As an important restoration substance in cells, reduced glutathione (GSH) plays an important physiological role in the human body. In this paper, GSH was determined by indirect Spectrophotometry. Through the optimization experimental conditions, the amount of Fe3+-1,10-phenanthroline chromogenic concentration was 0.8 mg/mL, and the optimum pH value was 4.0 mL, and the amount of buffer solution was 5.0 mL, and the heating time and cooling time were 10 min. In addition, the absorbance was measured at 510 nm, and GSH was in the range of 0~0.4 mg/mL. The linear regression equation was A=1.7981c+0.1348 (which A represents the absorbance and c represents the concentration of GSH), and the correlation coefficient was r2 = 0.9955. The method was applied to the determination of reduced GSH in three GSH samples. The indirect spectrophotometry method is simple, low cost and easy to operate.
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