SPE-HPLC-DAD同时测定大鼠血浆中斯皮诺素和6′′′-阿魏酰斯皮诺素的含量

Determination of Spinosin and 6′′′-Feruloylspinosin in Rat Plasma after Oral Administration of Flavonoid Extract from Ziziphi Spinosae Semen by SPE-HPLC-DAD

关键词:  酸枣仁黄酮提取物  血浆  代谢物  固相萃取-高效液相色谱-二极管阵列检测法

Key words:  flavonoid extract of Ziziphi Spinosae Semen  plasma  metabolite  SPE-HPLC-DAD

摘要:

目的 建立一种便捷、灵敏定量分析酸枣仁黄酮提取物在大鼠血浆中代谢成分的方法。方法 大鼠口服酸枣仁黄酮提取物后于0~24 h内分别自各组大鼠眼眶后窦采血，血样经固相萃取预处理后采用高效液相-二极管阵列检测器联用色谱法(SPE-HPLC-DAD)定量分析其主要代谢成分。结果 酸枣仁黄酮提取物在血浆中主要代谢成分仍以原形形式存在；斯皮诺素和6^′′′-阿魏酰斯皮诺素在血浆中的固相萃取平均回收率分别为84.76%和84.33%；斯皮诺素在0.52~119.31 mg·L^-1(r>0.999 8)、6^′′′-阿魏酰斯皮诺素在0.38~91.73 mg·L^-1(r>0.999 7)内线性关系良好，二者平均血浆药物浓度于给药后的6 h和4 h达峰值，并在体内缓慢代谢完毕。结论 SPE- HPLC-DAD可用于定量分析酸枣仁黄酮提取物在大鼠体内的代谢过程，为中药主要活性成分在生物体内药动学特征研究提供了一种有效、快捷途径。

Abstract:

OBJECTIVE To develop a convenient and sensitive method for determination of flavonoid metabolites in rat plasma after oral administration of Ziziphi Spinosae Semen extract. METHODS The rat blood samples were collected from each rat by puncture of the retro-orbital sinus from 0 to 24 h after oral administration of flavonoid extract of jujube seeds, then pretreated with a Waters OASISTM SPE column using cartridges. The plasma samples were analyzed by HPLC-DAD method to detect the main metabolites. RESULTS The original flavonoids were the major forms of flavonoid metabolites in rat plasma. The calibration curves were linear over the range of 0.52-119.3 μg·mL^-1(r>0.999 8) for spinosin and 0.38-91.7 μg·mL^-1 (r>0.999 7) for 6^′′′-feruloylspinosin with the mean extraction recoveries as 84.76% and 84.33%, respectively. The average plasma drug concentration of them reached the top level at the 12th and 24th hour after oral administration, and disappeared gradually in vivo. CONCLUSION This convenient SPE-HPLC-DAD method can be used to determinate the flavonoid metabolites of Ziziphi Spinosae Semen. The pharmacokinetic feature of main active components of traditional Chinese medicine in vivo can be characterized as well.