Journal
JOURNAL OF SEPARATION SCIENCE
Volume 36, Issue 12, Pages 1877-1882Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/jssc.201300202
Keywords
Hydrophilic interaction chromatography; Methyl propiolate; Postcolumn derivatization; Thiols; Yeast
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Funding
- European Union (European Social Fund, ESF)
- Greek national funds through the Operational Program Education and Lifelong Learning of the National Strategic Reference Framework (NSRF) - Research Funding Program: Heracleitus II
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In the present study, we report a new method for the determination of two primary thiols, cysteine (CYS) and glutathione (GSH), by hydrophilic interaction LC. The polar analytes are separated isocratically using a mobile phase consisting of 65% acetonitrile/35% ammonium acetate (15 mmol/L, pH 2.0) and are detected at 285 nm following on-line postcolumn derivatization by the thiol-selective reagent methyl propiolate. The main figures of merit included linearity in the range of 5-200mol/L and an LOD 0.6 mol/L for both compounds. The absence of matrix effect allowed the determination of CYS and GSH in various yeast samples. GSH was present in most of the samples at levels ranging between 0.9 and 3.1 mg/g, whereas CYS was determined in only one sample at a significantly lower concentration. In terms of accuracy, the percent recoveries ranged between 91.2 and 105.6% for GSH, and 91.6 and 106.9% for CYS.
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