Determination of Reduced Glutathione and Glutamic Acid in Brain Tissue by HPLC with Pre-column Derivatization Using O-Phthaldialdehyde
YANG Xinxia
WU Nanding
WANG Yutong
GUO Mengjie
ZHOU Ren
YANG Jing
Abstract：Objective To establish a high-performance liquid chromatography(HPLC)method for the detection of glutathione(GSH)and glutamate levels in brain tissue.Methods C18 column was used with a mobile phase of methanol-acetonitrile-water and 16 mM phosphate buffer(pH=8.2)under isothermal elution at 38 ℃.O-phthalaldehyde was used as a pre-column derivation agent,with a flow rate of 1 mL/min.The excitation wavelength was set to 338 nm and the emission wavelength to 425 nm to simultaneously detect GSH and glutamate levels.Results GSH and glutamate were baseline-separated,showing good linearity within the range of 0.25 to 4.00 μg/mL,with correlation coefficients of 0.9994 and 0.9996,respectively.The recovery rates were 99.71％and 99.27％.Conclusion The method is sensitive and reliable.The levels of GSH and glutamate in rat brain tissue were measured at(65.05±10.15)and(105.91±39.38)μg/g wet weight,respectively.
Keywords：pre-column derivatizationhigh-performance liquid chromatographyglutathioneglutamateo-Phthalaldehyde
Publication Date：2025-10-30
Online Publishing Date：2025-11-20(First online date of this platform, not the publication date of the document)
Pages：3( 15-17 )