UPLC-MS/MS method for analysis of sphingosine 1-phosphate in biological samples

A simple and sensitive liquid chromatography-tandem mass spectrometry (negative ion-electrospray ionization) methodology to determine sphingosine 1-phosphate (S1P) and sphinganine 1-phosphate (DH-S1P) in biological samples is described The method has been validated over the linearity range of 2-100 ng/ml (r > 0.999) using synthetic C(17)-sphingosine 1-phosphate (C17-S1P) as an internal standard In multiple reaction monitoring analysis (378 2 > 79 2), the lower limit of quantification for S1P was 5.0 ng/ml but the detection limit for the bioactive lipid was below 5 pg (12 fmol) Chromatographic sepal anon was achieved on a UPLC BEH Hilic column with a binary mobile phase consisting of 30 mM ammonium acetate (pH 4.0) and acetonitrile/MeOH/30 mM ammonium acetate buffer (pH 4 0) The methodology detected 176 7 +/- 540 ng/ml of SIP and 81 2 +/- 233 ng/ml of DH-S1P in human plasma, as well as 2010 +/- 72.0 ng/ml of SIP and 965 +/- 20 1 ng/ml of DH-S1P in mice plasma (C) 2010 Published by Elsevier Inc