雷公藤有效成分的提取分离和麻疯树籽的开发利用研究
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摘要
超临界流体萃取(SFE)是环境友好的化工分离技术,在天然产物和中药领域被视为提取分离现代化的关键技术之一。本文采用超临界流体萃取结合其它分离技术分别对雷公藤有效成分的提取分离和麻疯树籽的综合开发利用进行了研究。
雷公藤是我国传统中药,具有抗炎抗菌、免疫抑制和抗生育等药理作用,其中雷公藤甲素被公认为最主要的活性成分,雷公藤红素被认为具有较大毒副作用,但又具有明显的抗癌作用。
本文在实验室成员前期研究基础上,采用100升超临界流体萃取工业化装置分别对从雷公藤根芯和根皮中提取有效成分的SFE小试工艺的放大进行了研究。结果表明,当以雷公藤根芯为原料,以增加雷公藤甲素的提取率和降低雷公藤红素的提取率为研究目标,按优化的小试工艺条件,以75%乙醇水溶液为夹带剂进行超临界二氧化碳萃取时,放大工艺下的雷公藤甲素和雷公藤红素的提取率分别为传统提取方法的1.56和2.27倍;与SFE小试实验结果相比具有相同数量级,表明放大工艺可行。当以雷公藤根皮为原料,以提高雷公藤红素的提取率为研究目标,按优化的小试工艺条件,以乙醇为夹带剂进行超临界二氧化碳萃取时,放大工艺下的雷公藤红素的提取率是传统提取方法的1.28~2.56倍;是SFE小试实验的1.02倍,表明了放大实验与小试实验结果的一致性和放大的可行性。
本文还利用上述工业规模超临界流体萃取所得的雷公藤根芯和根皮的浸膏为原料,对其中的雷公藤甲素与雷公藤红素的进一步分离进行了研究,优化得到了“酸沉-碱化-溶剂萃取”的工艺路线和各步骤的适宜工艺条件。采用高效液相(HPLC)仪器对分离后的三种产品的检测结果表明,在优化条件下,雷公藤红素产品中检测不到雷公藤甲素;雷公藤甲素产品中检测不到雷公藤红素;总生物碱产品中也未检测出雷公藤甲素和雷公藤红素,表明优化的分离工艺路线可使雷公藤红素、雷公藤甲素、总生物碱三种有效成分均得到较好的分离和富集。
上述研究表明,本文建立的超临界流体萃取-酸沉-碱化-有机溶剂萃取的工艺路线既能高效提取雷公藤甲素和雷公藤红素又能有效地将雷公藤红素、雷公藤甲素和总生物碱三种有效成分分离,从而为中药制剂雷公藤根芯和根皮的合理利用提供了新的途径和技术支持。
麻疯树是一种籽油含量很高的非食用油料植物,为世界公认的最有可能成为未来替代化石能源的具有巨大开发潜力的树种。为实现对麻疯树籽资源的合理利用,本文采用超临界二氧化碳萃取技术对从麻疯树籽中提取籽油进行了研究,同时还对麻疯树籽中具有杀虫活性的皂苷的提取条件进行了考察。
首先采用超临界二氧化碳萃取技术从麻疯树籽中提取麻疯树籽油。对工艺参数(如原料粒度、提取温度和提取压力等)对麻疯树籽油提取率的影响进行了实验研究,结果表明,当麻疯树籽仁粉的粒度为40~60目、萃取压力40MPa、萃取温度55℃时,可使麻疯树籽油达到最大提取率为51.5%,回收率为92.1%。其次进行了中试放大实验研究,结果麻疯树籽油的提取率为47.25%,回收率为84.5%。虽然中试的提取率低于小试结果,但优于一般传统的提取方法,表明超临界二氧化碳萃取工艺的可行性。
另外,本文还以超临界萃取除油后的籽仁为原料,采用乙醇回流法对籽仁中皂苷成分的提取进行了研究。设计正交实验考察了水浴温度、乙醇浓度、料液比和提取次数对总皂苷提取率的影响。由极差分析知影响因素的排列顺序为:提取次数>料液比>乙醇浓度>水浴温度;最佳的提取条件为水浴温度80℃,乙醇浓度80%,料液比1:10,提取3次。在所选实验范围内,总皂苷的最大提取率为1.955%,回收率为88.64%。
最后,本文还基于超临界二氧化碳萃取麻疯树籽油的实验数据,计算了麻疯树籽油在超临界流体中的溶解度,并采用Chrastil方程和修正的Chrastil方程对溶解度数据进行关联,关联结果的平均相对误差分别为10.10%和3.468%,表明关联结果较好;采用破碎-完整细胞模型和两步扩散模型对SC-CO_2萃取麻疯树籽油的传质过程进行了模拟,拟合的平均相对误差分别为1.08%～3.67%和3.01%～10.24%,表明模拟结果良好。上述理论研究为麻疯树籽油的超临界二氧化碳萃取工艺的工业放大与设计提供基础数据和技术支持。
Supercritical fluid extraction (SFE) is an environmentally friendly chemical separation technology, which is considered as one of the key technologies in the modernization of extracting and separating natrual products and traditional Chinese medicine. In this paper, SFE combined with other advanced technologies was used to make both the extraction and separation of active components from Tripterygium Wilfordii Hook.f. (T. Wilfordii) and the utilization of the seeds of Jatropha curcas (J. curcas).
