Quantitative measurements of glutathione in yeast cell lysate using H NMR.

Rhieu, Steve; Urbas, Aaron; Lippa, Katrice; Reipa, Vytas

Methods for quantifying the level of glutathione (GSH) in yeast cell lysate are described using H NMR analysis. For quantification purposes, the H resonances corresponding to the Cys βCH of GSH were identified as having the fewest overlapping spectral interferences from lysate matrix components using GSH spiked yeast lysate samples. Two methods, standard addition based on peak integration and a spectral subtraction approach, were evaluated for quantifying GSH in lysate samples. The peak integration procedure required baseline estimation and a peak fitting step to correct for background interferences while the spectral subtraction procedure was comparatively straightforward. The level of GSH measured by H NMR was in good agreement with the concentration measured by the DTNB-GSSG reductase recycling assay. The proposed NMR method can lead to a reliable quantitation of GSH and could be applicable to a variety of other analytes of interest in complex biological matrices.

GLUTATHIONE; YEAST; NUCLEAR magnetic resonance; GLYCINE; HIGH performance liquid chromatography

Analytical & Bioanalytical Chemistry, 2013, Vol 405, Issue 14, p4963

1618-2642

Academic Journal

10.1007/s00216-013-6858-5