Characterization of sphingomyelins in lipid extracts using a HPLC-MS-Offline-NMR method.
Sphingomyelins were characterized using a combination of a novel isocratic reversed-phase HPLC method with electrospray time-of-flight mass spectrometric detection and optional online MS/MS. The constitution of the sphingomyelins is determined by MS/MS experiments. Baseline separation of 17 compounds of a bovine brain extract (2 main compounds and 15 minor or trace compounds) was achieved with a mobile phase consisting of methanol, 2-propanol, THF, and water on a RP-18-phenyl column. In parallel, the HPLC fraction were sampled to a 600-MHz NMR spectrometer to acquire 1D and 2D NMR spectra and to elucidate the molecular structure of individual sphingomyelin components.
Willmann
Jan
J
Institute of Organic Chemistry, University of Bremen, Leobener Strasse NW 2C, 28359 Bremen, Germany.
Mahlstedt
Kerstin
K
Leibfritz
Dieter
D
Spraul
Manfred
M
Thiele
Herbert
H
D000818:Animals; D001921:Brain; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D015394:Molecular Structure; D009862:Online Systems; D013109:Sphingomyelins / Q000737:chemistry*
D013109:Sphingomyelins