葛根素结晶工艺及溶解度测定
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摘要
天然产物有效成分是制药的重要原料。黄酮类活性成分一直是天然药物研究的热点。作为葛根的主要黄酮类活性成分,葛根素以其具有改善心脑血管循环、降低心肌耗氧量,降低血糖,防止高血压及动脉硬化等多种药理作用,日益受到广泛关注,其高纯度产品在国际市场上倍受青睐。
目前,葛根素纯化的工艺方法较多,而且各具优缺点,但均不能得到收率高的晶体。因此,研究开发葛根素新的纯化工艺路线是一个重要的研究方向。本文首先采用双注—底物结晶法对葛根素粗品进行初结晶,然后采用酸水解法除去葛根素衍生物。着重研究了结晶条件与酸解条件,筛选出了较优的工艺参数,并对葛根素在结晶溶剂中的溶解度进行了测定,为葛根素的工业化生产奠定了良好的基础。研究结果表明:无论是结晶工艺还是酸水解工艺与其它葛根素的纯化方法相比均有自己的独特之处,而且保证了较高的产品收率与纯度。为了提高葛根素的收率,本文还提出了一种回收结晶母液与酸水解母液中葛根素的工艺路线。
(1)采用双注—底物结晶法对纯度为45.94%的葛根素粗品进行结晶。通过研究结晶温度、搅拌速率、结晶时间、衬底液用量、溶析液与溶解液的用量比等因素对葛根素收率与纯度的影响规律,找出双注—底物结晶的最佳工艺条件。研究结果表明,结晶温度为50℃,搅拌速度为60r/min,衬底液用量为10mL,结晶时间为4h,溶析液与溶解液的用量比为0.5时,葛根素的收率为87.52%,纯度可达87.76%。
(2)采用酸水解法对纯度为72.52%的葛根素初结晶物进行酸解。通过研究不同强度的碱中和、中和至不同的pH值、不同的洗涤方法等因素对葛根素收率与纯度的影响规律,找出酸水解的最佳工艺条件。研究结果表明,采用NaOH中和至pH为4,过滤、滤饼干燥后采用方法—洗涤,葛根素的收率为62.18%,纯度可达98.19%。
(3)本文中提出一种溶剂法回收结晶母液与酸水解母液中葛根素的工艺路线,其回收率分别为43.17%与32.69%。
The effective ingredients of natural products are the important raw material of pharmacy. Active compounds, especially flavonoids, have been the focus of raw material drug. As the main active component of pueraria flavonoids, puerarin is paid more attention increasingly for it can improve the blood vessel circulation of heart and brain, reduce the oxygen consumption capacity of cardiac muscle, reduce the blood sugar, prevent the hypertension and artery teriosclerosis and so on many kinds of pharmacological action, the pure product is very popular in international market.
At present, there are many of purification techniques of puerarin which have different Advantages and disadvantages, but high yield can not be get. Therefore it is of great interest and significance to explore novel techniques to purify puerarin. In this paper, the purification techniques of puerarin are as follows: Coarse puerarin was crystallized by double-injection. Then, acid hydrolyzing process was imported to remove the ramification of pueraria. Emphatically, the condition of the crystal and acid hydrolyzing process were studied and the solubility of puerarin in crystal solvent was determined. The optimal operation parameters were chosen and consequently establish the favorable base for the industrialized production of puerarin. The experimental results indicate that, compared with some other methods purification, both the double-injection crystallization techniques and the acid hydrolyzing techniques have their inimitable traits. Furthermore, the techniques can ensure relative higher yield and purity of product. In order to get high yield, this paper bring forward a reclaim method of puerarin in crystal solution and acid hydrolyzing solution.
(1) The coarse puerarin in purity of 45.94% was crystallized by double-injection. The influences of crystal temperature, stirring velocity, crystal time, volume of substrate, ratio of the solventing-out solvent and the soluble solution were investigated in detail to find the optimal crystal techniques. It was found that the yield and purity of puerarin can reach 87.52% and 87.76% respectively, when the temperature is 45℃, the stirring velocity is 60r/min, the volume of substrate is 10mL, the crystal time is 4h, the ratio of the solventing-out solvent and the soluble solution is 0.5.
(2) The puerarin in purity of 75.52% after first step crystalization was hydrolyzed by acid. The influences of neutralization by different alkali neutralization, neutralizing to different pH, different washing mode were investigated in detail to find the optimal hydrolyzing techniques. The experimental results indicated that by taking NaOH to pH=4, filtrating, desiccating, washing by mode 1, the yield and purity of puerarin can reach 98.19% and 62.18% respectively.
(3) This paper establishes a reclaiming techniques of puerarin in crystal solution and acid hydrolyzing solution by solvent. The reclaiming yield of puerarin can reach 43.17% for the crystal solution and 32.69% for the acid hydrolyzing solution, respectively.
