 |
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.
Cited literature [20 references]
https://www.hal.inserm.fr/inserm-02380602
Contributor : Marc Poirot <>
Submitted on : Tuesday, November 26, 2019 - 12:19:41 PM
Last modification on : Tuesday, March 17, 2020 - 12:26:02 PM
Contributor : Marc Poirot <>
Submitted on : Tuesday, November 26, 2019 - 12:19:41 PM
Last modification on : Tuesday, March 17, 2020 - 12:26:02 PM
