基于谱效整合指纹的体外活性与体内活性的生物等效性研究

目的:基于谱效整合指纹图谱的药物体外生物活性与体内活性的生物等效性研究.方法:利用高效液相色谱-光电二极管阵列检测器-化学发光检测器(HPLC-DAD-CL)平台,建立9个不同产地山楂叶在线清除过氧化氢( H2O2)的活性整合指纹图谱,结合白藜芦醇清除H2O2的量效关系曲线方程,以1μg白藜芦醇的药效为一个效价单位,计算各指纹峰的效价值,用各指纹峰的效价值之和表示山楂叶提取物体外清除过氧化氢的总活性;130只小鼠随机分为13组,模型组与给药组皮下注射盐酸异丙肾上腺素,放入装有钠石灰的广口瓶中,密闭瓶口,考察缺氧状态下小鼠耐氧存活时间,计算山楂叶对心肌缺血小鼠的保护作用;对上述山楂叶抗氧化的体内外实验结果进行相关性研究,并采用药典方法测定了不同产地山楂叶总黄酮含量对上述结果进行佐证.结果:不同产地山楂叶基于HPLC-CL化学发光体系的体外清除H2O2的的总活性与其体内活性结果基本一致,具有很强的相关性.结论:本研究组已建立的在线活性筛选技术检测结果的具有一定的可靠性,本研究为其进一步推广应用提供科学依据.

Studies on Bioequivalence of Activity based on “Spectrum-Effect Integrated Fingerprint” and Activity In Vivo

This work was aimed to study the bioequivalence of activity based on "spectrum-effect integrated fingerprint" and activity in vivo.The hydrogen peroxide(H2O2) scavenging activity fingerprints of 9 batches of Crataegus leaves collected from different provinces were established through an on-line high-performance liquid chromatography-diode array detector combined with chemiluminescence detection(HPLC-DAD-CL) system.As the positive control,if resveratrol(1 μg) was presumed as a potency unit,the relative activities of active peaks were calculated as potency unit according to standard potency curves of resveratrol.And the relative total activities of Crataegus leaves should be the sum of potency of all characteristic peaks.A total of 130 mice were randomly divided into 13 groups,the acute ischemic anoxic myocardial injury was induced by subcutaneous injection isoproterenol in mice,the survival time in mice with hypoxic state was recorded and the life extension rate of the treated group was studied.The correlation between the H2O2 scavenging capacity of Crataegus leaves based on HPLC-DAD-CL and the protective effect in mice with myocardial hypoxia was investigated.And the content of total flavonoids was determined to make the results above more authentic.It is found the antioxidant activities in vitro and in vivo both confirm with each other,which means the on-line activity screening method and antioxidant-activity integrated fingerprint research platform can be further promoted.