As lipidomics has attracted increased attention in life science,advanced mass spectrometry(MS)technologies have been combined with other separation techniques to improve and expand the branch of study.This review inte...As lipidomics has attracted increased attention in life science,advanced mass spectrometry(MS)technologies have been combined with other separation techniques to improve and expand the branch of study.This review intends to provide general knowledge of offline and online coupling of flow field-flow fractionation(FlFFF)—a technique that encompasses the separation of nano-to micro-scale biomolecules—with MS for analysis of blood plasma lipoproteins,processes that are considered bottomup and top-down approaches,respectively.The first part of this review focuses on the bottom-up method using multiplexed hollow fiber FlFFF(MxHF5)and nanoflow liquid chromatography electrospray-ionization tandem mass spectrometry(nLC-ESI–MS/MS)for non-targeted identification of lipids.In this protocol,plasma lipoproteins of different types are collected using MxHF5,and the lipids within the lipoproteins are then extracted and analyzed via nLC-ESI–MS/MS.The second part of the review describes the top-down approach,which uses online coupling of miniaturized FlFFF to ESI–MS for a fast screening of targeted lipids.Here,the separation of lipoproteins and detection of their component lipids are achieved simultaneously.While both methods aim to quantify the lipids within lipoproteins,the bottom-up approach provides an extensive lipidome,whereas the top-down method is suitable for high-speed targeted lipidomic analysis.This review discusses variants of FlFFF-ESI–MS/MS that offer effective analytical technologies for lipidomics.展开更多
[目的]建立非对称场流分离检测鲍内脏多糖的方法。[方法]采用非对称场流分离系统与静态光散射、光电二极管阵列和示差折光检测器联用技术分离表征鲍内脏多糖。以0.05 mol/L Na NO3[含0.02%(W/V) Na N3]为流动相,研究横向流速和样品浓度...[目的]建立非对称场流分离检测鲍内脏多糖的方法。[方法]采用非对称场流分离系统与静态光散射、光电二极管阵列和示差折光检测器联用技术分离表征鲍内脏多糖。以0.05 mol/L Na NO3[含0.02%(W/V) Na N3]为流动相,研究横向流速和样品浓度对非对称场流分离多糖的影响,并利用动静态光散射测量鲍内脏多糖的分子特性(分子量、均方根旋转半径、分子构象、流体力学半径)。[结果]不同横向流速对多糖的分离表征有显著影响;一定范围内,不同多糖浓度对分离效果及分子特性结果无显著差异。鲍内脏多糖分子量为(25.40±1.78) k D,均方根旋转半径为(16.70±0.30) nm,流体力学半径为(143.23±15.49) nm,分子为无规则线团构象。[结论]非对称场流技术适用于鲍内脏多糖的分离检测。展开更多
基金Grant from the National Research Foundation of Korea(NRF-2015R1A2A1A01004677).
文摘As lipidomics has attracted increased attention in life science,advanced mass spectrometry(MS)technologies have been combined with other separation techniques to improve and expand the branch of study.This review intends to provide general knowledge of offline and online coupling of flow field-flow fractionation(FlFFF)—a technique that encompasses the separation of nano-to micro-scale biomolecules—with MS for analysis of blood plasma lipoproteins,processes that are considered bottomup and top-down approaches,respectively.The first part of this review focuses on the bottom-up method using multiplexed hollow fiber FlFFF(MxHF5)and nanoflow liquid chromatography electrospray-ionization tandem mass spectrometry(nLC-ESI–MS/MS)for non-targeted identification of lipids.In this protocol,plasma lipoproteins of different types are collected using MxHF5,and the lipids within the lipoproteins are then extracted and analyzed via nLC-ESI–MS/MS.The second part of the review describes the top-down approach,which uses online coupling of miniaturized FlFFF to ESI–MS for a fast screening of targeted lipids.Here,the separation of lipoproteins and detection of their component lipids are achieved simultaneously.While both methods aim to quantify the lipids within lipoproteins,the bottom-up approach provides an extensive lipidome,whereas the top-down method is suitable for high-speed targeted lipidomic analysis.This review discusses variants of FlFFF-ESI–MS/MS that offer effective analytical technologies for lipidomics.
文摘[目的]建立非对称场流分离检测鲍内脏多糖的方法。[方法]采用非对称场流分离系统与静态光散射、光电二极管阵列和示差折光检测器联用技术分离表征鲍内脏多糖。以0.05 mol/L Na NO3[含0.02%(W/V) Na N3]为流动相,研究横向流速和样品浓度对非对称场流分离多糖的影响,并利用动静态光散射测量鲍内脏多糖的分子特性(分子量、均方根旋转半径、分子构象、流体力学半径)。[结果]不同横向流速对多糖的分离表征有显著影响;一定范围内,不同多糖浓度对分离效果及分子特性结果无显著差异。鲍内脏多糖分子量为(25.40±1.78) k D,均方根旋转半径为(16.70±0.30) nm,流体力学半径为(143.23±15.49) nm,分子为无规则线团构象。[结论]非对称场流技术适用于鲍内脏多糖的分离检测。
