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Hydrophilic interaction liquid chromatography coupled with MS / MS to detect and quantify dicarboxyethyl glutathione, a metabolic biomarker of the fumarate hydratase deficient cancer cell
Author(s) -
Nguyen Hien P.,
Chandel Navdeep S.,
DeBerardinis Ralph J.,
Schug Kevin A.
Publication year - 2013
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201300602
Subject(s) - glutathione , chemistry , analyte , chromatography , dimethyl fumarate , glutathione disulfide , matrix (chemical analysis) , high performance liquid chromatography , hydrophilic interaction chromatography , conjugate , biochemistry , enzyme , psychology , mathematical analysis , mathematics , psychiatry , multiple sclerosis
The identification of a glutathione ( GSH ) fumarate conjugate, dicarboxyethyl glutathione, formed during the nonenzymatic succination of GSH by fumarate was confirmed in fumarate hydratase deficient cells using a product ion scan approach followed by hydrophilic interaction liquid chromatography coupled with MS/MS. GSH and its conjugates, including dicarboxyethyl glutathione and glutathione disulfide, were successfully separated on a zwitterionic stationary phase and detected by MS / MS operated under negative ESI mode. The relative quantitation of the analytes in cell extracts was carried out and a correction model was established to determine correction factors under matrix effects and the response mismatch between the analytes. These factors were calculated and iteratively used to measure all analytes in cell extracts, based on calibration curves constructed in neat solution. The model was a closed‐loop calculation, consisting of two sides with each side of the loop presenting a calculation pathway. Deviation of the correction factors obtained from these pathways manifested the model accuracy. The model was evaluated and there was no significant difference between the two pathways.