基于Au-MoS_2和hemin的双酚A电化学适配体传感器研究
|
摘要
本文基于双酚A(BPA)与其适配体互补链(cDNA)对适配体的竞争结合作用构建了检测BPA的电化学传感器。制备了金纳米粒子与二硫化钼的纳米复合物(Au-MoS_2),并将其修饰到玻碳电极(GCE)表面,制得修饰电极Au-MoS_2/GCE。通过巯基修饰的cDNA与适配体的杂交反应,生成双链DNA(dsDNA),利用其中cDNA的巯基在金纳米粒子表面生成Au-S键的化学吸附作用,将dsDNA修饰到电极表面。利用具有卟啉平面结构的氯化血红素(hemin)在dsDNA沟槽中的嵌插作用,制得电化学传感器Hemin-dsDNA/Au-MoS_2/GCE。基于hemin对于H_2O_2和对苯二酚(HQ)的化学反应的电催化作用,建立了差分脉冲伏安法(DPV)测定BPA的分析方法,由于BPA与适配体结合的亲和性较强,在一定实验条件下,溶液中BPA浓度越高,导致更多的hemin从电极表面脱落,结果表明,在优化的实验条件下,DPV峰电流值与BPA浓度在1.0 nmol·L~(-1)至10.0μmol·L~(-1)的范围内呈线性关系,检测限为0.8 nmol·L~(-1)。本文还考察了传感器的稳定性与选择性,并将其用于实际样品中BPA的检测,结果满意。
In this paper,an electrochemical aptamer sensor for bisphenol A(BPA)was fabricated based on the competition bindingof BPA and its aptamer complementary strand(cDNA)to the aptamers.A composite composed of gold nanoparticles and molybdenum disulfide was prepared and was denoted as Au-MoS_2.The composite was then modified onto the surface of a glassy carbon electrode(GCE).The thiol-modified cDNA was hybridized with the aptamer to form double-stranded DNA(dsDNA).The dsDNA was modified onto the surface of the electrode based on Au-S bonds produced by chemisorptions of thiol-modified cDNA on Au nanoparticles.Hemin which possesses the porphyrin plane structure was embedded into the dsDNA groove to prepare the electrochemical aptasensor(Hemin-dsDNA/Au-MoS_2/GCE).An analytical method for BPA was established by using differential pulse voltammetry(DPV).The reaction between H_2O_2 and hydroquinone(HQ)was electrochemically catalyzed by hemin.Because the affinity of BPA with aptamers is stronger,under certain experimental conditions,the higher concentration of BPA in solutioncan cause more hemin to fall off the surface of the electrode.The results showed that the peak current value of DPV in the range of 1.0 nmol·L~(-1) to 10.0 μmol·L~(-1) BPA was linearly decreased with the increase of BPA concentration under the optimized experimental conditions.The detection limit was 0.8 nmol·L~(-1).The stability and selectivity of the sensor were investigated.BPA in real sampleswasdetermined,and satisfactory results were obtained.
In this paper,an electrochemical aptamer sensor for bisphenol A(BPA)was fabricated based on the competition bindingof BPA and its aptamer complementary strand(cDNA)to the aptamers.A composite composed of gold nanoparticles and molybdenum disulfide was prepared and was denoted as Au-MoS_2.The composite was then modified onto the surface of a glassy carbon electrode(GCE).The thiol-modified cDNA was hybridized with the aptamer to form double-stranded DNA(dsDNA).The dsDNA was modified onto the surface of the electrode based on Au-S bonds produced by chemisorptions of thiol-modified cDNA on Au nanoparticles.Hemin which possesses the porphyrin plane structure was embedded into the dsDNA groove to prepare the electrochemical aptasensor(Hemin-dsDNA/Au-MoS_2/GCE).An analytical method for BPA was established by using differential pulse voltammetry(DPV).The reaction between H_2O_2 and hydroquinone(HQ)was electrochemically catalyzed by hemin.Because the affinity of BPA with aptamers is stronger,under certain experimental conditions,the higher concentration of BPA in solutioncan cause more hemin to fall off the surface of the electrode.The results showed that the peak current value of DPV in the range of 1.0 nmol·L~(-1) to 10.0 μmol·L~(-1) BPA was linearly decreased with the increase of BPA concentration under the optimized experimental conditions.The detection limit was 0.8 nmol·L~(-1).The stability and selectivity of the sensor were investigated.BPA in real sampleswasdetermined,and satisfactory results were obtained.