T. Wilfordii is a traditional Chinese medicine, which has many pharmacological actions such as anti-inflammatory, antibacterial, immunosuppressive and antifertility. Triptolide is considered as the main active component while tripterine as the main toxic component. Besides, tripterine is reported to be strongly anticancer.
Based on the results of preliminary studies, industrial SFE equipment was used for the sacle-up of SFE processes of T. Wilfordii. The peeled roots of T. Wilfordii were extracted with supercritical CO_2 (SC-CO_2) +75% ethanol, aiming to increase the yield of triptolide and decrease the yield of tripterine. The yields of triptolide and tripterine were 1.56 and 2.27 times as much as those of traditional methods, respectively. The yeilds were lower but still in the same magnitude compared to those of pilot experiments, indicating the scale-up process was feasible. The root bark of T. Wilfordii was extracted with SC-CO_2 + ethanol, aiming to increase the yield of tripterine. The yield of tripterine was 1.28-2.56 and 1.02 times as much as those of traditional methods and pilot experiments, respectively. The results were consistent to pilot experiments, indicating the feasibility of the scale-up process.
An“acidification–alkalization–organic solvent extraction”process was designed and investigated for further separation of triptolide and tripterine in the extracts obtained from the scale-up SFE experiments. High performance liquid chromatography was performed for the detection of triptolide and tripterine in the products. Under optimal separation condtions, the following results could be obtained: no triptolide was detectable in the product of tripterine, no tripterine was detectable in the product of triptolide, and neither triptolide nor tripterine was detectable in the product of total alkaloids. Therefore triptolide, tripterine and total alkaloids were well separated and enriched with the optimal separation process.
Above results showed the process of“SFE-acidification-alkalization-organic solvent extraction”can efficiently extract triptolide and tripterine, and well separate triptolide, tripterine and total alkaloids in the extracts. It provided a new approach and technology support for the appropriate utilization of T. Wilfordii. With a high content of non-edible seed oil, J. curcas is world recognized as the most potential plant to be the substitute of fossils. In this study, jatropha seed oil was extracted with SC-CO_2 and the extraction of molluscacidal saponins from the seeds was explored.
Firstly, seed oil was extracted from the seed kernels of J. curcas by SC-CO_2. The effect of some factors such as particle size, extraction temperature and pressure on the yield of seed oil was investigated. A maximum yield of 51.5% (recovery 92.1%) can be obtained under the optimal conditions (particle size: 250-380μm, 55℃and 40MPa). The scale-up experiment of the optimal conditions gave a yield of 47.25% (recovery 84.5%), which was lower than that of pilot experiemnts but higher than that of traditional methods, indicating the SC-CO_2 extraction process was feasible.
Secondly, saponins were extracted from the de-oiled seed kernels by heat reflux extraction with ethanol solution. The influences of water temperature, concentration of ethanol solution, solid-liquid ratio and extraction times on the yield of total saponins were investigated by an orthogonal design. The range analysis showed the factors were in an order of extraction times > solid-liquid ratio > concentration of ethanol solution > water temperature. And the optimal extraction conditions were extraction with 80% ethanol with a solid-liquid ratio of 1:10 under the water temperature of 80℃for 3 times. The maximum yield of total saponins was 1.955% (recovery 88.64%) among the experiments.
Lastly, the solubilities of jatropha seed oil in SC-CO_2 at definite temperatures and pressures were caculated from the initial slope of the extraction curve of yield versus volumes of SC-CO_2 and correlated with the Chrastil equation and modified Chrastil equation with AARDs of 10.10 % and 3.468 %, respectively. In addition, the SC-CO_2 extraction process of jatropha seed oil was simulated with the broken and intact cell model and two-stage diffusion model, respectively. The values of AARDs were in the range of 1.08-3.67 % and 3.01%-10.24%, respectively, indicating that both models had good simulation results. The above theoretical studies therefore provide basic data and technoglogy support for the industrial scale-up and process design of the SC-CO_2 extraction of jatropha seed oil.
