Hydrophilic interaction liquid chromatography coupled with MS/MS to detect and quantify dicarboxyethyl glutathione, a metabolic biomarker of the fumarate hydratase deficient cancer cell

Hien P. Nguyen, Navdeep S. Chandel, Ralph J. Deberardinis, Kevin A. Schug

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)3303-3309
Number of pages7
JournalJournal of Separation Science
Volume36
Issue number20
DOIs
StatePublished - Oct 17 2013

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Keywords

  • FH deficient cells
  • Fumarate hydratase
  • Glutathione
  • HILIC-MS/MS
  • Matrix effect

ASJC Scopus subject areas

  • Analytical Chemistry
  • Filtration and Separation

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