海洋微藻脂质高效色谱分离分析方法研究

. Microalgae were the primary producers and the main biological species in the ocean. Lipids were one of the most important components in the microalgae. Lipids had important biological functions and development values. Most of marine microalgae lipids were the amphipathic compounds. They contained both different hydrophobic fatty chains and polar groups. The lipids composition was very rich and their structures were diversity. The isomers formed were also numerous. They were difficult to be fully separated by chromatographic method before. By conventional GC and HPLC methods, there was selectivity, peak shape and the scope of the sample application problems. Thus, the comprehensive, accurate and rapid analysis of marine microalgae lipids could not be achieved. This project will mainly focus on developing new HPLC and SFC methods to solve the separation problems of the microalgae lipids. The nannochloropsis oculata and chrysophyta will be taken as examples. The non-aqueous RPLC method is developed to improve the hydrophobic selectivity of compounds with long aliphatic chain structures using solid phases containing long alkyl groups. Meanwhile, the polar group bonded materials are developed to improve the separation selectivity of the polar head. Then, the high-efficient HPLC analysis methods will be built. Besides, the NP and RP chromatographic materials will be applied in SFC to improve its separation speed and selectivity. The most appropriate materials will be designed. The multiple forces between stationary phases and the lipid compounds will be optimized. Finally, these multiple separation methods will be applied to the investigation of the lipid metabolisms regulation of the marine microalgae. Thus, the comprehensive, accurate and rapid analysis of marine microalgae lipids can be achieved. The developed methodologies have the advantages in both the in-depth biological function analysis of marine microalgae lipid metabolisms and further discovery of new activated lipid compounds. It is also the foundation for development and utilization of the microalgae lipids..

. 海洋微藻是海洋的初级生产者和生物种属的主体。脂质是微藻最重要的组成成分之一，具有重要的生物功能和应用开发价值。脂质的疏水脂肪链和极性基团千差万别，使得其组成丰富、结构多样且异构体多，是色谱分离的难点。常规的GC和HPLC方法，存在选择性、峰形及样品适用范围的问题，无法实现微藻脂质全面、准确、快速的分析。本项目以金藻和微拟球藻为对象，发展以HPLC和SFC为核心的分离分析方法，解决脂类尤其是类似物的高效分离问题。发展长烷基链的反相材料结合非水反相方法调控疏水选择性、发展极性键合相改善极性基团的选择性，建立高效的HPLC方法；筛选并设计分离材料，优化固定相与脂质的多种作用力，提供不同于HPLC的分离选择性，建立快速的SFC分离方法。最后，将发展的多种方法应用于微藻脂类代谢调控的监测中，实现其全面、准确、快速的分析，为微藻脂质的生物功能研究、应用开发及活性分子的发现提供高效分析方法基础和支撑。

海洋微藻是海洋的初级生产者和生物种属的主体。脂质是微藻最重要的组成成分之一，具有重要的生物功能和应用开发价值。脂质的疏水脂肪链和极性基团千差万别，使得其组成丰富、结构多样且异构体多，是色谱分离的难点。常规的GC和HPLC方法，存在选择性、峰形及样品适用范围的问题，无法实现微藻脂质全面、准确、快速的分析。本项目以金藻和微拟球藻为对象，发展了反相（C30，苯基/四氮唑）、亲水（两性离子聚合物、氨基酸）色谱分离材料，并用于微藻的分离分析中。发展了基质分散固相萃取的亲处理、液相色谱定量微藻中类胡萝卜素的方法。建立了微藻中甾醇的液相色谱-质谱及气相色谱-质谱方法。发展了藻油中不饱和脂肪酸如DHA的纯化方法，可制备得到高纯度的DHA。建立了超临界流体色谱分析微藻提取物的方法。为后续微藻脂质的生物功能研究、应用开发及活性分子的发现提供高效分析方法基础和支撑。项目共发表文章4篇，申请专利6项，1篇英文文章已投稿。