3. Quantitative Analysis of Underivatized 1,25-Dihydroxyvitamin D3 and D2 in Blood by UHPLC and Triple Quadrupole Mass Spectrometer Utilizing Ion Funnels Technology
Mon 12:06 PM - PosterSplash Track 1
Andre Szczesniewski
Agilent Technologies, Inc.
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A sensitive method was developed for the quantitation of underivitized 1,25-dihydroxyvitamin D3 and D2 in blood using UHPLC and Triple Quadruple equipped with ion funnels technology. The electrospray ionization interface in positive mode was used. The major ion for quantitative multiple reaction monitoring transition pair was the ammonium adduct at m/z 434.2, ([M+HN4]+) of 1,25 dihydroxyvitmin D3 ([M+H]+, m/z 417.2) that fragmented to m/z 399.2 [M-OH]+. The UHPLC system was setup with enrichment and analytical columns. The limits of detection for 1,25-dihydroxyvitamin D3 in serum extracted samples were found to be 8 pg/mL and limits of quantitation was 15 pg/mL.