【作者】 董蕊；

【导师】 郑毅男；

【作者基本信息】 吉林农业大学 ， 中药学， 2004， 硕士

【摘要】 本文以开发利用丹参的新资源，探明丹参的一个种下变种——白花丹参(Salviamiltiorrhiza Bunge var．alba C．Y．wu et H．W．Li)的化学成分及进行含量测定为主要目的。对白花丹参中的丹参酮进行提取分离，初步摸索了采用新的提取分离方法，着重分离得到了三种纯度较高的单体化合物。利用高效液相色谱法对白花丹参与紫花丹参中的丹参酮ⅡA进行含量测定，证明白花丹参在一定程度上可以替代丹参的药用价值，为开发利用白花丹参这一新的药用植物提供科学依据。本研究首次将复日FR-980生物电泳图像分析系统应用到对白花丹参与紫花丹参中丹参酮ⅡA的含量测定中，为扩大中药化学成分含量测定方法开拓新的研究领域。 正品丹参来源于唇形科鼠尾草植物丹参(Salvia miltiorrhiza Bunge)的干燥根及根茎，主产于陕西随州(现湖北省境内)，山东和河南等地。随着野生资源的减少，70年代中期，药材丹参已经供不应求，目前发现鼠尾草属Salvia共有40多个种(含变种、变型)的根及其根茎可作丹参使用。白花丹参为丹参的种下变种，白花丹参的花冠为白色或淡黄色，不同于正品丹参的紫色或紫红色花冠，为山东特产。据文献报道，白花丹参为优良品种，其有效成分二萜醌类成分高于正品丹参(紫花丹参)，被誉为妇科良药，因具有降血脂、降血压、延年益寿之功效，长久以来，民间得此为宝，常以礼品相赠。白花丹参野生物种极为稀少，已濒临绝迹。为挽救和开发利用这一名贵药材，1987年，国务院发布的《中国珍稀濒危保护植物名录》中，将具有活血化瘀作用的丹参Salviamiltiorrhiza和白花丹参S．miltiorrhiza Bunge var．alba包括在内。 近年来，对正品丹参的化学成分及药理作用的研究报道较多，但对白花丹参(S．miltiorrhiza．Bunge var alba C．Y．Wu et H．W．Li)的化学成分和含量测定的报道较少且不深入。到目前为止从白花丹参中仅分离鉴定了14种脂溶性单体成分，而山东省有关部门自1990年以来，对白花丹参的野生驯化、种植栽培、生药质量标准、药理药效分析、毒理试验等方面进行了研究。药理、药效研究证明，白花丹参有较强的血管扩张作用，对高血脂症和动脉粥样硬化具有显著疗效，对血栓闭塞性脉管炎具有独特疗效，临床应用前景十分广阔。吉林农业大学硕士学位论文白花丹参中丹参酮成分的提取分离与含量测定 为了给系统研究白花丹参增添新的内容，进一步证实白花丹参的药用范围，为正确评价白花丹参的质量提供理论依据，本文对白花丹参中的三个疗效显著的生物活性成分采用了新的提取分离方法及含量测定的方法，进行了研究。 本研究以山东省莱阳市天意药材基地出产的白花丹参为原料，改变了文献中记载的，以硅胶、氧化铝等柱层析，用苯、石油醚、丙酮、乙酸乙醋等极性小、毒性大、污染性强的有机溶剂进行提取分离的方法，而使用新的方法，即使用Sephadex LH一20柱，以乙醇、甲醇等为洗脱剂，进行提取分离。本文用此方法着重分离了三个二菇醒类单体成分，操作方法如下:取丹参根Ikg，洗净晾干，粉碎成粗粉，过40目筛，用1 0L95%乙醇浸提24小时，浸提3次，过滤后合并浸提液，减压浓缩成浓浸膏，用1的%甲醇溶解，浓缩甲醇溶解液成浸膏，即得总丹参酮。将总丹参酮过sephadex LH一20柱，用1 00%甲醇洗脱，分别得4个颜色组分段(黄色段、橙红色段、紫红色段、棕红色段)，各色段分别用高效硅胶板薄层层析，展开剂:苯一乙酸乙醋(19:1)，层析后挥干，在自然光下可清晰看见不同颜色斑点。根据薄层层析检测结果，按不同部分进行二次分离，再上SephadexLH一20柱，分别用50%，70%甲醇洗脱，用自动部分收集器部分收集。即可通过上述两次分离得到单体化合物。此方法操作简便，分离得率高，得到的3个单体纯度均达到92%以上。该方法不仅对操作人员的毒害性小，对环境的污染性也很小，而且成本低廉，易于操作，柱填料及洗脱剂均可反复回收利用，此方法可为工业化生产提供参考依据。 实验分离得到的三个单体化合物，通过化学反应、理化常数测定及波谱解析(紫外光谱、质谱和核磁共振)，确定它们的化学结构均为二裕类化合物，分别为:丹参酮1 IA，丹参酮I，甘西鼠尾新酮A。其中的甘西鼠尾新酮A是次甲基丹参醒的二聚体，在白花丹参中首次发现。 本文利用高效液相色谱法，检测了产自于山东同地，同年种植，同年采收的白花丹参与紫花丹参中丹参酮IIA的含量，结果表明:白花丹参中丹参酮IIA的含量高于紫花丹参，在一定程度上说明白花丹参可以替代丹参在临床上使用。在丹参的不同部位上:丹参侧根中丹参酮IIA含量最高，其次为丹参主根中丹参酮工IA含量，含量较低的为丹参茎部的丹参酮IIA含量，从而更好地确定丹参的有效药用部位。吉林农业大学硕士学位论文白花丹参中丹参酮成分的提取分离与含量测定 本实验首次使用复日FR一980生物电泳图像分析系统对白花丹参与紫花丹参中丹参酮I以的含量进行测定。根据该分析系统中数据处理器的像素定量分析原理，作者对白花丹参与紫花丹参中不同部位的丹参酮工IA的含量进行测定。此分析系统普遍应用于电泳的定量定性分析上，作者更多还原

