Hypericum perforatum is a widely used medicinal plant known that it has high bioactive performance due to its special biochemical compounds in its structure. The current study aimed to determine the optimum processing parameters of extraction conditions of bioactive constituents in H. perforatum by ultrasound application and its some bioactive performance. For this purpose, different processing variables namely sonication period (min), liquid/solid ratio and ethanol concentration (%) were selected to characterize the best conditions for extraction of H. perforatum. The statistical analysis results revealed that the most effective factor on the studied parameters was ethanol concentration and increment of ethanol level increased the characterized responses until a constant value. The constructed regression models fit very well with quite high determination of coefficients (R2>0.906). Total phenolic content was in the range of 39.54-63.55 mg GAE/g sample while total hypericin concentration ranged between 211.5-858.7 mg/kg. Multiple response optimization results revealed that the optimum extraction conditions would be at 47.6% ethanol concentration and 22.2 mL of liquid/solid ratio for 60 min ultrasonic process application. The results revealed that the optimized extraction conditions could be utilized for ultrasound extraction system by industrial application.