【作者】 王英姿；

【导师】 张兆旺；

【作者基本信息】 山东中医药大学 ， 中药学， 2003， 博士

【摘要】 目的：以复壮胶囊剂为例，探讨中药复方药效物质用“半仿生提取法（SBE法）”研究的模式。方法：以淫羊藿苷、β-谷甾醇、甜菜碱、人参二醇、人参三醇、总糖、干浸膏为综合评判指标。首先在药材粒度、提取温度、提取用水量、滤过、浓缩等条件相同的前提下，用均匀设计U₁（9¹×3³）优选出半仿生法提取用水的pH值和提取时间；再以上述指标及无水乙醇精制液的高效液相梯度洗脱总积分面积为综合评判指标，对该方中4味药材的15种组合方式进行优选，并用指纹图谱-模式识别法（主成分分析、聚类分析、逐步判别分析）对优选出的SBE法条件和提取时药材最佳组合方式进行验证性研究；然后采用此条件提取，用比例分割法优选SBE液较佳醇沉浓度；最后作SBE法、WE法（水提法）、SBAE法（半仿生提取醇沉法）、WAE法（水提醇沉法）4种方法提取液的成分、药效及兔血清指纹图谱的比较。在上述研究工作的基础上，进行了复壮胶囊制剂的研制，建立了质量控制方法和标准，并对制剂的稳定性进行了考察。结果：复壮胶囊药效物质用SBE法提取3次，水的pH值依次为：4.0、7.5、8.7，提取时间依次为：150、75、75min。以淫羊藿（A）、肉苁蓉（B）、枸杞（C）、人参（D）混合提取为最佳。指纹图谱—模式识别验证了该方药优选出的SBE法条件和提取时药材的组合方式为最优。方药SBAE法5种醇沉浓度的综合评判值Y为：Y_61.25％＞Y_55％＞Y_67.5％＞Y_30％＞Y_80％，结合生产实际，以调含醇量60％为最佳。方药4种方法提取液的综合评判值Y为：Y_SBE液＞Y_WE液＞Y_SBAE液＞Y_WAE液；为较全面地比较SBE液及WE液的化学成分的异同，进行了HPLC多成分分析比较，结果发现，在此分析范围内，SBE液及WE液的色谱峰峰数及峰位基本一致，两者对应色谱峰的积分值除1个峰外SBE液均大于WE液。说明2种提取液的成分没有质的改变，但SBE液中成分的含量显著大于WE液，提示本制剂的方药提取用SBE法为佳。4种方法提取液均能提高老年大鼠血清睾丸酮含量和下丘脑、额叶皮质中β-内啡肽含量，以SBE法3个剂量组作用最为显著，接近或超过青年鼠水平，说明SBE液比WE液的强壮和免疫功能调节作用强；兔血清HPLC、HPCE指纹图谱均表明，SBE液在体内的吸收速度整体快于WE液。对复壮胶囊中的3种植物性药材进行了薄层定性鉴别，动物性药材进行了高效毛细管电泳鉴别；初步制定了指标成分的含量限度：淫羊g＃不得少于 0．85mg·g”’，尸－谷笛醇不得少于 12．8 u g·g“1，甜菜碱不得少于 8．gmg·g”l，人参H醇不得少于 8．9 n g亿’1，人参三醇不得少于3．2。gl＼根据制剂的稳定性考察结果，规定该制剂有效期可暂定为二年。结论：按拟定的“ SBE法”研究模式研制的复壮胶囊是一种技术含量高，能保持原方特有疗效，质量稳定可控、使用方便的口服制剂。本文建立的“ SBE法研究模式”，为中药方剂研究和中药新药研究提供了新的思路与方法，具有较高的科学性和实用性。更多还原

【Abstract】 Objective: To probe into the basic research on extraction of available compound substances by semi-bionic extraction (SBE) and take example for Fuzhuang Capsule.Methods: Using the even designment to optimize the best extraction conditions of the semi-bionic extraction for Fuzhuang Capsule, withicariine, /?-sitosterol, betarine, panaxadiol, panaxitriol, total saccharine and dried extract weight as the indexes, in addition, using total integral calculus area of absolute alcohol solution by HPLC to optimize the extraction methods of combined ingredients of Fuzhuang Capsule. HPLC has been used for the finger print chromatograms of different samples. Principal components analysis, cluster analysis and stepwise recognition analysis have been used for the pattern recognition. Optimizing the alcohol precipitation concentration using proportional method. According to the former conditions, the four kinds of extract methods, including the semi-bionic extraction (SBE),the water extraction (WE), the semi-bionic extraction by precipitation with alcohol (SBAE) and the water extraction by precipitation with alcohol (WAE), were compared in the contents of the index ingredients, in the pharmacological activity and in the fingerprinting of the rabbit’s blood serum. Based on the former research, Fuzhuang Capsule was prepared.The quality control standards were established and the stability of the preparation was studied.Results: The technological conditions for SBE: the pH values of the water for the 1st, 2nd, and 3rd decoctions are in order of 4.0, 7.5, and 8.7, and the extraction times are 150min, 75min, and 75min respectively. Mixing herb Epimedii(A), herb Cistanchis(B), fructus Lycii(C) and radix Ginseng(D) together to extract is the best combination.The results of technological conditions and the extraction methods of combined ingredients of Fuzhuang Capsule by SBE are tested and found to be correct byfingerprint chromatograms and pattern recognition. The sequence of total index Y value of five kinds of alcohol precipitation concentration is:Y61.25%>Y55%>Y67.5%>Y30%.>Y80%. Combined with the production, the best alcohol precipitation concentration is 60%.The sequence of total index Y value of the four extraction technology is:YsBE>YwE> YSBAE>YWAE- Using HPLC for completely comparing the ingredients of SBE and WE, the result shows that the number and the place of the peaks in SBE and WE basically are the same, the integral calculus of SBE is all above WE besides one.It indicates that there has been no change in the nature of SBE and We, but the content in SBE is obviously high than in WE, SBE is the best extraction method in preparation of Fuzhuang Capsule.The four kinds of extractions especially SBE can raise the concentrations of testosterone in blood and lowered levels of both bata-EP in the hypothalamus and frontal contex in senile rats. SBE method can obviously promote the content near to young animal’s level.It is indicated that SBE can effectively improve the immunity function and make body stronger than WE. The HPLC fingerprinting and the HPCE fingerprinting of the rabbit blood serum show that the SBE decoction’s metabolic rate is more rapid than the WE decoction in vivo, SBE is easier to be absorbed than WE in the human body.Fuzhuang Capsule was prepared. Using TLC and HPCE to identify the medicinal materials.The quality control standards were established and the content range of the index ingredients are: icariine above 0.85mg ?g"1, ^-sitosterol above 12.8 ng ?g"1, betarine above 8.9mg ?g"1, panaxadiol above 8.9 P g ?g"1, panaxitriol above 3.2 u g ?g"1. The study of the preparation’s stability shows that the expiration date of the capsule is 2 year.Conclusion: Prepared in accordance with semi-bionic extraction patterns, Fuzhuang Capsule has high contents of chemical compositions , good curative effects, stability and being controlled quality and taken convenience. The established semi-bionic extraction patterns provide a new way of thinking and method for compound substances and development for更多还原