引文
[1]Kavlock R J,Daston G P,DeRosa C,et al.Research needs for the risk assessment of health and environmental effects of endocrine disruptors:a report of the U.S.EPA-sponsored workshop[J].Environ Health Perspect,1996,104(Suppl 4):715-740.
[2]Le HH,Carlson E M,Chua J P,et al.Bisphenol A is released from polycarbonate drinking bottles and mimics the neurotoxic actions of estrogen in developing cerebellar neurons[J].Toxicol Lett,2008,176(2):149-156.
[3]陈宗保,刘林海,董洪霞.基于双重增敏剂荧光法测定食品包装材料中痕量双酚A[J].化学研究与应用,2016,28(6):829-832.
[4]杨丹,李丹丹,刘姗姗,等.双酚A对机体的影响及其作用机制[J].现代预防医学,2008,35(17):3280-3282,3287.
[5]Sun H,Xu LC,Chen JF,et al.Effect of bisphenol A,tetrachlorobisphenol A and pentachlorophenol on the transcriptional activities of androgen receptor-mediated reporter gene[J].Food Chem Toxicol,2006,44(11):1916-1921.
[6]Kitamura S,JinnoN,OhtaS,et al.Thyroid hormonal activity of the flame retardants tetrabromobisphenol A and tetrachlorobisphenol A[J].Biochem Biophys Res Commun,2002,293(1):554-559.
[7]张杰,许家胜,崔岩.高效液相色谱法检测一次性纸杯中的双酚A[J].化学研究与应用,2014,26(3):454-456.
[8]Adachi T,Yasuda K,Mori C,et al.Promoting insulin secretion in pancreatic islets by means of bisphenol A and nonylphenol via intracellular estrogen receptors[J].Food ChemToxicol,2005,43:713-719.
[9]Szymański A,Rykowska I,Wasiak W.Determination of bisphenol A in water and milk by micellar liquid chromatography[J].ActaChromatogr,2006,(17):161-172.
[10]Yao Q H,Feng Y F,Tan C,et al.An on-line solid-phase extraction disc packed with a phytic acid induced 3D graphene-based foam for the sensitive HPLC-PDA determination of bisphenol A migration in disposable syringes[J].Talanta,2018,179:153-158.
[11]Eliseo H H,Rita C M,Encarnacion R G.Behavior of phenols and phenoxyacids on a bisphenol-A imprinted polymer.Application for selective solid-phase extraction from water and urine samples[J].Int J Mol SCI,2011,12(12):3322-3339.
[12]Sambe H,Hoshina K,et al,et al.Simultaneous determination ofbisphenol A and its halogenated derivatives in river water by combination of isotopeimprinting and liquid chromatography-mass spectrometry[J].J Chromatogr A,2006,1134:16-23.
[13]Zhao MP,Li YZ,GuoZ Q,et al.A new competitive enzyme-linked immunosorbent assay(ELISA)for determination of estrogenic bisphenols[J].Talanta,2002,57(6):1205-1210.
[14]Wang S H,Wei X T,Du L Y,et al.Determination of bisphenol A using a flow injection inhibitory chemiluminescence method[J].Lumin,2005,20:46-50.
[15]Lu Y,Li X C,Zhang L M,et al.Aptamer-based electrochemical sensors with aptamer-complementary DNA oligonucleotides as probe[J].Anal Chem,2008,80(6):1883-1890.
[16]Sharma A K,Ken A D,HeemstraJ M.Enzyme-linked small-molecule detection using split aptamer ligation[J].Anal Chem,2012,84(14):6104-6109.
[17]Mei Z L,Chu H Q,Chen W,et al.Ultrasensitive one-step rapid visual detection of bisphenol A in water samples by label-free aptasensor[J].BiosensBioelectron,2013,39(1):26-30.
[18]Li Y,Xu J Y,Wang L K,et al.Aptamer-based fluorescent detection of bisphenol A using nonconjugated gold nanoparticles and CdTe quantum dots[J].Sens Actuators,B:Chemical,2016,222:815-822.
[19]Wang C X,Lin H H,Xu Z Z,et al.One-step hydrothermal synthesis of flowerlike MoS2/CdSheterostructures for enhanced visible-light photocatalytic activities[J].RSC Adv,2015,5(20):15621-15626.
