研究生: |
林立心 LiHsin Lin |
---|---|
論文名稱: |
碳六十固定化酵素電化學葡萄糖感測器研製與應用 Preparation and Application of Immobilized Fullerence-Enzyme Based electrochemical glucose Sensor |
指導教授: |
施正雄
Shih, Jeng-Shong |
學位類別: |
碩士 Master |
系所名稱: |
化學系 Department of Chemistry |
論文出版年: | 2002 |
畢業學年度: | 90 |
語文別: | 中文 |
論文頁數: | 115 |
中文關鍵詞: | 葡萄糖 、碳六十固定化酵素 |
英文關鍵詞: | glucose, Immobilized Fullerence-Enzyme |
論文種類: | 學術論文 |
相關次數: | 點閱:224 下載:1 |
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本研究利用具混價性質之鈷赤血鹽和碳六十固定化酵素致被化學修飾電極來發展葡萄糖感測器,本實驗在0.1MKCl,pH6之磷酸鹽緩衝液中,施加電位0.0V(相對於Ag/AgCl參比電極)偵測葡萄糖,藉由含鈷赤血鹽對H2O2之催化能力,可將偵測電位降低至0.0V,且在還原模式下進行,此電位有效防止血液中電活性物質,如維生素C(Ascorbic acid),尿酸(Uric acid),乙醯基酚(Acetaminophen),半胱銨酸(Cysteine),酪銨酸(Tyrosine),半乳糖(Galactose)之干擾.H2O2氧化還原態的鈷赤血鹽而自身被還原,此還原電流和溶液中的葡萄糖濃度成正比關係.此碳六十固定化酵素感測器對葡萄糖具有不錯之線性範圍(8mM),快速的反應時間(5秒),以及不錯之靈敏度,其偵測極限可達1.6×10-6M,在20次連續操作下相對標準偏差維4.26%,使用100天後,訊號仍維持最初之80%,實驗顯示碳六十固定化酵素確實能增加訊號和延長感測器壽命.本研究也對此葡萄糖感測器探討了溫度,酸鹼度等效應.
A mixed-valence cluster of cobalt(Ⅱ)hexacyanoferrate and fullerene-enzyme based electrochemical glucose sensor was developed.A water insoluble fullerene-glucose oxidase(C60-GOD)was synthesized and applied as an immobilized enzyme on a glassy carbon electrode with cobalt(Ⅱ)hexacyanoferrate for analysis of glucose.The glucose was measured in 0.1MKCl/phosphate buffer solution with a pH of 6.0 When the electrode potential was applied at 0.0mV(v.s Ag/AgCl reference electrode).The glucose sensor exhobited efficient electro-catalytic activity toward the liberated hydrogen peroxide and allowed cathodic detection of the glucose,with no interferences from easily oxidizable constitutes such as uric acid,ascorbic acid,cysteine,tyrosine,acetaminophen and glactose.The H2O2 oxidized the reduced cobalt(Ⅱ)hexacyanoferrate and was reduced itself,and the reduced current was proportional to the concentration of glucose in aqueous solutions.The immobilized C60-GOD based glucose sensor showed a good linear response up to 8mM and a quite short response time of 5 sec.The glucose sensor also exhibited good sensitivity with a detection limit of 1.6×10-6M and a high reproducibility with a relative standard deviation(RSD) of 4.26%.The signal decreased to 80% of the initial value after continuous operations for 100 days.The immobilized Fullerene/glucose oxidase obiously enhanced the response singal and extends the lifetime of the glucose enzyme sensor.Effects of pH and temperature on the responses of the immobilized C60-GOD based glucose sensor were also studied and discussed.
參考資料
1. Guilbault,G.G. and Sadar,M.H.,Accounts of Chemical Research. 1979,12,344.
2.Clark,L.C.;Lyons,C.and Ann,N.Y.,Acad.Sci.1962,102,29.
3.Updike,S.and Hicks,G.,Nature(London).1967,214,986.
4.Olsson,B.;Lundback,H.;Jphansson,G.;Scheller,F.and Nentwig, J.,Anal.Chem.1986,58,1046.
5.Takahiko Kaku,;Hiroko I.Karan and Yoshiyuki Okamoto,Anal.Chem. 1994,66,1231.
6.Cespedes,F.;Martinez-Fabregas,E.and Alegret,S., Anal.Chem.Acta.1993,284,21.
7.Hale,P.D.;Boguslavsky,L.I.;Inagaki,T.;Karan,H.I.;Lee,H.S.;Skotheim,Y.A.and Okamoto,Y.,Anal.Chem.1991,63,677.
8.Sylvia,V.S.; Raymond,J.P.and Alexander,M.Y.,Anal.Chem. 1990,62,1111.
9.Cleland,N.and Sven-OlofEnfors,Anal.Chem.1984,56,1880.
10.Iannlello,R.M.and Yacynych,A.M.,Anal.Chem. 1981,53,2090
11.Iannlello,R.M.;Lindsay,T.J.and Yacynych,A.M.,Anal.Chem. 1982,54,1098.
