【作者】 吴泽强；

【导师】 余龙江； 赵春芳；

【作者基本信息】 华中科技大学 ， 生物化工， 2007， 硕士

【摘要】 复杂天然产物单体分离纯化以及工业化制备技术,是目前制约我国植物源天然产物单体制备产业化的瓶颈性问题,而将多种分离纯化方法有机组合是实现高纯度单体工业化制备的关键技术之一。本文以结构复杂而具高生物活性的长春碱的化学半合成产物——长春瑞滨终产物及重要中间体的分离纯化工艺研究为例,探讨其规模化制备中的分离纯化技术,并为其它天然产物单体工业化制备的创新技术研究提供技术和方法上的参考。整个研究主要包括以下内容:1.中间体脱水长春碱的分离纯化。在系统的研究结晶法和硅胶柱层析法分离纯化脱水长春碱的工艺后,通过对比,最终确定了结晶法为适合推向生产的纯化脱水长春碱的适合的工艺方法,并确定了具体的工艺参数。实验表明:纯度为30%左右的脱水长春碱经过一次结晶后纯度可以达到65%,结晶得率为70%左右。2.从缩环水解液中初步分离长春瑞滨。采用Sephadex LH-20柱层析和结晶法,从缩环水解液中初步分离长春瑞滨。结果显示虽然Sephadex LH-20柱层析有较高的固体回收率,但长春瑞滨与杂质未有效分离,最终确定用结晶法从缩环水解液中初步纯化长春瑞滨,该方法简便易行,可使瑞滨含量从30%提高到55%左右,适于工业化放大。3.通过中度分离与精细分离两个阶段进一步纯化与精制长春瑞滨。使用自填的硅胶干柱,直接套用薄层层析的溶剂展开系统,可使长春瑞滨纯度从55%提高到80%以上,硅胶的负载量为12 mg样品∕g硅胶,干柱的最佳径∕高比为1∕10,产品回收率在82%;利用MCI-GEL CHP-20填料进一步对难分离的长春瑞滨结构类似物进行反相分离,确定了反相分离的最佳条件为甲醇-水(V :V =80:20)流动相常压洗脱,反相柱的载样量为10 mg样品∕g填料,经过一次MCI-GEL CHP-20柱色谱纯化再结晶后,长春瑞滨纯度可达98%以上,该反相填料能反复利用,反相柱目标物收率达97%以上。表明采取这种正相硅胶干柱与反相MCI-GEL CHP-20柱色谱相组合的新方法,大大缩短了操作时间,提高了产品回收率,可快速高效的分离纯化长春瑞滨粗产品。本课题研究了生产长春瑞滨的高效分离与制备技术,确立了一套适合于产业化的产品分离与纯化方法。并为其它复杂天然产物单体的工业化制备提供了方法和技术参考。更多还原

【Abstract】 Technology in mass purification and industrialized production of complex natural resources monomer is bottleneck problem, which restricts the industrialization of activated natural resources from plants. Combination of different effective technologies is one of key technologies to produce complex natural resources monomer with high purity .This paper took Vinorelbine and its intermediate Anhydrovinblastine as example, which were semi-synthesized from vinblastine with complex and activated structure, studying technology about its mass purification and prudction, to provide academic and practical reference for mass purification and industrialized production of other natural resources. Here are the main studying content:1. Purification and separation of intermediate product Anhydrovinblastine. Comparison of different purification techniques for the Anhydrovinblastine was presented. Crystallography was selected out as the best method used to purify anhydrovinblastine in industry. Results showed that the purification of crud Anhydrovinblastine can get from 30% to 65% after the process of crystallization with the yield of 70%.2. Preliminary separating Vinorelbine from liquid hydrolysate. Sephadex LH-20 chromatography and crystallography were taken to preliminary separate Vinorelbine in liquid hydrolysate. Results indicated that the reclaim rate of solid matter is high ,but Sephadex LH-20 chromatography could not separate Vinorelbine from impurities effectively. At last, crystallography was selected out as the best method used to purify anhydrovinblastine in liquid hydrolysate. It can heighten the content of Vinorelbine from 30% to 50%. This process is simple and convenient, and suitable to be magnified in industrialization.3. Further purification of Vinorelbine, which include middling purification and high purification two steps. Normal dry-column manufactured by our-self was used to purify Vinorelbine with developer the same to TLC. After purified by dry-column flash chromatography, the purity of vinorelbine can get to 80% from 55% at the very beginning. The load of dry-column was 12 mg specimen per 1 g silica gel, the best diameter/tallness was 1/10, and the yield of vinorelbine can get to 82%. MCI-GEL CHP-20 reversed-phase chromatography was used in tandem to separate vinorelbine from impurities which were difficult to be separated. The best purification conditions of crud vinorelbine were methanol/water(V:V=80:20), The load of reversed-phase column was 10 mg specimen per 1 g MCI-GEL CHP-20 stuffing. The purity of vinorelbine can get to 98% when MCI-GEL CHP-20 reversed-phase chromatography and crystallization in sequence were used.Technique for high efficient purification of Vinorelbine was investigated in this thesis. A series of feasible process conditions for Purification and separation of Vinorelbine and Anhydrovinblastine were obtained, which are suitable to be used in synthesis of vinorelbine in industry. It gives both academic and practical reference for mass production and purification of other natural resources monomer.更多还原