Study of the antioxidant mechanisms of Trolox and eugenol with 2,2′-azobis (2-amidinepropane)dihydrochloride using ultra-high performance liquid chromatography coupled with tandem mass spectrometry
Study of the antioxidant mechanisms of Trolox and eugenol with 2,2′-azobis (2-amidinepropane)dihydrochloride using ultra-high performance liquid chromatography coupled with tandem mass spectrometry
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DOI：
10.1039/c1an15505a
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发表时间：
2012-01-01
期刊：
影响因子：
4.2
通讯作者：
Nerin, Cristina
中科院分区：
文献类型：
--
作者：
Bentayeb, Karim;Rubio, Carlos;Nerin, Cristina
抗氧化机制的研究是一项艰巨的任务，涉及到未知的中间和最终产品的监测和鉴定。大多数情况下，中间体的寿命太短，无法通过核磁共振（NMR）对其进行分离和随后的鉴定。随着超高效液相色谱技术的发展和质谱技术的进步，可以为抗氧化机制的研究提供新的手段。这项工作是基于参与氧自由基抗氧化能力（ORAC）和总自由基捕获抗氧化参数（TRAP）测定的反应。因此，在色谱装置的恒温自动进样器中进行2，2 '-偶氮二（2-脒丙烷）二盐酸盐（AAPH）自由基与抗氧化剂之间的反应。然后，每六分钟注入反应介质，并通过UHPLC分离化合物，并通过质谱法以扫描模式检测。在唯一文件中记录9次连续进样，然后监测单次运行中1小时的反应演变。以这种方式，研究了Trolox和丁香酚与AAPH的反应机理，分别检测到9种和13种不同的化合物。在产物离子扫描模式中获得的光谱的详尽分析导致化合物的鉴定。其中大多数是以前在文献中发现的物种，但其他从未报道过，因此建议了暂定结构。所有这些都允许在Trolox和丁香酚的抗氧化机制中提出几个步骤，显示UHPLC-MS/MS的出色性能，以补充NMR在抗氧化机制研究中的使用。
The study of antioxidant mechanisms is a difficult task that involves the monitoring and identification of unknown intermediate and final products. Most of the time, the lifetime of intermediates is too short to allow their isolation and subsequent identification by nuclear magnetic resonance (NMR). The developments of ultra-high performance liquid chromatography (UHPLC) coupled with the advances in the acquisition rates of mass spectrometry could facilitate the research on antioxidant mechanisms. This work is based on the reaction involved in the Oxygen Radical Antioxidant Capacity (ORAC) and Total Radical trapping Antioxidant Parameter (TRAP) assays. Hence, the reaction between 2,2'-azobis (2-amidinepropane)dihydrochloride (AAPH) radicals and an antioxidant was carried out in the thermostatized autosampler of a chromatographic device. Then, the reaction media were injected every six minutes, and the compounds were separated by UHPLC and detected by mass spectrometry in scan mode. Nine consecutive injections were registered in a unique file, then the evolution of the reaction for one hour in a single run was monitored. In this way, the reaction mechanisms of Trolox and eugenol with AAPH were studied, leading to the detection of nine and thirteen different compounds, respectively. An exhaustive analysis of the spectra obtained in product ion scan mode led to the identification of the compounds. Most of them were species previously found in the literature, but others have never been reported, so tentative structures were suggested. All this allowed the proposal of several steps within the antioxidant mechanisms of Trolox and eugenol, showing the great performance of UHPLC-MS/MS to complement the use of NMR in antioxidant mechanistic studies.