摘要
南极磷虾中含有丰富的磷脂资源,是磷脂的生物富集库。采用蛋白酶辅助Bligh&Dyer法提取南极磷虾磷脂技术,研究温度、pH值、时间以及酶的种类对南极磷虾磷脂提取率的影响;采用气相色谱法研究磷脂脂肪酸链结构组成,建立亲水色谱串联质谱法(HILIC-QTOF/MS)法对磷脂进行定量和鉴定。研究发现胰蛋白酶为最优提取酶,通过Box-Benhnken方法优化提取南极磷虾磷脂的最佳条件为:提取温度41℃,pH 9,提取时间3 h。该条件下得到的磷脂提取量为(3.66±0.03)%。经测定,本试验中提取的南极磷虾磷脂的脂肪酸链结构主要为C16:0(31.58%)、EPA(31.94%)、DHA(20.64%)等。采用酶辅助Bligh&Dyer法提取得到的脂肪酸中不饱和脂肪酸所占比例比较高,EPA和DHA达到52.58%。通过HILIC-QTOF/MS脂质组学研究成功分离磷脂酰胆碱(PC)、磷脂酰乙醇胺(PE)和磷脂酰肌醇(PI)三大类磷脂。对磷脂类中各磷脂分子实现离子源内分离后成功测定59种磷脂分子,经数据库比对和二级质谱佐证得到磷脂脂肪酸链结构信息:南极磷虾中存在大量含有EPA和DHA链的磷脂分子。本研究结果为南极磷虾磷脂的开发和综合利用提供了试验依据。
Antarctic krill is an important source of phospholipids,which is also a natural storage for phospholipid bioaccumulation.In this study,the phospholipids from Antarctic krill was extracted by enzyme-assisted Bligh&Dyer extraction.The influcing factors of the extraction rate,such as temperature,pH value,enzymatic hydrolysis time and the kind of enzyme were investigated.The structure composition of fatty acids was assayed by gas chromatography with the quantification and analytical identification of phospholipids by HILIC-QTOF/MS.An optimum phospholipid extractants of(3.66±0.03)%was obtained by the typsin,the extraction temperature of 41℃,the pH value of 9 and the extraction time of 3 h.The main structure of fatty acid chains were C16:0(31.58%),EPA(31.94%),DHA(20.64%).Under the enzyme-assisted Bligh&Dyer extraction,the content of EPA and DHA chains were up to 52.58%with the higher percentage of unsaturated fatty acids.Three kinds of phospholipids,including Phosphatidyl cholines(PC),Phosphatidyl ethanolamine(PE)and Phosphatidylinositol(PI),were successfully separated by HILIC-QTOF/MS.After the ion source separation,59 kinds of phospholipid molecules were totally detected,while the structure information of fatty acid chains were identified by database and secondary mass spectrometry.The results revealed that an abundance of EPA and DHA chains were existed in phospholipid molecules from Antarctic krill.In conclusion,this study provided the foundation for the development and utilization of Antarctic krill phospholipids.
作者
俞喜娜
陈康
张燕平
崔益玮
汪艺
戴志远
王萍亚
赵巧灵
沈清
Yu Xina;Chen Kang;Zhang Yanping;Cui Yiwei;Wang Yi;Dai Zhiyuan;Wang Pingya;Zhao Qiaoling;Shen Qing(Zhejiang Province Joint Key Laboratory of Aquatic Products Processing,Institute of Seafood,Zhejiang Gongshang University,Hangzhou 310012;Zhoushan Institute for Food and Drug Inspection and Testing,Zhoushan 316000,Zhejiang)
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2020年第11期97-106,共10页
Journal of Chinese Institute Of Food Science and Technology
基金
浙江省公益技术研究计划项目(LGN18C200001)
国家自然科学基金青年科学基金项目(31601542)
浙江省重点研发计划项目(2017C03041)。
关键词
南极磷虾
磷脂
酶辅助提取
脂质
脂肪酸
Antarctic krill
phospholipids
enzyme-assisted extraction
lipid
fatty acid
