Quantitative analysis of the tumor suppressor dendrogenin A using liquid chromatography tandem mass spectrometry. (October 2017)

• Record Type: Journal Article
• Authors: Noguer, Emmanuel; Soules, Régis; Netter, Claude; Nagarathinam, Citra; Leignadier, Julie; Huc-Claustre, Emilie; Serhan, Nizar; Rives, Arnaud; de Medina, Philippe; Silvente-Poirot, Sandrine; Poirot, Marc
• Is Part Of: Chemistry and physics of lipids. Volume 207(2017)Part B
• Page Start: 81
• Page End: 86
• Publication Date: 2017-10
• Subjects: 5, 6α-EC 5α, 6α-epoxy-cholestan-3β-ol -- 5, 6β-EC 5β, 6β-epoxy-cholestan-3β-ol -- CT cholestane-3β, 5α, 6β-triol -- OCDO 6-oxo-cholestan-3β, 5α-diol -- CDO-SG 3β, 5α-dihydroxycholestan-6β-yl-S-glutathione -- Dendrogenin A (DDA) 5α-hydroxy-6β[2-(1H-imidazol-4-yl)ethylamino] cholestan-3β-ol -- TFA trifluoroacteic acid -- OS oxysterol; Steroidal alkaloid -- Dendrogenin -- Cholesterol -- Phytosterol -- Histamine -- Polyamine; Lipids -- Periodicals; Lipids -- Periodicals; Lipides -- Périodiques; Lipids; Periodicals; Electronic journals; 547.77
• DOI: 10.1016/j.chemphyslip.2017.06.010
• Languages: English
• ISSNs: 0009-3084

Abstract

Highlights: Dendrogenin A is a polycationic amino-oxysterol. We have developed a new RP LC/MS method for DDA analysis that is fast, sensitive and selective. This novel technique efficiently separates DDA from its C17 isomer and other steroidal alkaloids from the same family. 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 × 4.6 mm 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 (10 min), resolving (peak width <4 s) 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.