臺灣博碩士論文加值系統

為了瞭解台灣市售飲料中咖啡因含量是否符合標準，本研究於2012年4月至11月間調查市售包裝及非包裝飲料中咖啡因含量，依據中華民國國家標準CNS-9432號檢驗法進行檢測，並將結果與電化學方法結果進行比對。本次檢測市售包裝飲料檢體共91件(其中咖啡飲料41件、茶類飲料39件和提神飲料11件)及市售非包裝飲料檢體23件(包括咖啡飲料8件及茶類飲料15件)。以高效能液相層析儀(High Performance Liquid Chromatography；HPLC)分析樣品結果顯示包裝咖啡飲料、茶類飲料和提神飲料中咖啡因含量分別介於116～863 ppm、0～264 ppm和0～595 ppm及非包裝咖啡飲料和茶類飲料中咖啡因含量分別介於378~1287 ppm和154~333 ppm。市售包裝咖啡飲料、茶類飲料和提神飲料中咖啡因含量超過標示10 %以上的分別有10件、1件和2件，其咖啡飲料最高超過包裝標示含量的106 %。市售包裝與非包裝的咖啡飲料其咖啡因含量相近，而市售包裝與非包裝的茶類飲料相比，非包裝茶類飲料咖啡因平均的含量比包裝茶類飲料高。此外，本研究電化學方法是利用奈米金修飾網版電極，以循環伏安法(Cyclic voltammetry，CV) 進行上述檢體咖啡因含量之檢測比對，所獲得的咖啡因含量與傳統方法比對結果顯示，其相對百分比達92 %以上，因此此方法可開發作為咖啡因濃度之檢測，未來可改良檢測條件，以提昇方法的靈敏度，進而提高樣品檢測的精準度。

To ensure that caffeine content of packaged and unpackaged beverages comply with their food labeling requirements, this study was to investigate the caffeine content of 91 commercially packaged beverages including 41 coffee drinks, 39 tea drinks, 11 energy drinks, and 23 unpackaged drinks including 8 coffee drinks and 15 tea drinks using the high performance liquid chromatography (HPLC) method in accordance with the Chinese National Standard inspection (CNS)-9432, as compared with the electrochemical method during the period of April to November, 2012. The results showed that the caffeine content of packaged coffee, tea and energy drinks ranged from 116 to 863 ppm, 0 to 264 ppm and 0 to 595 ppm, respectively, and the caffeine content of unpackaged coffee and tea drinks ranged from 360 to 1293 ppm and 154 to 333 ppm, respectively. The caffeine content in 10 packaged coffee drinks, 1 tea drink and 2 energy drinks were found exceed 10 % of their caffeine content labeling in which one coffee drink exceeded 106 % labeled caffeine content. The caffeine contents in commercially packaged coffee drinks were similar to that of unpackaged coffee drinks whereas the caffeine contents in commercially unpackaged tea drinks were more than that of packaged tea drinks. In addition, the electrochemical method based on Cyclic voltammetry with Gold-nanostructured screen-printed electrode was used to detect the caffeine content of those drinks above and compared the accuracy to the HPLC method. The electrochemical method showed above 92 % relative percentage as compared to the HPLC method. The electrochemical detection could be an alternative method for determining the caffeine content. The parameters of the detection conditions may be required to revise in order to improve the accuracy and sensitivity of the method in the further study.

誌謝 I
中文摘要 III
英文摘要 V
目錄 VII
圖目錄 IX
表目錄 X
第一章 緒論 1
1.1 咖啡因對人體的影響 1
1.2 咖啡因標示現況說明 3
1.3 咖啡因的檢測方法 5
1.4 研究動機與目的 7
第二章 利用電化學法測定咖啡因 8
2.1 電化學生物感測器 8
2.2 電化學方法於咖啡因含量測定上的應用 10
2.3 咖啡因在電化學的反應機制 12
2.4 奈米金修飾電極 14
2.5 電化學條件探討 15
第三章 材料與方法 16
3.1 實驗架構 16
3.2 試驗材料 17
3.3 儀器設備與裝置 17
3.4 試藥 19
3.5 各種溶液的配製 20
3.6 HPLC之檢測方法 21
3.6.1 檢液製備 21
3.6.2 標準品檢量線的製備 21
3.6.3 檢測方法與條件 22
3.6.4 鑑別試驗及含量測定 22
3.6.5 回收試驗 22
3.6.6 檢出極限偵測 23
3.7 電化學之檢測方法 23
3.7.1 電化學法之檢液製備 23
3.7.2 標準品檢量線的製備 23
3.7.3 電化學檢測方法及條件 23
3.7.4 奈米金修飾電極之製備 24
3.7.5 檢測系統之條件探討 24
第四章 結果與討論 26
4.1 HPLC檢測分析 26
4.1.1 HPLC檢量線製備與檢出極限 26
4.1.2 HPLC回收率試驗 27
4.1.3 以HPLC檢測各種飲料中咖啡因含量 27
4.2 循環伏安法檢測分析 29
4.2.1 利用不同的電解液於電流訊號之比較 29
4.2.2 測定條件的探討 29
4.2.3 檢量線製作 31
4.2.4 循環伏安法與HPLC檢量線之比較 31
4.2.5 循環伏安法與HPLC法之檢測咖啡因濃度比對 32
第五章 結論 34
第六章 參考文獻 36
6.1英文部分 36
6.2中文部分 39

