TY - JOUR
T1 - Top-down lipidomic analysis of human lipoproteins by chip-type asymmetrical flow field-flow fractionation-electrospray ionization-tandem mass spectrometry
AU - Kim, Ki Hun
AU - Lee, Ju Yong
AU - Lim, Sangsoo
AU - Moon, Myeong Hee
PY - 2013/3/8
Y1 - 2013/3/8
N2 - This study demonstrates the potential utility of on-line chip-type asymmetrical flow field-flow fractionation (cAF4) and electrospray ionization tandem mass spectrometry (ESI-MS-MS) for the top-down lipidomic analysis of human lipoproteins. Utilizing a cAF4, which is a miniaturized AF4 channel operated with a micro flow rate regime, enabled high density lipoprotein (HDL) and low density lipoprotein (LDL) to be separated by hydrodynamic diameter in an aqueous solution with the simultaneous desalting of lipoproteins. On-line desalting was found to enhance the ionization of lipoproteinic lipid molecules during the feeding of cAF4 effluent to ESI-MS when compared to the direct infusion of lipoproteins to MS. An evaluation of top-down lipidomic analysis was performed to test the efficiency of in-source fragmentation during cAF4-ESI-MS in the dissociation of lipoprotein particles into individual lipid molecules. This study demonstrates the structural identification of the following lipid classes: phosphatidylcholines (PCs), cholesteryl esters (CEs), and regioisomers of triacylglycerols (TAGs) having an identical mass but different acyl chains and dimeric forms of TAGs in the positive ion mode, and phosphatidylglycerols (PGs), phosphatidic acids (PAs), phosphatidyinositols (PIs), and their lyso species in the negative ion mode. The developed method was applied to plasma samples from patients with coronary artery disease (CAD) for the separation of HDL and LDL and for the simultaneous analysis of lipoproteinic lipids, resulting in the identification of 11 PCs, 9 PGs, 4 PAs, 2 PIs, 2 PEs, 18 TAGs, and 6 CEs.
AB - This study demonstrates the potential utility of on-line chip-type asymmetrical flow field-flow fractionation (cAF4) and electrospray ionization tandem mass spectrometry (ESI-MS-MS) for the top-down lipidomic analysis of human lipoproteins. Utilizing a cAF4, which is a miniaturized AF4 channel operated with a micro flow rate regime, enabled high density lipoprotein (HDL) and low density lipoprotein (LDL) to be separated by hydrodynamic diameter in an aqueous solution with the simultaneous desalting of lipoproteins. On-line desalting was found to enhance the ionization of lipoproteinic lipid molecules during the feeding of cAF4 effluent to ESI-MS when compared to the direct infusion of lipoproteins to MS. An evaluation of top-down lipidomic analysis was performed to test the efficiency of in-source fragmentation during cAF4-ESI-MS in the dissociation of lipoprotein particles into individual lipid molecules. This study demonstrates the structural identification of the following lipid classes: phosphatidylcholines (PCs), cholesteryl esters (CEs), and regioisomers of triacylglycerols (TAGs) having an identical mass but different acyl chains and dimeric forms of TAGs in the positive ion mode, and phosphatidylglycerols (PGs), phosphatidic acids (PAs), phosphatidyinositols (PIs), and their lyso species in the negative ion mode. The developed method was applied to plasma samples from patients with coronary artery disease (CAD) for the separation of HDL and LDL and for the simultaneous analysis of lipoproteinic lipids, resulting in the identification of 11 PCs, 9 PGs, 4 PAs, 2 PIs, 2 PEs, 18 TAGs, and 6 CEs.
KW - Asymmetrical flow field-flow fractionation (AF4)
KW - CAF4-ESI-MS-MS
KW - Chip-type AF4
KW - LDL
KW - Lipoproteins
KW - Top-down lipidomics
UR - http://www.scopus.com/inward/record.url?scp=84873525917&partnerID=8YFLogxK
U2 - 10.1016/j.chroma.2013.01.025
DO - 10.1016/j.chroma.2013.01.025
M3 - Article
C2 - 23375771
AN - SCOPUS:84873525917
SN - 0021-9673
VL - 1280
SP - 92
EP - 97
JO - Journal of Chromatography A
JF - Journal of Chromatography A
ER -