Yunnan Baiyao is a famous Chinese patent medicine in Yunnan Province. However, its mechanism for promoting blood circulation and removing blood stasis is not fully explained. Our study used metabonomics technology to reveal the regulatory effect of Yunnan Baiyao on small molecular metabolites in promoting blood circulation and removing blood stasis, and exploring the related urine biomarkers. The coagulation function, blood rheology, and pathological results demonstrated that after Yunnan Baiyao treatment, the pathological indexes in rats with epinephrine hydrochloride-induced blood stasis syndrome improved and returned to normal levels. This is the basis for the effectiveness of Yunnan Baiyao. UPLC-G2Si-HDMS was used in combination with multivariate statistical analysis to conduct metabonomic analysis of urine samples. Finally, using mass spectrometry technology, 28 urine biomarkers were identified, clarifying the relevant metabolic pathways that play a vital role in the Yunnan Baiyao treatment. These were used as the target for Yunnan Baiyao to promote blood circulation and remove blood stasis. This study showed that metabolomics strategies provide opportunities and conditions for a deep and systematic understanding of the mechanism of action of prescriptions.