1. Simultaneous quantitation of sphingoid bases and their phosphates in biological samples by liquid chromatography/electrospray ionization tandem mass spectrometry
- Author
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Kaori Suzuki, Takanori Hishinuma, Kotaro Hama, Nariyasu Mano, Masayoshi Saito, Hiroaki Yamaguchi, Junichi Goto, Kanako Shiba, Junken Aoki, Yoshihisa Tomioka, Daisuke Saigusa, Nagisa Iida, Naoto Suzuki, Michiyo Okutani, Asuka Inoue, and Tohru Kaneko
- Subjects
Spectrometry, Mass, Electrospray Ionization ,Chromatography ,Calibration curve ,Electrospray ionization ,Selected reaction monitoring ,Reproducibility of Results ,Tandem mass spectrometry ,Biochemistry ,Quantitative determination ,Analytical Chemistry ,Mice ,chemistry.chemical_compound ,Liquid chromatography electrospray ionization tandem mass spectrometry ,chemistry ,Sphingosine ,Tandem Mass Spectrometry ,Calibration ,Lc ms ms ,Animals ,Tissue Distribution ,lipids (amino acids, peptides, and proteins) ,Methanol ,Chromatography, Liquid - 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.
- Published
- 2012
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