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目的:建立同时测定大鼠血浆中葛根素、阿魏酸和川芎嗪质量浓度的高效液相色谱-二极管阵列检测器(HPLC-DAD)方法,为养阴通脑颗粒药动学研究提供准确可靠的方法。方法:制备大鼠大脑中动脉栓塞(MCAO)模型,再灌注同时灌胃养阴通脑颗粒4个主要有效部位后眼眶取血,血样预处理后,采用HPLC-DAD检测血样中葛根素、阿魏酸和川芎嗪的含量。结果:葛根素、阿魏酸和川芎嗪的进样质量浓度分别在(0.4000-80.0000)mg/L(R2=0.9998),(1.2000-100.0000)mg/L(R2=0.9999),0.04000-20.0000mg/L(R2=0.9990)范围内呈良好的线性关系,平均回收率均在95%-110%之间,日内、日间RSD均<10%,稳定性符合体内药物分析要求。药动学结果显示阿魏酸入血最多,保留最久,T1/2时间最短;葛根素表达峰时间最快;川芎嗪的体内分布最广。结论:所建立的HPLC-DAD测定方法专属性强,分离度好,分析时长适宜,操作简单快速,可用于MCAO大鼠体内葛根素、阿魏酸、川芎嗪含量的同时测定及其药代动力学研究。
OBJECTIVE: To establish a HPLC-DAD method for simultaneous determination of puerarin, ferulic acid and tetramethylpyrazine in rat plasma and to provide accurate and reliable data for the pharmacokinetic study of Yangyintongnao Granules Methods. Methods: The middle cerebral artery occlusion (MCAO) model was established in rats. At the same time, four major effective sites of Yangyintongnao particles were reperfused, and the blood was collected from orbital orbit. After the blood samples were pretreated, the puerarin was detected by HPLC-DAD Weic acid and ligustrazine content. Results: The injection quality of puerarin, ferulic acid and tetramethylpyrazine were (0.4000-80.0000) mg / L (R2 = 0.9998), (1.2000-100.0000) mg / L /L(R2=0.9990) showed a good linear relationship between the average recovery of 95% -110%, day and day RSD were <10%, the stability of in line with the requirements of in vivo drug analysis. Pharmacokinetics results show that ferulic acid into the blood up to retain the longest, T1 / 2 the shortest time; puerarin peak expression time; ligustrazine most widely distributed in vivo. Conclusion: The established HPLC-DAD method has the advantages of high specificity, good resolution, suitable analysis time, simple and quick operation, simultaneous determination of puerarin, ferulic acid and ligustrazine in MCAO rats and their pharmacokinetics Study.