High-performance liquid chromatography-mass spectrometry of glycosphingolipids: I. Structural characterization of molecular species of GlcCer and IV3 beta Gal-Gb4Cer.
A method of high-performance liquid chromatography-fast atom bombardment mass spectrometry (HPLC/FAB/MS) for the structural characterization of glycosphingolipids was developed, which involves a frit interface between the HPLC and the MS. The molecular species of glucosylceramide (GlcCer) purified from the spleen of a patient with Gaucher's disease and galactosylglobotetraosylceramide (IV3 beta Gal-Gb4Cer) from mouse kidney were analyzed using this system on a reversed-phase column, with methanol containing 1% glycerol as the elution solvent. The injection of 1 microgram of GlcCer gave the mass spectra of seven major molecular species, the pseudo-molecular ion for each of the seven molecular species being observed at m/z 698, 726, 754, 782, 808, 796, and 810, respectively. The injection of 200 pg of synthetic N-stearoyl glucosylsphingosine (d18:1) gave a clear peak with the single ion monitoring method detecting the pseudo-molecular ion at m/z 726. The injection of 5 micrograms of IV3 beta Gal-Gb4Cer gave the mass spectra of six major molecular species, the pseudo-molecular ions being observed at m/z 1,489, 1,471, 1,515, 1,497, 1,517, and 1,499. This report deals with a new HPLC/FAB/MS system, which was successfully applied to the structural characterization of the molecular species of neutral glycosphingolipids, and the system is a quite promising for development into a quantitative method for glycosphingolipids with high sensitivity and specificity.
Suzuki
M
M
Department of Membrane Biochemistry, Tokyo Metropolitan Institute of Medical Science.
Sekine
M
M
Yamakawa
T
T
Suzuki
A
A
D000818:Animals; D015295:Antigens, Tumor-Associated, Carbohydrate; D002554:Cerebrosides / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D005963:Glucosylceramides / Q000032:analysis*; D006017:Glycolipids / Q000032:analysis*; D006028:Glycosphingolipids / Q000032:analysis*; D007668:Kidney / Q000378:metabolism; D013058:Mass Spectrometry; D051379:Mice; D055746:Stage-Specific Embryonic Antigens
D015295:Antigens, Tumor-Associated, Carbohydrate; D002554:Cerebrosides; D005963:Glucosylceramides; D006017:Glycolipids; D006028:Glycosphingolipids; D055746:Stage-Specific Embryonic Antigens; C052924:stage-specific embryonic antigen-3