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Automated evaluation of the inhibition of glutathione reductase activity: application to the prediction of ionic liquids' toxicity

机译：自动评估谷胱甘肽还原酶活性的抑制作用：在预测离子液体毒性中的应用

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摘要

This work is based on the automated evaluation of the influence of several ionic liquids (ILs) on the activity of immobilized glutathione reductase (GR), as an indicator of their toxicity. The reactions involved are the reduction of oxidized glutathione (GSSG) by GR, followed by the oxidation of the formed product (reduced glutathione -GSH) by the sulfhydryl reagent 5,50-dithio-bis(2-nitrobenzoic acid) (DTNB) to form a yellow derivate 2-nitro-5-thiobenzoic acid (TNB), measurable at 412 nm. The bioassay was performed in a SIA system where the ILs were submitted to a simple and precise on-line dilution strategy before evaluation of their toxicity. The activity assays were implemented in strictly aqueous media and in the presence of increasing concentrations of seven commercially available ILs. EC50 values between 0.24 and 2.55 M were obtained for the tested compounds, revealing distinct inhibitory effects. The least and most toxic ILs found in this study were emim[Ms] and emim[BF4], respectively. The automated methodology is an interesting analytical tool for the evaluation of GR activity in ILs proving to be robust while presenting good repeatability, with RSD less than 5%, in all the assay conditions, leading also to a reduction of the consumption of solvents as well as of effluent production (2 mL per cycle).
机译：这项工作基于对几种离子液体（ILs）对固定化谷胱甘肽还原酶（GR）活性的影响的自动评估，以此作为其毒性的指标。所涉及的反应是GR还原氧化型谷胱甘肽（GSSG），然后用巯基试剂5,50-二硫代双（2-硝基苯甲酸）（DTNB）将生成的产物（还原型谷胱甘肽-GSH）氧化为形成黄色衍生物2-硝基-5-硫代苯甲酸（TNB），可在412 nm处测量。生物测定在SIA系统中进行，其中ILs在评估其毒性之前要经过简单而精确的在线稀释策略。活性测定是在严格的水性介质中，并且在浓度越来越高的七个市售IL的存在下进行的。对于测试化合物，获得的EC50值为0.24至2.55 M，显示出明显的抑制作用。在这项研究中发现的毒性最小和毒性最大的ILs分别是emim [Ms]和emim [BF4]。自动化方法学是用于评估IL中GR活性的有趣分析工具，被证明在所有测定条件下均具有较强的鲁棒性，同时具有良好的可重复性，RSD小于5％，因此也减少了溶剂的消耗从废水生产开始（每个周期2 mL）。

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