内部沸腾法强化提取若干中药有效成分的研究及评价
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摘要
我国药用植物资源丰富,中药提取的产业纷繁众多。然而,由于生产工艺的不同,许多企业在生产效率和产品质量上良莠不齐。多数企业仍沿用传统煎煮法和乙醇回流提取法作为主要的提取方法,这些方法虽然在设备投资环节上节约了生产成本,但由于生产效率低下、产品质量不高及在生产过程中能耗需求和有机溶剂消耗量过大等原因,使他们的发展进入了瓶颈。少数企业通过改良生产工艺,引进了新型的提取分离技术,如：超临界CO2萃取、微波辅助、超声波辅助等,但由于设备投资及能耗的增加,无形中增加了产品的成本,降低了企业的竞争力。针对这些问题,本课题组于2004年提出了内部沸腾提取的方法,该法在部分药用植物有效成分的提取分离中取得了较好的效果。然而,对于该法的研究尚需进一步完善。
本文针对内部沸腾法能实现低温、快速提取的特点,进一步扩大了内部沸腾法在药用植物提取上的应用范围。首先选取了香菇、灵芝、枸杞三种有效成分为多糖类天然高分子的特色植物,通过采用单因素实验对提取工艺条件的优化及与传统煎煮法的比较,评价了常压和减压内部沸腾法用于多糖类天然大分子提取的适应性；同时,也分别以匙羹藤和穿心莲两种植物为代表,研究了内部沸腾法在高色素含量植物提取中的应用。继而,通过单因素实验优化提取工艺及与后续纯化工艺的联用,探索了内部沸腾法对大孔吸附树脂再生性能的影响,也进一步扩充内部沸腾法在中药提取领域；其次,通过研究丹参、葛根、三七三种植物中的五种有效成分的提取动力学为例,依据内部沸腾法提取过程是对流扩散特点,推导并选取了合适的动力学模型,拟合了该提取过程的动力学方程,计算传质系数,并以此为基础对内部沸腾法提取过程中的热力学参数进行探讨,为该法提供理论支持；最后,分别以金银花、三七和穿心莲三种植物为例,选取指标成分为评价指标,结合构建地HPLC指纹图谱,对内部沸腾法和其他几种常见的提取技术进行了综合比较,并以模糊数学的思想对其进行了综合评判,为中草药提取工艺的优选提供了方法上的借鉴。
结果表明,在最佳的提取工艺条件下,内部沸腾能够适应于多糖类高分子物质的提取,尤其减压内部沸腾法可以对有效成分为酯溶性多糖,而单糖含量较高的药用植物实现选择性提取,并能大大降低提取过程中有机溶剂的消耗量。另一方而,对于色素含量较高的植物来说,内部沸腾法可以降低提取过程中色素的浸出量,减轻对后续纯化工艺上的压力,有助于提高大孔树脂的使用寿命。从提取动力学上来看,减压内部沸腾法的传质系数受温度影响较小,在0.07～0.14s-1,且在植物细胞破碎区和细胞完整区的传质系数基本接近,体现了对流传质的特征。与超声波辅助相比,内部沸腾法大大强化了内扩散的速度,使其传质系数比超声波辅助法大100倍以上。在提取过程中,内部沸腾法有效地降低了提取过程中的能垒,使得提取活化能较低,有利于提取地进行。整个过程的焓变△H和熵变△S均大于零,而提取自由能△G小于零,表明内部沸腾法的提取过程为吸热熵增加的过程,为自发过程。通过采用HPLC指纹图谱及相似度评价系统对几种不同提取方法的比较,结果发现内部沸腾法提取过程中不仅仅能在提取率上保证了指标成分的浸出,而且能够保障多种有效成分的同时浸出,与传统的甲醇冷浸法接近。对于维护原药有效成分的全貌,体现中医理论“君、臣、佐、使”的思想有着重要的意义。另外,通过借鉴模糊数学的思想,综合考虑了提取产品的质量、提取效率及扩大生产所需的设备投资等因素的影响,构建了评价指标模糊矩阵和权重矩阵,以数学评判的思想对减压内部沸腾法、传统煎煮法、甲醇冷浸法、超声波辅助法、微波辅助法和常压内部沸腾法进行了综合比较,结果表明内部沸腾法综合得分较高,具有巨大的推广潜力。
以上结果表明,内部沸腾法能够在较为广泛的药用植物资源中,实现中药有效成分的快速、绿色提取分离。相信在不久的将来,该法必将因其巨大的优势而被广泛投入到工业中。
In China, the medicinal plant resources are abundant, and numerous industries are utilizing the superiority of resources to produce Chinese traditional medicine. However, due to different production processes, many companies had large disparities in the efficiency and the quality of production. By far, decocting method and alcohol refluxing extraction are popular in most of the factories. Although the cost on the investment of equipment is saved, some disadvantages are clear:such as low efficiency, poor quality of the products and large consumptions of energy as well as organic solvents, which restricted the development of the companies. A few companies improved their manufacturing techniques and applied new extraction and separation technologies, for instance, extractions with supercritical CO2, microwave-assist and ultrasonic-assist, etc. But the operational cost increases with the higher equipment investment and energy consumption. For the purpose of solving these problems, a method called inner ebullition was proposed by our research team in2004. It is approved to achieve good results in separation of active constituents from some medicinal plants. The study for inner ebullition still needs to be improved, though.