12.Anthony E,G.C.;Graham,D.;Graeme,D.F.and Allen , H.O.Hill,Anal.Chem.1984,56,667.
13.Gough,D.A.;Lucisano,J.Y.and Tse,P.H.S.,Anal.Chem. 1985,57,2351.
14.Narasimhan,K.and Wingard,L.B.Jr.,Anal.Chem. 1986,58,2984.
15.Wang,J.;Angnes,L.,Anal.Chem.1992,64,456.
16.Wittstock,G.;Schuhmann,W.Formation and Imaging of Microscopic Enzymatically Active Spots on an Alkanethiolate-Covered Gold Electrode by Scanning Electrochemical Microscopy. Anal. Chem.1997, 69, 5059-5066.
17.Bulmus,V.;Ayhan,H.;Piskin,E.Modified PMMA monosize microbeads for glucose oxidase immobilization. The Chemical Engineering Journal. 1997, 65, 71-76.
18.Kang,E.T.;Neoh,K.G.;Huang.,S.W.;Lim,S.L.;Tan,K.L. Surface-Functionalized Polyaniline Films. J. Phys. Chem. B. 1997, 101, 10744-10750.
19.Ukeda,H.;Fujita,Y.;Ohira,M.;Sawamura,M.Immobilized Enzyme-based Microtiter Plate Assay for Glucose in Foods. J. Agric. Food Chem. 1996, 44, 3858-3863.
20.Turmanova,S.;Trifonov,A.;Kalaijiev,O.;Kostov,G. Radiation grafting of acrylic acid onto polytetrafluoroethylene films for glucose oxidase immobilization and its application in membrane biosensor. Journal of membrane Science. 1997, 127, 1-7.
21.Alves Da Silva,M.Helena Gil,J.S.Redinha, Ana M. Oliveira Brett, J. L. Costa Pereira. Immobilization of glucose oxidase on Nylon Membranes and Its Application in a Flow-Through Glucose Reactor. Journal of Polymer Science :Part A: Polymer Chemistry. 1991, 29, 275-279.
22.Audebert,P.;Demaille,C.Electrochemical Probing of the Activity of Glucose Oxidase Embedded Sol-Gel Matrices. Chem. Mater. 1993, 5, 911-913.
23.Künzelmann,U.;Böttcher,H.Biosensor properties of glucose oxidase immobilized within SiO2 gels. Sensors and Actuators B. 1997, 38-39, 222-228.
24.Higuchi,A.;Hara,M.;Yun,K.S.;Tak,T.M.Recognition of Substrates by Membrane Potential of Immobilized Glucose Oxidase Membranes. Journal of Applied Polymer Science. 1994, 51,1735-1739.
25.Godjevargova,Ts.;Dimov,A.;Vassileva,N.Effects of chemical modifications and immobilization of glucose oxidase onto acrylonitrile copolymer membranes on membrance potential and membrance charge density Journal of Membrane Science. 1996 , 116, 273-278.
26.Liu,B.;Hu,R;Deng,J.Fabrication of an Amperometric Biosensor Based on the Immobilization of Glucose oxidase in a modified Molecular Sieve Matrix. Analyst. 1997, 122, 821-826.
27.呂鋒洲; 林仁混. 基礎酵素學. 1991.
28.陳國誠. 微生物酵素工程學. 1989.
29.Chang,H.C.;Chen,P.Y.;Cheng,A.J.Enzyme Immobilization and Polymer Material. Chemistry. 1994, 52, 237-243.
30.陳治誠. 生化感測器技術簡介. 科儀新知. 1993, 15(2), 71-81.
31.Bowers,Larry D. Application of immobilized biocatalysts in chemical analysis. Anal. Chem. 1986, 58(4), 513A, 514A, 516A, 518A, 520A, 522A, 524A, 526A, 528A, 530A.
32.Herdan,J.M.;Balulescu,M.;Cira,O.Enantioselective hydrolysis of racemic esters using pig liver esterase. Journal of Molecular Catalysis A:Chemical. 1996, 107, 409-414.
33.Spagna,G.;Pifferi,P.G.;Gilioli E.Immobilization of a pectinlyase from Aspergillus niger for application in food technology. Enzyme Microb. Technol. 1995, 17, 729-738.
34.Alkorta,I.;Garbisu,C.;Llama,M.J.;Serra,J.L.Industrial applications of pectic enzymes : a review. Process Biochemistry. 1998, 33(1), 21-28.
35.Xu,C.X.;Marzouk,S.A.M.;Cosofret,V.V.;Buck,R.P.;Neuman,M. R.;Sprinkle,R.H.Development of a diamine biosensor. Talanta. 1997, 44, 1625-1632.
36.Mazzei,F.;Botrè,F.;Botrè,C.Acid phosphatase/glucose oxidase-based biosensors for the determination of pesticides. Analytica Chimica Acta. 1996, 336, 67-75.
37.Reiss,M.;Heibges,A.;Metzger,J.;Hartmeier,W. Determination of BOD-values of starch-containing waste water by a BOD-biosensor. Biosensors &Bioelectronics. 1998, 13, 1083-1090.