英文部分
1.Aikens, D.A. Electrochemical methods, fundamentals and applications. Journal of Chemical Education 1983；60：25.
2.Ajaero, C.; Abdelrahim, M.Y.M.; M-Palacios-Santander, J.; Almoraima, Gil, M.L.; Naranjo-Rodriguezc, I.; Cisnerosc, J.L.H.H.; M-Cubillana-Aguilerac, L. Comparative study of the electrocatalytic activity of different types of gold nanoparticles using Sonogel-Carbon material as supporting electrode. Sensors and Actuators B 2012；171-2：1244-56.
3.Akyilmaz, E. and Turemis, M. An inhibition type alkaline phosphatase biosensor for amperometric determination of caffeine. Electrochimica Acta 2010；55：5195-9.
4.Alizadeh, T.; Ganjali, M.R.; Zare, M. and Norouzi, P. Development of a voltammetric sensor based on a molecularly imprinted polymer (MIP) for caffeine measurement. Electrochimica Acta 2010；55：1568-74.
5.Andrews, K.W.; Schweitzer, A.; Zhao, C.; Holden, J.M.; Roseland, J.M.; Brandt, M.; Dwyer, J.T.; Picciano, M.F.; Saldanha, L.G.; Fisher, K. D.; Yetley, E.; Betz, J. M. and Douglass, L. The caffeine contents of dietary supplements commonly purchased in the US: analysis of 53 products with caffeine-containing ingredients. Anal Bioanal Chem 2007；389:231-9.
6.Ashihara, H. and Crozier, A. Caffeine: a well known but little mentioned compound in plant science. TRENDS in Plant Science 2001；6：407-13.
7.Brunetti, B.; Desimoni, E. and Casati, P. Determination of Caffeine at a Nafion-Covered Glassy Carbon Electrode. Electroanalysis 2006；19：285-88.
8.Datta, M. and Landolt, D. Fundamental aspects and applications of electrochemical microfabrication. Electrochimica Acta 2000；45：2535-58.
9.EI-Shahawi, M.S.; Hamza, A.; Bahaffi, S.O.; Al-Sibaai, A.A. and Abduljabbar, T.N. Analysis of some selected catechins and caffeine in green tea by high performance liquid chromatography. Food Chemistry 2012；134：2268-75.
10.Guo, S. and Wang, E. Synthesis and electrochemical applications of gold nanoparticles. Analytica Chimica Acta 2007；598：181-92.
11.Guo, S.; Zhu, Q.; Yang, B.; Wang, J. and Ye, B. Determination of caffeine content in tea based on poly(safranine T) electroactive film modified electrode. Food Chemistry 2011；129：1311-14.
12.Ho, J.A.; Chiu, J.K.; Chiu, J.K.; Hong, J.C.; Lin, C.C.; Hwang, K.C. and Hwu, J.R. Gold-Nanostructured Immunosensor for the Electrochemical Sensing of Biotin Based on Liposomal Competitive Assay. Journal of Nanoscience and Nanotechnology 2009；9：2324-29.
13.Huang, H.; Ran, P. and Liu, Z. Impedance sensing of allergen–antibody interaction on glassy carbon electrode modified by gold electrodeposition. Bioelectronics 2007；70：275-62.
14.Jeevagan, A.J. and John, S.A. Electrochemical determination of caffeine in the presence of paracetamol using a self-assembled monolayer of non-peripheral amine substituted copper(II) phthalocyanine. Electrochimica Acta 2012；77：137-42.
15.Juliano, L.M. and Griffiths, R.R. A critical review of caffeine withdrawal: empirical validation of symptoms and signs, incidence, severity, and associated features. Psychopharmacology 2004；176: 1-29.
16.Kerrigan, S. and Lindsey, T. Fatal caffeine overdose: two case reports. Foren Sci Internat 2005；153：67-9.
17.Liao, W.C. and Ho, J.A. Attomole DNA Electrochemical Sensor for the Detection of Escherichia coli O157. Analytical chemistry 2009；81：2470-6.
18.Martínez-Huitle, C.A.; Fernandes, N.S.; Ferro, S.; Battisti, A.D. and Quiroz, M.A. Fabrication and application of Nafion®-modified boron-doped diamond electrode as sensor for detecting caffeine. Diamond &; Related Materials 2010；19：1188-93.
19.Mersal, G.A.M. Experimental and Computational Studies on the Electrochemical Oxidation of Caffeine at Pseudo Carbon Paste Electrode and Its Voltammetric Determination in Different Real Samples. Food Anal. Methods 2012；5：520-9.
20.Nurminen, M.L.; Niittynen, L.; Korpela, R. and Vapaatalo, H. Coffee, caffeine and blood pressure: a critical review. European Journal of Clinical Nutrition 1999；53：831-9.
21.Wang, M.; Wang, L.; Wang, G.; Ji, X.; Bai, Y.; Li, T.; Gong, S. and Li, J. Application of impedance spectroscopy for monitoring colloid Au-enhanced antibody immobilization and antibody–antigen reactions. Biosensors and Bioelectronics 2004；19：575-82.
22.Weng, X.; Odouli, R. and Li, D.K. Maternal caffeine consumption during pregnancyand the risk of miscarriage: a prospective cohort study. Am J Obstet Gynecol 2008；198：279 e1-8.
23.Santos, W.R.; Santhiago, M.; Yoshida, I.V.P. and Kubota, L.T. Electrochemical sensor based on imprinted sol–gel and nanomaterial for determination of caffeine. Sensors and Actuators B 2012；166-167：739-45.
24.Smith, A. Effects of caffeine on human behavior. Food and Chemical Toxicology 2002；40：1243-55.
25.Sun, J.Y.; Huang, K.J.; Wei, S.Y.; Wu, Z.W. and Ren, F.P. A graphene-based electrochemical sensor for sensitive determination of caffeine. Colloids and Surfaces B: Biointerfaces 2011；84：421-6.
26.Švorc, L.; Tomcˇik, P.; Svitkova, J.; Rievaj, M. and Bustin, D. Voltammetric determination of caffeine in beverage samples on bare boron-doped diamond electrode. Food Chemistry 2012；135：1198-1204.
27.Yamauchi, Y.; Nakamura, A.; Kohno, I.; Kitai, M.; Hatanaka, K. and Tanimoto, T. Simple and Rapid UV Spectrophotometry of Caffeine in Tea Coupled with Sample Pre-treatment Using a Cartridge Column Filled with Polyvinylpolypyrrolidone (PVPP). Chem. Pharm. Bull. 2008；56(2)185-8.

