Azevedo Ana M O, Pereira Sarah A P, Passos Marieta L C, Costa Susana P F, Pinto Paula C A G, Araujo André R T S, Saraiva M Lúcia M F S
LAQV, REQUIMTE, Departamento de Ciências Químicas, Faculdade de Farmácia, Universidade do Porto, Rua Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal.
LAQV, REQUIMTE, Departamento de Ciências Químicas, Faculdade de Farmácia, Universidade do Porto, Rua Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal; Escola Superior de Saúde, Instituto Politécnico da Guarda, Avenida Rainha D. Amélia, S/N, 6300-749 Guarda, Portugal.
Chemosphere. 2017 Apr;173:351-358. doi: 10.1016/j.chemosphere.2016.12.138. Epub 2017 Jan 16.
Acylase I (ACY I) plays a role in the detoxication and bioactivation of xenobiotics as well in other physiological functions. In this context, an automated ACY I assay for the evaluation of ionic liquids' (ILs) toxicity was developed. The assay was implemented in a sequential injection analysis (SIA) system and was applied to eight commercially available ILs. The SIA methodology was based on the deacetylation of N-acetyl-l-methionine with production of l-methionine, which was determined using fluorescamine. ACY I inhibition in the presence of ILs was monitored by the decrease of fluorescence intensity. The obtained results confirmed the influence of ILs' structural elements on its toxicity and revealed that pyridinium and phosphonium cations, longer alkyl side chains and tetrafluoroborate anion displayed higher toxic effect on enzyme activity. The developed methodology proved to be robust and exhibited good repeatability (RSD < 1.3%, n = 10), leading also to a reduction of reagents consumption and effluents production. Thus, it is expected that the proposed assay can be used as a novel tool for ILs' toxicity screening.
酰基转移酶I(ACY I)在异生物素的解毒和生物活化以及其他生理功能中发挥作用。在此背景下,开发了一种用于评估离子液体(ILs)毒性的自动化ACY I检测方法。该检测方法在顺序注射分析(SIA)系统中实施,并应用于八种市售离子液体。SIA方法基于N - 乙酰 - l - 甲硫氨酸的脱乙酰化反应生成l - 甲硫氨酸,后者使用荧光胺进行测定。通过荧光强度的降低来监测在离子液体存在下ACY I的抑制情况。所得结果证实了离子液体结构元素对其毒性的影响,并表明吡啶鎓和鏻阳离子、较长的烷基侧链以及四氟硼酸根阴离子对酶活性表现出更高的毒性作用。所开发的方法被证明是稳健的,具有良好的重复性(相对标准偏差<1.3%,n = 10),同时还减少了试剂消耗和废水产生。因此,预计所提出的检测方法可作为一种新型工具用于离子液体的毒性筛选。