臺灣博碩士論文加值系統

據研究指出，魚肉中的汞大部分都是以甲基汞的形態存在，
而甲基汞為所有汞化合物中毒性最為劇烈著，且經血液循環至腦部，
進而造成腦神經的嚴重傷害，並具有生物累積性，一旦長期食用受到汞污染的魚肉，則將對人體造成莫大的傷害；所以汞及其化合物在環境上所造成的污染實不可輕忽，因此本研究的目的是建立甲基汞的快速分析方法。
傳統偵測魚肉中甲基汞的方法一般源自於Westoo之為氣相層析法─填充管柱配合電子捕捉偵測器法(GC-ECD)或氣相層析質譜偵測器法(GC-MS)，而萃取魚肉中甲基汞的前處理方法，乃利用甲苯萃取並經離心後注入氣相層析儀分析，其過程不僅耗時、且耗費大量溶劑。本研究用微波萃取裝置(MES-1000)來萃取魚肉中甲基汞，經田口式直交表法L9(34)找尋一個最佳微波萃取魚肉中甲基汞的條件，並以氣相層析法─毛細管柱配合電子捕捉偵測器法分析(CGC-ECD)，研究發現在萃取劑甲苯15mL、溫度120℃、微波功率100％、所需時間5分鐘、萃取3克魚肉的條件下，回收率可達90％以上，且經ANOVA多變數分析的結果顯示發現此4因子對於魚肉中甲基汞的微波萃取皆有相當的影響力，儀器偵測極限為0.01pg/L，本研究所測得之真實樣品鱸魚、鮪魚、旗魚、鯊魚的濃度分別為0.31, 0.68, 0.75, 0.40g/g，魚肉參考真實樣品DORM-2的準確度分析回收率則可達98%以上。此法不僅快速、操作簡單、溶劑耗量小以外、回收率亦相當的高，再加上操作方便，價格不昂貴的氣相層析儀，將使魚肉中甲基汞的偵測達到更快速、更便利的目的。

It’s well known that mercury enters the environment through a number of natural and industrial processes and is found in several chemical forms. Because many microorganisms exhibit the ability to methylate the inorganic mercury. The Large amount of methylmercury, the most toxic species among them, has been found in fish.
A simple and rapid method is described for the extraction and determination of methylmercury in fish. The procedure was mainly based on the quantitative microwave-assisted extraction of methylmercury from fish sample using toluene as extract solvent.
Taguchi's orthogonal array design (L9) have been used to optimize the microwave extraction process. Factors such as temperature, microwave power, extraction time and solvent volume were considered. The results suggest that all four factors are significantly. The optimize microwave process was 120˚C, 15 mL Toluene, 5 minutes extraction time, and 100%(947W) microwave power.
The analysis of extracts has been carried out by capillary gas chromatography with electronic capture detector. The detection limit of the procedure was 10 pg/μL. And the methylmercury in marine fish such as tuna, swordfish, shark were found 0.68, 0.75, 0.40 μg/g respectively and the river fish such as Tilapia wasn’t found any methylmercury. Validation was carried out by certified reference material(DORM-2) of National Research Concil in Canada, and the recovery was more than 98%.

第一章　緒論
1-1 前言...............................................1
1-2 研究緣起...........................................1
1-3 研究目的...........................................2

第二章　文獻回顧
2-1 汞簡介.............................................4
2-2 汞的物理化學特性...................................4
2-3 環境中汞之分布.....................................7
2-3-1 地表含汞量
2-3-2 水域汞含量
2-3-3 底泥汞含量
2-3-4 農作物中汞含量
2-3-5 陸地動物體中汞含量
2-3-6 魚肉中汞含量
2-4 汞物種之毒性.......................................10
2-5 目前含汞化合物在台灣的用途及污染源.................12
2-6 汞物種間之轉換與傳播...............................13
2-7 甲基化動力學.......................................16
2-8 汞與甲基汞萃取、分析之方法.........................18
2-9 微波萃取簡介.......................................20
2-10 微波原理.....................................................21
2-11 電子捕捉偵測器.....................................................23

