Study On Separation And Purification Of Active Components From Radix Et Rhizoma Rhei And Other Chinese Medicinal Materials

Traditional Chinese medicine is an important magic weapon for traditional Chinese medicine prevention and control.However,Chinese medicine is difficult to enter the international market.The main reason is that the traditional Chinese medicine is complex,the purification is difficult,the product quality is unstable,and there is no quantitative standard.The first step in the use of traditional Chinese medicine is to extract,isolate and purify the active ingredients in Chinese herbal medicines by appropriate methods,and lay a foundation for the chemical structure determination,pharmacological research and industrialization of traditional Chinese medicines.The separation and purification process of traditional Chinese medicine includes two aspects:firstly,according to the nature of the crude extract,the corresponding separation methods and conditions are established,and the medicinal substances are extracted;second,the ineffective and harmful components are removed,and the active ingredients or parts are retained as much as possible.Therefore,the use of modern science and technology to separate and prepare high purity medicinal active ingredients is an important task in the modernization of traditional Chinese medicine.Macroporous Resin?MAR?,also known as fully porous resin,is an organic high polymer adsorbent with low cost and reusability,fast adsorption rate and strong adsorption,stable physicochemical properties and good selectivity,specific surface area.Large-scale features are commonly used in the production of natural products and pharmaceuticals to make up for the shortcomings of traditional separation techniques.The study selected several traditional Chinese medicines,such as Radix et Rhizoma Rhei,Nelumbinis Plumula,Stephania tetrandra S.Moor,Cimicifugae Rhizoma,Flos Lonicerea.,to optimize the separation and purification process of different Chinese medicine extracts by macroporous adsorption resin,so as to choose different pH.The combination of a buffer solution or a low concentration of ethanol and a buffer solution achieves the purpose of separating and purifying the active ingredient in the traditional Chinese medicine.After separation and purification by MAR,the purity was determined by HPLC area normalization method,which provides a reference for the pharmacological research and development of the above traditional Chinese medicine1.Separation and purification of free Anthraquinones from Radix et Rhizoma Rhei by macroporous adsorption resinA method for separation and purification of total free Anthraquinones and rhein monomers from Radix et Rhizoma Rhei by macroporous adsorption resin was established.Six different types of MAR were screened by static adsorption rate and desorption rate of total free anthraquinones and styrene acid in Radix et Rhizoma Rhei,and then the purification process was optimized by static and dynamic adsorption desorption experiments.The results showed that HPD-400 macroporous resin had good adsorption and desorption properties for free anthraquinonesand styrene acid in rhubarb,and its adsorption isotherm equation was consistent with Langmuir model.The optimal adsorption conditions were:pH 4.5.The concentration of the sample solution was 4mg/mL;the maximum sample volume was 7 BV.The optimal elution conditions for total free anthraquinones are:firstly,eluted with 70 BV of 0.2 mol/L sodium bicarbonate solution,and then eluted with 15 BV of 95%ethanol.The eluate was collected and the95%ethanol wash was calculated by HPLC area normalization.The purity of the free anthraquinones of the detached portion was increased from 41.17%to 82.60%.The optimal elution conditions for rhein acid were as follows:take equal proportion of saturated HPD-400 resin and blank HPD-400 resin column,first elute the styric acid in rhubarb with 80 BV 0.2 moL/L sodium bicarbonate,then use 15 BV 0.1 mol/L sodium carbonate and sodium hydroxide mixture was used to elute the rhein.The purity of rhein was calculated by HPLC area normalization to be 70.01%.2.Separation and purification of alkaloids from Nelumbinis Plumula using macroporous adsorption resinA method for separation and purification of isoliensinine from Nelumbinis Plumula by macroporous adsorption resin was established.The separation and purification process was optimized by the static adsorption rate and desorption rate of total alkaloids in Nelumbinis Plumula.The results showed that the X-5 MAR had good adsorption and desorption properties for the target components in Nelumbinis Plumula.The optimal adsorption conditions were as follows:adsorption time was 1 h;sample solution concentration was 9 mg/mL;maximum sample volume was 15 BV.The impurities were removed using 8 BV of 20%ethanol solution having a pH of 12,and 12 BV of 20%ethanol solution having a pH of 1 was used to elute the isoliensinine.The total purity of isochelatin was 66.95%.3.Separation and purification of alkaloids from Stephania tetrandra S.Moor by macroporous adsorption resinA method for separating and purifying tetrandrine and fangchinoline from Stephania tetrandra S.Moor by macroporous adsorption resin was established.The purification process was optimized by the static adsorption rate and desorption rate of the two total alkaloids in the powder.The results showed that the adsorption and desorption performance of HPD-400 MAR was better than that of the target component.The optimum adsorption conditions were as follows:adsorption time was 1 h;sample concentration was 10 mg/mL;maximum sample loading was 10 BV.The buffer solution containing a 12 BV ethanol content of 8%and a pH of 12 was used to remove impurities,and the 6 BV ethanol solution having a 10%ethanol content of 1 buffered the anti-chinoline and tetrandrine in the powder.The peak area normalization method was used to calculate the total purity of the two components to be 87.64%.4.Separation and purification of organic acids in Cimicifugae Rhizoma by macroporous adsorption resinA method for separating and purifying caffeic acid,cohomarin and isoferulic acid from Cimicifugae Rhizoma is established by macroporous adsorption resin.First,the caffeic acid was eluted with 0.2 mol/L NaHCO₃ solution,the optimum dosage was 30 BV;then the residual caffeic acid was further eluted with 14BV 0.2 mol/L Na₂CO₃ solution,using 16BV 0.1 mol/L Na₂CO₃ and 0.1 mol/L NaOH mixture solution.The isoferulic acid was eluted;the heparin was eluted with 12 BV 30%ethanol solution.The purity of caffeic acid,ascomycin and isoferulic acid was 77.62%,62.48%and 75.03%,respectively.5.Separation and purification of organic acid compounds from Flos Lonicerea using macroporous adsorption resinA new method for separation and purification of chlorogenic acid and 3,5-dicaffeoyl quinic acid from Flos Lonicerea by macroporous adsorption resin was established.The peak chlorogenic acid was first eluted using 18 BV of pH 6 buffer solution;the3,5-dicaffeoyl quinic acid was then eluted using 10 BV 50%ethanol.The purity of chlorogenic acid and 3,5-dicaffeoylquinic acid were 84.61% and 83.59%,respectively.