一种超灵敏检测17β-雌二醇电化学核酸适配体传感器构建方法
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摘要
采用石墨烯(GS)-Nafion-纳米金(AuNPs)复合膜修饰玻碳电极(GCE/GS-Nafion-AuNPs),构建一种灵敏度高且稳定性好的电化学核酸适配体传感器,应用于环境内分泌干扰物——17β-雌二醇(E2)的快速检测.运用透射扫描电子显微镜(TEM)、循环伏安(CV)法和差分脉冲伏安(DPV)法,对该传感器的形貌和电化学性质进行表征.研究表明,在GS、Nafion和AuNPs协同作用下,修饰电极的电化学性能明显提高.在优化试验条件下,传感器的电流信号(I_p)与17β-雌二醇(E2)的浓度(c)的对数在0.002 5~0.300 0μmol/L范围内呈现良好的线性关系,回归方程为I_p=8.899 4+1.049 4logc,相关系数为0.994 1,E2浓度检出限为0.83nmol/L.该传感器制作简单、灵敏度高、检出限极低以及具有良好的重现性和稳定性,有望应用于环境样品中E2的检测.
The graphene(GS)and nano-gold(AuNPs)were both introduced into the Nafion film to develop a new matrix as the aptasensor platform for ultrasensitive detection of typical endocrine pollutants-17β-estradiol(E2).The morphology and electrochemical properties of the proposed sensors were characterized by transmission electron microscopy(TEM),cyclic voltammetry(CV)and differential pulse voltammetry(DPV).Due to the synergistic effect of GS,Nafion and AuNPs,biosensor modified by GS-Nafion-AuNPs composites exhibits excellent electrochemical performance.Under the optimized conditions,the current response of the proposed aptasensor have good linearity with the logarithm concentration of 17β-estradiol from 0.002 5 to 0.300 0μmol/L,and the detection limit is 0.83 nmol/L.With the advantages of simple preparation,high sensitivity,low detection limit,excellent reproducibility and stability,the proposed method could be used in the determination of E2 in environmental sample.
The graphene(GS)and nano-gold(AuNPs)were both introduced into the Nafion film to develop a new matrix as the aptasensor platform for ultrasensitive detection of typical endocrine pollutants-17β-estradiol(E2).The morphology and electrochemical properties of the proposed sensors were characterized by transmission electron microscopy(TEM),cyclic voltammetry(CV)and differential pulse voltammetry(DPV).Due to the synergistic effect of GS,Nafion and AuNPs,biosensor modified by GS-Nafion-AuNPs composites exhibits excellent electrochemical performance.Under the optimized conditions,the current response of the proposed aptasensor have good linearity with the logarithm concentration of 17β-estradiol from 0.002 5 to 0.300 0μmol/L,and the detection limit is 0.83 nmol/L.With the advantages of simple preparation,high sensitivity,low detection limit,excellent reproducibility and stability,the proposed method could be used in the determination of E2 in environmental sample.
引文
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[4]张明辉,吴蔓莉,杨瑞,等.利用酶联免疫测定水中雌二醇[J].分析试验室,2015,34(1):27-30.
[5]马丽莎,戴晓欣,谢文平,等.QuEChERS/GC-MS法同时测定鱼、虾中的雌二醇与己烯雌酚残留[J].分析测试学报,2015,34(1):62-66+72.
[6]范志影,刘庆生,常碧影.液相色谱串联质谱法测定牛奶中的内源性雌激素[J].分析试验室,2015,34(7):751-755.
[7]田家英,谭睿婕,柳玉英,等.基于现场检测和批试验条件下双酚A和雌二醇在沉积物上的分配系数[J].环境工程技术学报,2017,7(2):140-145.
[8]姜会聪,任舒悦,王瑜,等.上转换发光免疫层析法快速检测牛奶中雌二醇[J].食品研究与开发,2017,38(3):119-123.
[9]王丹华,陈盼盼,王宏亮.水产品中15种避孕药的HPLC方法测定[J].食品工业,2017,38(1):310-314.
[10]周雪妍,陈仁国,魏群利,等.液相色谱串联质谱法检测人血清雌酮、雌二醇[J].临床检验杂志,2015,33(5):328-332.
[11]ELLINGTON A D,SZOSTAK J W.In vitro selection of RNA molecules that bind specific ligands[J].Nature,1990,346:818-822.
[12]TUERK C,GOLD L.Systematic evolution of ligands by exponential RNA ligands to bacteriophage T4DNA-polymerase[J].Science,1990,249:505-510.
[13]BRIDONNEAU P,CHANG Y F,BUVOLI A V,et al.Sitedirected selection of oligonucleotide antagonists by competitive elution[J].Antisense and Nucleic Acid Drug Development,1999,9:1-11.
[14]JENISON R D,GILL S C,PARDI A,et al.High-resolution molecular discrimination by RNA[J].Science,1994,263:1425-1429.
[15]周云,丁辛,胡吉永,等.化学聚合和电化学聚合对聚吡咯/棉导电织物电性能的影响[J].东华大学学报(自然科学版),2016,42(6):822-826.
[16]黄子龙,赵昕,李英芝,等.MnO2/碳布复合材料的制备及其电化学行为[J].东华大学学报(自然科学版),2016,42(2):167-172.
[17]LIU J,LU Y.Preparation of aptamer-linked gold nanoparticle purple aggregates for colorimetric sensing of analytes[J].Nature Protocols,2006,1(1):246-252.
[18]KIM Y S,JUNG H S,MATSUURA,T,et al.Electrochemical detection of 17β-estradiol using DNA aptamer immobilized gold electrode chip[J].Biosensors and Bioelectronics,2007,22(11):2525-2531.
[19]吴唯,史江红,陈庆彩,等.沉积物中雌激素及壬基酚、辛基酚、双酚A的测定[J].环境科学,2013,34(2):724-731.
[20]史江红,吴唯,张晖,等.高基质干扰污水中雌激素、双酚A和烷基酚的共检测研究[J].给水排水,2013,39(11):124-127.