Concerning the advantage of rapid extraction at a low temperature, inner ebullition was utilized for expanding its application fields:firstly, lentinus edodes, ganoderma lucidum fungus and fructus, which active constituents belong to polymer polysaccharides, were selected in this study. The evaluations of the adaptability on the extraction of polysaccharides were given by the results of single factor experiments and the comparison with the conventional decoction. Meanwhile, taking gymnema sylvestre and andrographis paniculata for example, the applications of inner ebullition in extraction of medicinal plants with high content of pigments were studied. The effects of inner ebullition on the regenerability for the macroporous absorbent resin were also investigated. In addition, the proper kinetics models were selected and the kinetics equations were simulated, according to the mass manner of convection-diffusion for inner ebullition. Then, Taking salvia miltiorrhiza, pueraria root and notoginseng for example, the kinetic equations were fitted and the mass transfer coefficient was calculated. On the basis of the extraction kinetics, the thermodynamic parameters were investigated, which provided theoretical support for inner ebullition. Finally, in order to compare with other extraction methods, HPLC fingerprints of honeysuckle, notoginseng and andrographis paniculata were constructed respectively, which were taken as tools with the specific constituents to make an evaluation. The fuzzy mathematics was also used to make a comprehensive evaluation for different extraction methods, which provides some useful information for selecting extraction technology of Chinese herbs.
The result shows the method of inner ebullition is able to be applied in the extraction of polysaccharide polymer. In particular, the decompressing inner ebullition can achieve the selective extraction for those medical plants of which active constituent belongs to acetate-soluble polysaccharides while have a high content of reducing sugar. This process can save the consumption of organic solvents. On the other hand, the inner ebullition can reduce the yield of pigment during the extraction process and relieve the pressure to the following purification process. Thus, inner ebullition contributes to prolong the service life of macroporous absorbent resin. With regard to the extraction kinetics, the mass transfer coefficient of inner ebullition was in the range of0.07～0.14s-1, which affected little by temperature. Apart from that, the mass transfer coefficients in the broken and integrity area of plant cells were very similar, which reflected the characteristic of convection. Compared with ultrasonic-assisted extraction, inner ebullition obviously accelerated the extraction rate of inner diffusion and made the mass transfer coefficient100times more than that of ultrasonic-assisted. Undoubtedly, the energy barriers were cut down and active energies were reduced during the extraction process of inner ebullition, which helped the extraction process. To be more specific, the inner ebullition is a spontaneous, endothermic process with entropy increase, which can be certified by the thermodynamic parameters as follows:ΔH, ΔS>0; ΔG<0., it can also be found in the HPLC fingerprint and the comparison with several similarity evaluation systems. The result demonstrates that inner ebullition not only ensured the extraction ratio of the specific constituents, but also several kinds of active constituents can be extracted simultaneously. The results were similar to the leaching with methanol. It was very important to preserve the active components as much as possible and to reflect the theory of Chinese medicine-"monarch, minister, assistant and guide". In addition, by applying the fuzzy mathematics, and considering some factors such as the quality of the products, extraction efficiency and required equipment investment, the following evaluation indexes such as fuzzy matrix and weight matrix were proposed. They were used to compare those different extraction methods, namely, inner ebullition extraction (under atmospheric), traditional decocting method, methanol cold-maceration, ultrasonic-assisted extraction, microwave-assisted extraction were compared. The result shows that inner ebullition is better than other processes, which has a great potential to promotion.
