title: Simultaneous quantitation of sphingoid bases and their phosphates in biological samples by liquid chromatography/electrospray ionization tandem mass spectrometry.
abstract: We developed a liquid chromatography/electrospray ionization tandem mass spectrometry method for the simultaneous quantitative determination of C18 sphingosine (Sph), C18 dihydrosphingosine (dhSph), C18 phytosphingosine (pSph), C18 sphingosine-1-phosphate (S1P), C18 dihydrosphingosine-1-phosphate (dhS1P), and C18 phytosphingosine-1-phosphate (pS1P). Samples were prepared by simple methanol deproteinization and analyzed in selected reaction monitoring modes. No peak tailing was observed on the chromatograms using a Capcell Pak ACR column (1.5 mm i.d. × 250 mm, 3 μm, Shiseido). The calibration curves of the sphingoids showed good linearity (r > 0.996) over the range of 0.050-5.00 pmol per injection. The accuracy and precision of this method were demonstrated using four representative biological samples (serum, brain, liver, and spleen) from mice that contained known amounts of the sphingoids. Samples of mice tissue such as plasma, brain, eye, testis, liver, kidney, lung, spleen, lymph node, and thymus were examined for their Sph, dhSph, pSph, S1P, dhS1P, and pS1P composition. The results confirmed the usefulness of this method for the physiological and pathological analysis of the composition of important sphingoids.
authors.affiliation: Laboratory of Oncology, Pharmacy Practice and Sciences, Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3 Aoba, Aramaki, Sendai 980-8578, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Saigusa
authors.forename: Kanako
authors.initials: K
authors.lastname: Shiba
authors.forename: Asuka
authors.initials: A
authors.lastname: Inoue
authors.forename: Kotaro
authors.initials: K
authors.lastname: Hama
authors.forename: Michiyo
authors.initials: M
authors.lastname: Okutani
authors.forename: Nagisa
authors.initials: N
authors.lastname: Iida
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Saito
authors.forename: Kaori
authors.initials: K
authors.lastname: Suzuki
authors.forename: Tohru
authors.initials: T
authors.lastname: Kaneko
authors.forename: Naoto
authors.initials: N
authors.lastname: Suzuki
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Yamaguchi
authors.forename: Nariyasu
authors.initials: N
authors.lastname: Mano
authors.forename: Junichi
authors.initials: J
authors.lastname: Goto
authors.forename: Takanori
authors.initials: T
authors.lastname: Hishinuma
authors.forename: Junken
authors.initials: J
authors.lastname: Aoki
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Tomioka
mesh_terms: D000818:Animals; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D051379:Mice; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013110:Sphingosine / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014018:Tissue Distribution
chemical_list: D013110:Sphingosine