羊栖菜抑制乙酰胆碱酯酶活性油脂成分的提取优化  

Extraction Optimization of Acetylcholinesterase Inhibitory Active Oils from Hizikia fusiforme

作　　者：黎燕媚 聂影影 刘亚月 周龙建 宋采 洪鹏志 张永平 胡雪琼 张翼 LI Yan-mei;NIE Ying-ying;LIU Ya-yue;ZHOU Long-jian;SONG Cai;HONG Peng-zhi;ZHANG Yong-ping;HU Xue-qiong;ZHANG Yi(College of Food Science and Technology,Guangdong Ocean University,Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety,Guangdong Province Engineering Laboratory for Marine Biological Products,Guangdong Provincial Engineering Technology Research Center of Seafood,Key Laboratory of Advanced Processing of Aquatic Product of Guangdong Higher Education Institution,Guangdong Modern Agricultural Science and Technology Innovation Center,Zhanjiang Municipal Key Laboratory of Marine Drugs and Nutrition for Brain Health,Research Institute for Marine Drugs and Nutrition,Guangdong Ocean University,Zhanjiang 524088,China;Center for Marine Drugs Research&Development,Shenzhen Institute of Guangdong Ocean University,Shenzhen 518120,China;Collaborative Innovation Center of Seafood Deep Processing,Dalian Polytechnic University,Dalian 116034,China;Suothern Marine Science and Engineering Guangdong Laboratory(Zhanjiang),Zhanjiang 524006,China)

机构地区：[1]广东海洋大学食品科技学院/广东省水产品加工与安全重点实验室/广东省海洋生物制品工程实验室/广东省海洋食品工程技术研究中心/水产品深加工广东普通高等学校重点实验室/广东省现代农业科技创新中心/湛江市脑健康海洋药物与营养品重点实验室/海洋药物研究所,广东湛江524088 [2]广东海洋大学深圳研究院海洋医药研发中心,广东深圳518120 [3]大连工业大学海洋食品精深加工关键技术省部共建协同创新中心,辽宁大连116034 [4]南方海洋科学与工程广东省实验室(湛江市),广东湛江524006

出　　处：《广东海洋大学学报》2022年第3期97-106,共10页Journal of Guangdong Ocean University

基　　金：广东省科技专项资金-基础与应用基础研究专题(2021A05240);广东省普通高校重点领域专项(生物医药与健康)(2021ZDZX2064);深圳市大鹏新区科技研发项目(KJYF202001-07);深圳市科创委基础研究面上项目(JCYJ20190813105005619);深圳市大鹏新区产业发展资金(KY20180203);国家自然科学基金(21807015);广东省扬帆计划引进紧缺拔尖人才项目(201433009);湛江市海洋经济创新发展示范市建设项目(湛海创XM-202008-01B1);广东省高等学校科技创新团队项目(2021KCXTD021)。

摘　　要：【目的】建立药食同源海藻羊栖菜抑制乙酰胆碱酯酶(Acetylcholinesterase,AChE)活性油脂成分的优化提取工艺,探明其主要活性组分的具体化学成分。【方法】以提取物质量浓度1 mg/mL下的乙酰胆碱酯酶抑制率×提取物质量为指标,比较各个溶剂比、料液质量体积比、超声时间和提取次数等四个单因素水平下的抑制AChE活性成分提取效果,确定各个单因素中的最优水平范围,进而设计正交实验优化提取工艺条件;按最优提取工艺条件下提取制备粗浸膏,运用硅胶柱色谱法、薄层层析等方法对粗浸膏进行分离纯化并追踪其活性组分,运用气相色谱-质谱联用GC-MS进行成分分离与鉴定。【结果】羊栖菜抑制AChE活性成分的最优提取条件为:溶剂乙酸乙酯、石油醚体积比为5∶5,料液质量(g)体积(mL)比为1∶10,超声时间30 min,提取次数3次。此条件下对应的提取物质量浓度1 mg/mL下乙酰胆碱酯酶抑制率(%)×提取物质量(g)的值为7.078±1.038。从该粗提物中分离得到两个具有抑制AChE活性的组分Fr2和Fr3,主要成分为花生四烯酸、亚油酸、油酸等脂肪酸,Fr3还含有十四烷等多种烷烃。其抑制IC_(50)值分别为2.69 mg/mL和5.31 mg/mL。【结论】本研究确定的基于石油醚-乙酸乙酯混合溶剂的超声提取工艺可得到较高产量的羊栖菜抑制AChE活性成分,其主要成分为脂肪酸,可能在改善记忆方面具有一定应用价值。【Objective】To establish an optimized extraction process for acetylcholinesterase(AChE)inhibitory components from Hizikia fusiforme and to analyze and identify the chemical constituents.【Methods】Taking the product of inhibition rate to AChE at the dose of 1 mg/mL and extract mass(PIREM)as indicator,the optimal level range of each single factor was determined by comparing the extraction effects under different ratios of solvents,ratios of solid to liquid,ultrasonic times,and extraction times.Then,the extraction process was optimized by orthogonal experiment design.Under the optimal extraction condition,H.fusiforme was extracted to obtain the crude extract,which was separated and purified to trace the active components by silica gel column chromatography and thin layer chromatography.Finally,the constituents of the product was analyzed by gas chromatography-mass spectrometry.【Results】The results showed that the optimal extraction condition was,volume ratio of ethyl acetate to petroleum ether is 5∶5 as solvent,solid-liquid ratio=1∶10(g/mL),ultrasonical extraction for 30 min and three times.The PIREM value was 7.078±1.038 for the crude extract obtained under this condition.From this extract,two main active components Fr2 and Fr3 were prepared with inhibitory IC_(50) values of 2.69 mg/mL and 5.31 mg/mL,respectively.Their main constituents were revealed to be fatty acids like arachidonic acid,linoleic acid,oleic acid and also rich alkanes like tetradecane in Fr3.【Conclusion】By the supersonic extraction process based on mixed solvent of petroleum ether and ethyl acetate in this study,high yield of AChE inhibitory components can be prepared from H.fusiforme.This product mainly contains fatty acids and may be applicable for improving memory.

关 键 词：羊栖菜 正交实验 阿尔茨海默症 抑制乙酰胆碱酯酶活性 脂肪酸 

分 类 号：Q547[生物学—生物化学]