TY  - JOUR
T1  - A Practical HPLC Method to Measure Reduced (GSH) and Oxidized (GSSG) Glutathione Concentrations in Animal Tissues
AU - , Okkes Yilmaz AU - , Serhat Keser AU - , Mehmet Tuzcu AU - , Mehmet Guvenc AU - , Buket Cetintas AU - , Sevgi Irtegun AU - , Hakki Tastan AU - , Kazim Sahin 
JO  - Journal of Animal and Veterinary Advances
VL  - 8
IS  - 2
SP  - 343
EP  - 347
PY  - 2009
DA  - 2001/08/19
SN  - 1680-5593
DO  - javaa.2009.343.347
UR  - https://makhillpublications.co/view-article.php?doi=javaa.2009.343.347
KW  - GSH
KW  -GSSG
KW  -HPLC method
KW  -liver
KW  -wistar rat
AB  - Glutathione is a tripeptide ubiquitously distributed in living cells, which play an important role in the intracellular protective mechanism against oxidative stress. There are many methods for glutathione analysis including spectrophotometric, fluorometric and bioluminometric assays, often applied to HPLC. In this study, an improved and modified method to measure reduced and oxidized glutathione (GSH and GSSG, respectively) concentrations in animal tissues has been developed. In the study, quail and rat tissue samples were homogenized with the mixture of 10 mM ETDA and 50 mM NaClO<SUB>4 </SUB>0.1% H<SUB>3</SUB>PO<SUB>4</SUB> buffer. Proteins were precipitated by the addition 5% metaphosphoric acid. 50 mM NaClO<SUB>4</SUB> 0.1% H<SUB>3</SUB>PO<SUB>4</SUB> was used as mobile phase and Discovery RP-Amide C16 was used as HPLC column. Glutathione molecules were separated at 215 nm by the UV detector. GSH molecule was eluded between 2.3 and 3 min and GSSG molecule was eluted between 3.5 and 4.5 min from the column. It was observed that in low level of glutathione molecules was detected by this process. Recovery rates of GSH and GSSG were 99.32 and 90.29%, respectively. In conclusion, this method is speed, sensitive and achieved since, it doesn’t need forward derivatization process in contrast to many HPLC methods used for glutathione analysis.
ER  -