【作者】 王彤；

【导师】 叶正良；

【作者基本信息】 天津医科大学 ， 药学（专业学位）， 2018， 硕士

【摘要】 目的:研究表明皂苷和多糖类成分具有丰富的药理活性,人参皂苷Rg₃和Rh₂具有显著增强免疫力的功效,但只存在于红参中且含量极低,本文旨在获得含有丰富皂苷、多糖类成分且人参皂苷Rg₃和Rh₂含量较高的参麦提取物,并对提取物进行含量测定研究。方法:通过对比红参、麦冬药材的单提和合提不同提取方式的总皂苷和总糖含量,确定提取方式;通过正交试验法研究提取溶剂倍量、提取时间和提取次数对总皂苷和总糖收率的影响,优选出最佳提取工艺;利用大孔树脂法分离红参麦冬合提液中的皂苷和糖,并考察相关参数纯化皂苷部分;全面考察人参茎叶提取过程中的溶剂用量、提取时间参数,通过大孔树脂法纯化最佳提取工艺药液中的人参皂苷作为转化工艺底物;转化工艺分别考察生物转化法中的酶转化和化学转化法中的酸转化过程中各参数,对比两种最优转化工艺效果,进而确定转化工艺的方式和方法;采用紫外可见分光光度计法、高碘酸钠滴定法和高效液相色谱法分别对提取物进行含量测定研究。结果:1.红参、麦冬采用合提方式,合提工艺的正交试验结果分析和验证样品结果显示红参麦冬合提最佳提取工艺为5倍纯化水提取3次,每次1.5 h,总皂苷含量约为2.90%,总糖含量约为52%,固形物约为53%。2.利用AB-8大孔树脂分离法,对红参和麦冬合提工艺的药液进行纯化工艺考察,结果为最大上样体积为0.7 BV(上样液约为0.5 g·mL^-1),水洗脱总糖体积为3 BV,最后95%乙醇洗脱皂苷体积为4.5 BV,纯化后总皂苷含量约为2.49%。3.人参茎叶最佳提取工艺为分别以10倍、8倍、8倍纯化水提取3次,每次1 h,最终得到含有0.34%的Rb₁的提取液;纯化工艺同样利用AB-8大孔树脂法,最大上样量为2 BV(上样药液约为0.2 g·mL^-1),纯化水冲洗2 BV除去糖类等物质,0.5 mol·L^-1 NaOH的20%乙醇溶液冲洗3 BV,结合纯化水冲洗3.5 BV,除去叶绿素等杂质并将pH调至中性,6 BV的70%乙醇洗脱皂苷,最终得到含有0.29%人参皂苷Rb₁的纯化药液。4.人参茎叶皂苷最佳转化工艺为醋酸转化法,当乙酸浓度为15%,反应温度在90℃¹00℃,反应时间为1.5 h时,可以得到约0.99%¹.10%的人参皂苷Rg₃和Rh₂。5.对所得的三份提取物进行含量测定研究,测得提取物中含有25.07%红参麦冬总皂苷、67.72%红参麦冬总糖和11.79%人参皂苷Rg₃和Rh₂。结论:1.通过提取方式对比实验和正交试验设计,以总皂苷和总糖为指标,综合考察,建立了红参和麦冬药材合提最佳提取工艺。2.利用AB-8大孔树脂法,确立了红参和麦冬的纯化工艺,纯化获得红参麦冬总皂苷提取物,且同时获得总糖部分提取物。3.建立了人参茎叶最佳提取工艺和最优纯化工艺,均以人参皂苷Rb₁为考察指标。4.建立了同时检测稀有皂苷20（S）-Rg₃、20（R）-Rg₃、20（S）-Rh₂、20（R）-Rh₂的高效液相色谱方法,系统地考察了转化工艺中的各项参数,确定了转化效果最好且适于生产的醋酸转化工艺。5.对所得提取物进行含量研究,为本项目提取物在之后生产上的含量控制提供了参考基础和科学依据。更多还原

