云南小粒咖啡化学成分的表征技术及活性成分发现

As one of the three most popular drinks in the world, coffee become one of the commercial crops being researched sufficiently on their chemical components and their bioactivities. Reported researches mainly focused on bioactivities and functions of major components in coffee, such as caffeine, gynesine, chlorogenic acid, but limited researches was carried on bioactivities and functions of the other constitutes in coffee. In the past few years, Yunnan Arabica Coffee became characteristic agriculture of plateau which developed rapidly and its planting area reached 1.2 billions square meters. However, researches on characters of components and evaluation studies of their bioactivities of Yunnan Arabica Coffee was still limited. Recently, a novel type of diterpenoids with cytotoxicities have been discovered by our group, which indicated a good prospect of research on active components in coffee. Some issues of the research on coffee need to be clarified further: (1) Any new bioactivities of known characteristic components in Yunnan Arabica Coffee? (2) Any novel active components with high activities in the Coffee? (3) Could modern spectroscopy technics be applied, such as NMR, to build up a sufficient analysis method for main active components in Coffee? The basis of this project is using modern technics with NMR spectroscopy to detect major components in coffee, in order to find novel high active structures by applying Activity Guidance Strategy. The aim of this project is to complete methods of Quality Control for Yunnan Arabica Coffee; to enrich basic data and technic for researches on active components of Yunnan Coffee Arabica; to provide scientific evidence and establish technical basis for healthy and rapid development of Yunnan Arabica Coffee Industry and related application of Coffee Consumption in China.

因咖啡列世界三大饮品之首，成为化学成分及其活性研究最为充分的经济作物之一。以往学者过度关注小粒咖啡中咖啡因、葫芦巴碱、绿原酸等主要成分的活性和功能，对其他化学成分的结构及其功能反而研究较少。近年来云南小粒咖啡成为高原特色农业，得到快速发展，种植基地已经接近超180万亩，但对云南小粒咖啡中化学成分的特点，以及生物活性评估研究报道较少。最近本研究组从云南产小粒咖啡中发现了一系列具有细胞毒活性的新二萜类化合物，显示咖啡活性化学成分具有很好的前景。云南产小粒咖啡中特征性成分是否还有其它新的生物活性；以及咖啡中是否还含有其他活性好，结构新颖的化学成分等；是否能利用NMR等现代波谱技术，如何建立咖啡的主要活性成分快速分析方法等等；有待进一步阐明。本项目拟基于NMR的现代技术方法对咖啡中主要成分进行表征技术为基础，并以活性指导方式发现咖啡中结构新颖且活性较好的新型分子结构。完善云南小粒咖啡质量控制方法，

云南咖啡是我国独占鳌头的区域性优势高原特色农业产业，但生产加工和开发利用十分落后，一切归因于基础研究薄弱。项目组在实施项目过程中，从不同产地不同时间采集的云南小粒咖啡的7个研究样品中，提取分离纯化，并鉴定了80多个化合物的化学结构，发现40多个新化合物。从云南咖啡中发现的一系列具有生物活性内含物质，新咖啡二萜苷化合物（Food Chem 2021），新型咖啡二萜内酯（Food Chem 2018）重排 Δ(4,18)的新型二萜（Fitoterapia 2019），咖啡二萜内酰胺型化合物Cafemide A-G，达玛烷型三萜（NPB 2018）等，均是云南咖啡中发现的新型萜类化合物，再次诠释云南咖啡“内含物质积累丰富”的独特品质。还发现了：咖啡二萜内酯的脂肪酸脂类化合物γ-Lactones 1-8中一些化合物具有一定的体外激活血小板聚集活性；新型二萜γ-Nors 1-2具有大麻素受体CB1和CB2激动活性；Cafemides A-C、E、G表现出较好的α-葡萄糖苷酶抑制作用，其活性优于糖尿病一线临床治疗药物阿卡波糖。本项目还发展了NMR快速分析咖啡豆和银皮等的化学成分的技术方法；运用基于LC-MS /MS的分子网络技术发现了痕量内酰胺型咖啡二萜。也阐明了云南咖啡烘焙度与社会关注都很高的丙烯酰胺含量的变化趋势（Food &Function，2019）。并尝试利用与云南咖啡醇厚度相关度较高的11种化学成分构建了咖啡风味醇厚度（Body）杯测得分的预测模型，同时通过成熟度标记物在不同成熟程度了鲜果中的积累，结合咖啡杯测，阐明了成熟度标记物的积累与咖啡品质之间的关系，确定了云南咖啡鲜果的最佳采摘时机。这些结果为保证云南咖啡优质品质的新型绿色加工工艺的开发，奠定了科学依据。本项目发表SCI论文8篇，申请国家专利3项，培养硕博生6名。