The turbulent flow in stirred tank is highly complicated and anisotropic, especially when the macro-instability (MI) are involved. In this work, the numerical simulation method of the eccentric agitation was established based on the detached eddy simulation (DES) model to study the MI in an eccentric stirred tank. The turbulent flow in the eccentrically located Rushton turbine stirred tank was numerically investigated. The rotation of the impeller was simulated by the transient sliding mesh (SM) method. The effect of eccentricity, impeller Reynolds number and impeller-tank diameter ratio were studied in order to quantify the MI frequency. PIV experiments were performed to validate the DES results and frequency analyses were applied to the obtained time series of the velocity recordings. It was found that the flow field in eccentrically stirred tank are highly unsteady and is subject to MI with varying period less than 10 blade passage period. Good agreements have been found between the DES and PIV results, both indicate that the dominant frequency of MI increases linearly with the Reynolds number, increases with the impeller-tank diameter ratio and decreases with the eccentricity. According to the agreements between the experimental and simulation results, it can be concluded that the combination of DES and SM is suitable for the prediction of the MI phenomenon in stirred tanks.