【作者】 郭涛；

【导师】 应国清； 王鸿；

【作者基本信息】 浙江工业大学 ， 微生物与生化药学， 2008， 硕士

【摘要】 本文以黄芪生脉饮为研究对象,通过查阅文献资料和初步的抗氧化清除自由基实验明确组成复方的四味中药有效部位。四味药石油醚、乙酸乙酯和水提取部分抗氧化半抑制率浓度IC₅₀值分别为黄芪:100.51,52.93,76.58μg/ml,党参:300.69,111.53,130.14μg/ml,麦冬:183.15,43.55,93.07μg/ml,五味子:15.79,17.54,39.0μg/ml。明确了五味子的有效部位为木脂素,黄芪有效部位为黄芪多糖和黄芪皂苷,麦冬有效部位为麦冬多糖,党参有效部位为党参皂苷。通过正交实验确定单味中药有效部位最优分离工艺。采用HPLC法来检测五味子中五味子醇甲和五味子乙素的含量;HPLC-ELSD法来检测黄芪皂苷含量;苯酚-硫酸法测定多糖含量;香草醛-硫酸法测定总皂苷含量。实验结果:黄芪多糖最优提取工艺为7倍水100℃提取2小时,2倍乙醇沉淀,所得的黄芪多糖含量为22.85%;黄芪甲苷最优提取工艺为8倍75%乙醇提取2小时,经过AB-8型大孔树脂4倍柱体积的80%乙醇液洗脱,黄芪甲苷的含量达到0.821%;麦冬多糖最优提取工艺为5倍水65℃提取2小时,2倍乙醇沉淀,所得的麦冬多糖含量为48.53%;党参多糖最优提取工艺为7倍水65℃提取2小时,3倍乙醇沉淀,所得的党参多糖含量为19.89%;五味子提取最优工艺定为4倍85%乙醇提取3小时,五味子醇甲和五味子乙素的含量分别为1.534%和44.8%。以黄芪生脉饮汤剂为参照,进行中药方剂有效部位和剂量重组。依据2005药典生脉饮成方配伍,添加黄芪,正交实验设计各药材配比,大鼠心肌抗氧化脂质过氧化抑制实验和TLC检测实验确定重组黄芪生脉饮最优工艺为分离后的黄芪:五味子:麦冬:党参四味中药的质量比为3:4:5:1,抑制率为58.9%,高于黄芪生脉饮标准品的抑制率,达到2005版药典的质量标准。更多还原

【Abstract】 Based on the Huangqi Shengmai Yin for the study,the four herbs were to determine effective parts through reference and evaluation of antioxidant active utilizes the stable 2,2-diphenyl-1- picrylhydrazyl （DPPH）radical.The petroleum ether,ethyl acetate and water extracts of the four herbs were to determined IC₅₀values（inhibitory concentration 50%）of antioxidant active that 100.51,52.93,76.58μg/ml of Astragalus membranacens,300.69,111.53,130.14μg/ml of Codonopsis pilosula, 183.15,43.55,93.07μg/ml of Ophiopogon japonicus,15.79,17.54,39.0μg/ml of Schisandra sphenanthera,respective.The effective parts of S. sphenanthera are lignans,the effective parts of A.membranacens are polysaccharides and saponins,the effective parts of O.japonicus are polysaccharides,the effective parts of C.pilosula are saponins.The optimal technology of separation of the four herbs is chosen by orthogonal experiments.HPLC method was used to determine the content of Schisandrin and Kadsuranin,HPLC-ELSD method was used to determine the content of Astragalus saponins,the method of Phenol-sulphuric acid determine the content of polysaccharides,the method of Vanillin-sulfuric acid determine the content of total saponins. The results:Extraction optimal technology of Astragalus polysaccharides is 7 times the amount of water in the 100℃to extract 2 hours and used 2 times the amount of alcohol to precipitate:the polysacchaides,the Astragalus polysaccharides content is 22.85 percent.Extraction optimal orthogonal technology of Astragalus saponins is 8 times the amount by 75 percent of the ethanol extracting for 2 hours.After AB-8 macroporous resin purification,With four times the amount of the column of 80 percent ethanol eluting,the content of Astragalus saponins is 0.821 percent. Extraction optimal orthogonal technology of Ophiopogonis tuber polysaccharides is 5 times the amount of water in 65℃to extract 2 hours and use 2 times the amount of alcohol to precipitate a polysaccharide,The content of Ophiopogonis tuber polysaccharides is 48.53 percent.Extraction optimal orthogonal technology of Codonopsis pilosula saponins is 7 times the amount of water in 65℃to extract 2 hours,and 3 times the amount of alcohol to precipitate a polysaccharide, the content of Codonopsis pilosula saponins is 19.89 percent.Extraction optimal orthogonal technology of Schisandrae lignans is 4 times the amount of 85 percent of the ethanol extract for 3 hours,the contents of schisandrin and Kadsuranin were 1.534 percent and 44.8 percent.The recombination of effective parts and dosage for the four herbs refer to Huangqi Shengmai Yin as a standard.Based on the Chinese Pharmacopoeia 2005 of Shengmai Yin,add A.membranacens,orthogonal design of experiments to determine the optimal ratio of the various ingredients.Finally,determining the optimal orthogonal technology of the recombination Huangqi Shengmai Yin is that A.membranacens.S sphenanthera:O.japonicus:C.pilosula is 3:4:5:1 by the ability of inhibit lipid peroxidation of mous heart and TLC,the clearance rate was 58.9%that higher than the Huangqi Shengmai Yin’s as a standard,and to achieve the quality standards of the Chinese Pharmacopoeia 2005.更多还原