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研究生:林崇文
論文名稱:檢測氨氣之生物感測技術開發
論文名稱(外文):Development of Biosensor in Detecting Ammonia
指導教授:鄭武順鄭武順引用關係呂幸江
學位類別:碩士
校院名稱:長庚大學
系所名稱:化學工程研究所
學門:工程學門
學類:化學工程學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:65
中文關鍵詞:石英壓電晶體生物感測器酵素專一性架橋連接法氨基戊二酸脫氫酵素濕度恆溫吸附曲線
外文關鍵詞:piezoelectric quartz crystalbiosensorenzymespecificitycrosslinking reagentL-Glutamic Dehydrogenasehumiditylangmuir isotherm
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利用石英壓電晶體與特定生物活性物質組成靈敏度極高的生物感測器,本實驗用的生物活性物質為酵素。利用酵素的專一性,以架橋連接法將酵素固定於晶片上,吸附氣相中特定氣體。將對氨氣具有專一性的氨基戊二酸脫氫酵素(GLDH)固定於晶片上。由於用架橋連接法固定酵素,石英晶片上除了酵素外還有PEI、Glutaraldehyde、Glycine、NADH。必須對其所產生的影響作探討,根據實驗找出固定GLDH酵素較佳條件。固定Glycine目的是要將未接酵素的Glutaraldehyde覆蓋,減少干擾。但根據實驗結果可看出,固定Glycine會造成酵素活性降低。將含有NADH與未含NADH之酵素固定於晶片上,加NADH並未增加酵素吸附能力,反而增強對氨的鍵結,造成不易脫附。PEI對氨氣無吸附現象,但Glutaraldehyde對氨有少許吸附。酵素是親水性,故對氣相中水氣吸附能力比對氨氣還大,使用canister裝氣體,控制濕度,因為canister密閉性比集氣袋佳。在相對濕度70下對氨氣吸附量最大。在最佳吸附的濕度情況下,探討其專一性,觀察對甲胺、乙胺、CO2、丙酮、乙醇、異戊二烯吸附情況。GLDH酵素對甲胺、乙胺有吸附,但對氨吸附量比對甲胺、乙胺多,對甲胺、乙胺吸附後較不易脫附。GLDH酵素對乙醇也有吸附,但吸附量比氨少。觀察吸附3分鐘內吸附曲線之斜率與氣相中氨氣濃度之關係。求出吸附速率 R=aCb(a;0.0027,b:0.78)。根據不同溫度下,吸附平衡時的頻率下降量求出恆溫吸附曲線。
Piezoelectric quartz crystal and biologically active enzyme were used in biosensor of high sensitivity.Utilizing enzyme’s specificity,it can adsorb specific gas.To immobilize the specific L-Glutamic Dehydrogenase of ammonia to quartz crystal,crosslinking reagents and others were used such as PEI、glutaraldehyde、glycine、NADH on the quartz crystal besides enzyme.The purpose of immobilizing glycine is that we want to cover exposed glutaraldehyde with glycine to reduce the interfering factor.However the addition of glycine reduced the activity of enzyme.We also immobilzed enzyme on the quartz crystal that contained NADH.NADH can’t increase enzyme’s adsorptive capacity but will increase the bonding of ammonia and make it difficult to desorb.PEI did not adsorb ammonia,but glutaraldehyde showed some adsorption of ammonia. Enzyme is hydrophilic.Its adsorption ability of moisture is more than ammonia.Because canister’s airtight function is better than sample bag, we use canister to fill gas and control humidity.Enzyme showed the largest quantity of adsorption of ammonia at RH=70.Under the best humid condition of adsorption,the specificity and enzyme’s adsorptive conditions for methylamine、ethylamine、CO2、acetone、ethanol、isoprene were studied.GLDH can adsob methylamine and ethylamine,but the quantity of adsorption of ammonia is more than methylamine and ethylamine. It’s not easy to desorb adsorbed methylamine and ethylamine.GLDH can also adsob a small amount of ethanol.The relation between the slope of adsorptive curve and ammonia’s concentration in gas phase within three minutes were studied. The following adsorption rate was obtainedR=aCb(a:0.0027,b:0.78).The quantity of adsorption for ammonia at different temperature was used to calculate the Langmuir adsorption isotherm.
