氧化铝柱层析纯化蛋黄多不饱和脂肪酸卵磷脂及其脂肪酸组成分析
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摘要
以实验室自制蛋黄卵磷脂(PC)粗品为原料,采用中性氧化铝柱层析分离技术,制备高纯度蛋黄多不饱和脂肪酸卵磷脂(PC≥90%)。分别考察固定相粒径、洗脱液浓度、上样浓度、洗脱液流速、上样量等对氧化铝层析柱纯化蛋黄卵磷脂过程中的影响,同时采用薄层层析定性和高效液相色谱定量跟踪分析,采用气相色谱对所得样品脂肪酸种类及组成进行分析。最佳工艺条件:氧化铝粒径为100～200目、95%乙醇洗脱液、上样浓度1∶4(m/V)、洗脱流速8 mL/min、色谱柱负载量0.0222 g/g氧化铝;在最优条件下,高纯度蛋黄卵磷脂(PC≥90%)得率可达84.43%,一段PC产品的DHA含量高达13.35%。本试验可为工业化制备高纯度多不饱和脂肪酸卵磷脂提供技术参考。
In this study,high purity polyunsaturated fatty acid lecithin(PC≥90%) was prepared from lecithin material by aluminum oxide column chromatography.With qualitative and quantitative tracing by TLC and HPLC,the influence factors,such as size of Al2 O3,concentration of PC solution,eluent component,flow rate of eluent,and the load capacity were discussed.The types and composition of the fatty acids were analyzed by GC.The results showed that the optimum purification condition was:100～200 mesh aluminum oxide,95% ethanol water,8 mL/min flow rate,1 ∶ 4(PC material/solvent,m/V),load capacity 0.0222 g/g Al2 O3.Under the optimal conditions,the yield of PC(≥90%) was84.43%.The content of DHA in the first part of lecithin was 13.35%.The result of the research can provide data support and reference for industrial preparation of high purity polyunsaturated fatty acid lecithin.
In this study,high purity polyunsaturated fatty acid lecithin(PC≥90%) was prepared from lecithin material by aluminum oxide column chromatography.With qualitative and quantitative tracing by TLC and HPLC,the influence factors,such as size of Al2 O3,concentration of PC solution,eluent component,flow rate of eluent,and the load capacity were discussed.The types and composition of the fatty acids were analyzed by GC.The results showed that the optimum purification condition was:100～200 mesh aluminum oxide,95% ethanol water,8 mL/min flow rate,1 ∶ 4(PC material/solvent,m/V),load capacity 0.0222 g/g Al2 O3.Under the optimal conditions,the yield of PC(≥90%) was84.43%.The content of DHA in the first part of lecithin was 13.35%.The result of the research can provide data support and reference for industrial preparation of high purity polyunsaturated fatty acid lecithin.
引文
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[2]SHINN S,LIYANAGE R,LAY J,et al.Improved fatty acid analysis of conjugated linoleic acid rich egg yolk triacylglycerols and phospholipid species[J].Journal of Agricultural&Food Chemistry,2014,62(28):6608-6615.
[3]ABEDI E,SAHARI M A.Long-chain polyunsaturated fatty acid sources and evaluation of their nutritional and functional properties[J].Food Science&Nutrition,2014,2(5):443-463.
[4]COOK C M,HALLARKER H,SBP C,et al.Bioavailability of long chain omega-3 polyunsaturated fatty acids from phospholipid-rich herring roe oil in men and women with mildly elevated triacylglycerols[J].Prostaglandins Leukotrienes&Essential Fatty Acids,2016(111):17-24.
[5]陆文达,郑卫琴,李剑英,等.注射用富含卵磷脂蛋黄磷脂制备研究[J].粮食与油脂,2010(3):39-41.
[6]MORI T A,BURKE V,PUDDEY I B,et al.Purified eicosapentaenoic and docosahexaenoic acids have differential effects on serum lipids and lipoproteins,LDL particle size,glucose,and insulin in mildly hyperlipidemic men[J].American Journal of Clinical Nutrition,2000,71(5):1085-1094.
[7]吕名蕊.磷脂酰胆碱富集及热原去除的研究[D].无锡:江南大学,2010.
[8]汪勇,王兴国,欧仕益,等.大豆磷脂酰胆碱分离、纯化和检测方法[J].粮食与油脂,2003(2):6-8.
[9]周玉权,刘丹,张燕,等.高纯度磷脂酰胆碱的制备技术研究[C].北京:2013全球华人保健大会,2013.
[10]李召妍.高纯大豆卵磷脂的制备工艺研究[D].北京:中国石油大学,2010.
[11]柳叶,吕秀阳.氧化铝柱层析分离磷脂工艺的洗脱研究[J].浙江化工,2006,37(9):3-4.
[12]陈文波,马忠朝,乐盛,等.大豆多烯磷脂酰胆碱分离纯化工艺的研究[J].求医问药:学术版,2012,10(8):163-164.
[13]秦孝敏,苏明华,李翔,等.活性氧化铝吸附对大豆卵磷脂品质的影响[J].中国油脂,2008,33(8):57-59.
[14]曹栋,裘爱泳,王兴国.超临界流体分离大豆磷脂酰胆碱[J].中国油脂,2002,27(3):72-73.
[15]张牧焓,王道营,卞欢,等.氧化铝柱层析纯化鸭肝卵磷脂的工艺研究[J].南方农业学报,2014,45(6):1054-1059.
[16]RENKONEN O.Breakdown of lecithin on aluminum oxide columns[J].Journal of Lipid Research,1962,3(2):181-183.
[17]ROMBAUT R,DEWETTINCK K,CAMP J VPhospho-and sphingolipid content of selected dairy products as determined by HPLC coupled to an evaporative light scattering detector(HPLC-ELSD)[J]Journal of Food Composition&Analysis,2007,20(3/4):308-312.
[18]龚雁.磷脂化合物的分离制备和测定方法研究[D]北京:北京化工大学,2006.
[19]邓兴照,齐广海,刘福柱,等.日粮多不饱和脂肪酸类型对蛋鸡生产性能和蛋黄脂肪酸富集的影响[J]中国畜牧杂志,2006,42(3):31-34.