This study shows that inner ebullition can be widely applied in the medicinal plants, achieving their active components with rapid and environment-friendly extraction. In the near future, inner ebullition is believed to be widely used in the industry because of its great advantages.
本文针对内部沸腾法能实现低温、快速提取的特点,进一步扩大了内部沸腾法在药用植物提取上的应用范围。首先选取了香菇、灵芝、枸杞三种有效成分为多糖类天然高分子的特色植物,通过采用单因素实验对提取工艺条件的优化及与传统煎煮法的比较,评价了常压和减压内部沸腾法用于多糖类天然大分子提取的适应性；同时,也分别以匙羹藤和穿心莲两种植物为代表,研究了内部沸腾法在高色素含量植物提取中的应用。继而,通过单因素实验优化提取工艺及与后续纯化工艺的联用,探索了内部沸腾法对大孔吸附树脂再生性能的影响,也进一步扩充内部沸腾法在中药提取领域；其次,通过研究丹参、葛根、三七三种植物中的五种有效成分的提取动力学为例,依据内部沸腾法提取过程是对流扩散特点,推导并选取了合适的动力学模型,拟合了该提取过程的动力学方程,计算传质系数,并以此为基础对内部沸腾法提取过程中的热力学参数进行探讨,为该法提供理论支持；最后,分别以金银花、三七和穿心莲三种植物为例,选取指标成分为评价指标,结合构建地HPLC指纹图谱,对内部沸腾法和其他几种常见的提取技术进行了综合比较,并以模糊数学的思想对其进行了综合评判,为中草药提取工艺的优选提供了方法上的借鉴。
结果表明,在最佳的提取工艺条件下,内部沸腾能够适应于多糖类高分子物质的提取,尤其减压内部沸腾法可以对有效成分为酯溶性多糖,而单糖含量较高的药用植物实现选择性提取,并能大大降低提取过程中有机溶剂的消耗量。另一方而,对于色素含量较高的植物来说,内部沸腾法可以降低提取过程中色素的浸出量,减轻对后续纯化工艺上的压力,有助于提高大孔树脂的使用寿命。从提取动力学上来看,减压内部沸腾法的传质系数受温度影响较小,在0.07～0.14s-1,且在植物细胞破碎区和细胞完整区的传质系数基本接近,体现了对流传质的特征。与超声波辅助相比,内部沸腾法大大强化了内扩散的速度,使其传质系数比超声波辅助法大100倍以上。在提取过程中,内部沸腾法有效地降低了提取过程中的能垒,使得提取活化能较低,有利于提取地进行。整个过程的焓变△H和熵变△S均大于零,而提取自由能△G小于零,表明内部沸腾法的提取过程为吸热熵增加的过程,为自发过程。通过采用HPLC指纹图谱及相似度评价系统对几种不同提取方法的比较,结果发现内部沸腾法提取过程中不仅仅能在提取率上保证了指标成分的浸出,而且能够保障多种有效成分的同时浸出,与传统的甲醇冷浸法接近。对于维护原药有效成分的全貌,体现中医理论“君、臣、佐、使”的思想有着重要的意义。另外,通过借鉴模糊数学的思想,综合考虑了提取产品的质量、提取效率及扩大生产所需的设备投资等因素的影响,构建了评价指标模糊矩阵和权重矩阵,以数学评判的思想对减压内部沸腾法、传统煎煮法、甲醇冷浸法、超声波辅助法、微波辅助法和常压内部沸腾法进行了综合比较,结果表明内部沸腾法综合得分较高,具有巨大的推广潜力。
以上结果表明,内部沸腾法能够在较为广泛的药用植物资源中,实现中药有效成分的快速、绿色提取分离。相信在不久的将来,该法必将因其巨大的优势而被广泛投入到工业中。
In China, the medicinal plant resources are abundant, and numerous industries are utilizing the superiority of resources to produce Chinese traditional medicine. However, due to different production processes, many companies had large disparities in the efficiency and the quality of production. By far, decocting method and alcohol refluxing extraction are popular in most of the factories. Although the cost on the investment of equipment is saved, some disadvantages are clear:such as low efficiency, poor quality of the products and large consumptions of energy as well as organic solvents, which restricted the development of the companies. A few companies improved their manufacturing techniques and applied new extraction and separation technologies, for instance, extractions with supercritical CO2, microwave-assist and ultrasonic-assist, etc. But the operational cost increases with the higher equipment investment and energy consumption. For the purpose of solving these problems, a method called inner ebullition was proposed by our research team in2004. It is approved to achieve good results in separation of active constituents from some medicinal plants. The study for inner ebullition still needs to be improved, though.
