宁波大学应用海洋生物技术教育部重点实验室
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杨怡, 徐盼盼, 宋悦, 等. 基于HPLC-MS技术的黄颡鱼中不同部位脂质成分分析[J]. 分析测试学报, 2016,35(7):785-791.
Analysis on Lipids in Different Parts of Pelteoobagrus fulvidraco by Ultra performance Liquid Chromatography Coupled with Mass Spectrometry[J]. 2016,35(7):785-791.
利用超高效液相色谱-飞行时间质谱技术(UPLC-Q-TOF-MS/MS)分析了黄颡鱼不同部位的脂质成分。采用甲醇-氯仿(1∶1,体积比)对血清、肌肉和肝脏进行脂质成分提取。液相色谱采用C8色谱柱,柱温40℃,流动相A为乙腈-甲醇-异丙醇(1∶1∶1),流动相B为乙腈-水(0.1% 甲酸,0.01%氯化锂)。根据高分辨质谱获得精确分子量,推测出元素组成,并结合脂质成分的二级质谱裂解规律,在血清、肌肉和肝脏中共鉴定出85个脂质成分,主要有PC,Lyso-PC,PE,Lyso-PE,PI,PS,DAG,TAG,SM和Cer。在这3个部位中有19个共有脂质成分,包括Lyso-PC 16∶0,9个 PC,3个 PI,4个TAG和2个SM。此外 PS和Lyso-PE仅在肌肉中鉴定出,分别为PS 18∶0/22∶6,Lyso-PE 16∶0和Lyso-PE 18∶1;神经酰胺仅在肝脏中观察到,为Cer(d18∶1/24∶1);而PE和DAG只在肌肉和肝脏中存在,在血清中未发现。该方法操作简单、灵敏、高效,为快速、全面了解黄颡鱼中的脂质分布提供了科学依据。
An analytical method for the qualitative analysis of lipids in different parts of Pelteoobagrus fulvidraco was developed by ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS).The dried tissue and serum were extracted with methanol-chloroform(1∶1).The separation of extract was performed on a C8 analytical column(100mm×2.1mm,1.7μm),using methanol-acetonitrile-isopropanol(1∶1∶1)(A) and acetonitrile-water(B) as mobile phases.Both A and B contained 0.1% formic acid and 0.01%sodium formate.The analysis of lipids was performed in positive and negative ionization modes.Under the optimized condition,mass-to-charge values of each lipid were obtained by high-resolution mass spectrometry,and the elemental composition could be evaluated.Based on the fragmentation pathways and characteristic fragment ions of lipids,totally 85 lipids were identified in serum,muscle and liver,which contain ten classes: phosphatidylcholines(PC),lysophosphatidylcholines(Lyso-PC),phosphatidylethanolamine(PE),lysophosphatidylethanolamine(Lyso-PE),phosphatidylinositol(PI),phosphatidylserine(PS),diradylglycerols(DAG),triradylglycerols(TAG),sphingomyelin(SM) and ceramides(Cer).There were 19 lipids,including Lyso-PC 16∶0,nine PCs,three PIs,four TAGs and two SMs found in muscle,liver and serum.While PS and Lyso-PE were only observed in muscle,such as PS 18∶0/22∶6,Lyso-PE 16∶0 and Lyso-PE 18∶1.Ceramides was identified in the extracts of liver tissue.PE and DAG were identified in the muscle and liver tissue.The method was demonstrated to be simple,sensitive and rapid,and was suitable for the analysis of lipid in different parts of Pelteoobagrus fulvidraco.
黄颡鱼超高效液相色谱-质谱(UPLC-MS)脂质成分
Pelteoobagrus fulvidracoultra-performance liquid chromatography coupled with mass spectrometry(UPLC-MS)lipids
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