Tsamesidis Ioannis, Egwu Chinedu O, Pério Pierre, Augereau Jean-Michel, Benoit-Vical Françoise, Reybier Karine
Pharmadev, UMR 152, Université de Toulouse, IRD, UPS, 31400 Toulouse, France.
CNRS, LCC, Laboratoire de Chimie de Coordination, Université de Toulouse, 31077 Toulouse CEDEX 4, France.
Metabolites. 2020 Apr 28;10(5):175. doi: 10.3390/metabo10050175.
Red blood cells are constantly exposed to reactive species under physiological or pathological conditions or during administration of xenobiotics. Regardless of the source, its accurate quantification is paramount in the area of theragnostics, which had been elusive up until now. Even if there are a lot of approaches to evaluate the oxidative stress, very sensitive methods are missing for the blood system. We therefore sought to apply a highly sensitive approach, by liquid chromatography coupled to mass spectrometry (UPLC-MS), for the quantification of reactive species such as superoxide radical and hydrogen peroxide using dihydroethidium (DHE) and coumarin boronic acid (CBA) probes respectively through the detection of 2-hydroxyethidium (2OH-E) and 7-hydroxycoumarin (COH). The use of the high-resolution mass spectrometry associated to UPLC ensured a selective detection of superoxide and hydrogen peroxide in the blood system under diverse conditions such as oxidized red blood cells (RBCs), untreated and treated parasitized RBCs. Moreover, this technique allowed the determination of reactive species in human plasma. This protocol provides a huge opportunity for in-depth study of several pathological conditions vis-a-vis their treatment in modern medicine.
在生理或病理条件下，或在给予外源性物质期间，红细胞不断暴露于反应性物质中。无论其来源如何，其准确定量在治疗诊断领域至关重要，而这一点至今仍难以实现。即使有很多评估氧化应激的方法，但血液系统仍缺少非常灵敏的方法。因此，我们试图应用一种高灵敏度的方法，即超高效液相色谱-质谱联用（UPLC-MS），分别使用二氢乙锭（DHE）和香豆素硼酸（CBA）探针，通过检测2-羟基乙锭（2OH-E）和7-羟基香豆素（COH）来定量超氧阴离子自由基和过氧化氢等反应性物质。与UPLC联用的高分辨率质谱可确保在多种条件下，如氧化红细胞（RBC）、未处理和处理过的寄生红细胞中，选择性检测血液系统中的超氧阴离子和过氧化氢。此外，该技术还能测定人血浆中的反应性物质。该方案为深入研究现代医学中几种病理状况及其治疗提供了巨大机遇。
