UPLC-MS/MS 法分析生物样品中的鞘氨醇 1-磷酸。
UPLC-MS/MS method for analysis of sphingosine 1-phosphate in biological samples.
机构信息
CNR - Institute of Biomolecular Chemistry, via Campi Flegrei 34, 80078 Pozzuoli (NA), Italy.
出版信息
Prostaglandins Other Lipid Mediat. 2010 Sep;93(1-2):25-9. doi: 10.1016/j.prostaglandins.2010.06.001. Epub 2010 Jul 1.
A simple and sensitive liquid chromatography-tandem mass spectrometry (negative ion-electrospray ionization) methodology to determine sphingosine 1-phosphate (S1P) and sphinganine 1-phosphate (DH-S1P) in biological samples is described. The method has been validated over the linearity range of 2-100ng/ml (r>0.999) using synthetic C(17)-sphingosine 1-phosphate (C17-S1P) as an internal standard. In multiple reaction monitoring analysis (378.2>79.2), the lower limit of quantification for S1P was 5.0ng/ml but the detection limit for the bioactive lipid was below 5pg (12fmol). Chromatographic separation was achieved on a UPLC BEH Hilic column with a binary mobile phase consisting of 30mM ammonium acetate (pH 4.0) and acetonitrile/MeOH/30mM ammonium acetate buffer (pH 4.0). The methodology detected 176.7+/-54.0ng/ml of S1P and 81.2+/-23.3ng/ml of DH-S1P in human plasma, as well as 201.0+/-72.0ng/ml of S1P and 96.5+/-20.1ng/ml of DH-S1P in mice plasma.
描述了一种用于在生物样品中测定鞘氨醇 1-磷酸（S1P）和二氢鞘氨醇 1-磷酸（DH-S1P）的简单且灵敏的液相色谱-串联质谱（负离子-电喷雾电离）方法。该方法使用合成的 C（17）-鞘氨醇 1-磷酸（C17-S1P）作为内标，在 2-100ng/ml 的线性范围内进行了验证（r>0.999）。在多重反应监测分析（378.2>79.2）中，S1P 的定量下限为 5.0ng/ml，但生物活性脂质的检测限低于 5pg（12fmol）。色谱分离在 UPLC BEH Hilic 柱上进行，采用由 30mM 乙酸铵（pH 4.0）和乙腈/甲醇/30mM 乙酸铵缓冲液（pH 4.0）组成的二元流动相。该方法检测到人血浆中 S1P 的含量为 176.7+/-54.0ng/ml，DH-S1P 的含量为 81.2+/-23.3ng/ml，小鼠血浆中 S1P 的含量为 201.0+/-72.0ng/ml，DH-S1P 的含量为 96.5+/-20.1ng/ml。
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