Nguyen Tam T T N, Østergaard Jesper, Gammelgaard Bente
Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen, Denmark.
Anal Bioanal Chem. 2015 Nov;407(28):8497-503. doi: 10.1007/s00216-015-8997-3. Epub 2015 Sep 2.
An analytical method based on capillary electrophoresis (CE) and inductively coupled plasma mass spectrometry (ICP-MS) detection was developed for studies on the interaction of gold-containing drugs and plasma proteins using auranofin as example. A detection limit of 18 ng/mL of auranofin corresponding to 5.2 ng/mL Au and a precision of 1.5 % were obtained. Kinetic studies of the interaction between auranofin and protein were performed by incubation in aqueous solutions as well as 20 % human plasma at 37 °C. The reaction of auranofin with human serum albumin (HSA) and plasma proceeded fast; 50 % of un-bound auranofin disappeared within 2 and 3 min, respectively. By blocking the free cysteine (Cys-34) by iodoacetamide on HSA, it was shown that Cys-34 was the main reaction site for auranofin. By selective labeling of HSA present in 20 % human plasma with iophenoxate, it was demonstrated that HSA was the major auranofin-interacting protein in plasma. The CE-ICP-MS method is proposed as a novel approach for kinetic studies of the interactions between gold-based drugs and plasma proteins. Graphical Abstract Development of a CE-ICP-MS based method allows for studies on interaction of the gold containing drug auranofin with plasma proteins.
以金诺芬为例，开发了一种基于毛细管电泳（CE）和电感耦合等离子体质谱（ICP-MS）检测的分析方法，用于研究含金药物与血浆蛋白的相互作用。获得了金诺芬的检测限为18 ng/mL（相当于5.2 ng/mL金），精密度为1.5%。通过在水溶液以及37℃的20%人血浆中孵育，对金诺芬与蛋白质之间的相互作用进行了动力学研究。金诺芬与人血清白蛋白（HSA）和血浆的反应进行得很快；未结合的金诺芬分别在2分钟和3分钟内消失了50%。通过用碘乙酰胺封闭HSA上的游离半胱氨酸（Cys-34），表明Cys-34是金诺芬的主要反应位点。通过用碘苯酯选择性标记20%人血浆中的HSA，证明HSA是血浆中与金诺芬相互作用的主要蛋白质。CE-ICP-MS方法被提议作为一种研究金基药物与血浆蛋白相互作用动力学的新方法。图形摘要 基于CE-ICP-MS方法的开发使得能够研究含金药物金诺芬与血浆蛋白的相互作用。
