Abstract: OBJECTIVE To explore the effective constituents of Danshen Chuanxiongqin injection against hypoxic-ischemic brain injury from the three levels of injection, components and ingredients. METHODS Firstly, oxygen-glucose deprivation/reoxygenation(OGD/R) model was constructed in SH-SY5Y neuroblastoma cells. The protective effect of Danshen Chuanxiongqin injection on this ODG/R model was then detected by cell viability. Secondly, standard components of Danshen Chuanxiongqin injection were prepared by preparative liquid chromatography. The effective components and constituents of Danshen Chuanxiongqin injection were identified by the OGD/R model in SH-SY5Y cells. Finally, the active components and ingredients were analyzed by LC-MS, aiming to identify the effective constituents of Danshen Chuanxiongqin injection against hypoxic-ischemic brain injury. RESULTS Compared with the model group, Danshen Chuanxiongqin injection had a markedly protective effect on OGD/R injured SH-SY5Y cells in the dose range of 2.5%-15%. Further screening found that the components I, J, L, M, O, P, Q and salvianolic acid C, salvianolic acid A, Danshensu and ligustrazine hydrochloride could significantly alleviate OGD/R-induced injury in SH-SY5Y cells. The results of LC-MS analysis indicated that the components O and P contained salvianolic acid C, the component O contained salvianolic acid A and ligustrazine hydrochloride, and the component P contained Danshensu. CONCLUSION Danshen Chuanxiongqin injection has a markedly protective effect on hypoxic-ischemic brain injury, and salvianolic acid C, salvianolic acid A, Danshensu and ligustrazine hydrochloride may be the effective constituents of Danshen Chuanxiongqin injection against hypoxic-ischemic brain injury.