尿酸（Uric acid，UA）是人體內嘌呤（Purine）物質經肝臟代謝後的最終產物，存在於血液與尿液中，若濃度過高則易引發高尿酸血症（Hyperuricemia）、痛風（Gout）等疾病，因此監測血液或尿液中尿酸的濃度是掌握病況的重要方法。由於分析儀器小巧靈敏，使得利用電化學方法偵測尿酸之研發與日俱增，但在測定血液與尿液中之尿酸時，遭遇到最主要的挑戰是維他命C（Ascorbic acid，AA）的干擾，其主因是維他命C與尿酸之氧化電位太過接近。
本研究將多壁奈米碳管(Multi-Walled Carbon Nanotube，MWCNT)修飾在可拋棄式之網版印刷碳（Screen-Printed Carbon，SPC）電極表面，並經循環伏安法(Cyclic Voltammetry，CV)於0∼2 V間掃瞄作電化學預處理後，使得電極表面氧化生成羧基與羰基等官能基而具有電催化效果，致使原本幾乎重疊的UA和AA氧化波峰訊號變成分離的兩波峰（波峰電位相差約0.34 V）而不會彼此干擾。此外，奈米碳管除了尺寸極小、多孔性可增加反應表面積與靈敏度外，更可提升電子轉移速率而強化其電催化的能力。電化學阻抗光譜法（Electrochemical Impedance Spectroscopy）量測結果亦證實，經過電化學預處理或修飾上MWCNT之電極皆可明顯提昇電子轉移速率（阻抗值皆大幅降低）；且在電極特性探討過程中發現，尿酸在本研究使用之電極上氧化是屬於吸附控制（Adsorption-controlled）。在0.1 M pH 5.5之檸檬酸緩衝溶液中，利用差式脈波伏安法（Differential Pulse Voltammetry）偵測尿酸，測得訊號之再現性良好（不同電極：RSD = 8.3%，n = 5；而同一電極RSD = 3.1%，n = 5）。其校正曲線之線性範圍約在2∼30 μM，線性關係R2 = 0.996，偵測極限為1 μM；此外可能干擾物中Cysteine在高濃度（20 μM以上）會與尿酸競爭吸附，而降低尿酸氧化訊號；另外Dopamine亦會在高濃度時（10 μM以上）產生干擾。以此修飾之SPC電極可測得尿液與血清樣品中的尿酸含量，且所需之樣品量極少（各需25 μL、200 μL）。

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摘 要 ............................................................................................................... I
謝 誌 .............................................................................................................. II
目 錄 ............................................................................................................. III
圖 目 ............................................................................................................ VI
表 目 ............................................................................................................ IX
壹、緒論 ............................................................................................................... 1
一、尿酸及痛風簡介 .................................................................................... 1
二、傳統測定尿酸之方法 ............................................................................ 7
三、結合電化學方法偵測尿酸 .................................................................. 10
四、奈米碳管的特性 .................................................................................. 16
五、差式脈波伏安法 .................................................................................. 19
六、電化學阻抗光譜法 .............................................................................. 21
七、研究目的 .............................................................................................. 23
貳、實驗部份 ..................................................................................................... 25
一、儀器設備 .............................................................................................. 25
二、藥品 ...................................................................................................... 27
三、溶液之製備 .......................................................................................... 32
四、奈米碳管修飾之網版印刷碳電極之製作.......................................... 35
五、實驗過程 .............................................................................................. 40
參、結果與討論 ................................................................................................. 43
一、修飾奈米碳管與電化學預處理的效果 .............................................. 43
1. 未修飾奈米碳管僅作電化學預處理 .............................................. 43
2. 修飾奈米碳管並經過電化學預處理 .............................................. 46
3. 修飾奈米碳管並經過電化學預處理後之SPC 電極特性 ............ 52
二、差式脈波伏安法參數探討 .................................................................. 61
三、除氧效果的探討 .................................................................................. 64
四、利用奈米碳管修飾SPC 電極之最佳化條件探討 ............................ 66
五、溶液pH 值對尿酸與維他命C 偵測訊號之影響 .............................. 70
六、最佳實驗條件 ...................................................................................... 77
七、電極再現性 .......................................................................................... 78
八、線性範圍與偵測極限 .......................................................................... 80
九、可能干擾物的影響 .............................................................................. 83
十、真實樣品分析 ...................................................................................... 86
肆、結論 ............................................................................................................. 92
伍、參考文獻 ..................................................................................................... 93
陸、附錄 ............................................................................................................. 98
一、不同電化學分析法測定尿酸之比較 .................................................. 98
二、不同DPV 參數測定尿酸之比較 ........................................................ 98
三、測定多巴胺的可行性 ........................................................................ 101
柒、簡歷 ........................................................................................................... 103

1. 何敏夫，臨床化學第二版，合記圖書，1999，pp.228-232。
2. 李信興，廖桂聲，中西醫會診：痛風，書泉出版社，2002。
3. 魯焰，痛風四季飲食，開明書店，2003。
4. Skoog, D.A.; Holler, F. J.; Crouch, S. R. Principle of Instrumental Analysis,
6thed.; Thomas Brooks/Cole. 2007, pp.10-11.