目前,葛根素纯化的工艺方法较多,而且各具优缺点,但均不能得到收率高的晶体。因此,研究开发葛根素新的纯化工艺路线是一个重要的研究方向。本文首先采用双注—底物结晶法对葛根素粗品进行初结晶,然后采用酸水解法除去葛根素衍生物。着重研究了结晶条件与酸解条件,筛选出了较优的工艺参数,并对葛根素在结晶溶剂中的溶解度进行了测定,为葛根素的工业化生产奠定了良好的基础。研究结果表明:无论是结晶工艺还是酸水解工艺与其它葛根素的纯化方法相比均有自己的独特之处,而且保证了较高的产品收率与纯度。为了提高葛根素的收率,本文还提出了一种回收结晶母液与酸水解母液中葛根素的工艺路线。
(1)采用双注—底物结晶法对纯度为45.94%的葛根素粗品进行结晶。通过研究结晶温度、搅拌速率、结晶时间、衬底液用量、溶析液与溶解液的用量比等因素对葛根素收率与纯度的影响规律,找出双注—底物结晶的最佳工艺条件。研究结果表明,结晶温度为50℃,搅拌速度为60r/min,衬底液用量为10mL,结晶时间为4h,溶析液与溶解液的用量比为0.5时,葛根素的收率为87.52%,纯度可达87.76%。
(2)采用酸水解法对纯度为72.52%的葛根素初结晶物进行酸解。通过研究不同强度的碱中和、中和至不同的pH值、不同的洗涤方法等因素对葛根素收率与纯度的影响规律,找出酸水解的最佳工艺条件。研究结果表明,采用NaOH中和至pH为4,过滤、滤饼干燥后采用方法—洗涤,葛根素的收率为62.18%,纯度可达98.19%。
(3)本文中提出一种溶剂法回收结晶母液与酸水解母液中葛根素的工艺路线,其回收率分别为43.17%与32.69%。
The effective ingredients of natural products are the important raw material of pharmacy. Active compounds, especially flavonoids, have been the focus of raw material drug. As the main active component of pueraria flavonoids, puerarin is paid more attention increasingly for it can improve the blood vessel circulation of heart and brain, reduce the oxygen consumption capacity of cardiac muscle, reduce the blood sugar, prevent the hypertension and artery teriosclerosis and so on many kinds of pharmacological action, the pure product is very popular in international market.
At present, there are many of purification techniques of puerarin which have different Advantages and disadvantages, but high yield can not be get. Therefore it is of great interest and significance to explore novel techniques to purify puerarin. In this paper, the purification techniques of puerarin are as follows: Coarse puerarin was crystallized by double-injection. Then, acid hydrolyzing process was imported to remove the ramification of pueraria. Emphatically, the condition of the crystal and acid hydrolyzing process were studied and the solubility of puerarin in crystal solvent was determined. The optimal operation parameters were chosen and consequently establish the favorable base for the industrialized production of puerarin. The experimental results indicate that, compared with some other methods purification, both the double-injection crystallization techniques and the acid hydrolyzing techniques have their inimitable traits. Furthermore, the techniques can ensure relative higher yield and purity of product. In order to get high yield, this paper bring forward a reclaim method of puerarin in crystal solution and acid hydrolyzing solution.
(1) The coarse puerarin in purity of 45.94% was crystallized by double-injection. The influences of crystal temperature, stirring velocity, crystal time, volume of substrate, ratio of the solventing-out solvent and the soluble solution were investigated in detail to find the optimal crystal techniques. It was found that the yield and purity of puerarin can reach 87.52% and 87.76% respectively, when the temperature is 45℃, the stirring velocity is 60r/min, the volume of substrate is 10mL, the crystal time is 4h, the ratio of the solventing-out solvent and the soluble solution is 0.5.
(2) The puerarin in purity of 75.52% after first step crystalization was hydrolyzed by acid. The influences of neutralization by different alkali neutralization, neutralizing to different pH, different washing mode were investigated in detail to find the optimal hydrolyzing techniques. The experimental results indicated that by taking NaOH to pH=4, filtrating, desiccating, washing by mode 1, the yield and purity of puerarin can reach 98.19% and 62.18% respectively.
(3) This paper establishes a reclaiming techniques of puerarin in crystal solution and acid hydrolyzing solution by solvent. The reclaiming yield of puerarin can reach 43.17% for the crystal solution and 32.69% for the acid hydrolyzing solution, respectively.
引文
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[2]Mira J.Antioxidative,anticancer and antiinflamrrmtory properties of isoflavones from kudzu(Pueraria lobata)root[D].Rutgers The State university of New Jersey-New Brunswick,2003
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[4]陶忠华.新药葛根素的研究[J].中国新药杂志,1993,2(6):1-3
[5]张来九.一种高纯度药用葛根素的提取方法[P].中国:CN1276378A,1999.06.03
[6]左春耀,刘家骐,吴成军,等.制取葛根素的方法[P].中国:CN1154849C,1996.01.28
[7]左春旭.葛根素及葛根素注射液的研究与开发[J].中国中药报,1997.01.13
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