基于磁性氧化石墨烯核酸适配体分离材料构建17β-雌二醇传感器  

Construction of a 17β-estradiol sensor based on a magnetic graphene oxide/aptamer separating material

作　　者：金心宇 陈乐源 刘艳娜 谢文菁 彭汉勇 JIN Xinyu;CHEN Leyuan;LIU Yanna;XIE Wenjing;PENG Hanyong(State Key Laboratory of Environmental Chemistry and Ecotoxicology,Research Center for Eco-Environmental Sciences,Chinese Academy of Sciences,Beijing 100085,China;University of Chinese Academy of Sciences,Beijing 100049,China;School of Environment,Hangzhou Institute for Advanced Study,University of Chinese Academy of Sciences,Hangzhou 310024,China)

机构地区：[1]中国科学院生态环境研究中心,环境化学与生态毒理学国家重点实验室,北京100085 [2]中国科学院大学,北京100049 [3]中国科学院大学杭州高等研究院,环境学院,浙江杭州310024

出　　处：《色谱》2025年第1期87-95,共9页Chinese Journal of Chromatography

基　　金：国家自然科学基金面上项目(22276199);国家重点研发计划(2022YFC3701302,2023YFA0915102);中国科学院先导专项(XDB0750100)。

摘　　要：17β-雌二醇(E2)是天然的甾体雌激素,在生物体的各项生理活动中发挥着至关重要的作用。然而,外源性E2也被归类为内分泌干扰物(EDC),即使在ng/L水平上也会干扰内分泌系统功能,且在医疗、畜牧业废水中均能检测到E2污染。目前,E2的检测方法主要以色谱-质谱联用方法为主,但受仪器等限制,难以应用于现场检测或大量样品检测。为了解决该难题,本工作用油包水微乳液法(W/O microemulsion)合成了一种基于磁性氧化石墨烯(MGO)的核酸分离材料,并进行了系统的表征,以开发高灵敏度、快速检测、高通量的荧光恢复(“turn-on”)型生物传感器,用于检测环境样品中的E2。传感器基于荧光共振能量转移原理(FRET),即MGO通过吸附荧光基团标记的E2核酸适配体发生FRET效应猝灭荧光信号,而在E2存在时,核酸适配体与E2结合,从MGO上脱附,实现荧光信号恢复,且信号响应与一定范围的E2质量浓度呈线性关系。本工作引入了磁性固相分离方法,降低了背景信号值,从而提高了方法的灵敏度。本方法中相对荧光恢复强度与E2质量浓度呈良好的线性关系,检出限低至1 ng/mL,对多种干扰离子有较好的抵抗性,且在同族雌激素类似物中有良好的选择性。与色谱-质谱检测方法相比,该方法检测时间短,成本低,操作简便,可应用于环境水样中E2的快速检测。17β-Estradiol(E2)is a natural steroidal estrogen essential for a variety of physiological functions in organisms.However,external E2,which is renowned for its potent biological effects,is also considered to be an endocrine-disrupting compound(EDC)capable of disturbing the normal operation of the endocrine system,even at nanogram-per-liter(ng/L)concentrations.Studies have revealed that medical and livestock wastewater can be contaminated with E2,which poses potential risks to human health.Currently,the primary method for detecting E2 relies on liquid chromatography-mass spectrometry,which is limited with regard to on-site or large-scale sample testing due to instrumental constraints.Herein,we developed a magnetic graphene oxide(MGO)/aptamer separating material.The MGO was synthesized by creating a water-in-oil microemulsion at 90℃,an agarose hydrogel to load the Fe 3O 4 nanoparticles,and layered graphene oxide(GO).In contrast to conventional methods,such as chemical co-precipitation and solvothermal approaches,this method is more time-efficient and does not require high temperature or pressure.Moreover,the use of a physical encapsulation technique for enwrapping the Fe 3O 4 nanoparticles and layered GO eliminates the need for chemical modification.This approach reduces the use of harmful chemicals,ensures complete loading,and results in highly efficient encapsulation.The MGO was characterized using Fourier-transform infrared(FT-IR)spectroscopy and transmission electron microscopy(TEM),as well as dynamic light scattering(DLS)and Zeta potential analyses,which revealed that the Fe 3O 4 nanoparticles had been successfully loaded onto the GO to produce MGO particles mainly around 5μm in size.Additionally,this study demonstrated that the aqueous MGO dispersion is highly stable.This substance was used to develop a fluorescent biosensor that uses a“turn-on”mechanism to rapidly and highly sensitively detect E2.MGO is capable of adsorbing a fluorescently labeled E2 aptamer(FAM-Apt)in solution,resulting in fluorescen

关 键 词：17Β-雌二醇 磁固相分离 荧光生物传感器 磁性氧化石墨烯 

分 类 号：O658[理学—分析化学]