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

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• 作者：Elena Mellado-Ortega ^{elena.mellado@irnasa.csic.es} ; Iñ ; igo Zabalgogeazcoa ^{i.zabalgo@irnasa.csic.es} ; Beatriz R. V&aacute ; zquez de Aldana ^{beatriz.dealdana@irnasa.csic.es} ; Juan B. Arellano ; ^{juan.arellano@irnasa.csic.es}
• 关键词：AAPH ; ORAC ; Peroxyl radical ; Total antioxidant capacity ; Trolox ; 96-Well plate reader
• 刊名：Analytical Biochemistry
• 出版年：2017
• 出版时间：15 February 2017
• 年：2017
• 卷：519
• 期：Complete
• 页码：27-29
• 全文大小：295 K
• 卷排序：519

文摘

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.