【作者】 郭荣辉；

【导师】 李晖；

【作者基本信息】 四川大学 ， 应用化学， 2004， 硕士

【摘要】 本文以具有免疫抑制作用的中药苏木为研究对象,在建立苏木中总酚含量分光光度测定方法的基础上,以水、35%、65%、95%乙醇为溶剂,通过渗漉法、煎煮法、索氏法、超声波法对苏木的提取工艺进行了优选。并利用水、65%和95%乙醇提取物进行了免疫药效学的动物实验研究,以及苏木中成分的分离纯化、鉴定等进行了初步探讨。首先以苏木精为标准物,建立了分光光度法测定总酚含量的分析方法, 回归方程为C=10.2569A-0.07901,相关系数为0.9999。研究结果表明,在0～16.0μg/ml范围内呈良好线性关系,方法简便、快速、灵敏度高,具有一定的推广和应用价值。以总酚含量为考察指标,分别以水、35%、65%和95%乙醇作溶剂,利用正交设计法对苏木的渗漉法、索氏法、超声波法提取工艺进行了优化。结果显示,用水提取时,最佳方法为索氏提取法,其工艺为加10倍量水,提取2次,每次4.5h;用35%乙醇提取时,最佳方法为索氏提取法,其工艺为加20倍量35%乙醇,提取2次,每次3h;用65%乙醇提取时,最佳方法为超声波提取法,其工艺为加20倍量65%乙醇,提取3次,每次1.5h;用95%乙醇提取时,最佳方法为索氏提取法,其工艺为加10倍量95%乙醇,提取3次,每次6h。为苏木的进一步开发提供了理论依据。利用水、65%、95%乙醇提取物进行了免疫药效学的动物实验研究,考察了上述提取物对红细胞所致的小鼠溶血素抗体生成的影响以及小鼠网状内皮系统对血流中惰性碳粒的吞噬功能的影响。结果表明,三种提取物均具有抑制溶血素抗体生成的趋势,但无统计学上的意义;高、中、低剂量的苏木水提取物和高剂量的65%乙醇提取物对网状内皮吞噬功能具有明显的抑制作用,从而进<WP=3>一步证实了苏木的水提取物具有免疫抑制作用。根据苏木免疫药效学动物实验的研究结果,以出膏率和总酚含量为考察指标,在原苏木水提取工艺基础上,采用煎煮法对水提取工艺进行了进一步的优选。结果表明,苏木水提取的最佳工艺为加10倍量水,煎煮3次,每次1.5h。利用柱层析法分离纯化,用薄层色谱法(TLC)进行纯度鉴定,对苏木水提取物中的成分进行了初步探讨。结果表明,经过四个步骤分离,得到4种纯物质,其中两种为酚类化合物,一种为羟肟酸。本研究中,有关苏木中总酚含量的测定方法、苏木提取工艺的正交优选法以及采用溶血素法和碳粒廓清法对苏木提取物进行的药效学研究,至今未见报道。所得结果为苏木免疫抑制作用成分及其它活性成分的研究奠定了理论基础,对进一步开发苏木的药用价值具有非常重要的意义。更多还原

【Abstract】 Caesalpinia Sappan L. that had immunosuppressive function was investigated in this paper. On the basis of establishing the method of determining content of hydroxybenzene in Caesalpinia Sappan L., extractive technology was investigated by orthogonal design. Permeation, soxhlet and ultrasonic extraction were surveyed with water, thirty-five, sixty-five and ninety-five percent of ethanol as solvents. The immuneological functions of watery extract, sixty-five and ninety-five percent of ethanol extracts of Caesalpinia Sappan L. in mice were studied. Separation and identification of Components were primarily researched.With haematoxylin as the standard, the determining method of content of total hydroxybenzene in Caesalpinia Sappan L. was established by spectrophotometry. The regression of haematoxylin equation proved the calibration curve: C=10.2569×A-0.07901 and the coefficient was R=0.9999. The result showed that there was a good linearity in the range of 0～16μg/ml. The method was simple and quick, so it worthed popularizing and applying. Taking content of total hydroxybenzene as parameters, the extractive process was investigated by orthogonal design. Permeation, soxhlet and ultrasonic extraction were surveyed with water, thirty-five, sixty-five and ninety-five percent of ethanol as solvents. The results indicated that the optimal extractive methods were soxhlet extraction; soxhlet extraction; ultrasonic extraction and soxhlet extraction each. Technologies were adding ten times of water, soxhlet extracting twice and four point five hour; adding twenty times of thirty-five percent of ethanol, soxhlet extracting <WP=5>three times and three hour per time; adding twenty times of sixty-five percent of ethanol, ultrasonic extracting three times and one point five hour per time; adding ten times of ninety-five percent of ethanol, soxhlet extracting three times and six hour per time each. It afforded important reference to open up Caesalpinia Sappan L. The immuneological functions of watery extract, sixty-five and ninety-five percent of ethanol extracts of Caesalpinia Sappan L. in mice were studied. It was reviewed that extracts had effects on the production of hemolysin antibody by CRBC and meshy endothelium phagocytosis function in mice. In conclusion, three extracts restrained the form of hemolysin antibody, but the result wasn’t significant in statistics. However, high, middle and low doses of the watery extract and high dose of sixty-five of ethanol extract of Caesalpinia Sappan L. markedly restrained endodermis phagocytosis function. Therfore, immunosuppressive function of watery extract was confirmed. Taking yield of dry extract and content of total hydroxybenzene as parameters, extractive technology was further surveyed with water as solvent. The results showed that the optimal extractive technology was adding ten times of water, decoction extracting three times and one point five hour per time.Components of Watery extract were separated with glass column and each part was inspected and identified with TLC. As a result, four compounds were obtained from Caesalpinia Sappan L. by four processes. Two compounds were hydroxybenzene and one compound was hydroximic acid. Determination of total hydroxybenzene, extractive technology by orthogonal design and immuneological effect in way of hemolysin and carbon particle clearance determinations of Caesalpinia Sappan L.were studied for the first time. The results established academic foundation for researching components of immunosuppressive functions and other effective components. In addition, they had important significance for further making use of Caesalpinia Sappan L.更多还原