摘要
微波辅助提取技术是近年来发展较快的一种新型提取技术,与传统提取方法相比,具有操作简便、速度快、效率高、耗能少等优点,在中草药有效成分的提取中得到了广泛应用。本文研究采用微波法提,取复方中药乳块消,主要研究内容和结论如下:
1、应用高效液相色谱法建立了复方中药乳块消的指纹图谱,对指纹图谱中的色谱峰进行了分析,确定了19个主要的共有色谱峰,并对共有色谱峰进行了相对保留时间(RRT),相对峰面积(RPA),以及相似度的计算。应用缺味药材(阴性对照)与单味药材(阳性对照)提取物分别制备分析的方法,对复方中药乳块消指纹图谱中的色谱峰进行了来源分析鉴定。结果表明色谱峰主要来源于丹参和橘叶,证明了丹参和橘叶为乳块消药材中的“君药”的中医理论学说。
2、应用已经建立的高效液相色谱条件,分别测定了27批乳块消传统工艺水提取液与23批乳块消传统工艺醇提取液的指纹图谱。应用中药指纹图谱分析软件确定了水提液中8个主要的共有色谱峰及醇提液中的3个主要的共有色谱峰,并对各色谱峰的峰面积变化范围作了计算与鉴定。以传统工艺为参照,应用指纹图谱考察微波提取复方中药乳块消液的可行性。结果表明,当微波水提取条件为:温度85℃,时间14min,提取功率500W时,主要共有色谱峰可落入传统工艺色谱峰变化范围内;当微波醇提取条件为:温度70℃,时间4min,提取功率600W时,主要共有色谱峰可落入传统工艺色谱峰变化范围内。
3、通过大孔树脂的静态吸附与解吸实验考察了7种不同极性,不同孔径的HPD系列大孔树脂对丹参酮ⅡA的吸附解吸作用。考察了树脂的吸附动力学曲线,并应用Langmuir方程对曲线进行了吸附速率常数的计算;考察了树脂于25℃时的吸附等温线,并应用Langmuir方程与Freundlich方程进行了吸附等温线的拟合。结果表明非极性的HPD-100树脂的吸附解吸效果较好,是用于吸附分离丹参酮ⅡA较理想树脂。用树脂柱层析及乙醇动态梯度洗脱实验,进行丹参酮ⅡA的动态分离。结果表明,80%乙醇可以较好的洗脱丹参酮ⅡA,洗脱体积达到10BV时洗脱过程基本结束,所得丹参酮ⅡA的纯度为42%左右,回收率为82%,与上样液相比,纯度提高了2.5倍左右。
Microwave-assisted extraction(MAE) technique has become a fast developing technique in recent years.Compared with conventional methods,microwave-assisted extraction has many advantages.Because of its process simplicity,fast speed,high efficiency,less energy consuming and other advantages,it is used widely in the extraction of effective components from traditional Chinese medicine.In this thesis, MAE was used to extract compound traditional Chinese medicine Ru-Kuai-Xiao(RKX),the main research contents and conclusions are as follows:
1、A simple and reproducible method was developed by high performance liquid chromatography to establish the fingerprint of compound traditional Chinese medicine RKX.Under the optimum chromatographic condition,19 main peaks were selected as the common peaks.The relative retention time(RRT),relative peak area(RPA) and similarities were calculated.In addition,both negative control samples and positive control samples were investigated to determine the origins of these common peaks.The results showed that these common peaks in RKX derived mainly from Radix Salviae Miltiorrhizae and Folium Citrus Reticulatae,which testified the TCM theory that these two herbs were the sovereign drugs in RKX and took main responsibility for the pharmaceutical activity.
2、27 batches RKX water extract and 23 batches RKX ethanol extract prepared according to conventional technology were tested by the HPLC fingerprint chromatogram method.8 main common peaks in water extract and 3 main common peaks in ethanol extract were selected and signed. The area range of these common peaks was calculated.According to fingerprint chromatogram obtained by conventional technology,the feasibility of using MAE to extract RKX was tested.The results showed that the main common peaks area were within the range when MAE process for water extract and ethanol extract were carrying out under 85℃for 14 minutes with power of 500W and 700℃for 4 minutes with power of 600W,respectively.
3、Seven macroporous resins with different polarities and pore sizes were evaluated to determine the static adsorption and desorption of tanshinoneⅡA.Adsorption kinetics curve was obtained and Langmuir equation was used to calculate the constant of adsorption speed.Both Langmuir and Frendlich isotherms were used to describe the interactions between solutes and resins at 25℃.The results showed that HPD-100 displayed higher adsorption and desorption performance for tanshinoneⅡA than other resins.Dynamic adsorption experiments showed that 80% ethanol could desorb tanshinoneⅡA completely,and the desorprion process ended when the desorption solution reached about 10BV.The tanshinoneⅡA content in the final product was increased 2.5-fold from 17%to 42%,with a recovery yield of 80%.
引文
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