高效液相色谱法测定生物样品中磷酸鞘氨醇的含量

Quantitative Analysis of Sphingosine-1-phosphate in Biological Samples by HPLC

目的建立生物样品中磷酸鞘氨醇(S1P)的定量方法。方法采用有机溶剂萃取、碱性磷酸酶脱磷酸、衍生化后经高效液相色谱仪测定动物血清和组织中S1P的含量。结果建立了一种快捷、灵敏的检测生物样品中S1P含量的方法——高效液相色谱法(HPLC),并测得小鼠血清和肝组织中S1P的含量分别为(1.72±0.40)μmol/L和(150±25)pmol/g。结论S1P定量检测方法的建立为进一步研究其重要生物学功能奠定了基础。

Objective Sphingosine-1-phosphate（S1P） is a sphingolipid metabolite which has dual actions of both an intracellular second messenger and a ligand for a family of G protein-coupled receptors. It has been implicated in many and diverse biological processes, such as cell growth, cell survival, angiogenesis, cell migration and differentiation, and more recently, in the regulation of immune function. In the present study, the aim is to establish a method for the measurement of SIP from biological samples. Methods The method consists of two steps of sample pretreatment： the enzymatic dephosphorylation of sphingid-1-phosphate by alkaline phosphatase and the subsequent analysis of o-phthalaldehyde （OPA） derivatives of the liberated sphingid by high performance liquid chromatography （HPLC）. Results We presented a very simple, fast, and sensitive method for determination of S1P in mice serum and liver tissue. The concentrations of S1P in these samples were （ 1.72 ± 0.40 ） μmol/L and （ 150±25 ） pmol/g, respectively. Conclusion The method for the quantification of S1P by HPLC is essential to further investigate the possible role S1P in health and disease.