银离子络合法提取纯化大西洋鲑鱼中二十二碳六烯酸(DHA)用于特定位点碳同位素分析(PSIA-C)  

An optimized Ag+complexation method for docosahexaenoic acid(DHA)isolation from Atlantic salmon for position-specificcarbon isotope analysis(PSIA-C)

作　　者：马艺 朱震宇 马然 王颖 赵雨 周友平 MA Yi;ZHU Zhen-yu;MA Ran;WANG Ying;ZHAO Yu;ZHOU You-ping(College of Chemistry and Chemical Engineering,Isotopomics in Chemical Biology,Shaanxi University of Science&Technology,Xi′an 710021,China;Shaanxi Key Laboratory of Chemical Additives for Industry,Shaanxi University of Science&Technology,Xi′an 710021,China)

机构地区：[1]陕西科技大学化学与化工学院天然产物稳定同位素组学实验室,陕西西安710021 [2]陕西科技大学陕西省轻化工助剂重点实验室,陕西西安710021

出　　处：《陕西科技大学学报》2021年第1期75-81,共7页Journal of Shaanxi University of Science & Technology

基　　金：国家自然科学基金项目(41773032);陕西省科技厅自然科学基础研究计划项目(2019JQ-232)。

摘　　要：尽管DHA(全顺式4,7,10,13,16,19-二十二碳六烯酸)在许多人类生物过程中被证明具有重要作用,但关于这种全顺式长链多不饱和脂肪酸(LC-PUFA)的来源和代谢仍然存在一定争议.作为利用核磁共振(NMR)测定分子内特定位点碳同位素分析(PSIA-C)进而解决人体DHA来源问题的第一步,本文报道了从大西洋鲑鱼(人类膳食DHA的主要来源之一)提取大量(>200 mg)高纯度(>95%)DHA的银离子络合法.采用有机溶剂提取鱼肉中的脂肪酸,用BF 3-CH 3OH进行甲基酯化,将甲基酯化脂肪酸混合物(脂肪酸甲酯,FAMEs)经硝酸银硅胶柱层析,按极性依次洗脱分离混合脂肪酸,获得高纯度的DHA.基于GC/MS和GC分析表明,采用银化硅胶柱层析法,AgNO 3和硅胶的重量比为1∶9,二氯甲烷和乙酸乙酯体积比3∶1洗脱,DHA纯度达到95%.Despite its proven important roles in many human biological processes,controversies still exist around the source and metabolism of DHA(all cis-4,7,10,13,16,19-docosahexaenoic acid),an all cis long chain polyunsaturated fatty acid(LC-PUFA).As the first step toward resolving the DHA source issue in the human body using NMR-based position-specific carbon isotope analysis(PSIA-C),we report the preparation of high purity DHA(>95%)from Atlantic salmon in large quantity(>200 mg),one of the major sources of human dietary DHA,using an optimized silver ion complexation(argentation column chromatography)method.The fatty acids were first FOLCH extracted(Vchloroform/Vmethanol=2/1),and methylated with BF 3-CH 3OH.The methylated fatty acid mixture(fatty acid methyl esters,FAMEs)was then subjected to AgNO 3-silica gel column chromatography.GC and GC/MS-based analyses of the fractions containing DHA showed that a weight ratio between AgNO 3 and silica gel of 10/90 and a volume ratio of 3/1 for the eluent mix dichloromethane/ethyl acetate is needed for the preparation of DHA with purity greater than 95%.

关 键 词：二十二碳六烯酸 制备规模纯化 银化硅胶柱层析 大西洋鲑鱼 

分 类 号：O652.7[理学—分析化学]