次氯酸根响应的荧光探针的合成及应用研究  

Synthesis and application of fluorescent probe for detecting hypochlorite

作　　者：聂青丽 陈璞 冯海涛 NIE Qing-li;CHEN Pu;FENG Hai-tao(AIE Research Center,Shaanxi Key Laboratory of Phytochemistry,College of Chemistry and Chemical Engineering,Baoji University of Arts and Sciences,Baoji 721013,Shaanxi,China)

机构地区：[1]宝鸡文理学院化学化工学院,陕西省植物化学重点实验室,AIE(聚集诱导发光)研究中心,陕西宝鸡721013

出　　处：《宝鸡文理学院学报(自然科学版)》2024年第4期23-30,36,I0003,I0004,共11页Journal of Baoji University of Arts and Sciences(Natural Science Edition)

基　　金：陕西省科技厅创新团队项目(2022TD-36);陕西化生基础科学研究项目(22JHQ078);国家自然科学基金项目(52173152)。

摘　　要：目的次氯酸根(ClO^(-))作为一种常见的活性氧物质,在日常生活和生物体内均发挥着至关重要的作用。然而,由于ClO^(-)的强氧化性,过量ClO^(-)的存在会造成财产的损失和机体的损害,因此准确检测ClO^(-)的浓度显得尤为重要。荧光检测法由于其成本低、灵敏度高和操作简便而被广泛应用于离子的检测。本研究尝试开发一种能够选择性、快速检测ClO^(-)的荧光探针。方法通过有机合成手段合成探针2-氨基-3-(((E)-(4'-二苯基氨基)-3-羟基-[1,1-二苯基]-4-亚甲基)氨基)顺丁烯腈(TPAN),经核磁共振波谱、高分辨质谱对其结构进行表征和确认,利用紫外吸收光谱仪、荧光光谱仪对探针的性质进行了测试,并将其应用于ClO^(-)的识别和细胞成像。结果在探针TPAN的二甲基亚砜(DMSO)溶液中加入不同阴离子和各类活性氧,仅ClO^(-)能使溶液的颜色由黄色变为无色,荧光颜色由微弱的红色变成明亮的蓝色。且在其他离子或自由基存在时,TPAN对ClO^(-)的识别效果不受影响。在Hela细胞中加入20μmol/L的TPAN,细胞活性仍然在60%以上,通过细胞成像和与细胞共定位,确定探针TPAN在内质网中。探针TPAN在Hela细胞中能进行ClO^(-)成像,可有效用于细胞内外源性ClO^(-)的检测。结论本文所制备探针TPAN对ClO^(-)表现出高选择性和强抗干扰性,可以通过荧光法和比色法进行识别,且TPAN具有较好的生物相容性,可应用于细胞内ClO^(-)的识别。Purposes—To explore a fluorescent probe for selective and fast detection of ClO^(-)on the basis of the following fact that hypochlorite(ClO^(-)),as a common reactive oxygen species,plays important roles in our daily life and organism,however,excess ClO^(-)often causes some financial losses and body injury own to its strong oxidability,so it is necessary to achieve the accurate detection of the concentration of ClO^(-),and in several analytical methods,fluorescent technique was widely used in detection of ions due to its low cost,high sensitivity and easy operability.Methods—A fluorescent probe named TPAN was synthesized by organic reaction and characterized during NMR,HRMS,UV-vis and fluorescence spectra.In addition,TPAN was used in the detection of ClO^(-)and cell imaging.Results—Adding different anion and various reactive oxygen in to the DMSO solution of TPAN,only ClO^(-)can make the color of the solution changed from yellow to colorless,and the fluorescence changed from faint red to bright blue,and the ClO^(-)recognition effect of TPAN was not affected by other ions or free radicals.The cell viability was more than 60%,when the Hela cells were co-incubated with TPAN for 24 h on the concentration of 20μmol/L.After the experiments of cell imaging and colocalization,TPAN was proved to target the endoplasmic reticulum.Moreover,TPAN could be used as a probe for the detection of exogenous ClO^(-)in Hela cells.Conclusions—The fluorescent probe TPAN could detect ClO^(-)with high selectivity and strong anti-jamming both in fluorescent technique and colorimetric method.Moreover,TPAN was successfully applied for imaging ClO^(-)in living cells due to its great biocompatibility.

关 键 词：次氯酸根 荧光探针 细胞成像 

分 类 号：O656.22[理学—分析化学]