Simultaneous profiling of vitamin D metabolites in serum by supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS)

Abstract

Simultaneous profiling of serum vitamin D (VD) metabolites with similar structures is a big challenge. Thus, we developed and validated a SFC-MS/MS method, which is capable of eluting hydrophobic molecules, for quantification of VD 2 /VD 3 , 25-OH VD 2 /VD 3 , 3-epi-25-OH VD 2 /VD 3 , 1,25-(OH) 2 VD 2 /VD 3 and 24,25-(OH) 2 VD 2 /VD 3 . VD metabolites were extracted from human serum using acetonitrile solvent. Column stationary phase, elution gradients, flow rate, column temperature, ion-source type and buffer system in post-column make-up solvent were optimized. Baseline separation of 10 VD metabolites can be achieved using PFP column within 10 min; and detection performed under positive el..