抄録
Recently we have reported that sphingosylphosphorylcholine (SPC), which is a sphingolipid mediator and generated by N-deacylation of sphingomyelin, causes Rho-kinase-mediated Ca2+-sensitization of vascular smooth muscle, thereby leading to abnormal vascular contraction, such as cerebral vasospasm after subarachnoid hemorrhage (SAH). Indeed, SPC concentration in cerebrospinal fluid is much higher in post-SAH patients than in non-SAH patients. In addition, various vascular diseases, such as acute cerebral and myocardial infarction, are also likely to involve the vasospastic activity. Therefore, in order to assess the relationship between SPC concentration and vascular diseases, we tried to develop a rapid and accurate method to measure the serum concentration, using a reversed phase high-performance liquid chromatography/electrospray ionization-tandem mass spectrometry (HPLC/ESI-MS/MS). Using deuterium-labeled SPC as an internal standard, the ratio of ion abundance was determined by multiple reaction monitoring mode. The optimized HPLC/ESI-MS/MS condition was sufficient for the construction of linear calibration curve and continuous analysis. Furthermore, we examined the influence of differences in the technical procedures for the blood collection and the sample preparation and measurement on the yield and stability of SPC. [J Physiol Sci. 2007;57 Suppl:S125]
