A simple, post-additional antioxidant capacity assay using adenosine triphosphate-stabilized 2,2鈥?azinobis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) radical cation in a G-quadruplex DNAzyme catalyzed ABTS-H₂O₂ system

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• 作者：Shu-Min Jia^a ; Xiao-Fei Liu^a ; De-Ming Kong^a ; ^b ; ^{kongdem@nankai.edu.cn} ; Han-Xi Shen^b
• 关键词：G-quadruplex DNAzyme ; Antioxidant capacity ; ATP ; ABTS ; Free radical
• 刊名：Biosensors and Bioelectronics
• 出版年：2012
• 期刊代码：12_09565663
• 类别：ch
• 出版时间：15 May, 2012
• 卷：35
• 期：1
• 页码：407-412
• 文件大小：670 K

摘要

The scavenging of 2,2鈥?azinobis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) radical cation (ABTS⁺) by antioxidants has been widely used in antioxidant capacity assay. Because of ABTS⁺ disproportionation, however, this radical cannot be prepared on a large scale and stored long-term, making it unsuitable for high-throughput detection and screening of antioxidants. We developed a modified 鈥減ost-additional鈥?antioxidant capacity assay. This method possessed two remarkable features: First, instead of natural peroxidases, an artificial enzyme, G-quadruplex DNAzyme, was used for the preparation of ABTS⁺, thus greatly reducing the cost of the assay, and eliminating the strict demand for the storage of enzymes. Second, an ABTS⁺ stabilizer, adenosine triphosphate (ATP), was used. In the presence of ATP, the disproportionation of ABTS⁺ was effectively inhibited, and the lifetime of this radical cation was prolonged about 6-fold (12 days versus 2 days), making the large-scale preparation of ABTS⁺ possible. Utilizing this method, the antioxidant capacities of individual antioxidants and real samples can be quantified and compared easily. In addition, this method can be developed as a high-throughput screening method for antioxidants. The screening results could even be judged by the naked eye, eliminating the need for expensive instruments.