title: Fluorous derivatization combined with liquid chromatography/tandem mass spectrometry: a method for the selective and sensitive determination of sialic acids in biological samples.
abstract: We have developed a novel method for selective and sensitive analysis of sialic acids (N-acetylneuraminic, N-glycolylneuraminic, and 2-keto-3-deoxy-D-glycero-D-galactonononic acid) utilizing liquid chromatography/tandem mass spectrometry (LC/MS/MS) combined with a fluorous derivatization technique. In this method, the carboxylic groups in the sialic acids are derivatized via amidation with heptadecafluoroundecylamine, a commercially available perfluoroalkylamine reagent. This reaction proceeds rapidly and readily at room temperature in the presence of a condensation reagent. Subsequently, the derivatives are retained specifically on an LC column with a perfluoroalkyl stationary phase by means of a fluorophilic or 'fluorous' interaction, and detected by positive electrospray ionization MS/MS. The detection limits of the examined sialic acids are in the range of 60-750 amol on column. We show that the proposed method can be used to analyze trace amounts of sialic acids in biological samples.
authors.affiliation: Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka 814-0180, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Hayama
authors.forename: Yohei
authors.initials: Y
authors.lastname: Sakaguchi
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: D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods*; D005466:Fluorocarbons / Q000737:chemistry; D006801:Humans; D016014:Linear Models; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012794:Sialic Acids / Q000097:blood / Q000737:chemistry / Q000652:urine; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*; D000509:alpha-Fetoproteins / Q000737:chemistry
chemical_list: D005466:Fluorocarbons; D012794:Sialic Acids; D000509:alpha-Fetoproteins