[20]Li B L,Luo H Q,LeiJ L,et al.Hemin-functionalized MoS2 nanosheets:enhanced peroxidase-like catalytic activity with a steady state in aqueous solution[J].RscAdv,2014,4:24256-24262.
[21]Zhou K Q,Jiang S H,BaoC L,et al.Preparation of poly(vinyl alcohol)nanocomposites with molybdenum disulfide(MoS2):structural characteristics and markedly enhanced properties[J].RscAdv,2012,2:11695-11703.
[22]Sun M Y,Adjaye J,Nelson A E.Theoretical investigations of the structures and properties of molybdenum-based sulfide catalysts[J].Appl Catal A,2004(263):131-143.
[23]Huang K J,Wang L,Li J,et al.Electrochemical sensing based on layered MoS2-grapheme composites[J].Sens Actuators B:Chemical,2013,178:671-677.
[24]Huang K J,Liu Y J,Liu Y M,et al.Molybdenum disulfide nanoflower-chitosan-Au nanoparticlescomposites based electrochemical sensing platform for bisphenol Adetermination[J].J Hazard Mater,2014,276:207-215.
[25]Wang Y H,Huang K J,Wu X.Recent advances in transition-metal dichalcogenides based electrochemicalbiosensors:A review[J].BiosensBioelectron,2017,97:305-316.
[26]Shu Y,Chen J Y,Xu Q,et al.MoS2nanosheet-Au nanorod hybrids for highly sensitive amperometric detection of H2O2 in living[J].J Mater Chem B,2017,5:1446-1453.
[27]Zhou X S,Wan L J,GuoY G.Synthesis of MoS2nanosheet-graphene nanosheet hybrid materials for stable lithium storage[J].ChemCommun,2013,49:1838-1840.
[28]Du X L,Kang T F,Lu L P,et al.An electrochemiluminescence sensor based on CdSe@CdS functionalized MoS2 and hemin/G-quadruplex-based DNAzymebiocatalytic precipitation for sensitive detection of Pb(II)[J].Anal Chem,2018,10:51-58.
[29]Jiang X Y,Chai Y Q,Wang H J,et al.Electrochemiluminescence of luminol enhanced by the synergetic catalysis of hemin and silver nanoparticles for sensitive protein detection[J].BiosensBioelectron,2014,54:20-26.
[30]Lu L P,Liu C,Kang T F,et al.In situ enhanced electrochemiluminescence based on co-reactant self-generated for sensitive detection of microRNA[J].Sens Actuators,B:Chemical,2018,225:35-41.
[31]Kara P,OzkanD,Kerman K,et al.DNA sensing on glassy carbon electrodes by using hemin as the electrochemical hybridization label[J].Anal Bioanal Chem,2002,373:710-716.
[32]Jo M,Ahn J,Lee J,et al.Development of single-stranded DNA aptamers for specific bisphenol A detection[J].Oligonucleotides,2011,21(2):85-91.
[33]Hu F,Chen S,Yuan R.Application of magnetic core-shell microspheres on reagentlessimmunosensor based on direct electrochemistry of glucose oxidase for detection of carbohydrate antigen 19-9[J].Sens Actuators B:Chemical,2013,176:713-722.
[34]Zhang S,Zhong H,Ding C.Ultrasensitive flow injection chemiluminescence detection of DNA hybridization using signal DNA probe modified with Au and CuS nanoparticles[J].Anal Chem, 2008,80(19):7206-7212.
[35]杜晓丽,康天放.基于Au-MoS2纳米复合物的DNA传感器检测Hg2+研究[J].中国科技论文在线精品论文,2017,10(24):2679-2687.
[36]Zhou J,Du L P,Wang P,et al.An ultrasensitive electrochemical immunosensor for carcinoembryonic antigen detection based on staphylococcal protein A-Au nanoparticle modified gold electrode[J].Sens Actuators B,2014,197:220-227.
[37]PoorahongS,ThammakhetC,ThavarungkulP,et al.Amperometric sensor for detection of bisphenol A using a pencil graphite electrode modified with polyaniline nanorods and multiwalled carbon nanotubes[J].Microchim Acta,2012,176:91-99.
