| Biomembrane chromatography is a new separation mode developed recent years. One of important brance is membrance chromatography, which integrates the screening and separation of active components and can imitate the process between active components and acceptors in vivo. With the development of the modern cell molecular biology and separation science, cell membrance chromatography has been made increasingly progress, while immobilized cell membrane extraction coupled with chromatography can make up drawbacks of cell membrance chromatography and present a bright future.There are two parts in the thesisThe first part: review1. Biomembrane chromatography and its implications in the screening of active components in Traditional Chinese medicines (TCM).Traditional chromatography, such as HPLC, is only a separative tool, used to study TCM.with disadvantages such as pharmacological parameters of the active components uncharactered and interferences. Biomembrane chromatography has concurrent merits of biology and chromatography. So it is suitable to screening active components in TCM. There are six branches of it,namly, MolecularBochromatography,ArtificialBiomembranechromatography,CellBiochromatography,Bota nical Biochromatography,cell membrane chromatography, membrane immobilized chromatography. Theorys of every branche and their implications in the screening of active components from TCM were introduced. Some problems of the Biomembrane chromatography and some solutions were also involved. Finally there was a prospect of Biomembrane chromatography.2. A survey of study on Polygonum Cillinerve (Nakai) OhwiChemical and pharmacological study advances on Polygonum Cillinerve (Nakai) Ohwi were proposed this part. There are anthraquinone. tannins, water-soluble proteins in Polygonum Cillinerve (Nakai) Ohwi. The active components are anthraquinones. Anthraquinones existing in Polygonum Cillinerve (Nakai) Ohwi include emodin, physcion, and the glucopyranosides of them. Polygonum Cillinerve (Nakai) Ohwi has mainly antitumo, anti-inflammatory, antivirus activity. 3. Advances study on anthraquinones in natural plantsAs effective compositions in much Chinese herbal medicine, anthraquinone has different bioactivities. Studies show that it functions very well in evacuating, lowering blood pressure and blood fat, antiviral, anticancer and coagulation. Methods of extraction and separation, quantitative analysis, are concluded.The second part: Research papersPrepaing of the Cardiac Cell Membrane and determining its physical and chemical constants. Based on flow injection stopped-flow and soft matter on-line sampling techniques, a molybdenum blue spectrometric method for determining the bioactivity of sarcolemma adenosine triphosphate enzyme (ATPase) was presented. Compared with the traditional method, the proposed method is 7 times faster, more manipulatively convenient with lower reagent consumption. The method was successfully applied to the study of rabbit cardiac muscle sarcolemma ATPase activity, the results show that ATPase activity decreases with increasing endurance time studied, which may supply important information for the further studies on sarcolemma purity and the activity of its receptors.Studying on immersion extraction of water-soluble components from Polygonum Cillinerve (Nakai) Ohwi and its technology. The work is aimed to studying on extraction technology of Polygonum Cillinerve (Nakai) Ohwi by single-factor experiments coupling to response surface experiments. The evaluation index is the peak area of one bound anthraquinone. From the propered work ,following conclusions were concluded : the extraction results are affected by socking time,socking temperature and ratio of material to liquid. A better extraction technology for water-soluble components from Polygonum Cillinerve (Nakai) Ohwi is:socking time for 42.8min,socking temperature at 81℃,ratio of material to liquid at 27.3: 1. The conclusions offer important information for next experiments.Studying on the interactions between Cardiac Cell Membrane and the water-soluble extracts from Polygonum Cillinerve (Nakai) Ohwi with HPLC. Based on Cell Membrance Chromatography, Membrane Immobilized Extract Techonoloy was built up by the following steps: fistly, a membrane extraction cartridge is made by packing live myocardial cellular membrane coated on activated macroporous silica gel. After water-soluble crude extracts of Polygonum Cillinerve (Nakai) Ohwi was injected into the membrane extraction cartridge, target compounds were retented on the cartridge. Unacting compounds were washed by a buffer of pH 7.4 Tris-HCl, repeatedly. The retented ones were eluted from the cartridge by another buffer of pH 4.0 phosphate. Results show there were five compounds in the water-soluble crude extracts of Polygonum Cillinerve (Nakai) Ohwi, two of them may not interact with cardiac cell membrane and the other three ones meybe interact with it. One main compound of the three was determined, it may be emodin monoglycocide. |
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