Concerning the advantage of rapid extraction at a low temperature, inner ebullition was utilized for expanding its application fields:firstly, lentinus edodes, ganoderma lucidum fungus and fructus, which active constituents belong to polymer polysaccharides, were selected in this study. The evaluations of the adaptability on the extraction of polysaccharides were given by the results of single factor experiments and the comparison with the conventional decoction. Meanwhile, taking gymnema sylvestre and andrographis paniculata for example, the applications of inner ebullition in extraction of medicinal plants with high content of pigments were studied. The effects of inner ebullition on the regenerability for the macroporous absorbent resin were also investigated. In addition, the proper kinetics models were selected and the kinetics equations were simulated, according to the mass manner of convection-diffusion for inner ebullition. Then, Taking salvia miltiorrhiza, pueraria root and notoginseng for example, the kinetic equations were fitted and the mass transfer coefficient was calculated. On the basis of the extraction kinetics, the thermodynamic parameters were investigated, which provided theoretical support for inner ebullition. Finally, in order to compare with other extraction methods, HPLC fingerprints of honeysuckle, notoginseng and andrographis paniculata were constructed respectively, which were taken as tools with the specific constituents to make an evaluation. The fuzzy mathematics was also used to make a comprehensive evaluation for different extraction methods, which provides some useful information for selecting extraction technology of Chinese herbs.
The result shows the method of inner ebullition is able to be applied in the extraction of polysaccharide polymer. In particular, the decompressing inner ebullition can achieve the selective extraction for those medical plants of which active constituent belongs to acetate-soluble polysaccharides while have a high content of reducing sugar. This process can save the consumption of organic solvents. On the other hand, the inner ebullition can reduce the yield of pigment during the extraction process and relieve the pressure to the following purification process. Thus, inner ebullition contributes to prolong the service life of macroporous absorbent resin. With regard to the extraction kinetics, the mass transfer coefficient of inner ebullition was in the range of0.07～0.14s-1, which affected little by temperature. Apart from that, the mass transfer coefficients in the broken and integrity area of plant cells were very similar, which reflected the characteristic of convection. Compared with ultrasonic-assisted extraction, inner ebullition obviously accelerated the extraction rate of inner diffusion and made the mass transfer coefficient100times more than that of ultrasonic-assisted. Undoubtedly, the energy barriers were cut down and active energies were reduced during the extraction process of inner ebullition, which helped the extraction process. To be more specific, the inner ebullition is a spontaneous, endothermic process with entropy increase, which can be certified by the thermodynamic parameters as follows:ΔH, ΔS>0; ΔG<0., it can also be found in the HPLC fingerprint and the comparison with several similarity evaluation systems. The result demonstrates that inner ebullition not only ensured the extraction ratio of the specific constituents, but also several kinds of active constituents can be extracted simultaneously. The results were similar to the leaching with methanol. It was very important to preserve the active components as much as possible and to reflect the theory of Chinese medicine-"monarch, minister, assistant and guide". In addition, by applying the fuzzy mathematics, and considering some factors such as the quality of the products, extraction efficiency and required equipment investment, the following evaluation indexes such as fuzzy matrix and weight matrix were proposed. They were used to compare those different extraction methods, namely, inner ebullition extraction (under atmospheric), traditional decocting method, methanol cold-maceration, ultrasonic-assisted extraction, microwave-assisted extraction were compared. The result shows that inner ebullition is better than other processes, which has a great potential to promotion.
This study shows that inner ebullition can be widely applied in the medicinal plants, achieving their active components with rapid and environment-friendly extraction. In the near future, inner ebullition is believed to be widely used in the industry because of its great advantages.
引文
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