中文部分
1.行政院衛生署。衛署食字第88027006號令發布: 食品添加物使用範圍及用量標準。1988年。行政院衛生署，臺灣。
2.行政院衛生署。行政院消費者保護委員會及行政院衛生署特函：咖啡因含量紅黃綠標示。2006年。行政院衛生署，臺灣。http://www.ntuh.gov.tw/DD/education/Shared%20Documents/%E5%92%96%E5%95%A1%E5%9B%A0.pdf
3.行政院衛生署食品藥物管理局。衛署食字第0960404584號令發布: 飲料類衛生標準。2007年。行政院衛生署食品藥物管理局，臺灣。
4.李蕙芳、鄭邱真和周薰修。市售飲料中咖啡因含量檢測方法之探討與調查.藥物食品檢驗局調查研究年報。1993；11：257-64.
5.吳朗。感測器原理與應用。1987。第24頁。全華出版社。新北市。
6.周淑芬和陳建源。免疫感測器之技術發展及其應用。中國化學學會，臺北。2001；59：263-71.
7.林偉炤、李淵博。分析化學。2009。第371頁。原出版者：W.H. Freeman and Company，發行者：歐亞書局有限公司。新北市。
8.胡啟章。電化學原理與方法。2002。第105頁。五南圖書出版股份有限公司。臺北市。
9.張崇新。具極短脈衝之三維電化學加工之實驗與分析。2009。國立中央大學機械工程學系碩士論文。
10.黃裕成。應用於生物感測器之可攜式多通道恆電位儀設計。2010。國立臺南大學通訊工程研究所碩士論文。
11.經濟部標準檢驗局。中華民國國家標準CNS-9432號：食品中咖啡因含量檢驗法。2009，經濟部標準檢驗局，臺灣。
12.蘇宏基。化學生物感測器。2009。 http://www.chem.ndhu.edu.tw/ezfiles/27/1027/img/828/su.pdf