Berthou Margaux, Pallotta Arnaud, Beurton Jordan, Chaigneau Thomas, Athanassiou Athanassia, Marcic Christophe, Marchioni Eric, Boudier Ariane, Clarot Igor
Université de Lorraine, CITHEFOR, F-54000 Nancy, France.
Université de Lorraine, CITHEFOR, F-54000 Nancy, France; Nanocontrol, F-54000 Nancy, France.
J Chromatogr B Analyt Technol Biomed Life Sci. 2022 May 15;1198:123244. doi: 10.1016/j.jchromb.2022.123244. Epub 2022 Apr 9.
Thiols are very important molecules in the biomedical field involved for example in redox homeostasis. Their detection and quantification remain difficult due to their poor stability (oxidation) linked to their strong reactivity towards other thiols (by the formation of S-S bonds) or other interfering molecules in the medium. Cellulose membranes with immobilized gold nanoparticles (AuNP) were developed to capture and quantify thiols in simple and complex matrices. This device was first optimized and characterized in terms of nanostructuration and thiol adsorption. N-Acetylcysteine (NAC) and reduced glutathione (GSH), chosen as model molecules, were filtered through the device demonstrating a maximal adsorption capacity of 270 and 60 nmol respectively. In a second step, the adsorbed species were subjected to ligand exchange using a more reactive thiol, dithiothreitol. The results showed release rates of approximately 90% for NAC and GSH. Finally, the amount of endogenous GSH in rat plasma was determined without any pretreatment. For the first time to our knowledge, a nanostructured device for the capture, selective and sensitive quantification of thiols is proposed. This device is easy to handle and overcomes matrix effects. Moreover, the very large concentration factor induced by this technology will be a valuable asset to decrease the quantification limits of analytical methods.
硫醇是生物医学领域中非常重要的分子,例如参与氧化还原稳态。由于其稳定性差(氧化),与其他硫醇(通过形成S-S键)或介质中的其他干扰分子具有很强的反应性,它们的检测和定量仍然很困难。开发了固定有金纳米颗粒(AuNP)的纤维素膜,用于捕获和定量简单和复杂基质中的硫醇。该装置首先在纳米结构和硫醇吸附方面进行了优化和表征。选择N-乙酰半胱氨酸(NAC)和还原型谷胱甘肽(GSH)作为模型分子,通过该装置过滤,显示最大吸附容量分别为270和60 nmol。第二步,使用反应性更强的硫醇二硫苏糖醇对吸附的物质进行配体交换。结果显示NAC和GSH的释放率约为90%。最后,在没有任何预处理的情况下测定了大鼠血浆中内源性GSH的含量。据我们所知,首次提出了一种用于捕获、选择性和灵敏定量硫醇的纳米结构装置。该装置易于操作,克服了基质效应。此外,该技术诱导的非常大的浓缩因子将是降低分析方法定量限的宝贵资产。