5. http://www.chemedu.ch.ntu.edu.tw/lecture/molecular/2.htm （2001）
6. Campbell, N. A.; Mitchell, L. G.; Reece, J. B. 彩色圖說生物學 : 槪念與
關聯性. 合記圖書，2003.
7. Tang, H.; Hu, G. Z.; Jiang, S. X. Selective determination of uric acid in the
presence of ascorbic acid at poly(p-aminobezene sulfonic acid)-modified
glassy carbon electrode. J. Appl. Electrochem., 2009, 39, 2323-2328.
8. Zhang, L.; Zhang, C. H; Lian, J. Y. Electrochemical synthesis of polyaniline
nano-networks on p-aminobenzene sulfonic acid functionalized glassy
carbon electrode Its use for the simultaneous determination of ascorbic acid
and uric acid. Biosens. Bioelectron., 2008, 24, 690-695.
9. Kalimuthu, P.; John, S. A. Simultaneous determination of ascorbic acid,
dopamine, uric acid and xanthine using a nanostructured polymer film
modified electrode. Talanta, 2010, 80, 1686-1691.
10. Kalimuthu, P.; John, S. A. Simultaneous determination of epinephrine, uric
acid and xanthine in the presence of ascorbic acid using an ultrathin polymer
film of 5-amino-1,3,4-thiadiazole-2-thiol modified electrode. Anal. Chim.
Acta, 2009, 647, 97-103.
11. Kuwabata, S.; Nakaminami, T.; Ito, S.; Yoneyama, H. Preparation and
properties of amperometric uric acid sensors. Sens. Actuators B Chem., 1998,
52, 72-77.
12. Wang, X. Y.; Yin, F.; Tu, Y. F. A uric acid biosensor based on
Langmuir-Blodgett film as an enzyme-immobilizing matrix. Anal Lett., 2010, 43, 1507-1515.
13. 趙辰濤，電流式尿酸生物感測器之製備及測試，碩士論文，國立台灣科
技大學化學工程系，2009。
14. Cete, S.; Yasar, A.; Arslan, F. An Amperometric Biosensor for Uric Acid
Determination Prepared from Uricase Immobilized in Polypyrrole Film. Artif.
Cells Blood Substit. Immobil. Biotechnol., 2006, (34), 367-380.
15. Kissinger, P. T.; Heineman, W. R. Laboratory Techniques in
Electroanalytical Chemistry, 2nded.; Marcel Dekker,Inc, 1996.
16. Zen, J. M.; Hsu, C. T. A selective voltammetric method for uric acid
detection at Nafion®-coated carbon paste electrodes. Talanta, 1998, 46,
1363-1369.
17. Shahrokhian, S.; Ghalkhani, M. Simultaneous voltammetric detection of
ascorbic acid and uric acid at a carbon-paste modified electrode
incorporating thionine–nafion ion-pair as an electron mediator. Electrochim.
Acta, 2006, 51, 2599-2606.
18. Noroozifar, M.; Motlagh, M. K.; Taheri, A. Preparation of silver
hexacyanoferrate nanoparticles and its applictation for the simultaneous
determination of ascorbic aicd, dopamine, and uric acid. Talanta, 2010, 80,
1657-1664.
19. Tehrani, R. M. A.; Ghani, S. A. Volatammetric analysis of uric acid by
zinc-nickel nanoalloy coated composite graphite. Sens. Actuators B Chem.,
2010, 145, 20-24.
20. Huang, X.; Li, Y.; Wang, P.; Wang, L. Sensitive Determination of
Dopamine and Uric Acid by the Use of a Glassy Carbon Electrode Modified
with Poly(3-methylthiophene)/Gold Nanoparticle Composites. Anal. Sci.,
2008, 24, 1563-1568.
21. Zare, H. R.; Nasirizadeh, N. Simultaneous determination of ascorbic acid,
adrenaline and uric acid at a hematoxylin multi-wall carbon nanotube modified glassy carbon electrode. Sens. Actuators B Chem., 2010, 143,
666-672.
22. Zhang, Y.; Pan, Y.; Su, S.; Zhang, L.; Li, M.S. A novel functionalized
single-wall carbon nanotube modified electrode and its application in
detection of dopamine and uric acid in the presence of high concentration of
ascorbic acid. Electroanalysis, 2007, 19, 1695-1701.
23. Manjunatha, R.; Suresh, G. S.; Melo, J. S.; D’Souza, S. F.; Venkatesha, T. V.
Simultaneous determination of ascorbic acid , dopamine and uric acid using
polystyrene sulfonate wrapped multiwalled carbon nanotubes bound to
graphite through layer-by-layer technique. Sens. Actuators B Chem., 2010,
145, 643-650.
24. Ardakani, M. M.; Beitollahi, H.; Ganjipour, B.; Naeimi, H.; Nejati, M.
Electrochemical and catalytic investigations of dopamine and uric acid by
mofdified carbon nanotube paste electrode. Bioelectrochemistry, 2009, 75,
1-8.
