氮掺杂石墨烯与发夹DNA修饰的电极为工作电极-差分脉冲伏安法用于测定多巴胺
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摘要
将玻碳电极(GCE)打磨至呈镜面,在其表面上滴加氮掺杂石墨烯悬浮液5.0μL,在50℃的红外灯下烘干,制得氮掺杂石墨烯修饰的GCE;然后取5.0μmol·L-1发夹DNA(H DNA)溶液10μL滴涂于氮掺杂石墨烯修饰电极表面,制得氮掺杂石墨烯和H DNA修饰的GCE。用此修饰电极作为工作电极,用差分脉冲伏安法(DPV)测定人体血清中多巴胺(DA)的含量。试验表明:氮掺杂石墨烯和H DNA修饰的电极对DA的电化学氧化具有更好的电催化作用。DA在此修饰电极上的氧化峰电流与其浓度在4.0×10-7～6.0×10-5 mol·L-1内呈线性关系,检出限(3s/k)为6.6×10-8 mol·L-1。测定时用pH 6.5磷酸盐缓冲溶液(PBS)作为支持电解质。分析血清样品时前处理如下:取血清样品2.0mL,加入甲醇4.0mL,离心沉淀。取上清液2.0mL,加入等体积的pH 6.5PBS,充分混匀后供测定。用pH 6.5的PBS配制DA标准溶液系列,利用DPV对DA标准溶液系列进行测定,记录其氧化峰电流值,制作工作曲线。应用此方法分析了人体血清样品并以此样品为基体进行加标回收试验,测得回收率在90.0%～110%之间,测定值的相对标准偏差(n=5)在1.7%～3.7%之间。
Glassy carbon electrode(GCE)was polished to give a mirrorlike surface,and 5.0μL of nitrogen doped graphene suspension were dropped onto the surface of GCE which was then dried at 50 ℃ radiated from an IR-lamp.Nitrogen doped graphene modified GCE was thus obtained.Furthermore,10μL of 5.0μmol·L-1 of H DNA solution were dropped onto the surface of nitrogen doped graphene modified GCE to obtain the nitrogen doped graphene and H DNA modified GCE.This compositly modified GCE was used as working electrode in DPV determination of dopamine(DA)in human blood serum.It was shown that better electrocatalytic action on electrooxidation of DA at the compositly modified GCE was obtained.Linear relationship was found between values of oxidation peak current and concentration of DA in the range of 4.0×10-7-6.0×10-5 mol·L-1,and the detection limit(3 s/k)was 6.6×10-8 mol·L-1.In DPV determinations,pH 6.5 PBS was used as supporting electrolyte.In the analysis of human blood serum,the sample was pretreated as follows:2.0 mL of the serum sample were taken and 4.0 mL of methanol was added and the mixture was centrifuged for precipitation.2.0 mL of the supernatant were taken,and an equal volume of pH 6.5 PBS was added and mixed thoroughly.The solution obtained was used for DPV determination.To prepare standard curve,a series of DA standard solutions with various concentrations were prepared by dilution with pH 6.5 PBS and analyzed by DPV,and data of various oxidation peak current given by the standard DA were recorded.The proposed method was used in analysis of 3 blood samples and test for recovery was made by standard addition method using these samples as matrixes,giving results of recovery in the range of 90.0%-110%,and RSDs(n=5)were found in the range of 1.7%-3.7%.
Glassy carbon electrode(GCE)was polished to give a mirrorlike surface,and 5.0μL of nitrogen doped graphene suspension were dropped onto the surface of GCE which was then dried at 50 ℃ radiated from an IR-lamp.Nitrogen doped graphene modified GCE was thus obtained.Furthermore,10μL of 5.0μmol·L-1 of H DNA solution were dropped onto the surface of nitrogen doped graphene modified GCE to obtain the nitrogen doped graphene and H DNA modified GCE.This compositly modified GCE was used as working electrode in DPV determination of dopamine(DA)in human blood serum.It was shown that better electrocatalytic action on electrooxidation of DA at the compositly modified GCE was obtained.Linear relationship was found between values of oxidation peak current and concentration of DA in the range of 4.0×10-7-6.0×10-5 mol·L-1,and the detection limit(3 s/k)was 6.6×10-8 mol·L-1.In DPV determinations,pH 6.5 PBS was used as supporting electrolyte.In the analysis of human blood serum,the sample was pretreated as follows:2.0 mL of the serum sample were taken and 4.0 mL of methanol was added and the mixture was centrifuged for precipitation.2.0 mL of the supernatant were taken,and an equal volume of pH 6.5 PBS was added and mixed thoroughly.The solution obtained was used for DPV determination.To prepare standard curve,a series of DA standard solutions with various concentrations were prepared by dilution with pH 6.5 PBS and analyzed by DPV,and data of various oxidation peak current given by the standard DA were recorded.The proposed method was used in analysis of 3 blood samples and test for recovery was made by standard addition method using these samples as matrixes,giving results of recovery in the range of 90.0%-110%,and RSDs(n=5)were found in the range of 1.7%-3.7%.
引文
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[21] NARAYANA P V,MADHUSUDANA REDDY T,GOPAL P,et al.Electrocatalytic boost up of epinephrine and its simultaneous resolution in the presence of serotonin and folic acid at poly(serine)/multiwalled carbon nanotubes composite modified electrode:A voltammetric study[J].Materials Science and Engineering:C,2015,56:57-65.
