title: Simultaneous Quantification of Sphingolipids in Small Quantities of Liver by LC-MS/MS.
abstract: Sph, S1P, and Cer, derived from the membrane sphingolipids, act as intracellular and intercellular mediators, involved in various (path) physiological functions. Accordingly, determining the distributions and concentrations of these sphingolipid mediators in body tissues is an important task. Consequently, a method for determination of sphingolipids in small quantities of tissue is required. Sphingolipids analysis has been dependent on improvements in mass spectrometry (MS) technology. Additionally, decomposition of sphingosine-1-phosphate (S1P) in the tissue samples before preparation for MS has hindered analysis. In the present study, a method for stabilization of liver samples before MS preparation was developed using a heat stabilizer (Stabilizor™ T1). Then, a LC-MS/MS method using a triple-quadrupole mass spectrometer with a C8 column was developed for simultaneous determination of sphingolipids in small quantities of liver specimens. This method showed good separation and validation results. Separation was performed with a gradient elution of solvent A (5 mmol L(-1) ammonium formate in water, pH 4.0) and solvent B (5 mmol L(-1) ammonium formate in 95% acetonitrile, pH 4.0) at 300 μL min(-1). The lower limit of quantification was less than 132 pmol L(-1), and this method was accurate (∼13.5%) and precise (∼7.13%) for S1P analysis. The method can be used to show the tissue distribution of sphingolipids.
authors.affiliation: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University; Tohoku University School of Medicine; CREST, Japan Science and Technology Corporation (JST).
authors.forename: Daisuke
authors.initials: D
authors.lastname: Saigusa
authors.affiliation: Department of Molecular and Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Tohoku University.
authors.forename: Michiyo
authors.initials: M
authors.lastname: Okudaira
authors.affiliation: Department of Molecular and Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Tohoku University.
authors.forename: Jiao
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Molecular and Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Tohoku University.
authors.forename: Kuniyuki
authors.initials: K
authors.lastname: Kano
authors.affiliation: Department of Clinical Laboratory, Medicine Graduate School of Medicine, The University of Tokyo.
authors.forename: Makoto
authors.initials: M
authors.lastname: Kurano
authors.affiliation: Department of Clinical Laboratory, Medicine Graduate School of Medicine, The University of Tokyo.
authors.forename: Baasanjav
authors.initials: B
authors.lastname: Uranbileg
authors.affiliation: Department of Clinical Laboratory, Medicine Graduate School of Medicine, The University of Tokyo.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Ikeda
authors.affiliation: Department of Clinical Laboratory, Medicine Graduate School of Medicine, The University of Tokyo.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Yatomi
authors.affiliation: Department of Integrative Genomics, Tohoku Medical Megabank Organization, Tohoku University; Department of Gene Expression Regulation, Institute of Development, Aging and Cancer, Tohoku University.
authors.forename: Hozumi
authors.initials: H
authors.lastname: Motohashi
authors.affiliation: CREST, Japan Science and Technology Corporation (JST); Department of Molecular and Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Tohoku University.
authors.forename: Junken
authors.initials: J
authors.lastname: Aoki
keywords: ceramide; heat stabilizer; liver; sphingosine; sphingosine-1-phosphates
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