| Objective The topic of this paper was solved by A549lung cancer screend tests to determine the active ingredients on Ganoderma lucidum extracted including total triterpenoids and polysaccharides, took them as the research objects, prepared with enhanced immunity and antioxidant function of Ganoderma lucidum guttate pills. Completed guttate pills preformulation study, to study the extraction process, molded technology researches, quality standards, preparation for evaluation and preliminary studies on pharmacodynamics.MethodsResearch on preformulation:This section firstly took total triterpenoids and polysaccharides regard them as the index components, to establish an UV vitro assay method of Ganoderma effective ingredient. Then based on the different sources of Ganoderma lucidum medicinal materials, with the contents of total triterpenes and polysaccharides as indexes, established of Ganoderma lucidum medicinal material sources.Research on extraction technology:Used A549cells screened tests to determine the active ingredient of Ganoderma extract included total triterpenoids and polysaccharides. Took ethanol as solvent extract the total triterpenoids, extraction of polysaccharide component of Ganoderma lucidum by water decocted method, the single factor investigation combined with orthogonal experiment method were to determine the optimum extraction process of Ganoderma lucidum polysaccharides and total triterpenoids.Researches on preparation model process:Used analytic experiment and different types of macroporous resin to adsorb triterpenoids of Ganoderma lucidum statically, screened ideal macroporous resin. Then combined single factor investigation with dynamic adsorption analytical method to optimized purification process of total triterpenoids with D101macroporous resin.Took pharmacodynamics experiment and triterpenoid content as indexes,determined by MTT method investigaed without purification and purification of ganoderma lucidum triterpenoids extractions on A549cells proliferation, enhanced the immunity function of the strongest component and optimum time and concentration, layed the foundation for the later researches work.Through a comprehensive analysis of the clinical demands, drug properties and formulation characteristics to determine the guttate pills formulation. On the basis of preliminary experiment, combinded the single factor investigation with orthogonal experiment methods, determined the optimum preparation forming process of Ganoderma lucidum guttate pills.Quality standards research:With referenced to the Chinese pharmacopoeia (2010edition),Accorded to the appearance characters, TLC identification, weight variation,disintegration time, content determination,and other aspects of the standard researches.Preliminary study of stability:According to clinical quality standards and Chinese Pharmacopoeia; three batches of guttate pills were investigated and accelerated stability study.Preliminary study of pharmacodynamics:Corrected phagocytic index a in mouse model carbon clearance test as the indexes, studied the thymus index and spleen index of immune system and the enhancement of immune ability; took animal models of serum superoxide dismutase enzyme activity and malondialdehydecontent as the indexes, investigated the antioxidant ability.ResultsThis paper determined the effective component of Ganoderma lucidum (total triterpenoids and polysaccharides), established in vitro analysis methods, good linear relationship, high precision, recovery rate meet the requirements for the determination Then based on the different sources of ganoderma lucidum medicinal materials, with the contents of total triterpenes and polysaccharides as indexes, established of Ganoderma lucidum medicinal material sources.The optimum extraction process of Ganoderma lucidum.Used lung cancer A549cells screened tests to determine the active ingredient of Ganoderma extract included total triterpenoids and polysaccharides.Added30times amount of90%alcohol, refluxed and extracted for2times heated at80℃,2h/time. Then Ganoderma lucidum after ethanol drugs, added30times amount of water, soaked for0.5h, water decocted and extracted for3times,2h/time.D101macroporous resin packed column by wet pretreatment, the best purification process of total triterpenoids:the concentration of sample was59mg/mL, the volume of sample was88mL, absorbed completely with3BV of distilled water at the speed of2BV/h impurity, then4BV80%ethanol as eluant, paralled three times to purified process validation, The results were feasibly.Ganoderma lucidum purifed triterpenoid on A549cells:Determined by MTT methods study resulted without the purification of triterpenes and polysaccharide were stronger inhibited effection on lung couldcer cells proliferation, inhibited48h rate could reach47.68%. Illustrated the Ganoderma lucidum polysaccharide extract triterpenoid and improved immunity function, the results of the preparation technology and the efficacy of the later study provided reliably experimental basis.The optimumed preparation process of guttate pills was drug:matrix=l:1.5, PEG2000: PEG6000=2:1, the melted temperature was80℃, and nozzled temperature was30℃.Three batch samples of Ganoderma lucidum guttate pills, the appearance was round, pills weight variation, dissolution time and other indicators comply with the provisions, TLC method was good specificity. In UV the determination of content of total diterpene in Ganoderma lucidum guttate pills were not less than20mg/g, polysaccharide content were not less than43mg/g, the method was simple, precise and reproducible.According to the systematic information of guttate pills stability influence factors tests and accelerated tests, we could determine the drug storage information, packaged materials and other relevant conditions, to improve the stability of the effective components of drugs.Research on preliminary pharmacodynamics:the successful preparation of immunocompromised and oxidative damage animal model, they were compared in each experimental group which given immune and antioxidant indices after administration, the conclusions were significant difference.Conclusions This paper used the model cells screened test to determine the active ingredient for Ganoderma lucidum, and combined with the advantages of guttate pills, preparated pills included polysaccharide components and total triterpenoids, completed the preformulation study, to study the extraction process of the total triterpenoids, purification of Ganoderma lucidum triterpenoids, molded process research, quality evaluation, preliminary pharmacodynamic and other contents researches, provided the quality control methods. It also layed a solid foundation for further developing of pharmacokinetic study and industrialization researches. |
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