第一章 緒 論
1.1 研究背景及目的…………………………………………… 1
第二章 文獻回顧………………………………………………………3
2-1呼氣檢測……………………………………………………………3
2-2 嗅覺作用機制……………………………………………………...10
2-3生物感測器…………………………………………………………14
第三章 實驗藥品設備及方法…………………………………………28
3.1 實驗藥品…………………………………………………………...28
3.2.實驗氣體……………………………………………………………28
3.3 實驗儀器…………………………………………………………...29
3.4.實驗裝置……………………………………………………………30
第四章 實驗方法………………………………………………………31
4.1 晶片上酵素之固定………………………………………………...33
4.1.1 胺基乙酸(Glycine)對酵素活性的影響………………………...35
4.1.2 PEI、Glutaraldehyde、GLDH酵素對氨氣吸附比較……………35
4.1.3 NADH影響……………………………………………………….37
4.1.4 比較物理吸附與化學法固定酵素……………………...……….37
4.1.5 酵素固定量與氨氣吸附量比較…………………………………38
4.2 GLDH酵素氣相吸附……………………………………………….39
4.2.1 濕度對GLDH酵素吸附能力之影響……………………………39
4.2.2 酵素專一性………………………………………………………39
第五章結果與討論……………………………………………………..40
5.1 酵素之固定………………………………………………………...40
5.1.1 胺基乙酸(Glycine)對酵素活性的影響………………………...40
5.1.2 PEI、Glutaraldehyde 影響………………………………………..40
5.1.3 NADH影響……………………………………………………….43
5.1.4 物理吸附與化學法固定酵素……………………………………43
5.1.5 酵素固定量與氨氣吸附量比較…………………………………46
5.2. GLDH酵素氣相吸附………………………………………………48
5.2.1 濕度對GLDH酵素吸附能力之影響…………………………….48
5.3 酵素吸收速率……………………………………………………...52
5.4 Langmuir isotherm………………………………………………….54
第六章 結論……………………………………………………………55
第七章 參考文獻………………………………………………………57
1. Krotoszynski, B. K., Garbriel, G., O''Neill, H. J., and Claudio, M. P. A., Charactrization of human expired air : A promising investigative and diagnostic technique. J. Chromatogr. Sci, 15, 239-244 (1977).
2. Krotoszynski,.B. K., Bruneau, G. M., and O''Neill, H. J., Measurement of chemical inhalation exposure in an urban population in the presence of endogenous effluents. J. Anal. Toxicol . 3, 225-234(1979).
3. Barkley, J., Bunch , J., and Bursey ,J.T., Gas chromatography -mass spectrometry-computer analysis of volatile halogenated hydrocarbons in man and his environmental-a multimedia environmental study . Biomed . Mass Sectrom.7, 139-147(1980).
4. Conkle, J. P., Camp, B. J., and Welch, B. E., Trace composition of human respiratory breath. Arch. Environ. Health. 30, 290-295 (1975).
5. Manolis, A. (1983) The diagnostic potential of breath analysis. Clin.Chem. 29:5-15.
6. Phillips, M. (1992) Breath tests in medicine. Sci. Am. 266:74-79.
7. Ping, W., Yi, T., Haibao, X. and Farong, S. (1997) A novel method for diabetes diagnosis based on electronic nose. Biosens. Bioelec. 12:1031-1036.
8. Ryabtsev, S. V., Shaposhnick, A. V., Lukin, A. N. and Domashevskaya, E.P. (1999) Application of semiconductor gas sensors for medical diagnostics. Sens. Actuators (B) 59:26-29.
9. Wallace, L., Buckley, T., Pellizzari, E. and Gordon, S. (1996) Breath measurements as volatile organic compound biomarkers . Environmental Health Perspectives. 104 (Supplement 5):861-869.
10. Phillips, M. (1997) Method for the collection and assay of volatile organic compounds in breath. Anal. Biochem. 247:272-278.
