School of Chemical Engineering, University of Science and Technology Liaoning, Anshan, 114044, China.
Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, 4072, Australia.
Chem Asian J. 2018 Sep 17;13(18):2611-2618. doi: 10.1002/asia.201800957. Epub 2018 Aug 6.
The development of effective bioanalytical methods for rapid, sensitive and specific detection of HOCl in vitro and in vivo plays a key role for better understanding the roles of this molecule in normal and diseased conditions, but remains challenging due to the highly reactive nature of HOCl and the complicated biological conditions. In this work, a new fluorescence probe, PQI, was developed for monitoring of the HOCl level in biological samples. PQI was easily synthesized by a one-step condensation reaction. Upon addition of HOCl, significant changes in the absorption spectra and the color of the solution were noticed, facilitating the "naked eye" detection of HOCl in PBS buffer. The fluorescence of PQI was found to be significantly increased within a few seconds, leading to "OFF-ON" fluorescence response towards HOCl. The sensing mechanism, oxidation of thioether by HOCl, was confirmed by HRMS titration analysis. PQI features a large Stokes shift, high sensitivity and selectivity, and rapid fluorescence response towards HOCl. Quantitative detection of HOCl in single live cells was demonstrated through fluorescence imaging and flow cytometry analysis. PQI was then successfully used in visualisation of HOCl in live zebrafish and nude mice.
开发用于快速、灵敏和特异性检测 HOCl 的有效生物分析方法在体外和体内对于更好地理解该分子在正常和疾病状态下的作用至关重要，但由于 HOCl 的高反应性和复杂的生物学条件，这仍然具有挑战性。在这项工作中，开发了一种新的荧光探针 PQI，用于监测生物样品中的 HOCl 水平。PQI 可通过一步缩合反应轻松合成。加入 HOCl 后，注意到吸收光谱和溶液颜色发生明显变化，便于在 PBS 缓冲液中“肉眼”检测 HOCl。发现 PQI 的荧光在几秒钟内显著增加，导致对 HOCl 的“关-开”荧光响应。通过高分辨率质谱滴定分析证实了传感机制，即 HOCl 对硫醚的氧化。PQI 具有大的斯托克斯位移、高灵敏度和选择性，以及对 HOCl 的快速荧光响应。通过荧光成像和流式细胞术分析成功实现了对单个活细胞中 HOCl 的定量检测。然后，PQI 成功用于活体斑马鱼和裸鼠中 HOCl 的可视化。
