| Marasmius androsaceus (L.ex Fr.) Fr.is called Gui-maozhen.It is a kind of fungus,it is belong to taxonomy,Agaricales,small white mushrooms Branch Marasmius.It grows on wood or bamboo sticks which are planted deep in the densely forested mountains where condition is cool and wet,acidic soil near the water.It is one of higher value traditional Chinese medicines.This paper mainly research on related literatures, systematic pre-experiment, screening the effeetive part, extraction and purified technology of total alkaloids and its quality standard.Literature Research:In this paper,biological characteristics, germplasm resources biological form, identification, chemical constituents, pharmacological and toxicological study, application research and research prospects of Marasmius androsaceus are summarized,also the research situation of alkaloids, including the extraction and purification methods, pharmacological activity research and application, in order to further design and implementation of the experimental scheme provides theory basis.Systematic pre-experiment: the components in Marasmius androsaceus contain amino acid, protein, alkaloid and phenols etc. which based on systematic pre-experiment, chemical identification and the extracts by different polarity solution (petroleum ether, chloroform, ethyl acetate, butanol, methanol, ethanol and water). Alkaloids especially soluble ones are mostly among these components. Combined with the result of research on pharmacology and pharmacodynamics of Marasmius androsaceus, the research content of this paper contains mainly aim at alkaloids compounds.Screening the effeetive part:To screen the effective fraction of analgesia in Marasmius androsaceus. Methods:Hot plate and acetic acid writhing test in mice were used to evaluate the pain releasing effect of Marasmius androsaceus. The fraction in 80% ethanol could significantly prolong the latencies of paw licking and decrease mice writhing. So the effective fraction of analgesia in Marasmius androsaceus is 80% ethanol and water extract.Extraction technology of total alkaloids:Single factor exploration determined single factor level, the five factors are as follows:concentration of ethanol, extraction temperature, extraction time, amount of solvent, extraction times. Select factor and level for orthogonal design L9 (34), study the four factors:extraction time, amount of solvent, extraction times and extraction temperature, and how they effect the extraction efficiency. Finally determine the optimum extraction factor of sweet tea:ten times 80% ethanol, extraction temperature is 80℃, extract three times, each 1.5 hour.Purified technology of total alkaloids:select fifteen macroporous resins that have different polarities, study the static and dynamic adsorption properties of macroporous resin and polyamide, also research on how these factors affect the enrichment of total alkaloids, the factors are the concentration of liquid filled column, flow velocity of upper samples, divulging curves, elution solvent and elution amount, according to which the optimal technology for enrichment is determined and then to verify it. The optimal technology parameter of macroporous resin type HP-20 is as nine kinds of resins were compared and the best one was chosen. Then study the technique condition for separating and purifying Total Alkaloids from Marasmius androsaceus. And HP-20 was selected for its excellent adsorption and desorption properties,80% ethanol was found to be the best elution solution. The best result was based on the followings:feed rate-2 BV/h, sample concentration-0.4 g/mL, the ratio of total alkaloids to HP-20 macroporous resin-139.76 mg/g, elution flow rate-4 BV/h, eluting agent amount-3 BV; then the purity reached 61.31% and the yield was 3.19. So the Total Alkaloids of Marasmius androsaceus can be effectively purificated and separated by HP-20 macroporous resin. |
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