金铃子滴丸有效部位及制剂工艺研究

【作者】 黄娴；

【导师】 张素斋；

【作者基本信息】 北京中医药大学 ， 中药学， 2007， 硕士

【摘要】 金铃子散方出自金元四大家之一刘完素的《素问病机气宜保命集》,由延胡索、金铃子两味药组成,其历史悠久,组方精简,疗效确切。本课题以多学科理论为基础,采用化学分析、分离、提取方法,建立有效组分、制剂的质量控制与评价方法,优化金铃子散的剂型,使金铃子散成为疗效确切,有效组分基本清楚,质量稳定可控,具有“三小”、“三效”、“五方便”等特点的现代方剂。课题分为以下几个研究部分:一、对金铃子散中川楝子以及延胡索的成分及药理活性、临床研究进行了综述,共引用193文献篇。二、对金铃子散进行了化学成分研究,从延胡索中共分离了7个生物碱类化合物,分别为紫堇碱(d-Corydaline)、四氢黄连碱(Tetrahydrocoptisine)、延胡索乙素(Tetrahydropalmatine)、四氢非洲防己胺(l-Tetrahydrocolumbamine)、原阿片碱(protopine)、四氢小檗碱(Tetrahydroberberine)、去氢海罂粟碱(dyhydroglaucine)。从川楝子中分离得到四个化合物,鉴定了其中3个,分别为β-谷甾醇(β-sitosterol)、菜油甾醇(campesterol)、川楝素(toosendanin)与异川楝素(isotoosendanin)的混合物。三、延胡索总生物碱有效部位研究首先优选了延胡索总生物碱的提取工艺,以总生物碱的转移率为指标,对延胡索提取工艺进行了考察,经数据分析,最终得出延胡索的最佳提取工艺为:60%盐酸乙醇(1:100)浸泡12h,12倍量,0.06ml/min/g渗漉提取。其次,考察了延胡索生物总碱有效部位的富集纯化工艺。最终确定延胡索脂溶性生物总碱有效部位的富集纯化工艺为:延胡索提取浸膏,水分散,1/3乙酸乙酯萃取6次,合并,回收溶剂,即得。四、川楝子总三萜有效部位研究以总三萜的含量位指标,筛选优化了川楝子总生物碱有效部位的提取纯化工艺,最后确定,川楝子总三萜有效部位的制备方法为:八倍量的乙酸乙酯提取3次,每次2小时。五、延胡索生物碱有效部位的质量标准研究1采用紫外法测定延胡索生物碱有效部位总生物碱的含量。以延胡索乙素为对照品,检测波长225nm,所得线性方程为:y=64.459x+0.0092,r=0.9996(n=5)。表明延胡索乙素在225nm检测波长下,浓度在0.001635～0.01475mg/ml之间与吸光度成良好线性关系。有效部位含量为50%以上,加样回收率为97.95%,RSD=2.69%(n=5)。2采用高效液相法对有效部位中延胡索乙素、四氢黄连碱、紫堇碱、四氢非洲防己胺进行了含量测定,延胡索乙素的标准曲线y=796.21x-0.2375,R=0.9999 (n=5),表明延胡索乙素在0.24～3.78μg范围内质量与峰面积呈良好线性;紫堇碱的标准曲线为y=828.08x-7.0042,R=0.9999(n=5),表明在0.20～3.24μg范围内质量与峰面积呈良好线性。四氢非洲防己胺的标准曲线为y=925.12x-0.8875,R=0.9999(n=5),表明四氢非洲防己胺在0.24～3.85μg范围内质量与峰面积呈良好线性。标准曲线为y=972.16x-0.8875,R=0.9999(n=5),表明四氢黄连碱在0.236～3.776μg范围内质量与峰面积呈良好线性。经测定,有效部位中含有延胡索乙素86.36mg/g,平均回收率为99.4%,RSD%=1.4%,含有紫堇碱101.12mg/g,由以上数据可知平均回收率为97.7%,RSD%=1.33%;含有四氢黄连碱20.78mg/g,由以上数据可知平均回收率为97.7%,RSD%=1.4%;四氢非洲防己胺43.74mg/g,平均回收率为102.4%,RSD%=0.72%六、川楝子总三萜有效部位的质量标准研究以川楝素为对照品,香草醛-高氯酸显色,在520nm下采用可见分光光度法测定川楝子有效部位总三萜的含量,所得标准曲线为:y=114.53x-0.3939,r= 0.9987(n=5)。表明川楝素在上述条件下,浓度在0.004-0.02mg/ml之间与吸光度呈良好线形关系。经测定,川楝子总三萜有效部位的含量为0.551g/g,平均回收率为103.2%,RSD%=2.3%七、金铃子散滴丸成型工艺研究及质量标准研究金铃子散有效部位滴丸的制剂成型工艺及质量标准研究。考察了滴丸制剂成型工艺,确定了基质种类以及基质与有效部位的混合比例。并建立了滴丸的定性与定量的质量标准。八、有效部位的指纹图谱的建立用HPLC法建立了延胡索生物碱有效部位的指纹图谱,通过考察五批有效部位指纹图谱,建立了有效部位的对照指纹图谱。并对其中的七个峰进行了指认。九、总结与讨论更多还原

