Profiling of phospholipids in lipoproteins by multiplexed hollow fiber flow field-flow fractionation and nanoflow liquid chromatography-tandem mass spectrometry.
This study describes a coupled analytical method to carry out the systematic profiling of phospholipids (PLs) in high-density lipoproteins (HDL) and low-density lipoproteins (LDL) from human blood plasma. HDL and LDL of healthy human plasma samples were separated by size and collected on a semi-preparative scale using multiplexed hollow fiber flow field-flow fractionation (MxHF5). Phospholipid mixtures contained in the resulting HDL and LDL fractions were analyzed by shotgun nanoflow liquid chromatography-tandem mass spectrometry (nLC-ESI-MS-MS). We utilized a dual scan method for the separation and simultaneous characterization of complicated PL mixtures by nLC-ESI-MS-MS, such that phosphatidylcholine (PC) and phosphatidylethanolamine (PE) molecules were detected in positive ion mode in a first LC run. In a second LC run, phosphatidylinositol (PI), phosphatidylglycerol (PG), and phosphatidic acid (PA) were detected in negative ion mode. In this study, a total of 56 PLs from HDL and 52 PLs from LDL particles were characterized by their molecular structures from data dependent collision-induced dissociation (CID) experiments, and their relative abundances were compared.
Lee
Ju Yong
JY
Department of Chemistry, Yonsei University, 134 Shinchon-dong, Seoul 120-749, South Korea.
Min
Hye Kyeong
HK
Choi
Donghoon
D
Moon
Myeong Hee
MH
D002853:Chromatography, Liquid / Q000379:methods*; D031521:Fractionation, Field Flow / Q000379:methods*; D006801:Humans; D008074:Lipoproteins / Q000097:blood* / Q000302:isolation & purification; D010743:Phospholipids / Q000097:blood* / Q000302:isolation & purification; D053719:Tandem Mass Spectrometry / Q000379:methods*
D008074:Lipoproteins; D010743:Phospholipids