LC-MS-based method for the qualitative and quantitative analysis of complex lipid mixtures.
A simple and robust LC-MS-based methodology for the investigation of lipid mixtures is described, and its application to the analysis of human lipoprotein-associated lipids is demonstrated. After an optional initial fractionation on Silica 60, normal-phase HPLC-MS on a YMC PVA-Sil column is used first for class separation, followed by reversed-phase LC-MS or LC-tandem mass spectrometry using an Atlantis dC18 capillary column, and/or nanospray MS, to fully characterize the individual lipids. The methodology is applied here for the analysis of human apolipoprotein B-associated lipids. This approach allows for the determination of even low percentages of lipids of each molecular species and showed clear differences between lipids associated with apolipoprotein B-100-LDL isolated from a normal individual and those associated with a truncated version, apolipoprotein B-67-containing lipoproteins, isolated from a homozygote patient with familial hypobetalipoproteinemia. The methods described should be easily adaptable to most modern MS instrumentation.
Sommer
Ulf
U
Mass Spectrometry Resource, Department of Biochemistry, Boston University School of Medicine, Boston, MA, USA.
Herscovitz
Haya
H
Welty
Francine K
FK
Costello
Catherine E
CE
D002853:Chromatography, Liquid / Q000379:methods*; D005732:Gangliosides / Q000032:analysis; D006801:Humans; D008055:Lipids / Q000032:analysis*; D008074:Lipoproteins / Q000032:analysis; D013058:Mass Spectrometry / Q000379:methods*
D005732:Gangliosides; D008055:Lipids; D008074:Lipoproteins