Lipid analysis of human HDL and LDL by MALDI-TOF mass spectrometry and (31)P-NMR.
The analysis of HDL and LDL is important for the further understanding of atherosclerosis because changes of the protein and lipid moieties occur under pathological conditions. Because destruction of lipids leads to the formation of well-defined products such as lysophospholipids or chlorohydrins, methods that allow their fast and reliable determination would be useful. In this study, matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) was applied for the analysis of the lipid composition of human lipoproteins. These data were compared with high resolution (31)P-NMR spectroscopy. Differences between LDL and HDL in sphingomyelin and phosphatidylcholine content could be monitored by NMR and mass spectrometry, and differences with respect to the extraction efficiency were found by MALDI-TOF MS. Additionally, treatment of LDL with hypochlorite and phospholipase A(2) resulted in marked changes (formation of chlorohydrines and lysolipids). Lysophosphatidylcholines were detectable by both methods, whereas MALDI-TOF MS failed to detect chlorohydrines of phospholipids. We conclude that MALDI-TOF MS provides rapidly a reliable lipid profile of lipoproteins. However, a previous lipid separation must be performed to detect lipid oxidation products. NMR can be directly applied, but suffers from lower sensitivity, and provides only limited information on fatty acid composition.
Schiller
J
J
Institute of Medical Physics and Biophysics, Medical Department, University of Leipzig, Liebigstr. 27, 04103 Leipzig, Germany. schij@medizin.uni-leipzig.de
Zschörnig
O
O
Petković
M
M
Müller
M
M
Arnhold
J
J
Arnold
K
K
D002728:Chlorohydrins / Q000032:analysis; D002788:Cholesterol Esters / Q000032:analysis; D006801:Humans; D006997:Hypochlorous Acid / Q000494:pharmacology; D008055:Lipids / Q000032:analysis*; D008075:Lipoproteins, HDL / Q000097:blood*; D008077:Lipoproteins, LDL / Q000097:blood*; D008244:Lysophosphatidylcholines / Q000032:analysis; D009682:Magnetic Resonance Spectroscopy; D010713:Phosphatidylcholines / Q000032:analysis; D010741:Phospholipases A / Q000494:pharmacology; D010743:Phospholipids / Q000032:analysis; D012997:Solvents; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013109:Sphingomyelins / Q000032:analysis; D014280:Triglycerides / Q000032:analysis
D002728:Chlorohydrins; D002788:Cholesterol Esters; D008055:Lipids; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D008244:Lysophosphatidylcholines; D010713:Phosphatidylcholines; D010743:Phospholipids; D012997:Solvents; D013109:Sphingomyelins; D014280:Triglycerides; D006997:Hypochlorous Acid; D010741:Phospholipases A