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

Chem Phys Lipids. 2017 Oct;207(Pt B):81-86. doi: 10.1016/j.chemphyslip.2017.06.010. Epub 2017 Jul 3.


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.

Keywords: Cholesterol; Dendrogenin; Histamine; Phytosterol; Polyamine; Steroidal alkaloid.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Breast Neoplasms / chemistry
  • Breast Neoplasms / metabolism*
  • Cholestanols / analysis*
  • Cholestanols / chemistry*
  • Cholestanols / isolation & purification
  • Chromatography, High Pressure Liquid
  • Female
  • Humans
  • Hydrogen-Ion Concentration
  • Imidazoles / analysis*
  • Imidazoles / chemistry*
  • Imidazoles / isolation & purification
  • Molecular Conformation
  • Tandem Mass Spectrometry


  • 5-hydroxy-6-(2-(1H-imidazol-4-yl)ethylamino)cholestan-3-ol
  • Cholestanols
  • Imidazoles