title: Liquid chromatography/tandem mass spectrometry with fluorous derivatization method for selective analysis of sialyl oligosaccharides.
abstract: BACKGROUND
  A separation-oriented derivatization method using a specific fluorous affinity between perfluoroalkyl-containing compounds was applied to selective liquid chromatography/tandem mass spectrometric (LC/MS/MS) analysis of sialyl oligosaccharides. The perfluoroalkyl-labeled sialyl oligosaccharides could be selectively retained on an LC column with the perfluoroalkyl-modified stationary phase and effectively distinguished from non-derivatized species.
  
  
  METHODS
  Sialyl oligosaccharides (3'-sialyllactose, 6'-sialyllactose, sialyllacto-N-tetraose a, sialyllacto-N-tetraose b, sialyllacto-N-tetraose c, and disialyllacto-N-tetraose) were derivatized with 4,4,5,5,6,6,7,7,8,8,9,9,10,10,11,11,11-heptadecafluoroundecylamine via amidation in the presence of 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (condensation reagent). The obtained derivatives were directly injected onto the fluorous LC column without any pretreatments and then detected by positive electrospray ionization MS/MS.
  
  
  RESULTS
  The method enabled accurate determination of the sialyl oligosaccharides in biological samples such as human urine and human milk, because there was no interference with matrix-induced effects during LC/MS/MS analysis. The limits of detection of the examined sialyl oligosaccharides, defined as signal-to-noise (S/N) = 3, were in the range 0.033-0.13 nM. Accuracy in the range 95.6-108% was achieved, and the precision (relative standard deviation) was within 9.4%.
  
  
  CONCLUSIONS
  This method enabled highly selective and sensitive analysis of sialyl oligosaccharides, enabling accurate measurement of even their trace amounts in biological matrices. The proposed method may prove to be a powerful tool for the analysis of various sialyl oligosaccharides.
authors.affiliation: Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka, 814-0180, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Sakaguchi
authors.forename: Tadashi
authors.initials: T
authors.lastname: Hayama
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Yoshida
authors.forename: Miki
authors.initials: M
authors.lastname: Itoyama
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Todoroki
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Yamaguchi
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Nohta
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D002675:Child, Preschool; D002853:Chromatography, Liquid / Q000379:methods*; D005260:Female; D005466:Fluorocarbons / Q000737:chemistry; D006801:Humans; D057230:Limit of Detection; D008297:Male; D008895:Milk, Human / Q000737:chemistry; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry* / Q000652:urine; D015203:Reproducibility of Results; D012794:Sialic Acids / Q000032:analysis / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult
chemical_list: C558756:4,4,5,5,6,6,7,7,8,8,9,9,10,10,11,11,11-heptadecafluoro-n-undecylamine; D005466:Fluorocarbons; D009844:Oligosaccharides; D012794:Sialic Acids