期刊
MOLECULES
卷 26, 期 21, 页码 -出版社
MDPI
DOI: 10.3390/molecules26216590
关键词
oxidative stress; glutathione; brain; liquid chromatography; diode array detector
资金
- George Emil Palade University of Medicine, Pharmacy, Science and Technology of Targu Mures [293/4/14.01.2020]
This study successfully developed an HPLC/DAD method for detecting and quantifying GSH and GSSG levels in the rat brain, meeting validation criteria. The method has been applied in a chronic toxicology study of haloperidol under different treatment regimens.
This study aimed to develop a HPLC/DAD method in order to determine and quantify the reduced glutathione (GSH) and oxidized glutathione (GSSG) levels in rat brain. Due to the presence of the thiol group (-SH), GSH can interact with the Ellman & PRIME;s reagent (DTNB), with which it forms a reaction product through which the level of GSH can be quantified, using the DAD detection system. Chromatographic separation was achieved after a derivatization process by using a mobile phase acetonitrile (A) and phosphate buffer (20 mM, pH = 2.5) (B). The compounds of interest were detected at 330 nm using a chromatographic C8 column. The method of determination met the validation criteria, specified by the regulatory bodies. The applicability of the method was demonstrated in a chronic toxicology study of central nervous system (CNS), following different treatment regimens with haloperidol.
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