A rapid and validated HPLC method to quantify sphingosine 1-phosphate in human plasma using solid-phase extraction followed by derivatization with fluorescence detection.
We describe the development and validation of analytical methodology for the determination of sphingosine 1-phosphate (S1P) in plasma. It uses solid-phase extraction (SPE) followed by an automated reversed-phase gradient HPLC column-switching system with a pre-column derivatization with o-phthalaldehyde (OPA) and fluorescence detection. The limit of quantification was determined at 100 ng/ml exogenous sphingosine 1-phosphate with a relative standard deviation for precision and accuracy <15%. The within- and between-day relative standard deviation for precision and accuracy were also less than 15%. This validated method should be suitable to quantify plasma concentration of sphingosine 1-phosphate in relatively large numbers of samples.
Butter
Jan J
JJ
Department of Pharmacology and Pharmacotherapy, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. j.j.butter@amc.uva.nl
Koopmans
Richard P
RP
Michel
Martin C
MC
D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D008246:Lysophospholipids / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D015203:Reproducibility of Results; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification; D009764:o-Phthalaldehyde / Q000737:chemistry
D008246:Lysophospholipids; C060506:sphingosine 1-phosphate; D009764:o-Phthalaldehyde; D013110:Sphingosine