Development of method to measure the serum concentration of sphingosylphosphorylcholine, an inducer of abnormal vascular contraction

Yuichi Takada; Hiroko Kishi; Hozumi Kawamichi; Katsuko Kajiya; Fengling Guo; Dan Xu; Chen Wang; Sayaka Matsuo; Saori Miwa; Sei Kobayashi

doi:10.14849/psjproc.2007.0.125.1

Proceedings of Annual Meeting of the Physiological Society of Japan

Session ID : 1PHP-028

DOI https://doi.org/10.14849/psjproc.2007.0.125.1

Conference information

Development of method to measure the serum concentration of sphingosylphosphorylcholine, an inducer of abnormal vascular contraction

*Yuichi Takada, Hiroko Kishi, Hozumi Kawamichi, Katsuko Kajiya, Fengling Guo, Dan Xu, Chen Wang, Sayaka Matsuo, Saori Miwa, Sei Kobayashi

Author information

*Yuichi Takada
Dept. of Molec. Physiol. and Med. Bioreg., Grad. Sch. of Med. Yamaguchi Univ, Ube, Japan
Innovation Plaza Hiroshima, JST, Japan
Hiroko Kishi
Dept. of Molec. Physiol. and Med. Bioreg., Grad. Sch. of Med. Yamaguchi Univ, Ube, Japan
Hozumi Kawamichi
Dept. of Molec. Physiol. and Med. Bioreg., Grad. Sch. of Med. Yamaguchi Univ, Ube, Japan
Katsuko Kajiya
Dept. of Molec. Physiol. and Med. Bioreg., Grad. Sch. of Med. Yamaguchi Univ, Ube, Japan
Fengling Guo
Dept. of Molec. Physiol. and Med. Bioreg., Grad. Sch. of Med. Yamaguchi Univ, Ube, Japan
Dan Xu
Dept. of Molec. Physiol. and Med. Bioreg., Grad. Sch. of Med. Yamaguchi Univ, Ube, Japan
Chen Wang
Dept. of Molec. Physiol. and Med. Bioreg., Grad. Sch. of Med. Yamaguchi Univ, Ube, Japan
Sayaka Matsuo
Dept. of Molec. Physiol. and Med. Bioreg., Grad. Sch. of Med. Yamaguchi Univ, Ube, Japan
Saori Miwa
Dept. of Molec. Physiol. and Med. Bioreg., Grad. Sch. of Med. Yamaguchi Univ, Ube, Japan
Sei Kobayashi
Dept. of Molec. Physiol. and Med. Bioreg., Grad. Sch. of Med. Yamaguchi Univ, Ube, Japan

Keywords: sphingosylphosphorylcholine, vascular disease, serum concentration, HPLC/ESI-MS/MS

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Abstract

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]

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