Evodiamine

Carbamazepine (CAS No.: 298–46-4), berberine (CAS No.: 2086-83-1), coptisine (CAS No.: 6020-18-4), epiberberine (CAS No.: 6873-09-2), palmatine (CAS No.: 3486-67-7), jatrorrhizine (CAS No.: 3621-38-3), columbamine (CAS No.: 483–34-1), evodiamine (CAS No.: 518–17-2) and rutaecarpine (CAS No.: 20575–76-2), all > 98% purity, were purchased from Shanghai Yuanye Bio-Technology Company. Acetonitrile and methanol (LC grade) were purchased from Fisher Chemical, USA. Formaldehyde (AR) was purchased from Beijing Tong Guang Fine Chemicals Company. Isoflurane was provided by HARVEYBIO Company.
TAZF was provided by the State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University. The concentrations of the compounds in TAZF were measured using high-performance liquid chromatography (HPLC), with berberine accounting for the majority of TAZF at a ratio of 14.73% (g/g), followed by palmatine (3.14%), coptisine (2.52%), evodiamine (1.41%), epiberberine (1.30%), rutaecarpine (1.27%), columbamine (0.90%) and jatrorrhizine (0.47%).

H9c2 cardiomyocytes (5,000/well) were cultured into 96-well plates for 24 h, and were exposed to a series concentration of evodiamine and rutaecarpine (Shanghai Yuanye Bio-Technology Co., Ltd., China) for another 24 h. Cell viability was measured using the cell counting kit-8 (CCK-8) solution assay (Biorigin Inc., China) at 450 nm. Subsequently, the absorbance values were applied to calculate the half maximal inhibitory concentration (IC₅₀ values) and select appropriate concentrations for further experiments. All experiments were performed independently in triplicate.