海洋生物脂肪酸不同提取方法比较——以斑节对虾为例
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摘要
本文以海南八门湾斑节对虾为研究对象,通过不同提取方式及转移试剂的结果对比,优化海洋生物样品脂肪酸的前处理方法。实验结果表明:采用微波提取,辅以正己烷和甲醇(含5%HCl)为转移试剂的提取效果优于其他方法,回收率可达到90%以上,且实验步骤简化,提取时间缩短。同时,该方法不会对单个脂肪酸产生选择性影响,可广泛用于海洋生物样品的脂肪酸提取。
This paper took Penaeus monodon as the study object to try to optimize fatty acids extraction of marine biological samples by comparing the extracting means and choosing different transfer regents. The results suggested that the method using microwave extraction with n-hexane and methanol( containing 5% HCl) as transfer reagents was the best method.The recovery of this method was higher than 90%.Moreover,the experiment procedure was simplyfied and the extraction time was shortened.Meanwhile,this method had no selective influence on the relative content of individual fatty acids,and could be widely used in the extraction of fatty acids of marine biological samples.
This paper took Penaeus monodon as the study object to try to optimize fatty acids extraction of marine biological samples by comparing the extracting means and choosing different transfer regents. The results suggested that the method using microwave extraction with n-hexane and methanol( containing 5% HCl) as transfer reagents was the best method.The recovery of this method was higher than 90%.Moreover,the experiment procedure was simplyfied and the extraction time was shortened.Meanwhile,this method had no selective influence on the relative content of individual fatty acids,and could be widely used in the extraction of fatty acids of marine biological samples.
引文
[1]BODE A,ALVAREZ-OSSORIO M T.Taxonomic versus trophic structure of mesozooplankton:a seasonal study of species succession and stable carbon and nitrogen isotopes in a coastal upwelling ecosystem[J].ICES Journal of Marine Science,2004,61(4):563-571.
[2]王娜.脂肪酸等生物标志物在海洋食物网研究中的应用-以长江口毗邻海域为例[D].上海:华东师范大学,2008.
[3]FOLCH J,LEES M,SLOANE STANLEY G H.A simple method for the isolation and purification of total lipides from animal tissues[J].The Journal of Biological Chemistry,1957,276(1):497-509.
[4]EDER K,REICHLMAYR-LAIS A M,KIRCHGESSNER M.Studies on themethanolysis of small amounts of purified phospholipids for gas chromatographic analysis of fatty acid methyl esters[J].Journal of Chromatography A,1992,607(1):55-67.
[5]MORRISON W R,SMITH L M.Preparation of fatty acid methyl esters anddimethylacetals from lipids with boron fluoridemethanol[J].Journal of Lipid Research,1964,5:600-608.
[6]LEPAGE G,ROY CC.Direct transesterification of all classes of lipids in a one-step reaction[J].Journal of Lipid Research,1986,27(1):114-120.
[7]BUTTE W.Rapid method for the determination of fatty acid profiles from fats and oils usingtrimethylsulphonium hydroxide for transesterification[J].Journal of Chromatography A,1983,261:142-145.
[8]YAO J Y,LIU Y,LI Y,et al.Effects of pretreatment procedures on fatty acid composition and stable carbon isotopes in the marine microalga Isochrysis zhanjiangenisis[J].Acta Oceanologica Sinica,2016,35(6):1-6.
[9]黄建立,张青龄,许世泉,等.微波辅助合成/萃取-气相色谱法测定植物黄油中反式脂肪酸[J].中国油脂,2010,35(3):59-62.
[10]YU D H,TIAN L,MA D X,et al.Microwave-assisted fatty acid methyl ester production from soybean oil by Novozym 435[J].Green Chemistry,2010,12(5):844-850.
[11]ZU Y G,ZHANG S,FU Y J,et al.Rapid microwave-assisted transesterification for the preparation of fatty acid methyl esters from the oil of yellow horn(Xanthoceras sorbifolia Bunge.)[J].European Food Research and Technology,2009,229(1):43-49.
[12]马宁,赵雷,刘国琴.微波处理对油脂脂肪酸组成影响研究进展[J].粮食与油脂,2009(1):17-18.
[13]刘国琴,李琳,胡松青,等.功率超声和微波处理对花生油脂肪酸组成的影响[J].中国油脂,2005,30(7):46-49.
[14]CUI Y,WU Y,ZHANG J,et al.Potential dietary influence on the stable isotopes and fatty acid compositions of jellyfishes in the Yellow Sea[J].Journal of the Marine Biological Association of the United Kingdom,2012,92(6):1325-1333.
[15]IVERSON S J,LANG S L C,COOPER M H.Comparison of thebligh and dyer and folch methods for total lipid determination in a broad range of marine tissue[J].Lipids,2001,36(11):1283-1287.
[16]EDER K.Gas chromatographic analysis of fatty acid methyl esters[J].Journal of Chromatography B:Biomedical Sciences and Applications,1995,671(1/2):113-131.
[17]CUI Y,WU Y,XU Z L,et al.Potential dietary influence on the stable isotopes and fatty acid composition of migratory anchovy(Coilia mystus)around the Changjiang Estuary[J].Journal of the Marine Biological Association of the United Kingdom,2015,95(1):193-205.
[18]BUDGE S M,IVERSON S J,KOOPMAN H N.Studying trophic ecology in marine ecosystems using fatty acids:a primer on analysis and interpretation[J].Marine Mammal Science,2006,22(4):759-801.