25. Liu, A.; Honma, I.; Zhou, H. Simultaneous voltammetric detection of
dopamine and uric acid at their physiological level in the presence of
ascorbic acid using poly(acrylic acid)-multiwalled carbon-nanotube
composite-covered glassy-carbon electrode. Biosens. Bioelectron., 2007, 23,
74-80.
26. Zhang, Y.; Pan, Y.; Su, S.; Zhang, L.; Li, S.; Shao, M. A Novel
Functionalized Single-Wall Carbon Nanotube Modified Electrode and Its
Application in Determination of Dopamine and Uric Acid in the Presence of
High Concentrations of Ascorbic Acid. Electroanalysis, 2007, 19,
1695-1701.
27. http://www.nsc.gov.tw/scicircus/public/Attachment/731912574671.pdf
28. Zhou, O. F., R. M.; Murphy, D. W.; Chen, C. H.; Haddon, R. C.; Ramirez, A.
P.; Glarum, S. H Defects in carbon nanostructures. Science, 1994, 163, 1744-1747.
29. Amelinckx, S. B., D; Zhang, X. B.; Tendeloo, V.; Landyut, J. V. A Structure
model and Growth Mechanism for Multishell Carbon Nanotubes. Science,
1995, 267, 1334-1338.
30. Ebbesen, T. W. L., H. J.; Hiura, H. J.; Bennett, W.; Ghaemi, H. F.; Thio, T.
Electrical conductivity of individual carbon nanotubes. Nature, 1996, 382,
54-56.
31. Dai, H. J. W., E. W.; Lieber, C. M. Probing electrical transport in
nanomaterials: Conductivity of individual carbon nanotubes Science, 1996,
272, 523-526.
32. 成會明，奈米碳管，五南出版社，2004。
33. Saito, R. Physical properties of carbon nanotubes; Imeprial college press,
1998.
34. Unwin, P. R. Instrumentation and electroanalytical chemistry；Weinheim :
Wiley-VCH, 2003.
35. Park, S.M.; Yoo, J.S. Peer Reviewed: Electrochemical Impedance
Spectroscopy for Better Electrochemical Measurements. Anal. Chem., 2003,
75, 455A-461A.
36. Thiagarajan, S.; Tsai, T. H.; Chen, S. M. Easy modification of glassy carbon
electrode for simultaneous determination of ascorbic acid, dopamine and uric
acid. Biosens. Bioelectron., 2009, 24, 2712-2715.
37. Chang, C. W.; Tseng, W. L. Gold Nanoparticle Extraction Followed by
Capillary Electrophoresis to Determine the Total, Free, and Protein-Bound
Aminothiols in Plasma. Anal. Chem., 2010, 82, 2696-2702.
38. 鍾協訓，曾志明，網版印刷電極再分析化學上的製作與應用，科儀新知，
1998，第119期，72-82。
39. Rezaei, B.; Zare, S. Z. M. Modified glassy carbon electrode with multiwall
carbon nanotubes as a voltammetric sensor for determination of noscapine in biological and pharmaceutical samples. Sens. Actuators B Chem., 2008, 134,
292-299.
40. Yano, J.; Hirayama, H.; Harima, Y.; Kitanib, A. Electrochemical and
UV-Visible Spectroscopic Study on Direct Oxidation of Ascorbic Acid on
Polyaniline for Fuel Cells. J. Electrochem. Soc., 2010, 157, 506-511.
41. Alwarappan, S.; Liu, G.; Li, C.-Z. Simultaneous detection of dopamine,
ascorbic acid, and uric acid at electrochemically pretreated carbon nanotube
biosensors. Nanomedicine., 2010, 6, 52-57.
42. Ensafi, A. A.; Taei, M.; Khayamian, T. Differential Pulse Voltammetric
Method for Simultaneous Determination of Ascorbic Acid, Dopamine and
Uric Acid Using Poly(3-(5-Chloro-2-Hydroxyphenylazo)-4,5-
Dihydroxynaphthalene-2,7-Disulfonic Acid) Film Modified Glassy Carbon
Electrode. J. Electroanal. Chem., 2009, 633, 212-220.
43. Thiagarajan, S.; Chen, S. M. Preparation and characterization of PtAu hybrid
film modified electrodes and their use in simultaneous determination of
dopamine, ascorbic acid and uric acid. Talanta, 2007, 74, 212-222.
44. Liu, Y.; Huang, J.; Hou, H.; You, T. Simultaneous determination of
dopamine, ascorbic acid and uric acid with electrospun carbon nanofibers
modified electrode. Electrochem. Commun, 2008, 10, 1431-1434.
45. Zheng, D.; Ye, J.; Zhou, L.; Yu, Y. Z. C. Simultaneous determination of
dopamine, ascorbic acid and uric acid on ordered mesoporous carbon/Nafion
composite film. J. Electroanal. Chem., 2009, 625, 82-87.
46. Li, Y. X.; Lin, X. Q. Simultaneous electroanalysis of dopamine, ascorbic
acid and uric acid by poly (vinyl alcohol) covalently modified glassy carbon
electrode. Sens. Actuators B Chem., 2006, 115, 134-139.