chinese medicine Gekko swinhonis was done from the aspects of name, variety, habitat, medicinal part, harvest, process and flavor of herbal medicines based on consulting material medica of the past dynasties and modern literatures. At the same time, literatures of Gekko swinhonis at home and abroad were reviewed from the aspects of animal resources, pharmacognostic studies, chemical constituents, pharmacological action and clinical application.2. Pharmacognostic study---Variety identification was conducted for Gekko swinhonis medicinal materials from10habitats with application of animal taxonomy. And character analysis was conducted from the aspects of external morphology, size, color, smell and so on. Moreover, microscopic identification and TLC were used in pharmacognosy identification of Gekko swinhonis.3. Components determination---Orthogonal test was employed to optimize the extraction techniques of protein and polysaccharide, and the content of water-soluble protein and polysaccharide of Gekko swinhonis medicinal materials from10habitats were determined using the technology of Coomassie brilliant blue stained and phenol-sulfuric acid method, respectively. Contents of crude fat and total phospholipid were also determined by molybdenum blue colorimetry.4. Examination items determination---Content of water, ash and extract of Gekko swinhonis from different habitats were determined according to the Chinese Pharmacopoeia in2010.5. Fingerprint study---Establishing the protein fingerprint of Gekko swinhonis by using polyacrylamide gel electrophoresis. The ultraviolet fingerprint spectrums group draft of Gekko swinhonis were established with the different multi-polar solvent system extract. The infrared spectrum of Gekko swinhonis was established with pressing potassium bromide troche. The chromatographic fingerprint of Gekko swinhonis was established by high performance liquid chromatography.6. Pharmacodynamic study---Polypeptide and polysaccharide components of Gekko swinhonis medicinal materials were extracted to detect the killing effect of the active extracts on bel-7402(human hepatocellular carcinoma cell line) and MCF-7(human breast cancer cell line) by MTT assay.Results1. The medicinal materials in this study are all Gekko swinhonis, the morphological and microscopical characters are similar, and the difference of external morphology maybe due to different living environment, breeding way and processing method, etc.2. The TLC identification method and detection index of Gekko swinhonis medicinal materials were developed.3. The optimized extraction technique of protein is:pH12, solid-liquid ratio1’.20and80min for extraction at the temperature of50℃. The content of water-soluble protein of Gekko swinhonis medicinal materials showed obvious difference with habitat, so it can be served as an index of quality evaluation of Gekko swinhonis.4. The optimized extraction technique of polysaccharide is:20min for extraction, solid-liquid ratio1:30and alcohol concentration of90%. The content of polysaccharide of Gekko swinhonis medicinal materials also showed obvious difference with habitat, so it can be served as another index of quality evaluation of Gekko swinhonis.5. The content of crude fat and total phospholipid showed obvious difference with habitat, which suggested that there was great difference in the quality of Gekko swinhonis medicinal materials. 6. Reference standards for the content of water, ash and extract of Gekko swinhonis are constructed and the concentration of alcohol for alcohol extract determination is70%according to the results of examination items determination.7. The infrared spectrum draft of Gekko swinhonis was established for the first time.8. The ultraviolet fingerprint spectrums group drafts of Gekko swinhonis were established for the first time.9. The protein fingerprints of total body, body without head and tail of Gekko swinhonis were established for the first time.10. The HPLC fingerprints of Gekko swinhonis were established for the first time and8common peaks of fingerprints were marked.11. The results of pharmacodynamic study showed that the Polypeptide and polysaccharide components of Gekko swinhonis had relatively high antineoplastic activity.12. The quality standards (draft) of Gekko swinhonis were established and foundation for quality control was supplied.ConclusionChinese medicine Gekko swinhonis has relatively high antineoplastic activity, which has been widely used in clinical practice. It hasn’t been embodied in Chinese Pharmacopoeia, and there’s also no quality evaluation standard, research on the quality control and evaluation hasn’t been reported too. In this sense, this research has an initiating significance, which would promote the study on animal drug Gekko swinhonis. This research carried out a systemic and all-around study on the key techniques of quality control and evaluation standard of Chinese medicine Gekko swinhonis for the first time from the aspects of morphological character, microscopic identification and TLC identification, components determination, fingerprinting establishment, examination items determination, pharmacodynamic study, etc. The protein, spectrum and chromatographic fingerprint and the quality standards (draft) of Gekko swinhonis were established for the first time, which laid a foundation for the quality control and evaluation of Chinese medicine Gekko swinhonis. |
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