Premium
A high‐throughput liquid chromatography/tandem mass spectrometry method for screening glutathione conjugates using exact mass neutral loss acquisition
Author(s) -
CastroPerez Jose,
Plumb Robert,
Liang Li,
Yang Eric
Publication year - 2005
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.1855
Subject(s) - chemistry , mass spectrometry , chromatography , triple quadrupole mass spectrometer , tandem mass spectrometry , selected reaction monitoring , metabolite , liquid chromatography–mass spectrometry , glutathione , reagent , quadrupole mass analyzer , mass , mass spectrum , biochemistry , organic chemistry , enzyme
Chemically reactive metabolites may cause hepatotoxicity and as a result liver failure or other adverse side reactions. Therefore, this is a vital topic of interest because early reactive metabolite screening may prevent compound failure at a later stage. In order to address this issue, a screening assay has been developed to detect the formation of reactive metabolites by using glutathione as a trapping reagent, which will allow us to search for phase I metabolites and also glutathiones during in vitro metabolite screening using liquid chromatography/tandem mass spectrometry (LC/MS/MS) with exact mass. Glutathione conjugations when fragmented by the mass spectrometer give a common loss corresponding to the pyroglutamic acid moiety, which can be monitored. Until recently, this work has been carried out with triple quadrupole technology using nominal mass. The advantage of the hybrid quadrupole time‐of‐flight mass spectrometer is the selectivity and sensitivity that can be achieved. Exact neutral loss detection is achieved via sequential low‐ and high‐energy MS acquisitions. After detection of the loss of the pyroglutamic acid moiety, using a window of ±20 mDa on the high‐energy scan, MS/MS is carried out on the parent mass of interest to confirm the common neutral loss. Copyright © 2005 John Wiley & Sons, Ltd.