目的 建立隔山香药材中异芹菜脑（Isoapide，Iso）的高效液相色谱法（ HPLC）， 测定方法并初步探讨异芹菜脑对体外培养乳鼠心肌细胞缺 氧/复氧损伤的保护作用。方法 采用高效液相色谱法，色谱柱：Kromasil C18柱（4.6 mm×250 mm，5 μm）；流动相：以甲醇-水（70 ∶ 30）洗脱，流速为 1.0 mL·min-1，检测波长为 280 nm，柱温室温，进样量：10 μL。并用原代培养的乳鼠心肌细胞建立缺氧复/氧损伤模型，异芹菜脑在建模前进行预干预，心肌细胞随机分为5 组，空白对照组（Control，CON），缺氧/复氧组（hypoxia/reoxygenation，H/R），异芹菜脑低、中、高剂量干预组（IsoL、IsoM、IsoH）。缺氧4 h/复氧4 h后，采用 CCK-8法检测心肌细胞存活率。结果 异芹菜脑在0.02096~0.2096 mg·mL-1线性关系良好（r=0.9999），平均回收率为 98.09 %，RSD=1.25 %。与空白对照组比较，缺氧/复 氧组心肌细胞存活率显著降低（P ＜ 0.01）；与缺氧/复氧组比较，IsoL、IsoM、IsoH剂量干预组心肌细胞存活率均显著升高。结论 异芹菜脑对体外培养乳鼠心肌细胞缺氧 /复氧损伤具有一定的保护作用，隔山香药材中异芹菜脑的 HPLC 测定方法准确、简便、重现性好，有助于完善隔山香药材的质量控制。

Objective To establish a high performance liquid chromatography（HPLC） method for the determination of isoapiole in Lemonfragrant Angelica root and to investigate the protective effect of isoapiole on cultured neonatal rat cardiomyocytes with hypoxia/reoxygenation injury. Methods HPLC method was performed on Kromasil C18 column（4.60 mm×250 mm，5 μm）with a mobile phase of methanol-water（70∶30） at a flow rate of 1.0 mL·min-1 and at room temperature. The detective wavelength was set at 280 nm and the injection volume was 10 μL. Primary cultured neonatal rat cardiomyocytes were exposed to hypoxia/reoxygenation and then were randomly divided into 5 groups，namely blank control group， hypoxia/reoxygenation（H/R） group，and low-，middle-and high-dose of isoapiole groups. After hypoxia for 4h and reoxygenation for 4 h，the survival rate of cardiomyocytes was detected by CCK-8 method. Result The calibration curve of isoapiole showed a good linearity relationship in the range of 0.02096~0.2096 mg·mL-1（r=0.9999）. The average recovery was 98.09 %（n=6），and the relative standard deviation was 1.25 %. Compared with the blank control group，cardiomyocytes survival rate in H/R group was decreased significantly（45.41±9.73 % vs 100.00±10.05 %，P＜ 0.01）. Compared with H/R group，cardiomyocytes viability in isoapiole groups was increased significantly. Conclusion Isoapiole has protective effect on hypoxia/reoxygenation injuried cardiomyocytes in vitro. The developed method is accurate，simple，and reproducible，which can be used for the quality control of Lemonfragrant Angelica root.

R284.1；285.5

国家自然科学基金项目（81274191）。