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 2-100ng/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 5.0ng/ml but the detection limit for the bioactive lipid was below 5pg (12fmol). Chromatographic separation was achieved on a UPLC BEH Hilic column with a binary mobile phase consisting of 30mM ammonium acetate (pH 4.0) and acetonitrile/MeOH/30mM ammonium acetate buffer (pH 4.0). The methodology detected 176.7+/-54.0ng/ml of S1P and 81.2+/-23.3ng/ml of DH-S1P in human plasma, as well as 201.0+/-72.0ng/ml of S1P and 96.5+/-20.1ng/ml of DH-S1P in mice plasma.
Cutignano
Adele
A
CNR - Institute of Biomolecular Chemistry, via Campi Flegrei 34, 80078 Pozzuoli (NA), Italy. acutignano@icb.cnr.it
Chiuminatto
Ugo
U
Petruzziello
Filomena
F
Vella
Filomena Monica
FM
Fontana
Angelo
A
D000328:Adult; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D008246:Lysophospholipids / Q000097:blood*; D051379:Mice; D008815:Mice, Inbred Strains; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000097:blood; D053719:Tandem Mass Spectrometry / Q000379:methods*
D008246:Lysophospholipids; C060504:dihydrosphingosine 1-phosphate; C060506:sphingosine 1-phosphate; D013110:Sphingosine