以微生物为对象设计了研究性实验,以核磁共振氢谱(1HNMR)为导向优选代谢产物产量高且有特色化学位移的菌株,用反相十八烷基硅烷键合硅胶(ODS)减压柱层析和制备高效液相色谱分别进行提取物分段和单体纯化,成功跟踪分离到一个环四肽化合物,鉴定了结构.综合运用了超声波辅助提取、核磁共振测试、减压柱层析,制备高效液相分离等操作,并通过结构式查阅网络数据库和用核磁共振鉴定结构的方法,有助于学生全面认识天然药物化学的发展趋势和科研流程、增加学习兴趣、提升操作技能和创新能力.实验的整个流程可推广到微生物代谢产物分离纯化的其他研究性实验当中.%By taking micro-organisms as an object, a research-oriented experiment is designed.Guided by 1H nuclear magnetic resonance spectroscopy (1HNMR), the optimal production of metabolites is high with the decompression column chromatography of bacterial strain for reversed phase eighteen alkyl silane bonded silica gel (ODS) of unique chemical shifts.The high performance liquid chromatography is prepared to carry out the object segmentation and monomer purification of extracts respectively.A cyclotetrapeptide is successfully traced and isolated, and its structure is identified.By comprehensive application of the ultrasonic assisted extraction of nuclear magnetic resonance and vacuum column chromatography, the operations such as high performance liquid phase separation are prepared.By the method for checking network database with the structure type and identifying the structure by nuclear magnetic resonance, it is helpful for the students to fully understand the development trend and research process of natural medicinal chemistry, stir up their interest in learning, and improve their operational skills and innovative ability.The whole experimental process can be expended to other research-oriented experiments for the isolation and purification of the metabolites of microorganisms.
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