Abstract:

Objective     To establish a high performance liquid chromatography with UV-absorbance detection method for determination of the level of reduced glutathione in rat livers. Methods     After the liver tissue homogenate was made under the condition of ice bathing and precipitated tissue proteins by acetonitrile, the derivative reaction of the supernatant and fluorescent 4-fluoro-7-nitrobenzofuraza was carried out in borate buffer. The mobile phase was acetonitrile-0.02mol/L potassium dihydrogen phosphate (23∶77, V/V), the flow rate was 1.0mL/min, the detection wave-length was 470nm and the injection volume was 10μL. The reaction mixture was quantified with the external standard method. Results     The linear range of reduced glutathione was 0.200-125μg/mL (r=0.9999, n=5), and the lowest limit of determination was 0.0400μg/mL(S/N=3). The recovery rate was 96.2%-104.5%, the intra-day and inter-day relative standard deviations were 1.1％-4.6％ and 1.9％-7.5％, respectively. Conclusion     This method is simple， rapid, accurate, sensitive and stable, which is suitable for determination of reduced glutathione in the liver.

Key words: Reduced glutathione; 4-fluoro-7-nitrobenzofuraza; Liver tissue; High performance liquid chromatography