使用N，N-二甲基-对苯二苯二胺/ Nafion比色传感器同时检测超氧阴离子自由基和测定抗氧剂的超氧清除活性。,Analytical Methods

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使用N，N-二甲基-对苯二苯二胺/ Nafion比色传感器同时检测超氧阴离子自由基和测定抗氧剂的超氧清除活性。
Analytical Methods ( IF 2.7 ) Pub Date : 2017-10-04 00:00:00 , DOI: 10.1039/c7ay02132a
Ferda Dondurmacıoğlu _{1,

2,

3,

4,

5} , Aslı Neslihan Avan _{1,

2,

3,

4,

5} , Reşat Apak _{1,

2,

3,

4,

5}

Affiliation

广泛使用的氮蓝四唑（NBT）为超氧阴离子自由基比色法（O ₂ ˙ ^-）检测从诸如从类似的电子供体的甲的溶解性和干扰的限制受到影响。这项工作报告一个新的分析为超氧化物的发展，其中，O ₂ ˙ ^-由NADH / PMS / O生成₂系统的氧化Ñ，Ñ二甲基p-亚苯基二胺（DMPD）试剂可将粉红色的DMPD-半醌（DMPDQ）阳离子自由基保留在阳离子交换膜上，并进行比色测定。抗氧化剂显示出超氧阴离子自由基清除活性（SRSA）引起更少DMPDQ自由基的产生，导致在膜的Nafion减毒颜色强度，吸光度差（Δ甲在514nm）的成比例的抗氧化剂浓度。该方法适用于14种抗氧化剂化合物，包括trolox，酚酸和羟基肉桂酸，类黄酮和硫醇，以及线性浓度范围，校正方程式（ΔA与浓度），并建立了这些抗氧化剂的trolox等效抗氧化剂能力。根据结构-活性关系讨论了结果。该方法用于测量抗氧化剂混合物和绿茶提取物的SRSA。将与Nafion®膜接触的绿茶提取物转移到在线HPLC柱后CUPRAC系统中，以测量未消耗的儿茶素。将通过开发的方法发现的绿茶抑制百分比与通过参考NBT方法测得的抑制百分比进行了比较。

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Simultaneous detection of superoxide anion radicals and determination of the superoxide scavenging activity of antioxidants using a N,N-dimethyl-p-phenylene diamine/Nafion colorimetric sensor

The widely used nitroblue tetrazolium (NBT) colorimetric method for superoxide anion radical (O₂˙⁻) detection suffers from limitations such as formazan solubility and interference from similar electron donors. This work reports the development of a novel assay for superoxides, where O₂˙⁻ generated by a NADH/PMS/O₂ system oxidizes the N,N-dimethyl-p-phenylene diamine (DMPD) reagent to pink-colored DMPD-semiquinone (DMPDQ) cationic radicals retained by a cation exchange Nafion® membrane and measured colorimetrically. Antioxidants exhibiting superoxide radical scavenging activity (SRSA) cause less DMPDQ radical production, resulting in an attenuated color intensity on the Nafion® membrane, the absorbance difference (ΔA) at 514 nm being proportional to the antioxidant concentration. The developed method was applied to 14 antioxidant compounds including trolox, phenolic and hydroxycinnamic acids, flavonoids, and thiols, and the linear concentration ranges, calibration equations (as ΔA versus concentration), and trolox-equivalent antioxidant capacities of these antioxidants were established. The results were discussed in the light of structure–activity relationships. This method was used for measuring the SRSA of antioxidant mixtures and a green tea extract. The Nafion® membrane-contacted green tea extract was transferred to an online-HPLC post-column CUPRAC system to measure unconsumed catechins. The percentage inhibitions for green tea found with the developed method were compared to those measured by the reference NBT method.

更新日期：2017-10-30

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