Department of Pharmacognosy, Lithuanian University of Health Sciences, Mickeviciaus Str. 9, LT-44307, Lithuania.
J Chromatogr A. 2012 Apr 13;1233:8-15. doi: 10.1016/j.chroma.2012.02.019. Epub 2012 Feb 15.
ABTS and FRAP post-column techniques evaluate the antioxidant characteristics of HPLC separated compounds with specific reagents. ABTS characterize their ability to scavenge free radicals by electron-donating antioxidants, resulting in the absorbance decrease of the chromophoric radical. FRAP - is based on the reduction of Fe(III)-tripyridyltriazine complex to Fe(II)-tripyridyltriazine at low pH by electron-donating antioxidants, resulting in an absorbance increase. Both post-column assays were evaluated and compared according to the following validation parameters: specificity, precision, limit of detection (LoD), limit of quantitation (LoQ) and linearity. ABTS and FRAP post-column assays were specific, repeatable and sensitive and thus can be used for the evaluation of antioxidant active compounds. Antioxidant active compounds were quantified according to TEAC for each assay and ABTS/FRAP ratio was derived. No previous records of antioxidative activity of leaves and fruits of strawberries (Fragaria viridis, Fragaria moschata) research have been found. The research results confirm the reliability of ABTS and FRAP post-column assays for screening of antioxidants in complex mixtures and the determination of radical scavenging and ferric reducing ability by their TEAC values.
ABTS 和 FRAP 柱后衍生技术使用特定试剂评估 HPLC 分离化合物的抗氧化特性。ABTS 通过电子供体抗氧化剂来表征其清除自由基的能力,导致发色自由基的吸光度降低。FRAP 基于电子供体抗氧化剂在低 pH 下将 Fe(III)-三吡啶三嗪络合物还原为 Fe(II)-三吡啶三嗪,导致吸光度增加。根据以下验证参数对两种柱后检测方法进行了评估和比较:特异性、精密度、检测限 (LoD)、定量限 (LoQ) 和线性。ABTS 和 FRAP 柱后检测方法具有特异性、可重复性和灵敏度,因此可用于评估抗氧化活性化合物。根据每种测定方法的 TEAC 对抗氧化活性化合物进行定量,并得出 ABTS/FRAP 比值。没有发现关于草莓(Fragaria viridis、Fragaria moschata)叶和果实抗氧化活性的先前研究记录。研究结果证实了 ABTS 和 FRAP 柱后检测方法在复杂混合物中筛选抗氧化剂以及通过其 TEAC 值测定自由基清除和铁还原能力的可靠性。