Abstract:
Free iron in human serum or non-transferrin-bound iron (NTBI) can generate free radicals and lead to oxidative damage. Moreover, it is highly toxic to various tissues and a vital biomarker related to the iron-loading status of thalassemia and Alzheimer\'s patients. In NTBI in healthy individuals, NTBI levels are typically less than 1 µM; current NTBI analysis usually requires advanced instrumentation and many-step sample pretreatment. To address this issue, we employed our invented BODIPY derivative, BODIPY-PH, as a fluorescence probe and trapped it onto the microcentrifuge tube lid using tapioca starch. The fluorescence intensity of BODIPY-PH increased with increasing NTBI concentration (turn-on). The developed portable reaction chamber facilitates rapid analysis (∼5 min) using small sample volumes (10 μL sample in a total volume of 600 μL). Under optimum conditions, using the sample-developed portable fluorescence device and fluorescence spectrometer, we achieved impressive limits of detection (LOD) of 0.003 and 0.0015 μM, respectively. Furthermore, the developed sensors show relatively high selectivity toward Fe3+ over other metal ions and biomolecules (i.e., Fe2+, Cr3+, Cu2+, and glucose). The sensor performance in serum samples of thalassemia patients exhibited no significant difference compared to the labeled value (obtained from standard methods). Overall, the developed fluorescence sensor is suitable for determining NTBI and offers high sensitivity, high selectivity, and a short incubation time (5 min). Moreover, the method requires a limited number of reagents, is simple to use, and uses low-cost equipment to determine NTBI in human serum samples.
摘要:
人血清中的游离铁或非转铁蛋白结合铁(NTBI)可产生自由基并导致氧化损伤。此外,它对各种组织都有很强的毒性,并且是与地中海贫血和阿尔茨海默症患者的铁负荷状态有关的重要生物标志物。在健康个体的NTBI中,NTBI水平通常小于1µM;当前的NTBI分析通常需要先进的仪器和多步骤样品预处理。为了解决这个问题,我们使用了我们发明的BODIPY导数,BODIPY-PH,作为荧光探针,并用木薯淀粉将其捕获到微量离心管盖上。BODIPY-PH的荧光强度随着NTBI浓度的增加(开启)而增加。开发的便携式反应室有助于使用小样品体积(总体积为600μL的10μL样品)进行快速分析(〜5分钟)。在最佳条件下,使用样品开发的便携式荧光装置和荧光光谱仪,我们实现了令人印象深刻的检测限(LOD)0.003和0.0015μM,分别。此外,与其他金属离子和生物分子相比,开发的传感器对Fe3+显示出相对较高的选择性(即,Fe2+,Cr3+,Cu2+,和葡萄糖)。与标记值(从标准方法获得)相比,地中海贫血患者血清样品中的传感器性能没有显着差异。总的来说,开发的荧光传感器适用于测定NTBI,并提供高灵敏度,高选择性,和短的孵育时间(5分钟)。此外,该方法需要有限数量的试剂,使用简单,并使用低成本设备测定人血清样品中的NTBI。