The Comparison of Spectrophotometric and HPLC Methods in Reduced Glutathione Measurements
Abstract
Objective: Reduced glutathione is the most important antioxidant molecule of intracellular medium and a good marker for investigating antioxidant defence mechanism in diseases which are caused by oxidative stress. In this study we compared the spectrophotometric measurement method developed by Beutler and colleagues and High Performance Liquid Chromatography (HPLC) method for measuring the levels of reduced glutathione in erythrocyte.
Materials and Methods: For this aim we measured reduced glutathione levels of the whole blood samples collected from fifty individuals (15 male, 35 female) by spectrophotometric and HPLC methods.
Results: The mean and SD values of the reduced glutathione were calculated as 953.15±215.91 µmol/L by spectrophotometric method and 972.58±230.19 µmol/L by HPLC method and we determined statistically meaningful correlation between both methods (r = 0.9499, 95% CI 0.913 Ğ 0.971; y= 1.018x).
Conclusion: We concluded that in reduced glutathione measurement HPLC method may be more suitable than spectrophotometric method for busy clinical laboratory when easy application and cost are considered.