降低基于荧光的氧自由基吸收能力（ORAC）测定中与偶氮二异丁腈盐酸盐（AAPH）添加相关的系统误差的解决方案。

Solutions to decrease a systematic error related to AAPH addition in the fluorescence-based ORAC assay.

作者信息

Mellado-Ortega Elena, Zabalgogeazcoa Iñigo, Vázquez de Aldana Beatriz R, Arellano Juan B

机构信息

Departamento de Estrés Abiótico, Instituto de Recursos Naturales y Agrobiología de Salamanca (IRNASA-CSIC), Cordel de Merinas 52, 37008 Salamanca, Spain.

出版信息

Anal Biochem. 2017 Feb 15;519:27-29. doi: 10.1016/j.ab.2016.12.009. Epub 2016 Dec 11.

DOI:10.1016/j.ab.2016.12.009

PMID:27965064

Abstract

Oxygen radical absorbance capacity (ORAC) assay in 96-well multi-detection plate readers is a rapid method to determine total antioxidant capacity (TAC) in biological samples. A disadvantage of this method is that the antioxidant inhibition reaction does not start in all of the 96 wells at the same time due to technical limitations when dispensing the free radical-generating azo initiator 2,2'-azobis (2-methyl-propanimidamide) dihydrochloride (AAPH). The time delay between wells yields a systematic error that causes statistically significant differences in TAC determination of antioxidant solutions depending on their plate position. We propose two alternative solutions to avoid this AAPH-dependent error in ORAC assays.

摘要

在96孔多检测板读数仪中进行的氧自由基吸收能力（ORAC）测定是一种用于确定生物样品中总抗氧化能力（TAC）的快速方法。该方法的一个缺点是，由于在分配产生自由基的偶氮引发剂2,2'-偶氮二（2-甲基丙脒）二盐酸盐（AAPH）时存在技术限制，抗氧化剂抑制反应不会在所有96个孔中同时开始。孔之间的时间延迟会产生系统误差，这会导致根据抗氧化剂溶液在板上的位置不同，其TAC测定结果出现统计学上的显著差异。我们提出了两种替代解决方案，以避免ORAC测定中这种与AAPH相关的误差。