薄层色谱-酶抑制反应评价橄榄叶提取物的黄嘌呤氧化酶抑制活性

Thin layer chromatography-enzyme inhibition assay for assessing xanthine oxidase inhibitory activity of olive leaf extract

研究以薄层色谱为平台,联用黄嘌呤氧化酶催化黄嘌呤氧化反应,以反应产物(尿酸)的紫外吸光特征变化为指标建立了一种可靠性好的黄嘌呤氧化酶活性抑制能力定量评价方法,并应用到橄榄叶提取物的非靶向活性评价。在方法建立过程中,重点研究了光密度检测参数和酶-底物浓度对检测的影响,并提出了一种以抑制指数为定量评价计算方法的评价模式。

Thin-layer chromatography was employed as a platform to mediate xanthine oxidation reaction catalyzed by xanthine oxidase. By monitoring the UV absorption change caused by the formation of uric acid produced by the reaction, a quantitative method tailored for assessing xanthine oxidase inhibition activity was established with satisfactory reliability. Then the established method was applied in non-targeted bioactivity assessment of olive leaf extract. In the method establishment, effort was focused on the effect of densitometry parameters, enzymatic concentrations on visualization time on the optimization of detection. Meanwhile, an inhibition-index based calculation mode was proposed for quantitative evaluation.