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Determination of glutathione and cysteine in yeasts by hydrophilic interaction liquid chromatography followed by on‐line postcolumn derivatization
Author(s) -
Karakosta Theano D.,
Tzanavaras Paraskevas D.,
Themelis Demetrius G.
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.201300202
Subject(s) - derivatization , chemistry , chromatography , glutathione , cysteine , thiol , ammonium acetate , reagent , analyte , high performance liquid chromatography , quantitative analysis (chemistry) , acetonitrile , hydrophilic interaction chromatography , ammonium , reversed phase chromatography , matrix (chemical analysis) , organic chemistry , enzyme
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 5–200 μmol/L and an LOD 0.6 μmol/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 .