Reduced glutathione (GSH) is a biologically active peptide compound with a variety of physiological functions, produced mainly by fermentation. In this study, macroporous resin A was used for purification of the GSH fermentation broth. The experimental results showed that the macroporous resin A had a good adsorption on GSH. The adsorption reached equilibrium state when the process was carried out for 150 minutes. At pH 3.0, the macroporous resin A had the strongest adsorption of GSH with the maximum adsorption capacity and adsorption rate. Then, feasible pH, eluting agent species, concentration and flow rate were determined and validated by dynamic adsorption. Under the best absorption conditions (pH 3.0, 0.2 mol/L phosphate buffer as eluting agent and the eluting agent rate 2 mL/min), the purity of glutathione can be increased from 58.5% to 95.3%, and the glutathione recovery ratio in the eluting agent was > 87%, indicating that this method was effective for purification of GSH fermentation broth.