雷公藤中有效成分的提取分离与微球制剂的制备
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摘要
雷公藤(Tripterygium Wilfordii Hook.f.)是一种传统的中草药,研究发现具有抗炎、抗菌、免疫抑制、抗生育、抗肿瘤等药理作用。目前雷公藤中药制剂被用于治疗类风湿性关节炎,治疗效果显著。但该中药制剂多是粗提物,其中雷公藤甲素被公认为最主要的活性成分,而红素雷公藤红素则被认为具有较大毒副作用。因此,如何有效提高雷公藤甲素的含量和降低雷公藤红素的含量,达到增效减毒的目的,一直是雷公藤制剂研究中急待解决的问题。近年来由于又发现雷公藤红素具有很好的抗肿瘤作用而受到高度重视,因此,本文对雷公藤甲素和雷公藤红素的提取与分离技术进行了探讨,该研究对增效减毒问题的解决和新药的开发均具有重要意义。
本文首先对生药中雷公藤红素分析方法中的前处理进行了改进,通过甲醇超声提取-酸沉-乙酸乙酯萃取的前处理方法,提取率与文献报道值相比,提高了34%,RSD为2.65%。该方法操作简单、快速、准确。
本文采用先进的超临界CO2萃取技术从雷公藤药材中高效地提取出同时含有雷公藤甲素和雷公藤红素的粗提物,然后首次结合酸沉-碱化-有机溶剂萃取等分离纯化手段将雷公藤甲素和雷公藤红素进行了有效分离。采用HPLC仪器检测的结果表明,雷公藤甲素产品中已检测不到的雷公藤红素;而雷公藤红素产品中雷公藤甲素含量小于0.05%。超临界CO2萃取技术与后续分离方法的合理集成也提高了雷公藤甲素和雷公藤红素在各自产品中的含量,雷公藤甲素的提取率为2.71×10-3%,雷公藤甲素在产品中含量为0.75%,均远高于文献报道值。雷公藤红素在产品中含量高达35.76%。
本文还以含有雷公藤红素的提取物为原料,对微球制剂的制备进行了研究,考察了PLGA浓度、PVA浓度、药与PLGA的比例、搅拌转速四种因素对微球制备的影响,确定了微球的适宜制备工艺条件,由此制备的微球包封率高达约80%,微球球形圆整,表层光滑,粒径比较均匀,为进一步开发缓释制剂提供基础数据。
Tripterygium Wilfordii Hook.f. (T. Wilfordii) is a traditional Chinese herb which has many pharmacological effects such as anti-inflammation, antibacterial effect, immunosuppression, antifertility, anti-tumor and so on. So far most of products from T. Wilfordiii are crude extracts,in which triptolide is considered to have great curative effect on rheumatoid arthritis, but tripterine has been considered to have toxicity and side effects. It is a problem all long that how to reduce the tripterine and how to increase the content of effective triptolide in the extracts. Recently tripterine was found to be a effective anti-tumor in spite of its toxicity. The purpose of this paper is to extract and isolate triptolide and tripterine by advanced extraction and separation technologies, which has important values for developing new medicines.
The pretreatment method for analysis of tripterine in raw material was improved. With the raw material was first carried out by ultrasonic extraction in methanol, and then treated by acid precipitation and extraction with ethyl acetate, the yield can be increased by 34% with RSD of 2.65%. This improved method is simple, rapid and accurate.
Supercritical carbon dioxide extraction was used to extract both triptolide and tripterine effectively under the investigated optimal conditions. Then a valid purification process of acidification–alkalization–organic solvent extraction was further employed to separate triptolide and tripterine. The analytical results show that the content of tripterine in the product of triptolide is close to zero, while the content of triptolide in the product of tripterine is also reduced to 0.05%. The yield of triptolide is 2.71×10-3% and the content of triptolide in its products is 0.75%, both of which are much higher than the literature reported values. The content of tripterine in its product is high up to 35.76%.
The preparation of microspheres of the tripterine product was studied. The influences of the concentration of PLGA, the concentration of PVA, the ratio of drug to PLGA and the rotate speed on the encapsulation efficiency were investigated. Under the reasonable conditions, round smooth and even particle size microspheres with the encapsulation efficiency of about 80% can be obtained.
本文首先对生药中雷公藤红素分析方法中的前处理进行了改进,通过甲醇超声提取-酸沉-乙酸乙酯萃取的前处理方法,提取率与文献报道值相比,提高了34%,RSD为2.65%。该方法操作简单、快速、准确。
本文采用先进的超临界CO2萃取技术从雷公藤药材中高效地提取出同时含有雷公藤甲素和雷公藤红素的粗提物,然后首次结合酸沉-碱化-有机溶剂萃取等分离纯化手段将雷公藤甲素和雷公藤红素进行了有效分离。采用HPLC仪器检测的结果表明,雷公藤甲素产品中已检测不到的雷公藤红素;而雷公藤红素产品中雷公藤甲素含量小于0.05%。超临界CO2萃取技术与后续分离方法的合理集成也提高了雷公藤甲素和雷公藤红素在各自产品中的含量,雷公藤甲素的提取率为2.71×10-3%,雷公藤甲素在产品中含量为0.75%,均远高于文献报道值。雷公藤红素在产品中含量高达35.76%。
本文还以含有雷公藤红素的提取物为原料,对微球制剂的制备进行了研究,考察了PLGA浓度、PVA浓度、药与PLGA的比例、搅拌转速四种因素对微球制备的影响,确定了微球的适宜制备工艺条件,由此制备的微球包封率高达约80%,微球球形圆整,表层光滑,粒径比较均匀,为进一步开发缓释制剂提供基础数据。
Tripterygium Wilfordii Hook.f. (T. Wilfordii) is a traditional Chinese herb which has many pharmacological effects such as anti-inflammation, antibacterial effect, immunosuppression, antifertility, anti-tumor and so on. So far most of products from T. Wilfordiii are crude extracts,in which triptolide is considered to have great curative effect on rheumatoid arthritis, but tripterine has been considered to have toxicity and side effects. It is a problem all long that how to reduce the tripterine and how to increase the content of effective triptolide in the extracts. Recently tripterine was found to be a effective anti-tumor in spite of its toxicity. The purpose of this paper is to extract and isolate triptolide and tripterine by advanced extraction and separation technologies, which has important values for developing new medicines.
The pretreatment method for analysis of tripterine in raw material was improved. With the raw material was first carried out by ultrasonic extraction in methanol, and then treated by acid precipitation and extraction with ethyl acetate, the yield can be increased by 34% with RSD of 2.65%. This improved method is simple, rapid and accurate.
Supercritical carbon dioxide extraction was used to extract both triptolide and tripterine effectively under the investigated optimal conditions. Then a valid purification process of acidification–alkalization–organic solvent extraction was further employed to separate triptolide and tripterine. The analytical results show that the content of tripterine in the product of triptolide is close to zero, while the content of triptolide in the product of tripterine is also reduced to 0.05%. The yield of triptolide is 2.71×10-3% and the content of triptolide in its products is 0.75%, both of which are much higher than the literature reported values. The content of tripterine in its product is high up to 35.76%.
The preparation of microspheres of the tripterine product was studied. The influences of the concentration of PLGA, the concentration of PVA, the ratio of drug to PLGA and the rotate speed on the encapsulation efficiency were investigated. Under the reasonable conditions, round smooth and even particle size microspheres with the encapsulation efficiency of about 80% can be obtained.
引文
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