The aim of this study was to assess inhibitory effect of safranin on human erythrocyte glutathione reductase (GR) and glucose 6-phosphate dehydrogenase (G6PD). For this purpose, human erythrocyte glutathione reductase was purified 2175-fold (31.35% yield) using 2’,5’-ADP Sepharose 4B affinity gel and Sephadex G-200 gel filtration chromatography and erythrocyte glucose 6-phosphate dehydrogenase was purified 8681.3-fold (24.77% yield) using 2’,5’-ADP Sepharose 4B affinity gel.
Safranin exhibited inhibitory effects on GR and G6PD enzymes in vitro. The IC50 values of safranin were 1.15 mM for GR and 3.22 mM for G6PD, respectively, and the Ki constants were 2.63 ± 0.13 and 4.53 ± 0.17 mM, respectively. Safranin displayed noncompetitive inhibition for GR and G6PD.