Rapid profiling of bovine and human milk gangliosides by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry

Hyeyoung Lee, Hyun Joo An, Larry A. Lerno, J. Bruce German, Carlito B Lebrilla

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Gangliosides are anionic glycosphingolipids widely distributed in vertebrate tissues and fluids. Their structural and quantitative expression patterns depend on phylogeny and are distinct down to the species level. In milk, gangliosides are exclusively associated with the milk fat globule membrane. They may participate in diverse biological processes but more specifically to host-pathogen interactions. However, due to the molecular complexities, the analysis needs extensive sample preparation, chromatographic separation, and even chemical reaction, which makes the process very complex and time-consuming. Here, we describe a rapid profiling method for bovine and human milk gangliosides employing matrix-assisted desorption/ionization (MALDI) Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS). Prior to the analyses of biological samples, milk ganglioside standards GM3 and GD3 fractions were first analyzed in order to validate this method. High mass accuracy and high resolution obtained from MALDI FTICR MS allow for the confident assignment of chain length and degree of unsaturation of the ceramide. For the structural elucidation, tandem mass spectrometry (MS/MS), specifically as collision-induced dissociation (CID) and infrared multiphoton dissociation (IRMPD) were employed. Complex ganglioside mixtures from bovine and human milk were further analyzed with this method. The samples were prepared by two consecutive chloroform/methanol extraction and solid phase extraction. We observed a number of differences between bovine milk and human milk. The common gangliosides in bovine and human milk are 2NeuAc-2Hex-Cer (GD3) and NeuAc-2Hex-Cer (GM3); whereas, the ion intensities of ganglioside species are different between two milk samples. Kendrick mass defect plot yields grouping of ganglioside peaks according to their structural similarities. Gangliosides were further probed by tandem MS to confirm the compositional and structural assignments. We found that only in human milk ganglioside was the ceramide carbon always even numbered, which is consistent with the notion that differences in the oligosaccharide and the ceramide moieties confer to their physiological distinctions.

Original languageEnglish (US)
Pages (from-to)138-150
Number of pages13
JournalInternational Journal of Mass Spectrometry
Volume305
Issue number2-3
DOIs
StatePublished - Aug 15 2011

Fingerprint

Cyclotron resonance
milk
Gangliosides
cyclotron resonance
Ionization
Mass spectrometry
Desorption
Fourier transforms
mass spectroscopy
Ceramides
desorption
Ions
ionization
Lasers
matrices
lasers
ions
Oligosaccharides
Pathogens
Chlorine compounds

Keywords

  • Bovine milk
  • Fourier transform ion cyclotron resonance
  • Ganglioside
  • Human milk
  • Kendrick mass defect
  • Matrix-assisted laser desorption/ionization

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Spectroscopy
  • Condensed Matter Physics
  • Instrumentation

Cite this

Rapid profiling of bovine and human milk gangliosides by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry. / Lee, Hyeyoung; An, Hyun Joo; Lerno, Larry A.; German, J. Bruce; Lebrilla, Carlito B.

In: International Journal of Mass Spectrometry, Vol. 305, No. 2-3, 15.08.2011, p. 138-150.

Research output: Contribution to journalArticle

Lee, Hyeyoung ; An, Hyun Joo ; Lerno, Larry A. ; German, J. Bruce ; Lebrilla, Carlito B. / Rapid profiling of bovine and human milk gangliosides by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry. In: International Journal of Mass Spectrometry. 2011 ; Vol. 305, No. 2-3. pp. 138-150.
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