[38]Zhou L,Wang J P,Li D J,et al.An electrochemical aptasensor based on gold nanoparticles dotted graphene modified glassy carbon electrode for label-free detection of bisphenol A in milk samples[J].Food Chem,2014,162(1):34-40.
[2]Le HH,Carlson E M,Chua J P,et al.Bisphenol A is released from polycarbonate drinking bottles and mimics the neurotoxic actions of estrogen in developing cerebellar neurons[J].Toxicol Lett,2008,176(2):149-156.
[3]陈宗保,刘林海,董洪霞.基于双重增敏剂荧光法测定食品包装材料中痕量双酚A[J].化学研究与应用,2016,28(6):829-832.
[4]杨丹,李丹丹,刘姗姗,等.双酚A对机体的影响及其作用机制[J].现代预防医学,2008,35(17):3280-3282,3287.
[5]Sun H,Xu LC,Chen JF,et al.Effect of bisphenol A,tetrachlorobisphenol A and pentachlorophenol on the transcriptional activities of androgen receptor-mediated reporter gene[J].Food Chem Toxicol,2006,44(11):1916-1921.
[6]Kitamura S,JinnoN,OhtaS,et al.Thyroid hormonal activity of the flame retardants tetrabromobisphenol A and tetrachlorobisphenol A[J].Biochem Biophys Res Commun,2002,293(1):554-559.
[7]张杰,许家胜,崔岩.高效液相色谱法检测一次性纸杯中的双酚A[J].化学研究与应用,2014,26(3):454-456.
[8]Adachi T,Yasuda K,Mori C,et al.Promoting insulin secretion in pancreatic islets by means of bisphenol A and nonylphenol via intracellular estrogen receptors[J].Food ChemToxicol,2005,43:713-719.
[9]Szymański A,Rykowska I,Wasiak W.Determination of bisphenol A in water and milk by micellar liquid chromatography[J].ActaChromatogr,2006,(17):161-172.
[10]Yao Q H,Feng Y F,Tan C,et al.An on-line solid-phase extraction disc packed with a phytic acid induced 3D graphene-based foam for the sensitive HPLC-PDA determination of bisphenol A migration in disposable syringes[J].Talanta,2018,179:153-158.
[11]Eliseo H H,Rita C M,Encarnacion R G.Behavior of phenols and phenoxyacids on a bisphenol-A imprinted polymer.Application for selective solid-phase extraction from water and urine samples[J].Int J Mol SCI,2011,12(12):3322-3339.
[12]Sambe H,Hoshina K,et al,et al.Simultaneous determination ofbisphenol A and its halogenated derivatives in river water by combination of isotopeimprinting and liquid chromatography-mass spectrometry[J].J Chromatogr A,2006,1134:16-23.
[13]Zhao MP,Li YZ,GuoZ Q,et al.A new competitive enzyme-linked immunosorbent assay(ELISA)for determination of estrogenic bisphenols[J].Talanta,2002,57(6):1205-1210.
[14]Wang S H,Wei X T,Du L Y,et al.Determination of bisphenol A using a flow injection inhibitory chemiluminescence method[J].Lumin,2005,20:46-50.
[15]Lu Y,Li X C,Zhang L M,et al.Aptamer-based electrochemical sensors with aptamer-complementary DNA oligonucleotides as probe[J].Anal Chem,2008,80(6):1883-1890.
[16]Sharma A K,Ken A D,HeemstraJ M.Enzyme-linked small-molecule detection using split aptamer ligation[J].Anal Chem,2012,84(14):6104-6109.
[17]Mei Z L,Chu H Q,Chen W,et al.Ultrasensitive one-step rapid visual detection of bisphenol A in water samples by label-free aptasensor[J].BiosensBioelectron,2013,39(1):26-30.
[18]Li Y,Xu J Y,Wang L K,et al.Aptamer-based fluorescent detection of bisphenol A using nonconjugated gold nanoparticles and CdTe quantum dots[J].Sens Actuators,B:Chemical,2016,222:815-822.
[19]Wang C X,Lin H H,Xu Z Z,et al.One-step hydrothermal synthesis of flowerlike MoS2/CdSheterostructures for enhanced visible-light photocatalytic activities[J].RSC Adv,2015,5(20):15621-15626.