[22] ESKANDARI K, GHOURCHIAN H.Structural changes of glucose oxidase upon interaction with goldcoated magnetic nano-particles[J].International Journal of Biological Macromolecules,2012,51(5):998-1002.
[2] DU J,YUE R R,REN F F,et al.Novel graphene flowers modified carbon fibers for simultaneous determination of ascorbic acid,dopamine and uric acid[J].Biosensors and Bioelectronics,2014,53:220-224.
[3] NOROOZIFAR M, KHORASANI-MOTLAGH M,AKBARI R,et al.Simultaneous and sensitive determination of a quaternary mixture of AA,DA,UA and Trp using a modified GCE by iron ion-doped natrolite zeolite-multiwall carbon nanotube[J].Biosensors and Bioelectronics,2011,28(1):56-63.
[4] HU S R,HUANG Q T,LIN Y,et al.Reduced graphene oxide-carbon dots composite as an enhanced material for electrochemical determination of dopamine[J].Electrochimica Acta,2014,130:805-809.
[5] AHN M,KIM J.Electrochemical behavior of dopamine and ascorbic acid at dendritic Au rod surfaces:Selective detection of dopamine in the presence of high concentration of ascorbic acid[J].Journal of Electroanalytical Chemistry,2012,683:75-79.
[6] JIN L Y,GAO X,WANG L S,et al.Electrochemical activation of polyethyleneimine-wrapped carbon nanotubes/in situ formed gold nanoparticles functionalised nanocomposite sensor for high sensitive and selective determination of dopamine[J].Journal of Electroanalytical Chemistry,2013,692:1-8.
[7] HYMAN S E,MALENKA R C.Addiction and the brain:The neurobiology of compulsion and its persistence[J].Nature Reviews Neuroscience,2001,2(10):695-703.
[8] TOBLER P N,FIORILLO C D,SCHULTZ W.Adaptive coding of reward value by dopamine neurons[J].Science,2005,307(5715):1642-1645.
[9] LI X Y,LU X J,KAN X W.3Delectrochemical sensor based on poly(hydroquinone)/gold nanoparticles/nickel foam for dopamine sensitive detection[J].Journal of Electroanalytical Chemistry,2017,799:451-458.
[10]孙登明,田相星,马伟.银掺杂聚L-酪氨酸修饰电极同时测定多巴胺、肾上腺素和抗坏血酸[J].分析化学,2010,38(12):1742-1746.
[11] GU H,VARNER E L,GROSKREUTZ S R,et al.In vivo monitoring of dopamine by microdialysis with1min temporal resolution using online capillary liquid chromatography with electrochemical detection[J].Analytical Chemistry,2015,87(12):6088-6094.
[12] WANG H Y,SUN Y,TANG B.Study on fluorescence property of dopamine and determination of dopamine by fluorimetry[J].Talanta,2002,57(5):899-907.
[13] ZHOU Y,YAN H,XIE Q,et al.Simultaneous analysis of dopamine and homovanillic acid by highperformance liquid chromatography with wall-jet/thin-layer electrochemical detection[J].The Analyst,2013,138(23):7246-7253.
[14] BRUMFIEL G.Graphene gets ready for the big time[J].Nature,2009,458(7237):390-391.
[15] SYKES E C H.Surface assembly:Graphene goes undercover[J].Nature Chemistry,2009,1(3):175-176.
[16] WU D Q,ZHANG F,LIANG H W,et al.Nanocomposites and macroscopic materials:assembly of chemically modified graphene sheets[J].Chemical Society Reviews,2012,41(18):6160-6177.
[17] O′NEIL M A,BARTON J K.DNA charge transport:conformationally gated hopping through stacked domains[J].Journal of the American Chemical Society,2004,126(37):11471-11483.
[18] LIU X G,PENG Y H,QU X J,et al.Multi-walled carbon nanotube-chitosan/poly(amidoamine)/DNA nanocomposite modified gold electrode for determination of dopamine and uric acid under coexistence of ascorbic acid[J].Journal of Electroanalytical Chemistry,2011,654(1/2):72-78.
[19] O′NEILL M A, BARTON J K. DNA-mediated charge transport requires conformational motion of the DNA bases:Elimination of charge transport in rigid glasses at 77 K[J].Journal of the American Chemical Society,2004,126(41):13234-13235.
[20] WANG X F,YOU Z,SHA H L,et al.Sensitive electrochemical detection of dopamine with a DNA/graphene bi-layer modified carbon ionic liquid electrode[J].Talanta,2014,128:373-378.
[21] NARAYANA P V,MADHUSUDANA REDDY T,GOPAL P,et al.Electrocatalytic boost up of epinephrine and its simultaneous resolution in the presence of serotonin and folic acid at poly(serine)/multiwalled carbon nanotubes composite modified electrode:A voltammetric study[J].Materials Science and Engineering:C,2015,56:57-65.
[22] ESKANDARI K, GHOURCHIAN H.Structural changes of glucose oxidase upon interaction with goldcoated magnetic nano-particles[J].International Journal of Biological Macromolecules,2012,51(5):998-1002.
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