【作者】 龚明贵；

【导师】 尉芹； 马希汉；

【作者基本信息】 西北农林科技大学 ， 野生动植物保护与利用， 2005， 硕士

【摘要】 本文以板蓝根、大青叶及其指标性成分(靛蓝、靛玉红)为研究对象,系统研究了高效液相色谱法、分光光度法和薄层扫描法测定靛蓝、靛玉红含量的方法,对3 种方法的优缺点进行了比较分析;研究了不同炮制方法和不同回流水浴温度对板蓝根、大青叶中靛蓝、靛玉红含量的影响;研究了不同气候带板蓝根、大青叶有效成分合成积累动态规律、生长积累动态规律和不同产地板蓝根、大青叶有效成分含量差异性。结果如下: 1. 通过对高效液相色谱法、双波长分光光度法和薄层扫描法测定靛蓝、靛玉红含量的方法研究,可以看出,3 种分析方法各有优缺点,可用于不同精度下对靛蓝、靛玉红含量测定。对3 种方法稳定性、精密度、重复性和加样回收率等实验结果对比、分析表明,高效液相色谱法测定靛蓝、靛玉红各项指标均优于其他方法。双波长分光光度法由于无法克服提取液中其他成分对靛蓝、靛玉红的干扰,比其他方法测得的含量要高。薄层扫描法可用于大青叶中靛蓝、靛玉红含量的测定。如果原料中成分含量较低,薄层板展开后斑点显现不明显,影响测定结果的准确性,故该法不适宜于板蓝根中这两种成分的测定。2. 不同炮制方法对板蓝根、大青叶有效成分含量影响结果表明:60℃下烘箱中烘干板蓝根、大青叶有效成分损失最少,以60 ℃烘干为标准,高温炮制使板蓝根有效成分的损失高达40%~60%、大青叶的有效成分损失高达30%~60%;结合药材性状、有效成分含量等指标认为,50~80℃烘干及晒干处理板蓝根、大青叶是其适宜的炮制方法,高温不适宜于板蓝根和大青叶的炮制。3. 采用索氏提取方法、以氯仿为提取溶剂时,水浴温度对板蓝根、大青叶有效成分提取率影响很大,80℃水浴温度下提取率最高。4. 板蓝根、大青叶生长年限与有效成分合成积累有一定关系,当年生大青叶有效成分含量比次年生的高,板蓝根有效成分含量比次年生的低。5. 杨凌产区板蓝根不同月份有效成分含量在当年10 月至11 月中旬及次年4 月积累达到高峰。大青叶有效成分含量在当年7 月和10 月分别达到高峰,8 月份最低。板蓝根、大青叶有效成分积累高峰期出现在植株的营养生长缓慢期或停滞期。6. 北亚热带板蓝根有效成分积累高峰期比南温带早1 个月,北亚热带高峰期出现在9～10 月,南温带出现在10 月中旬～11 月中旬;南温带大青叶有效成分积累量在7 月和10 月份出现两次高峰,北亚热带8 月初、9 月初和10 月中旬出现三次积累高峰。7. 板蓝根应在当年生长量趋于稳定、外观、性状符合要求和有效成分含量相对较高的时期进行采收。不同气候带的采收期不一致,南温带板蓝根采收期应集中在当年10 月下旬至11 月上旬,北亚热带板蓝根采收集中在当年10 月中旬。大青叶应在当年分次采收。北亚热带大青叶最佳采收期可在8 月初、9 月初和10 月下旬分次进行,南更多还原

【Abstract】 Isatis indigotica is a traditional medicinal plant, belonging to the family of Cruciferae. Its roots can be prepared as the crude drug, radix isatidis, and the leaves can either be dried as another crude drug folium isatidis or be processed to the third crude drug, indigo naturalis. Indigo and indirubin both are bioactive constituents in thses three crude drugs. Three methods are systematically studied to measure these two bioactive constituents to probe their advantages and disadvantages, i.e., high performance liquid chromatography (HPLC), thin layer chromatographic scanning (TLCS) and spectrophotometry with double wavelength (DWSP) measurement. Different preparing methods are compared while processing folium isatidis and radix isatidis. The influence of the temperature of water bath on the contents of Indigo and indirubin was studied while measuring their contest according to the method described by “The Pharmacopoeia of the People’s Republic of China”. Rules of the dynamic accumulation of both index constituents in different climate zones and different growth periods and different locations are investigated. The results are as follows. 1. Based on the methodological study of three analysis methods, HPLC, TLCS, DWSP, it has been found that there exist advantages and disadvantages among them. They can be used within different scope of precision. Comparison of three methods on stabilities, precisions, respectabilities and recover rates show that HPLC is the best method. DWSP gets higher analytical results due to the interference of other impurities. TLCS is suitable to analyze indigo and indirubin contained in folium isatidis and indigo naturalis because of the higher contents, not suitable to radix isatidis for its low contents of the two constituents. 2. Different drying methods for processing radix isatidis and folium isatidis show that drying raw materials at the temperature of 60℃in an oven is the best method for its lowest losses of bioactive constituents. Drying raw materials at higher temperatures can result the losses of bioactive constituents as high as 40%~60% for the preparation of radix isatidis and 30%~60% for folium isatidis. Considering the appearance of the crude drugs, contents of bioactive contents, the suitable methods for processing crude drugs are drying at temperatures between 50~80℃or sun drying. 3. To determine the contents of indigo and indirubin in crude drugs by soxhlet extraction methods according to the pharmacopoeia, the suitable temperature of water bath is 80℃. 4. The contents of bioactive constituents are closely related to growth duration. The contents are higher in folium isatidis harvested in the first growth year than those in the second year, and reversely in radix isatidis. 5. The peak periods of the accumulation of bioactive constituents in the roots appear in July, October to the middle of November and April in the next year; in the leaves however, appear in July, and October, August shows the lowest contents. The peak periods of the accumulation of bioactive constituents are oppositely related to the growth of the plant. 6. The peak of dynamic accumulation of bioactive constituents in northern semitropical zone advances 2 weeks compared to northern temperate zone. The peak periods in the former appear in September to October; and in the middle of October to the middle of November. 7. Roots used for radix isatidis should be harvested when the contents of bioactive constituents are higher and while the growth of the plant is stable, and the appearance of the root can meet the standard. The harvest time should be varied in different climate zones. 8. The contents of bioactive constituents vary with locations where the medicinal plant grow, indicating it plays an important role for the quality of crude drugs.更多还原