【Abstract】 The paper is for the exploration and exploitation of a new source of tanshin, Salvia miltiorrhiza Bunge var. alba C. Y. Wu et H. W. Li, a variation of tanshin. The focus is on analyzing its chemical compositions and identifying relevant contents. Since new approach of composition isolation is applied, three compounds in tanshintone are isolated from Salvia miltiorrhiza Bunge var. alba C. Y. Wu et H. W. Li with high standard of purity. This provides key parameters for industrial manufacture. The content measure of tanshinone IIA from Salvia miltiorrhiza Bunge var. alba C. Y. Wu et H. W. Li and Salvia miltiorrhiza Bunge by HPLC proves that Salvia miltiorrhiza Bunge var. alba C. Y. Wu et H. W. Li is a substitute for tanshin to some extent. In the research, FR-980 BIAS is applied for the first time in record to measure the content of tanshinone IIA. Therefore, the field of chemical composition measure for Chinese herbal medicines has been enlarged.Tanshin, of the Salvia miltiorrhiza Bunge, is mainly found in Hubei, Shandong, and Henan Province. Since more and more wild plants gradually die out, it cannot meet demands after 1970s. 40 species of variation belonging to Salvia miltiorrhiza Bunge are being used as substitutes for tanshin. Salvia miltiorrhiza Bunge var. alba C. Y. Wu et H. W. Li, one of the variations, is only found in Shandong Province, with white or light yellow coronal, different from tanshin of purple coronal. In it, diterpenoid tanshinones with higher content than that of Salvia miltiorrhiza is always regarded effective in treating women diseases, decreasing blood liquid, lowering blood pressure and prolonging longevity. The wild species of Salvia miltiorrhiza and Salvia miltiorrhiza Bunge var. alba are so scarce that they were listed as among the rare and endangered plants to be preserved in 1987 by the State Council.In recent years scientific reports about Salvia miltiorrhiza can be frequently found, while the measure of Salvia miltiorrhiza Bunge’s compositions and content is not people’s attention. Up to now, only 14 compounds have been isolated from Salvia miltiorrhiza Bunge var. alba. Since 1990, systematic research has been conducted in Shandong Province for its taming, cultivation, quality criteria, effectiveness analysis, and toxicity tests. It proves thatSalvia miltiorrhiza Bunge var. alba is specially effective in vascular ectasis, with prosperous clinical use.In order to explore the untracked research field of Salvia miltiorrhiza Bunge var. alba, testify its medical value, and provide theoretical basis for quality appraisal, the paper makes a systematic research on three compounds by conducting novel isolation and identification approaches.The Salvia miltiorrhiza Bunge var. alba purchased from Tianyi Herbal Market in Laiyang City, Shandong Province was adapted as raw material in the research. In literature, all the approaches mentioned include alumina and silica gel column chromatography, and the poisonous solvents of benzene, ether, acetone, chloroform, and dichloromethane as well. The approach of Sephadex LH-20 column and ethanol or methanol as material is proved to be better than the above-mentioned methods and materials. My work focused on isolating three diterpenoid compounds by this means, with detailed flow as follows: take 1 kg dry tanshin root; ground and filter of 40 mm; then put into 10 L 95% ethanol for 24 hours a time, 3 repeats all together. After filtration, put the outcome into 100 % methanol, that is tanshinone. Put it to the Sephadex LH-20 column, with eluting liquid of 100% methanol, four-color sections (yellow, salmon pink, mauve, and brown-red) come into being. Filtered by silica gel plate, different color spots can be identified in the sunlight. Once more eluted by Sephadex LH-20 column, with eluting liquid of 50% and 70% methanol respectively, compounds of 92% purity can be extracted. The approach is easy to handle with low pollution and cost, meanwhile, the filling stuff in the column and the eluting liquid can be recycled. This approach is feasible for mass pr更多还原