38.Kroto,H.W.;Hoath,J.R.;Brich,S.C.;Curl,R.F.,Smalley,R.E.,Nature.1985,318,162.
39.Krätschmer,W.;Lamb,L.D.;Fostiropoulos,K.;Huffman,D.R. Solid C60 : a new form of carbon. Nature(London). 1990, 347, 354-358.
40.Hawkins,J.M.;Meyer,A.;Lewis,T.A.;Loren,S.;Hollander,F.J. Crystal Structure of Osmylated C60 : Confirmation of the Soccer Ball Framework. Science. 1991, 252, 312-313.
41.Withers,J.C.;Loutfy,R.O.;Lowe,T.P.Fullerene Commercial Vision. Fullerence Science And Technology. 1997, 5(1), 1-31.
42.Scrivens,W.A.;Bedworth,P.V.;Tour,J.M.Purification of Gram Quantities of C60. A New Inexpensive and Facile Method. J. Am. Chem. Soc. 1992, 114, 7917-7919.
43.Kroto,H.W.;Allaf,A.W.;Balm,S.P.C60: Buckminsterfullerene. Chem. Rev. 1991, 91, 1213-1235.
44. Chen,W.;Xu,Z.Temperature Dependence of C60 Solubility in Different Solvent. Fullerence Science And Technology. 1998, 6(4), 695-705.
45.Haufler,R.E.;Conceicao,J.;Chibante,L.P.F.; Chai,Y.; Byrne, N.E.;Flanagan,S.;Haley,M.M.; O’Brien,S.C.;Pan,C.;Xiao,Z.; Billups,W.E.;Ciufolini,M.A.;Smalley,R.H.Efficient production of C60 (buckminsterfullerene), C60H36, and the solvated buckide ion. J. Phys. Chem. 1990, 94(24), 8634-8636.
46.Taylor,R.;Walton,D.R.M.The chemistry of fullerenes. Nature. 1993, 363, 685-693.
47.Moriguchi,T.;Yano,K.;Hokari,S.; Sonoda,M.Effect of repeated application of C60 combined with UVA radiation onto hairless mouse back skin. Fullerene Sci. Technol. 1999, 7(2), 195-209.
48.Al-Mohamad,A.;Allaf,A.W.,Fullerene-60 thin films for electronic applications. Synthetic Metals. 1999, 104, 39-44.
49.Haddon,R.C.; Hebard,A.F.Conducting Films of C60 and C70 by alkai-metal doping.Nature,1991,350,320-322.
50.Yi-Cong,Wu,;Meng-Shen,Lin,Tamkang University.
51.Lowry,J.P.and O,Neill,R.D.,Anal.Chem.1992,64,453.
52.Vanos, P.J.H.J.; Bult,A.andBennekom, W.P.V., Anal.Chem.Acta. 1995,305,18.
53.Wang,J.and Chen,Q.,Anal.Chem.1994,66,1007.
54.Creutzin, C.; Lippard Ed,S.J.,Progress in Inorganic Chemistry,1983,John Wiley,New York.
55.Keggin J.F.and Miles,F.D.,Nature.1936,137,577.
56.Marcus,R.A. andSutin,N,.Biochimica et Biophysica Acta.1985,811,265.
57.Shriver, D.F.; W.Atleins,P. and Langford,C.H.,Inorganic Chemistry.1994,Oxford University Press,Oxford Melbourne,Tokyo,611.
58.Wyckoff, R.W.G.Crystal Structures.Vol.111,2nd ed, Interscience,New York, N.Y.,1965,382.
59.Ludi,A.;Hans-Ulrich Gudel and Ruegg,M.,Inor.Chem. 1970,9,2224.
60.Buser H.J.; Schwarzenbach,D.; Petter,W.and Ludi,A., Inor.Chem.1977,16,2704.
61.Itaya,K.; Shoji,N.and Uchida,I.,J.Am.Chem.Soc. 1984,106,3423.
62.Feldman,B.J.and Melroy,O.R.,J.Electroanal.Chem. 1987,234,213.
63.Neff,V.D.,J.Electrochem.Soc.1978,125,886.
64.Itaya,K.;Akahoshi,H.and Toshima,S.J.Electrochem.Soc. 1982,129,1498.
65.Pawel,J.K.and Karl,D.J.Electroanal.Chem.1989,274,95.
66.Karyakin,A.A.; Gitelmacher,O.V. and Karyakina,E.E., Anal.Chem.1995,67,2419.
67.Karyakin, A. A.; Karyakina, E. E.; Gorton, L.; Bobrova, O.A.; Lukachova, L.; Gladilim, A. K.; Jevashov, A. V., Anal. Chem. 1996,68(24),4335.
68. Karyakin, A. A.; Karyakina, E. E.; Gorton, L., Anal. Chem. 2000,72(7),1720.
69.Liang,C.;Waldemar,G.,Anal.Chem.2001,73,2862.
70.Ghosh,S.N.,J.Inorg.Nuel.Chem,1974,36,2465.