雷公藤是我国传统中药,具有抗炎抗菌、免疫抑制和抗生育等药理作用,其中雷公藤甲素被公认为最主要的活性成分,雷公藤红素被认为具有较大毒副作用,但又具有明显的抗癌作用。
本文在实验室成员前期研究基础上,采用100升超临界流体萃取工业化装置分别对从雷公藤根芯和根皮中提取有效成分的SFE小试工艺的放大进行了研究。结果表明,当以雷公藤根芯为原料,以增加雷公藤甲素的提取率和降低雷公藤红素的提取率为研究目标,按优化的小试工艺条件,以75%乙醇水溶液为夹带剂进行超临界二氧化碳萃取时,放大工艺下的雷公藤甲素和雷公藤红素的提取率分别为传统提取方法的1.56和2.27倍;与SFE小试实验结果相比具有相同数量级,表明放大工艺可行。当以雷公藤根皮为原料,以提高雷公藤红素的提取率为研究目标,按优化的小试工艺条件,以乙醇为夹带剂进行超临界二氧化碳萃取时,放大工艺下的雷公藤红素的提取率是传统提取方法的1.28~2.56倍;是SFE小试实验的1.02倍,表明了放大实验与小试实验结果的一致性和放大的可行性。
本文还利用上述工业规模超临界流体萃取所得的雷公藤根芯和根皮的浸膏为原料,对其中的雷公藤甲素与雷公藤红素的进一步分离进行了研究,优化得到了“酸沉-碱化-溶剂萃取”的工艺路线和各步骤的适宜工艺条件。采用高效液相(HPLC)仪器对分离后的三种产品的检测结果表明,在优化条件下,雷公藤红素产品中检测不到雷公藤甲素;雷公藤甲素产品中检测不到雷公藤红素;总生物碱产品中也未检测出雷公藤甲素和雷公藤红素,表明优化的分离工艺路线可使雷公藤红素、雷公藤甲素、总生物碱三种有效成分均得到较好的分离和富集。
上述研究表明,本文建立的超临界流体萃取-酸沉-碱化-有机溶剂萃取的工艺路线既能高效提取雷公藤甲素和雷公藤红素又能有效地将雷公藤红素、雷公藤甲素和总生物碱三种有效成分分离,从而为中药制剂雷公藤根芯和根皮的合理利用提供了新的途径和技术支持。
麻疯树是一种籽油含量很高的非食用油料植物,为世界公认的最有可能成为未来替代化石能源的具有巨大开发潜力的树种。为实现对麻疯树籽资源的合理利用,本文采用超临界二氧化碳萃取技术对从麻疯树籽中提取籽油进行了研究,同时还对麻疯树籽中具有杀虫活性的皂苷的提取条件进行了考察。
首先采用超临界二氧化碳萃取技术从麻疯树籽中提取麻疯树籽油。对工艺参数(如原料粒度、提取温度和提取压力等)对麻疯树籽油提取率的影响进行了实验研究,结果表明,当麻疯树籽仁粉的粒度为40~60目、萃取压力40MPa、萃取温度55℃时,可使麻疯树籽油达到最大提取率为51.5%,回收率为92.1%。其次进行了中试放大实验研究,结果麻疯树籽油的提取率为47.25%,回收率为84.5%。虽然中试的提取率低于小试结果,但优于一般传统的提取方法,表明超临界二氧化碳萃取工艺的可行性。
另外,本文还以超临界萃取除油后的籽仁为原料,采用乙醇回流法对籽仁中皂苷成分的提取进行了研究。设计正交实验考察了水浴温度、乙醇浓度、料液比和提取次数对总皂苷提取率的影响。由极差分析知影响因素的排列顺序为:提取次数>料液比>乙醇浓度>水浴温度;最佳的提取条件为水浴温度80℃,乙醇浓度80%,料液比1:10,提取3次。在所选实验范围内,总皂苷的最大提取率为1.955%,回收率为88.64%。
最后,本文还基于超临界二氧化碳萃取麻疯树籽油的实验数据,计算了麻疯树籽油在超临界流体中的溶解度,并采用Chrastil方程和修正的Chrastil方程对溶解度数据进行关联,关联结果的平均相对误差分别为10.10%和3.468%,表明关联结果较好;采用破碎-完整细胞模型和两步扩散模型对SC-CO_2萃取麻疯树籽油的传质过程进行了模拟,拟合的平均相对误差分别为1.08%～3.67%和3.01%～10.24%,表明模拟结果良好。上述理论研究为麻疯树籽油的超临界二氧化碳萃取工艺的工业放大与设计提供基础数据和技术支持。
Supercritical fluid extraction (SFE) is an environmentally friendly chemical separation technology, which is considered as one of the key technologies in the modernization of extracting and separating natrual products and traditional Chinese medicine. In this paper, SFE combined with other advanced technologies was used to make both the extraction and separation of active components from Tripterygium Wilfordii Hook.f. (T. Wilfordii) and the utilization of the seeds of Jatropha curcas (J. curcas).
