Simultaneous quantitation of sphingoid bases and their phosphates in biological samples by liquid chromatography/electrospray ionization tandem mass spectrometry
Simultaneous quantitation of sphingoid bases and their phosphates in biological samples by liquid chromatography/electrospray ionization tandem mass spectrometry
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DOI：
10.1007/s00216-012-6004-9
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发表时间：
2012-06-01
影响因子：
4.3
通讯作者：
Tomioka, Yoshihisa
中科院分区：
文献类型：
--
作者：
Saigusa, Daisuke;Shiba, Kanako;Tomioka, Yoshihisa
建立了C18鞘氨醇(Sph)、C18二氢鞘氨醇(DhSph)、C18植酸鞘氨醇(PSph)、C18鞘氨醇-1-磷酸(S1P)、C18二氢鞘氨醇-1-磷酸(DhS1P)和C18-1-磷酸植鞘糖苷(PS1P)的同时定量测定方法。样品通过简单的甲醇脱蛋白制备，并在选定的反应监测模式下进行分析。使用Capcell Pak ACR柱(1.5 mm I.D.)在色谱图上没有观察到峰拖尾。X 250毫米，3微米，资生堂)。狮身人面肌的校正曲线在每次注射0.050-5.00pmol范围内呈良好的线性关系(r>；0.996)。使用四个具有代表性的生物样本(血清、脑、肝脏和脾)证明了该方法的准确性和精密度，这些样本来自含有已知数量的狮身人面像的小鼠。对小鼠血浆、脑、眼、睾丸、肝、肾、肺、脾、淋巴和胸腺等组织的sph、dhSph、pSph、S1P、dhS1P和pS1P组成进行了检测。结果证实了该方法在重要狮身人面像的生理和病理分析中的有效性。
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. x 250 mm, 3 mu m, 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.