TY - JOUR
T1 - Pitfalls in the analysis of the physiological antioxidant glutathione (GSH) and its disulfide (GSSG) in biological samples
T2 - An elephant in the room
AU - Giustarini, Daniela
AU - Tsikas, Dimitrios
AU - Colombo, Graziano
AU - Milzani, Aldo
AU - Dalle-Donne, Isabella
AU - Fanti, Paolo
AU - Rossi, Ranieri
N1 - Funding Information:
This work was supported by grants to PF from US National Institutes of Health (NIH), National Center for Complementary and Alternative Medicine (NCCAM) (grant no. AT004490) and US Department of Veterans Affairs (Merit Review no. 1I01CX000264).
Funding Information:
This work was supported by grants to PF from US National Institutes of Health (NIH) , National Center for Complementary and Alternative Medicine (NCCAM) (grant no. AT004490 ) and US Department of Veterans Affairs (Merit Review no. 1I01CX000264).
PY - 2016/4/15
Y1 - 2016/4/15
N2 - Glutathione (GSH) is the most abundant low-molecular-mass thiol within cells and one of the major antioxidant compounds in body fluids. Under pro-oxidant conditions, two GSH molecules donate one electron each and are converted into glutathione disulfide (GSSG). The GSH/GSSG molar ratio is considered a powerful index of oxidative stress and disease risk. Despite high interest in GSH/GSSG titration as measures of thiol redox balance, no broad agreement has yet been reached as to the best pre-analytical and analytical methods for the quantitation of these molecules in biological samples. Consequently, measured concentrations of GSH and GSSG and calculated GSH/GSSG molar ratios vary widely among laboratories. Here, we describe in detail the main analytical and pre-analytical problems related to the artificial oxidation of the sulfhydryl (SH) group of GSH that occur during sample manipulation. We underline how this aspect has been neglected for long time after its first description more than fifty years ago. Finally, selected reliable procedures and methods to measure GSH and GSSG in biological samples are discussed.
AB - Glutathione (GSH) is the most abundant low-molecular-mass thiol within cells and one of the major antioxidant compounds in body fluids. Under pro-oxidant conditions, two GSH molecules donate one electron each and are converted into glutathione disulfide (GSSG). The GSH/GSSG molar ratio is considered a powerful index of oxidative stress and disease risk. Despite high interest in GSH/GSSG titration as measures of thiol redox balance, no broad agreement has yet been reached as to the best pre-analytical and analytical methods for the quantitation of these molecules in biological samples. Consequently, measured concentrations of GSH and GSSG and calculated GSH/GSSG molar ratios vary widely among laboratories. Here, we describe in detail the main analytical and pre-analytical problems related to the artificial oxidation of the sulfhydryl (SH) group of GSH that occur during sample manipulation. We underline how this aspect has been neglected for long time after its first description more than fifty years ago. Finally, selected reliable procedures and methods to measure GSH and GSSG in biological samples are discussed.
KW - Glutathione
KW - Glutathione disulfide
KW - Oxidative stress
KW - Protocols
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U2 - 10.1016/j.jchromb.2016.02.015
DO - 10.1016/j.jchromb.2016.02.015
M3 - Article
C2 - 26905452
AN - SCOPUS:84969344761
VL - 1019
SP - 21
EP - 28
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
SN - 1570-0232
ER -