Zingiberis Rhizoma is a typical Chinese herb in 'hot' property. Nowadays, increasing attention has been aroused for its essential role in both health care and treatment of 'cold' syndrome. And research about characteristics of Zingiberis Rhizoma is able to provide an important reference for its deeper understand and successful clinic application. However, cognition level about the 'hot' property of Zingiberis Rhizoma is superficial, and mechanism behind the 'hot' presentation is still elusive and unclear. In this study, the 'hot' property of Zingiberis Rhizoma was investigated at a monomer level with assistance of microcalorimetry, an objective and sensitive method in evaluating biological activity. Three bioactive compounds including 6-gingerol, 8-gingerol and 6-shogaol were selected, and their thermal activities were explored with mitochondria as target. The power-time curves were recorded, and five thermal parameters (k, P max , T max , Q and P av ) were obtained. With PCA, P av was calculated to be the main parameter to measure the bioactivities of 6-gingerol, 8-gingerol and 6-shogaol. Finally, the bioactivities of three subjects were compared with sequence to be 6-gingerol [ 8-gingerol [ 6-shogaol. Generally, our study is promising to offer a reference to the research about characteristics of Zingiberis Rhizoma or other herbs and is of great interest for clinic practice.