Profiling the Concentration of Reduced and Oxidized Glutathione in Rat Brain Using HPLC/DAD Chromatographic System.
George JîtcăErzsébet MájaiBianca-Eugenia ŐszCamil Eugen VariIbolya FülöpMircea Dumitru CroitoruCarmen-Maria RuszMaria Titica DogaruPublished in: Molecules (Basel, Switzerland) (2021)
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'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.
Keyphrases
- simultaneous determination
- ms ms
- solid phase extraction
- liquid chromatography tandem mass spectrometry
- liquid chromatography
- high performance liquid chromatography
- tandem mass spectrometry
- ultra high performance liquid chromatography
- mass spectrometry
- fluorescent probe
- molecularly imprinted
- gas chromatography mass spectrometry
- transcription factor
- blood brain barrier
- single cell
- tyrosine kinase
- drug induced
- quantum dots
- cerebrospinal fluid