[20]Li B L,Luo H Q,LeiJ L,et al.Hemin-functionalized MoS2 nanosheets:enhanced peroxidase-like catalytic activity with a steady state in aqueous solution[J].RscAdv,2014,4:24256-24262.
[21]Zhou K Q,Jiang S H,BaoC L,et al.Preparation of poly(vinyl alcohol)nanocomposites with molybdenum disulfide(MoS2):structural characteristics and markedly enhanced properties[J].RscAdv,2012,2:11695-11703.
[22]Sun M Y,Adjaye J,Nelson A E.Theoretical investigations of the structures and properties of molybdenum-based sulfide catalysts[J].Appl Catal A,2004(263):131-143.
[23]Huang K J,Wang L,Li J,et al.Electrochemical sensing based on layered MoS2-grapheme composites[J].Sens Actuators B:Chemical,2013,178:671-677.
[24]Huang K J,Liu Y J,Liu Y M,et al.Molybdenum disulfide nanoflower-chitosan-Au nanoparticlescomposites based electrochemical sensing platform for bisphenol Adetermination[J].J Hazard Mater,2014,276:207-215.
[25]Wang Y H,Huang K J,Wu X.Recent advances in transition-metal dichalcogenides based electrochemicalbiosensors:A review[J].BiosensBioelectron,2017,97:305-316.
[26]Shu Y,Chen J Y,Xu Q,et al.MoS2nanosheet-Au nanorod hybrids for highly sensitive amperometric detection of H2O2 in living[J].J Mater Chem B,2017,5:1446-1453.
[27]Zhou X S,Wan L J,GuoY G.Synthesis of MoS2nanosheet-graphene nanosheet hybrid materials for stable lithium storage[J].ChemCommun,2013,49:1838-1840.
[28]Du X L,Kang T F,Lu L P,et al.An electrochemiluminescence sensor based on CdSe@CdS functionalized MoS2 and hemin/G-quadruplex-based DNAzymebiocatalytic precipitation for sensitive detection of Pb(II)[J].Anal Chem,2018,10:51-58.
[29]Jiang X Y,Chai Y Q,Wang H J,et al.Electrochemiluminescence of luminol enhanced by the synergetic catalysis of hemin and silver nanoparticles for sensitive protein detection[J].BiosensBioelectron,2014,54:20-26.
[30]Lu L P,Liu C,Kang T F,et al.In situ enhanced electrochemiluminescence based on co-reactant self-generated for sensitive detection of microRNA[J].Sens Actuators,B:Chemical,2018,225:35-41.
[31]Kara P,OzkanD,Kerman K,et al.DNA sensing on glassy carbon electrodes by using hemin as the electrochemical hybridization label[J].Anal Bioanal Chem,2002,373:710-716.
[32]Jo M,Ahn J,Lee J,et al.Development of single-stranded DNA aptamers for specific bisphenol A detection[J].Oligonucleotides,2011,21(2):85-91.
[33]Hu F,Chen S,Yuan R.Application of magnetic core-shell microspheres on reagentlessimmunosensor based on direct electrochemistry of glucose oxidase for detection of carbohydrate antigen 19-9[J].Sens Actuators B:Chemical,2013,176:713-722.
[34]Zhang S,Zhong H,Ding C.Ultrasensitive flow injection chemiluminescence detection of DNA hybridization using signal DNA probe modified with Au and CuS nanoparticles[J].Anal Chem, 2008,80(19):7206-7212.
[35]杜晓丽,康天放.基于Au-MoS2纳米复合物的DNA传感器检测Hg2+研究[J].中国科技论文在线精品论文,2017,10(24):2679-2687.
[36]Zhou J,Du L P,Wang P,et al.An ultrasensitive electrochemical immunosensor for carcinoembryonic antigen detection based on staphylococcal protein A-Au nanoparticle modified gold electrode[J].Sens Actuators B,2014,197:220-227.
[37]PoorahongS,ThammakhetC,ThavarungkulP,et al.Amperometric sensor for detection of bisphenol A using a pencil graphite electrode modified with polyaniline nanorods and multiwalled carbon nanotubes[J].Microchim Acta,2012,176:91-99.
[38]Zhou L,Wang J P,Li D J,et al.An electrochemical aptasensor based on gold nanoparticles dotted graphene modified glassy carbon electrode for label-free detection of bisphenol A in milk samples[J].Food Chem,2014,162(1):34-40.