Plasma sphingosine-1-phosphate measurement in healthy subjects: close correlation with red blood cell parameters

Ann Clin Biochem 2008;45:356-363
© 2008 Association for Clinical Biochemistry


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Original Articles

Ryunosuke Ohkawa1,
Kazuhiro Nakamura1,
Shigeo Okubo1,
Shigemi Hosogaya2,
Yukio Ozaki2,
Minoru Tozuka1,
Noriko Osima3,
Hiromitsu Yokota1,
Hitoshi Ikeda1,4 and
Yutaka Yatomi1,4

1 Department of Clinical Laboratory, The University of Tokyo Hospital, Tokyo, Japan;
2 Department of Clinical and Laboratory Medicine, Faculty of Medicine, University of Yamanashi, Yamanashi, Japan;
3 Department of Biomolecular Science, Faculty of Science, Toho University, Chiba, Japan;
4 Department of Clinical Laboratory Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan

Corresponding author: Dr Yutaka Yatomi. Email: yatoyuta-tky{at}

Background: Since sphingosine-1-phosphate (Sph-1-P) plays an important roleas an extracellular mediator through interaction with specificcell surface receptors, especially in the area of vascular biologyand immunology/haematology, determination of its plasma concentrationmay become important from the clinical viewpoint. Thus, we attemptedto develop a method of measuring the plasma Sph-1-P concentrationfor use in the clinical laboratory setting.

Methods: After two-step lipid extraction, Sph-1-P was coupled with o-phthaldialdehyde, and the resultant fluorescent derivative was separated by high-performance liquid chromatography. C17-Sph-1-P was used as the internalstandard, instead of dihydrosphingosine-1-phosphate, which hadbeen used previously for the same purpose but was actually detectedin plasma.

Results: Our procedures for preparing the plasma samples and assay Sph-1-Pwere found to be satisfactory for clinical laboratory testing.The plasma Sph-1-P concentrations were significantly higherin men (413.1 ± 52.0 nmol/L; mean ± SD) than inwomen (352.4 ± 39.7 nmol/L). Unexpectedly, strong positivecorrelations were found between the plasma Sph-1-P concentrationand red blood cell (RBC)-related parameters, rather than platelet-relatedparameters.

Conclusions: Our present study confirmed the possibility of the clinicalintroduction of plasma Sph-1-P measurement, and in addition,suggested that RBCs may be involved in the regulation of plasmaSph-1-P concentrations.

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