银杏花粉黄酮组分分离纯化及其清除DPPH自由基能力  

Isolation and purification of flavonoids from Ginkgo biloba pollen and their DPPH scavenging activities

作　　者：裘纪莹 陈相艳[1] 闫慧娇 王岱杰[2] 陈蕾蕾[1] Qiu Jiying;Chen Xiangyan;Yan Huijiao;Wang Daijie;Chen Leilei(Institute of Agro-Food Science and Technology, Shandong Academy of Agricultural Sciences/Key Laboratory of Agro-Products Processing Technology of Shandong Province/Key Laboratory of Novel Food Resources Processing, Ministry of Agriculture, Jinan 250100, China;Shandong Analysis and Test Center /Key Laboratory of TMC Quality Control Technology, Jinan 250014, China)

机构地区：[1]山东省农业科学院农产品研究所/山东省农产品精深加工技术重点实验室/农业部新食品资源加工重点实验室,济南250100 [2]山东省分析测试中心/山东省中药质量控制技术重点实验室,济南250014

出　　处：《农业工程学报》2018年第10期289-295,共7页Transactions of the Chinese Society of Agricultural Engineering

基　　金：山东省自然科学基金(ZR2016YL022;ZR2014YL020);山东省农业科学院青年英才培养计划项目;泰山学者专项工程项目;国家自然科学基金(21506119)联合资助

摘　　要：为了分离纯化银杏花粉中具有潜在抗氧化活性的黄酮组分及研究其潜在抗氧化活性,该文对银杏花粉的总黄酮进行提取和分步萃取,采用1,1-diphenyl-2-picrylhydrazyl(DPPH)自由基清除率检测和组分分析锁定待纯化萃取相。通过DPPH-HPLC-PAD技术筛选目标清除自由基组分,并采用制备型液相色谱技术分离纯化这些组分,然后采用紫外可见吸收光谱、质谱、核磁共振氢谱、碳谱等技术进行结构鉴定,最后通过测定DPPH自由基清除率初步研究其潜在抗氧化活性。结果显示,银杏花粉粗提物具有清除DPPH自由基活性,对DPPH的半抑制率(IC50值)为1.32 mg/m L,乙酸乙酯相和正丁醇相的IC50值为0.46、0.84 mg/m L,分别是粗提物的34.85%和63.64%。可见,乙酸乙酯相的清除DPPH活性最高。进一步研究显示,乙酸乙酯相富集了6种主要银杏花粉黄酮,而且均具有清除DPPH活性。经纯化和结构鉴定分别为山奈酚-3,4’-双-O-β-D-葡萄糖苷(1)、山奈酚-3-O-β-D-葡萄糖基-7-O-α-L-鼠李糖苷(2)、山奈酚-3-O-β-D-葡萄糖苷(3)、山奈酚-3-O-α-L-鼠李糖苷(4)、柚皮素(5)和山奈酚(6)。其清除DPPH自由基的活性由高到低依次为:化合物6>化合物2>化合物3>化合物4>化合物1>化合物5。其中山奈酚清除DPPH的IC50值为0.017 mg/m L,与维生素C及芦丁相比没有显著性差异(P>0.05)。上述研究为银杏花粉功能成分的深入研究奠定了基础。Ginkgo biloba pollens are pollens from the male Ginkgo biloba trees, and contain many active components represented by flavonoids, terpene lactones, and polysaccharides. The total flavonoid content of Ginkgo biloba pollens was approximately 21.4 mg/g,much higher than that of Ginkgo biloba leaves.Previous analyses of flavonoids in Ginkgo biloba pollens were based on the results from Ginkgo biloba leaves,and the flavonoids of pollens and leaves were found to be very different in type and content.For example,rutin was reported to be abundant in Ginkgo biloba leaves,while the rutin content in Ginkgo biloba pollens was less than 6% of that in Ginkgo biloba leaves. Now the research on chemical constituents of Ginkgo biloba pollens was still in its infancy.Thus,studying the chemical composition of Ginkgo biloba pollens by indirectly measuring the flavonoids in Ginkgo biloba leaves was meaningless,and it was necessary to systematically analyze and purify the flavonoids from Ginkgo biloba pollens. In this study, crude extract of total flavonoids from Ginkgo biloba pollens was obtained by microwave-assisted extraction, and was separated into 4 fractions, petroleum ether, ethyl acetate, n-butanol and water by a fractional extraction method. The extract fraction used for further isolation and purification was selected through an assay of 1,1-diphenyl-2-picrylhydrazyl （DPPH） scavenging activities and an analysis of flavonoid components. The target flavonoids with DPPH scavenging activities in the selected fraction were screened by a 1,1-diphenyl-2-picrylhydrazyl - high performance liquid chromatography - photodiode array detection （DPPH-HPLC-PAD） method, then were isolated and purified by a preparative liquid chromatography （preparative LC）, and their chemical structures were identified using ultraviolet-visible spectroscopy, mass spectrometry, hydrogen-1 nuclear magnetic resonance （1H NMR） spectroscopy and carbon-13 nuclear magnetic resonance （13C NMR） spectroscopy. In addition, the DPPH scavenging activi

关 键 词：纯化 农产品 加工 银杏花粉 黄酮 DPPH自由基 抗氧化 

分 类 号：TS201.2[轻工技术与工程—食品科学]