T. Wilfordii is a traditional Chinese medicine, which has many pharmacological actions such as anti-inflammatory, antibacterial, immunosuppressive and antifertility. Triptolide is considered as the main active component while tripterine as the main toxic component. Besides, tripterine is reported to be strongly anticancer.
Based on the results of preliminary studies, industrial SFE equipment was used for the sacle-up of SFE processes of T. Wilfordii. The peeled roots of T. Wilfordii were extracted with supercritical CO_2 (SC-CO_2) +75% ethanol, aiming to increase the yield of triptolide and decrease the yield of tripterine. The yields of triptolide and tripterine were 1.56 and 2.27 times as much as those of traditional methods, respectively. The yeilds were lower but still in the same magnitude compared to those of pilot experiments, indicating the scale-up process was feasible. The root bark of T. Wilfordii was extracted with SC-CO_2 + ethanol, aiming to increase the yield of tripterine. The yield of tripterine was 1.28-2.56 and 1.02 times as much as those of traditional methods and pilot experiments, respectively. The results were consistent to pilot experiments, indicating the feasibility of the scale-up process.
An“acidification–alkalization–organic solvent extraction”process was designed and investigated for further separation of triptolide and tripterine in the extracts obtained from the scale-up SFE experiments. High performance liquid chromatography was performed for the detection of triptolide and tripterine in the products. Under optimal separation condtions, the following results could be obtained: no triptolide was detectable in the product of tripterine, no tripterine was detectable in the product of triptolide, and neither triptolide nor tripterine was detectable in the product of total alkaloids. Therefore triptolide, tripterine and total alkaloids were well separated and enriched with the optimal separation process.
Above results showed the process of“SFE-acidification-alkalization-organic solvent extraction”can efficiently extract triptolide and tripterine, and well separate triptolide, tripterine and total alkaloids in the extracts. It provided a new approach and technology support for the appropriate utilization of T. Wilfordii. With a high content of non-edible seed oil, J. curcas is world recognized as the most potential plant to be the substitute of fossils. In this study, jatropha seed oil was extracted with SC-CO_2 and the extraction of molluscacidal saponins from the seeds was explored.
Firstly, seed oil was extracted from the seed kernels of J. curcas by SC-CO_2. The effect of some factors such as particle size, extraction temperature and pressure on the yield of seed oil was investigated. A maximum yield of 51.5% (recovery 92.1%) can be obtained under the optimal conditions (particle size: 250-380μm, 55℃and 40MPa). The scale-up experiment of the optimal conditions gave a yield of 47.25% (recovery 84.5%), which was lower than that of pilot experiemnts but higher than that of traditional methods, indicating the SC-CO_2 extraction process was feasible.
Secondly, saponins were extracted from the de-oiled seed kernels by heat reflux extraction with ethanol solution. The influences of water temperature, concentration of ethanol solution, solid-liquid ratio and extraction times on the yield of total saponins were investigated by an orthogonal design. The range analysis showed the factors were in an order of extraction times > solid-liquid ratio > concentration of ethanol solution > water temperature. And the optimal extraction conditions were extraction with 80% ethanol with a solid-liquid ratio of 1:10 under the water temperature of 80℃for 3 times. The maximum yield of total saponins was 1.955% (recovery 88.64%) among the experiments.
Lastly, the solubilities of jatropha seed oil in SC-CO_2 at definite temperatures and pressures were caculated from the initial slope of the extraction curve of yield versus volumes of SC-CO_2 and correlated with the Chrastil equation and modified Chrastil equation with AARDs of 10.10 % and 3.468 %, respectively. In addition, the SC-CO_2 extraction process of jatropha seed oil was simulated with the broken and intact cell model and two-stage diffusion model, respectively. The values of AARDs were in the range of 1.08-3.67 % and 3.01%-10.24%, respectively, indicating that both models had good simulation results. The above theoretical studies therefore provide basic data and technoglogy support for the industrial scale-up and process design of the SC-CO_2 extraction of jatropha seed oil.
引文
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