Quantitative analysis of sphingosine-1-phosphate by HPLC after napthalene-2,3-dicarboxaldehyde (NDA) derivatization

Xingxuan He, Chien-Ling Huang, Edward H. Schuchman

Research output: Journal article publicationJournal articleAcademic researchpeer-review

24 Citations (Scopus)

Abstract

Sphingosine-1-phosphate (S1P) is an important sphingolipid signaling molecule that regulates numerous cellular processes. In this paper we report a new method to quantify the levels of S1P in biological samples that relies on derivatization with naphthalene-2,3-dicarboxaldehyde (NDA) and quantification by reverse-phase high performance liquid chromatography (HPLC). The limit of detection (LOD) using S1P standards was 20.9 fmol (12.6 nM), and the limit of quantification (LOQ) was 69.6 fmol (41.7 nM). The recovery of S1P standards was up to 97.5%. The mean relative standard deviations (RSD) for the intra- and inter-day assay were 4.1% and 7.7%, respectively. To validate this procedure, we quantified the S1P levels in plasma from human, horse, and mouse (mean values of 0.45, 0.25, and 1.23 μM, respectively). We also used this technique to evaluate the S1P content in mouse tissues, as well as in rat neuronal cell cultures before and after sphingosine treatment. The results demonstrate that this new procedure can provide fast, sensitive, and reproducible S1P quantification, and offers several advantages over existing methods. The technique also may be used for determining the activity, as well as the inhibition, of sphingosine kinase. In the future it could be an important tool for investigators studying the role of S1P in signal transduction, cell growth and differentiation, and disease pathogenesis and treatment.
Original languageEnglish
Pages (from-to)983-990
Number of pages8
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume877
Issue number10
DOIs
Publication statusPublished - 1 Apr 2009
Externally publishedYes

Keywords

  • Biological samples
  • HPLC
  • Naphthalene-2,3-dicarboxaldehyde
  • Sphingosine-1-phosphate

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

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