Liquid chromatography coupled to negative electrospray/ion trap mass spectrometry for the identification of isomeric glutathione conjugates of catechol estrogens

Conjugation to glutathione (GSH) represents an important detoxification pathway for preventing DNA damage due to oxidation products of catechol estrogens. In order to identify isomeric GSH conjugates of catechol estrogens, liquid chromatography coupled to electrospray/ion trap mass spectrometry was used. For this purpose, both positive and negative ion modes were applied, generating protonated and deprotonated species, respectively. Energy-resolved fragmentation of each isomeric quasi-molecular ion was achieved in two regions of the mass spectrometer: (i) the mass analyzer (ion trap mass spectrometer) and (ii) the interface region of the electrospray ionization source. The resonance excitation of [M + H](+) protonated ions carried out into the ion trap did not show any isomeric differentiation. Although ion source fragmentation of these same species allowed identification of each isomer, this method requires a good chromatographic separation, making it inefficient for the analysis of low sample amounts from in vitro or in vivo sources. Conversely, using resonance excitation of deprotonated ions, isomer distinction could be achieved. Thus, this technique should yield the best data for the direct characterization of isomers of catechol estrogen-GSH conjugates from biological samples. (C) 2004 Elsevier B.V. All rights reserved.