Quantitative analysis of the tumor suppressor dendrogenin A using liquid chromatography tandem mass spectrometry

Abstract : Dendrogenin A (DDA) was recently identified as a mammalian cholesterol metabolite that displays tumor suppressor and neurostimulating properties at low doses. In breast tumors, DDA levels were found to be decreased compared to normal tissues, evidencing a metabolic deregulation of DDA production in cancers. DDA is an amino-oxysterol that contains three protonatable nitrogen atoms. This makes it physico-chemically different from other oxysterols and it therefore requires specific analytical methods We have previously used a two-step method for the quantification of DDA in biological samples: 1) DDA purification from a Bligh and Dyer extract by RP-HPLC using a 250×4.6mm column, followed by 2) nano-electrospray ionization mass spectrometry (MS) fragmentation to analyze the HPLC fraction of interest. We report here the development a liquid chromatography tandem mass spectrometry method for the analysis of DDA and its analogues. This new method is fast (10min), resolving (peak width <4s) and has a weak carryover (<0.01%). We show that this technique efficiently separates DDA from its C17 isomer and other steroidal alkaloids from the same family establishing a proof of concept for the analysis of this family of amino-oxysterols.
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Emmanuel Noguer, Régis Soules, Claude Netter, Citra Nagarathinam, Julie Leignadier, et al.. Quantitative analysis of the tumor suppressor dendrogenin A using liquid chromatography tandem mass spectrometry. Chemistry and Physics of Lipids, Elsevier, 2017, 207, pp.81-86. ⟨10.1016/j.chemphyslip.2017.06.010⟩. ⟨inserm-02380602⟩

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