11. Pleil, J. D. and Lindstrom, A.B. (1997) Exhaled human breath measurement method for assessing exposure to lalogenated volatile organic compounds. Clin. Chem. 43:723-730.
12. Groves, W. A., Zellers, E. T. and Frye, G. C. (1998) Analyzing organic vapors in exhaled breath using a surface acoustic wave sensor array with preconcentration: Selection and characterization of the preconcentrator adsorbent. Anal. Chim. Acta. 371:131-143.
13. Parker, K. M. and Green, J.L. (1990) Delayed ethanol analysis of breath specimens: long-term field experience with commercial silica gel tubes and breathalyzer collection. J. Forensic Sci. 35:1353-1359.
14. Michael Phillips and Joel Greenberg,Ion-Trap Detection of Volatile Organic Compounds in Alverlar Breath,Clinical Chemistry, Vol.38, No.160-65 (1992).
15. 黃崇民(1996)認識傳統醫學的真貌。台灣書店印行。台灣。
16. Rating, D. and Langhans, C.D. (1997) Breath tests: concepts, applications and limitations. Eur. J. Pediatr. 156[Suppl 1]:S18-S23.
17. Bell, G.D. (1998) Clinical practice - breath tests. British Medical Bulletin 54:187-193.
18. Smith, D. and Spanel, P. (1996) The novel selected-ion flow tube approach to trace gas analysis of air and breath. Rapid Commun. In Mass Spectro. 10:1183-1198.
19. Spanel, P. and Smith, D. (1996) Review-Selected ion flow tube: a technique for quantitative trace gas analysis of air and breath. Med. & Biol. Eng. & Comput. 34:409-419.
20. Smith, M. (1982) The use of smell in differential diagnosis. Lancet, Dec.25:1452.
21. Cheng, W. H. and Lee, W. J. (1999) Review articles: Technology development in breath microanalysis for clinical diagnosis. J. Lab.Clin. Med. 133:218-228.
22. Ebeler, S.E., Clifford, A.J. and Shibamoto, T. (1997) Quantitative analysis by gas chromatography of volatile carbonyl compounds in expired air from mice and human. J. of chroma. B. 702:211-215.
23. Davies, S., Spanel, P. and Smith, D. (1997) Quantitative analysis of ammonia on the breath of patients in end-stage renal failure. Kidney International. 52:223-228.
24. Amoore,J.E.(1963)Stereochemical theory of olfaction, Nature,198:271-272.
25. Davis,J.T.(1965)A theory of the quality of odours.J.Theor.Biol.,8:1-7.
26. Lancet,D.(1986)Vertebrate olfactory reception,Annu.Rev. Neurosci., 9:329-355.
27. Heinz,B.(1997)Sense of smell:Signal recognition and transduction in olfactory receptor neurons,Chapter 22,CRC Press,INC.,pp521-592.
28. Byfield,M.P.,and Abuknesha,R.A..Biochemical aspects of biosensors.Biosen.Bioelec.9,373-400(1994).
29. Shthi,R.S.,Transducer aspects biosensors. Biosensors & Bioelectronics ,.9,243-264( 1994 )
30. Scheller, F.,.Research and development of biosensors. Analyst. 114:653-662(1989).
31. Waston,L.D.,P.Maynard,D.C.Cullen,R.S.Sethi J.Brettle and C.R.Lower,”A microelectronic conductimetric biosensor.” Biosensor,3,101-115 (1987).
32. Koryta,JIRI,’Theory and application of ion selective electrodes.” Anal. Chem., 159,1-46(1984 ).
33. Turner,A.P.F.,I.Karube and G.S.Wilson, ”Biosensor Fund- amentals and Application.” Oxford University Press.(1987)
34. 陳美琴,”PZ 生物感測器在HIV-1抗體檢測之應用”;,台灣大學農業化學研究所碩士論文。
35. 吳宗正,”壓電晶體生物感測器之研究與其應用”.台灣大學農業化學研究所碩士論文。
36. 呂信賢,〝生物電子鼻對臭異味分子的偵測及應用〞, 國立東華大學生物技術所,碩士論文(2000)。
37. Sauerbray,G.Z.,Use of quartz crystal vibrator for weighting thin flim on a microbalance.Z.Phys.155,206-210 ( 1959 ).
38. Kurosawa,S.,Oscillating frequency of piezoelectric quartz crystal in solutions. Anal .Chim.Acta 230,41-49(1990).
39. Kanazawa,K., and Gordon,J.G.,The oscillation frequency of aquartz resonator in contact with a liquid. Anal. Chim. Acta 175,99-105 ( 1985 ).
