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Journal of Biochemistry 2006 139(4):771-777; doi:10.1093/jb/mvj090
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© 2006 The Japanese Biochemical Society.

Regular Paper

Convenient Structural Analysis of Glycosphingolipids Using MALDI-QIT-TOF Mass Spectrometry with Increased Laser Power and Cooling Gas Flow

Yusuke Suzuki1, Minoru Suzuki1, Emi Ito1, Naoko Goto-Inoue1, Kozo Miseki2, Junko Iida2, Yuzo Yamazaki2, Masaki Yamada2 and Akemi Suzuki1,*

1 Sphingolipid Expression Laboratory, RIKEN Frontier Research System, Wako, Saitama 351-0198; and 2 Shimadzu Corporation, Nakagyo-ku, Kyoto 604-8511

* To whom correspondence should be addressed. Tel: +81-48-467-9615, Fax: +81-48-462-4692; E-mail: aksuzuki{at}riken.jp

Matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry (MALDI-QIT-TOF MS) was applied to the structural characterization of neutral glycosphingolipids. Lithium adduct ions of glycosphingolipids were analyzed using MALDI-QIT-TOF MS under strong conditions of increased laser power and cooling gas flow. The relative intensities of fragment ions were increased under the strong conditions, and the resulting spectra revealed the presence of oligosaccharide ions fragmented from the glycosphingolipids. Consequently, the oligosaccharide sequences of the glycosphingolipids were readily obtained. To obtain more detailed structural information, MS/MS (MS2) and MS/MS/MS (MS3) analyses were performed with selection of the lactosylceramide and ceramide ions, respectively. The resulting data were sufficient to determine the structures of both the oligosaccharide and the ceramide moiety of each glycosphingolipid. The fragmentation patterns of MS2 and MS3 for Forssman glycolipid under the strong conditions were comparable to those of MS3 and MS4 obtained under standard conditions, respectively. Thus, MALDI-QIT-TOF MS with increased laser power and cooling gas flow is a convenient method for glycosphingolipid analysis.


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