[19]褚武菁,张俊,陆胜民.胡柚果肉不同极性提取物对3T3-L1脂肪细胞葡萄糖摄取率、白细胞介素-6及游离脂肪酸的影响[J].中医药学报,2014,42(3):14-18.
[20]PARRISH C C,ABRAJANO T A,BUDGE S M,et al.Lipid and phenolic biomarkers in marine ecosystems:analysis and applications[M]//Wangersky P J.Marine Chemistry.Berlin,Heidelberg:Springer,2000:193-233.
[21]KHOTIMCHENKO S V.Fatty acids and polar lipids of seagrasses from the Sea of Japan[J].Phytochemistry,1993,33(2):369-372.
[22]Li G P,SINCLAIR A J,LI D.Comparison of lipid content and fatty acid composition in the edible meat of wild and cultured freshwater and marine fish and shrimps from China[J].Journal of Agricultural and Food Chemistry,2011,59(5):1871-1881.
[2]王娜.脂肪酸等生物标志物在海洋食物网研究中的应用-以长江口毗邻海域为例[D].上海:华东师范大学,2008.
[3]FOLCH J,LEES M,SLOANE STANLEY G H.A simple method for the isolation and purification of total lipides from animal tissues[J].The Journal of Biological Chemistry,1957,276(1):497-509.
[4]EDER K,REICHLMAYR-LAIS A M,KIRCHGESSNER M.Studies on themethanolysis of small amounts of purified phospholipids for gas chromatographic analysis of fatty acid methyl esters[J].Journal of Chromatography A,1992,607(1):55-67.
[5]MORRISON W R,SMITH L M.Preparation of fatty acid methyl esters anddimethylacetals from lipids with boron fluoridemethanol[J].Journal of Lipid Research,1964,5:600-608.
[6]LEPAGE G,ROY CC.Direct transesterification of all classes of lipids in a one-step reaction[J].Journal of Lipid Research,1986,27(1):114-120.
[7]BUTTE W.Rapid method for the determination of fatty acid profiles from fats and oils usingtrimethylsulphonium hydroxide for transesterification[J].Journal of Chromatography A,1983,261:142-145.
[8]YAO J Y,LIU Y,LI Y,et al.Effects of pretreatment procedures on fatty acid composition and stable carbon isotopes in the marine microalga Isochrysis zhanjiangenisis[J].Acta Oceanologica Sinica,2016,35(6):1-6.
[9]黄建立,张青龄,许世泉,等.微波辅助合成/萃取-气相色谱法测定植物黄油中反式脂肪酸[J].中国油脂,2010,35(3):59-62.
[10]YU D H,TIAN L,MA D X,et al.Microwave-assisted fatty acid methyl ester production from soybean oil by Novozym 435[J].Green Chemistry,2010,12(5):844-850.
[11]ZU Y G,ZHANG S,FU Y J,et al.Rapid microwave-assisted transesterification for the preparation of fatty acid methyl esters from the oil of yellow horn(Xanthoceras sorbifolia Bunge.)[J].European Food Research and Technology,2009,229(1):43-49.
[12]马宁,赵雷,刘国琴.微波处理对油脂脂肪酸组成影响研究进展[J].粮食与油脂,2009(1):17-18.
[13]刘国琴,李琳,胡松青,等.功率超声和微波处理对花生油脂肪酸组成的影响[J].中国油脂,2005,30(7):46-49.
[14]CUI Y,WU Y,ZHANG J,et al.Potential dietary influence on the stable isotopes and fatty acid compositions of jellyfishes in the Yellow Sea[J].Journal of the Marine Biological Association of the United Kingdom,2012,92(6):1325-1333.
[15]IVERSON S J,LANG S L C,COOPER M H.Comparison of thebligh and dyer and folch methods for total lipid determination in a broad range of marine tissue[J].Lipids,2001,36(11):1283-1287.
[16]EDER K.Gas chromatographic analysis of fatty acid methyl esters[J].Journal of Chromatography B:Biomedical Sciences and Applications,1995,671(1/2):113-131.
[17]CUI Y,WU Y,XU Z L,et al.Potential dietary influence on the stable isotopes and fatty acid composition of migratory anchovy(Coilia mystus)around the Changjiang Estuary[J].Journal of the Marine Biological Association of the United Kingdom,2015,95(1):193-205.
[18]BUDGE S M,IVERSON S J,KOOPMAN H N.Studying trophic ecology in marine ecosystems using fatty acids:a primer on analysis and interpretation[J].Marine Mammal Science,2006,22(4):759-801.
[19]褚武菁,张俊,陆胜民.胡柚果肉不同极性提取物对3T3-L1脂肪细胞葡萄糖摄取率、白细胞介素-6及游离脂肪酸的影响[J].中医药学报,2014,42(3):14-18.
[20]PARRISH C C,ABRAJANO T A,BUDGE S M,et al.Lipid and phenolic biomarkers in marine ecosystems:analysis and applications[M]//Wangersky P J.Marine Chemistry.Berlin,Heidelberg:Springer,2000:193-233.
[21]KHOTIMCHENKO S V.Fatty acids and polar lipids of seagrasses from the Sea of Japan[J].Phytochemistry,1993,33(2):369-372.
[22]Li G P,SINCLAIR A J,LI D.Comparison of lipid content and fatty acid composition in the edible meat of wild and cultured freshwater and marine fish and shrimps from China[J].Journal of Agricultural and Food Chemistry,2011,59(5):1871-1881.