40. Bao. L.,,A rapid method for determination of Proteus Vulgaris with a piezoelectric quartz crystal sensor coated with a thin liquid film.Biosen.Bioelec.11 ,1193-1198 ( 1996 ).
41. Lai,C.S.I.,Moody,G.J., and Thomas, J.D.R., Piezoelectric quartz crystal detection of ammonia using pyridoxine hydrochloride supported on a polyethoxylate mateix . Analyst 111,511-515 (1986 ).
42. Hlavay, J., and Guilbault, G. G., Detection of ammonia in ambient air with coated piezoelectric crystal detector. Anal. Chem. 50,1044-1046 ( 1978).
43. Bell,G.,Halling,P.J., Moore, B. D., Partridge, J.,and Rees, D. G., Biocatalyst behaviour in low-water systems.Tibtech.13, 468-473 ( 1995 ) .
44. Robert, H., Lamare, S., Parvaresh, F., and Legoy, M. D., The role of water in gaseous biocatalysis. Biocatalysis in Non-Convent. Media 85-92 ( 1992 ).
45. Yang, F., and Russell, A. J., The role of hydration in enzyme activity and stability : 2.Alcohol dehydrogenase activity and stability In a continuous gas phase reactor. Biotech. Bioeng.49,700-716 (1996).
46. 高志達,〝固定化丙三醇脫氫酵素吸附微量一,二-丙二醇之研究〞, 國立台灣科技大學化工所,碩士論文(1998)。
47. 林于生,應用於腎臟病變呼氣檢測模組晶片之研究,國立東華大學生物技術研究所碩士論文。
48. M..Aizawa ,T. Seyama , K.Fueki,J.Shiokawa and S. Suzuki. 1983. Proceedings of the International Meeting of Chemical Sensors , Fukuoka, Elsevier, Amsterdam. pp. 683.
49. V.Glaser and K. G. Krul. 1994. Biosensors: Analytical devices for biomedical application. Specrum. Decision Resources, Inc. p 57-1~57-11.
50. Krotoszynski,.B. K., Bruneau, G. M., and O''Neill, H. J., Measurement of chemical inhalation exposure in an urban population in the presence of endogenous effluents. J. Anal. Toxicol . 3, 225-234(1979).
51. 鄭宗記,”石英晶體微天平研製與其在生化感測功能應用上之開發”.成功大學醫學工程研究所碩士論文。
52. Bodian,D.(1967)Neurons,circuits,and neuroglia.In the Neuroscience:A study program(Quarton,G.C.,Melnechuk,T. and Schmitt,F.O.ets.),New York:Rockefeller,pp.6-24.
53. Schild,D.(1988)Principles of ordor coding and a neural network for odor discrimination,Biophys.J.,54:1001-1011.
54. Philips,M.and Greenberg,J.,Ion-trap detection of volatile organic compounds in alveolar breath.Clin.Chem.38,60-65 (1992).
55. 林俊成,”氣相層析質譜儀與生物感測器在消化性潰瘍呼氣撿測技術之開發”.長庚大學化學工程研究所碩士論文。(1998)
56. 53.張怡南〝生物感測器(Biosensor )〞生物技術的發展與應用, p303-320(1997 ).
57. Eggins, B. (1996) Biosensors: an introduction. Wiley Press, New York.Chapter 3: Immobilisation of biological component. pp. 31-50.
58. Guilbault, G. G. and Jordan, J. M. (1988) Analytical uses of piezoelectric crystals. CRC Rev. 19:1-28.
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