【Abstract】 Jinlingzi San is a classical and efficient formula, presented in SU WEN BING JI QI YI BAO MIN JI, simply composed of Fructus Toosenedan 9g and Rhizoma Corydalis 9g and mainly used for disease of the digestive system. Based on multi-disciplinary theory, the study is fisrtly on the quality evaluation method of the effective components in Fructus Toosenedan and Rhizoma Corydalis as well as Jinlingzi San and then on the preparing technology of Jinlingzi San, in order to make Jinlingzi San a modern formula with characteristics of“3 small”,“3 effective”and“5 convenient”.This dissertation has nine parts:Chapter 1 In this chapter, separately overview and summarize the chemical composition of Fructus Toodensin, Rhizoma Corydalis and Jinlingzi san, as well as its pharmacology and clinical research. 193 references were cited.Chapter 2 The effective fraction was researched in this part. Seven Alkaloids were isolated from the Rhizoma Corydalis. The structures of them all are identified: d-Corydaline, Tetrahydrocoptisine, Tetrahydropalmatine, l-Tetrahydrocolumbamine, Protopine, Tetrahydroberberine and Dyhydroglaucine. Four compounds were isolated from Fructus Toodensin, three of whose structures are identified:β-Sitosterol, Campesterol, mixture of Toosendanin and Isotoosendanin.Chapter 3 In this part, the preparation of the effective fraction of Rhizoma Corydalis.was studied. Percolate. Firstly the extraction of Rhizoma Corydalis was optimized with the content of Alkaloids as selecting index. The optimum extraction was studied by orthogonal test and test with single factor. As a result, the optimum technology is that Rhizoma Corydalis is soaked by 2.5times 60% alcohol (containing 1%HCl), then percolated with flow rate 0.06ml/min/g for 12times volume Secondly the technology of purifying and enriching the effective fraction of Rhizoma Corydalis is optimized The optimum technology. is followed: Rhizoma Corydalis is that the extractive by alcohol, is diluted by ten times water with Ph 11to 12, Etac (three times) extract 6 times. To get all extract together, dry them. That is the effective fraction of Rhizoma Corydalis.Chapter 4 Optimization of the technology of preparing the effective fraction of Fructus Toosenedan. with the total content of Triterpene as reference substance As a result, the optimum technology is that Fructus Toosenedan is extracted by eight times acetoacetate for three times,two hours each time.Chapter 5 establishes methods of determining the effective fraction of Fructus Toosenedan.Firstly, the content of total alkaloid is determined with UV method (225nm), with Tetrahydropalmatine as reference substance. The calibration curve(y=64.459x+0.0092) is linear at ranges of 0.001635～0.01475mg/ml, r=0.9996(n=5). And the content is more than 50 %, the average recoveries were 97.95 %( RSD=2.69%, n=5).Secondly, the contents of index compounds including Tetrahydropalmatine, l-Tetrahydrocolumbamine, d-Corydaline, Tetrahydrocoptisine are determined by HPLC. The calibration curves(y=796.21x-0.2375) are linear at ranges of 0.24～3.78ug (for Tetrahydropalmatine , r=0.9999). And the content is 86.36mg/g, the average recoveries are 99.4% (RSD=1.4%, n=5). The calibration curves(y=828.08x-7.0042) are linear at ranges of 0.20ug～3.24ug (for d-Corydaline, r=0.9999), The content is 101.12mg/g, the average recoveries are 97.7% (RSD=1.33%, n=5). The calibration curves(y=972.16x-0.8875) are linear at ranges of 0.24ug～3.85ug( for l-Tetrahydrocolumbamine, r=0.9999). The content is 43.74mg/g, the average recoveries were 102.4%(RSD=0.72%, n=5). The calibration curves(y=972.16x-0.8875), are linear at ranges of 0.236ug～3.776ug( for Tetrahydrocoptisine, r=0.9999), The content was 20.78mg/g, the average recoveries are 97.7%(RSD=1.4%, n=5).Chapter 6 The content of total alkaloid are determined with UV method (520nm), with Toosendanin as reference substance. The calibration curve (y=114.53x-0.3939) is linear at ranges of 0.004～0.02mg/ml, r=0.9987(n=5). And the content is more than 0.551g/g, the average recoveries are 103.2 %( RSD=2.3%, n=5).Chapter 7 In this part, we research on preparation technology of dropping pills made of effective fraction of Jinlingzi San. And we establish the quality standard of the dropping pill using the method of HPLC.Chapter 8 In this part we establish HPLC fingerprint of the effective fraction of Fructus Toosenedan and Rhizoma Corydalis. So we can accurately control the overall quality of effective fraction with fingerprint.Chapter 9 analysis and summary更多还原