【Abstract】 Objective:Studies have shown that ginsenosides and polysaccharides are rich in pharmacological activities.Ginsenosides Rg₃ and Rh₂ have significant immunity b-oosting effects,but they only exist in red ginseng and are extremely low in content.T-his article aims to obtain the Shenmai extract,which has rich saponins,polysaccharid-es and high content ginsenosides Rg₃ and Rh₂,and the extract content of research.Methods:The extraction method was determined by comparing the total ginsenosides and total saccharides content of different extraction methods of red ginseng and Ophiopogon japonicum.The effects of extraction methods of red ginseng and Ophiopogon japonicum.The effects of extracting solvents,extraction time and extraction times on total ginsenosides and total saccharides yield were studied by ortho gonal test method,and the best extraction process was optimized.The ginsenosides and saccharides were separated by the macroporous resin method,and the related parameters were used to purify the saponin.The solvent amount and extraction time parameters in the extraction process of the best extraction process liquid ginsenosides by the macroporous resin method as the substrate for the conversion process.Examine the parameters of transformation process in the biotransformation method and the acid transformation method and comparing the effects of the two optimal conversion processes,then determine the ways and methods of the conversion process.The content of extracts was determined by spectrophotometric method,sodium periodate titration and high performance liquid chromatography.Results:1.Red ginseng and Ophiopogon japonicus were extracted by combine-d method,the results showed that the best extracting process was 5 times purified wa-ter for 3 times,1.5 h each time.The content of total ginsenosides is about 2.90%,the content of total polysaccharides is about 52%,and the solids content is about 53%.2.Using AB-8 macroporous resin separation method to examine the purification process of red ginseng and Ophiopogon japonicus extracted liquid.The maximum sample volume was 0.7 BV(about 0.5 g·mL^-1of the sample solution).The sugar volume was 3 BV that washed with water,the final 95%ethanol eluted saponin volume was 4.5 BV,and the total ginsenosides content after purification was approximately 2.49%.3.The optimum extraction process of ginseng stems and leaves was extracted with 10 times,8 times,and 8 times purified water for 3 times,each time for 1 hour,and which includes 0.34%of Rb₁ extract was finally obtained.The purification process also used AB-8 macroporous resin method.The maximum sample volume is2 BV(on the sample solution is about 0.2 g·mL^-1),2 BV of purified water is used to remove sugar and other substances,3 BV is washed with 20%ethanol solution of 0.5mol·L^-1 NaOH,and 3.5 BV purified water adjusts the pH to neutral,6 BV of 70%ethanol eluted the ginsenosides,finally obtaining a purified liquid containing 0.29%of ginsenoside Rb₁.4.The optimum transform process of ginsenosides is acetic acid transformation.When the concentration of acetic acid is 15%,the reaction temperature is between90°C and 100℃,and the reaction time is 1.5h,about 0.99%to 1.10%of ginsenosides Rg₃ and Rh₂ can be obtained.5.The contents of the three extracts were determined.The results showed that it contained 25.07%total ginsenosides,67.72%total polysaccharides and 11.79%ginsenosides Rg₃ and Rh₂.Conclusion:1.Through the comparison of extraction method and orthogonal test method,total ginsenosides and total polysaccharides were taken as indexes to comprehensively examine and establish the optimal extrac tion process for red ginseng and Ophiopogon japonicus.2.Using AB-8 macroporous resin method,the purification process of red ginseng and Ophiopogon japonicus was established,and the total ginsenosides extract and the total polysaccharides extract were obtained.3.The optimum extraction process and the optimal purification process of ginseng stems and leaves were established,and ginsenoside Rb₁ was used as the inspection index.4.A high performance liquid chromatographic method for the simultaneous detection of rare saponins 20（S）-Rg₃,20（R）-Rg₃,20（S）-Rh₂,and 20（R）-Rh₂ was established and systematically examined in the transformation process.The parameters determine the acetic acid conversion process with the best transform effect and suitable for production.5.The contents of the obtained extracts were studied to provide a reference basis and scientific basis for the content control of the extracts in the project.更多还原