第三章　實驗材料與方法
3-1 實驗藥品與試劑.....................................................26
3-1-1氯化甲基汞標準品.....................................................26
3-1-2內標準品，氯化乙基汞.....................................................26
3-1-3氯化苯基汞.....................................................26
3-1-4魚類驗證參考物質..................................26
3-1-5配製氯化甲基汞標準品使用之溶劑與清洗器皿之有機溶劑
........................................26
3-1-6淨化及其他物質....................................26
3-2 實驗設備與條件.....................................................27
3-2-1　氣相層析儀器及分析操作條件......................27
3-2-2　氣體............................................28
3-2-3　氣體純化裝置....................................28
3-2-4　層析管柱........................................28
3-2-5　資料處理系統....................................29
3-2-6　微波萃取裝置....................................29
3-3 實驗方法...........................................37
3-3-1　容器之清洗......................................37
3-3-2　魚肉前處理與儲存................................37
3-3-3　添加試驗........................................37
3-3-4　管柱調理步驟及傳統魚肉中萃取甲基汞之流程
......................................37
3-3-5　微波萃取－微波功率測定步驟......................38
3-3-6　微波萃取－微波萃取魚肉中甲基汞流程.......................39
3-3-7　萃取溶劑探討....................................40
3-3-8　甲基汞萃取液之管柱乾燥程序......................40
3-3-9　魚肉均質化樣品製作流程圖........................40

第四章　結果與討論
4-1 氯化甲基汞的鑑定與停留時間的建立...................46
4-1-1　停滯時間(Retention time)........................46
4-1-2　檢量線..........................................46
4-2 微波功率測試結果...................................49
4-3 管柱靜相厚度的影響.................................50
4-4 萃取溶劑的探討.....................................52
4-5 傳統離心法萃取魚肉鐘甲基汞結果.....................52
4-6 最佳微波萃取條件─第一部份.........................58
4-7 最佳微波萃取條件─第二部份.........................62
4-8 魚肉中甲基汞均質化驗證.............................67
4-9 儀器偵測極限.......................................69
4-10 方法偵測極限......................................71
4-11 準確度............................................71
4-12 精密度............................................72
4-12-1 參考物質的精密度驗證............................72
4-12-2 微波萃取之精密度................................72
4-13 微波萃取魚肉中甲基汞之真實樣品分析................73
4-14 空白實驗..........................................80
4-14-1 溶劑空白實驗....................................80
4-14-2 無水硫酸鈉空白實驗..............................80
4-14-3 方法空白試驗....................................80
第五章　結論與建議.....................................84
第六章　參考文獻.......................................86

王文亮，「魚介類含汞問題與合理衛生標準之探討」，漁業推廣，第70期，第11∼16頁(1992)

Lindlberg, S. E." Introduction:The fourth international conference on mercury as a global pollutant?,Atmospheric Environment ,Vol:32 , pp.807∼808 (1998).

潘子明、吳順振，「魚貝類中甲基汞含量檢測方法之研究」，中國環境工程學刊，第三卷、第二期(1993)

阮國棟，「汞之污染特性及處理技術」，工業污染防治，第15期，第161∼183頁(1985)

「魚蝦類衛生標準」，衛署藥字第103816號公告，衛署食字第8143635號公告修正，81年8月26日

「放流水標準」，環署水字第七八八０四號令修正發布，86年12月24日

王仁澤，「食品衛生與安全學」，供學出版社，第681∼721頁，台南(1991)

高思懷，「汞在垃圾掩埋廠及環境中污染之研究」，台灣大學土木工程學研究所碩士論文(1986)

Ali, I. B.; Joiris, C. R.; Holsbeek, L.涆otal and Organic Mercury in the Starfish Ctenodiscus Crispatus and the Polychaete Maldanes Sarsi From the Barents Sea?The Science of the Total Environment , pp.189∼194 (1997).
Holsbeek, L.; Das, H. K.; Joiris, C. R."Mercury speciation and accumulation in Bangladesh freshwater and anadromous fish? The science of the total environment ,Vol:198, pp.201∼210 (1997).

Wagemann, R.; Trebacz, E.; Boila, G.; Lockhart, W.L."Methylmercury and total mercury in tissue of arctic marine mammals?,The Science of the Total Environment, Vol:218, pp.19∼31 (1998).

Dickman, M.D ; Leung K.M.C."Mercury and organochlorine exposure from fish consumption in Hong Kong", Chemosphere, Vol:37, pp.991∼1015 (1998).

張木彬、吳秀東，「汞在環境中的傳播與宿命」， 國立中央大學環境工程學刊，第四期(1997).

Tao, G. ; Willie, S.C ; Sturgeon R.E " Detremination of total mercury in biological tissues by flow injection cold vapor generation atomic absorption spectrometry following tetramethylammonium hydroxide digestion.?,Analyst , Vol:123, pp.1215∼1218(1998).

葉錫溶等“以中子活化分析法定量魚肉中汞含量“Chemistry(The Chinese Chem. Soc.,Taiwan, China)No.2, pp.33~37(1973)

Yoshinaga, J.; Morita, M."Determination of mercuryin biological and Environmental samples by inductively coupled plasma mass spectrometry with the isotope dilution technique" ,Journal of Analytical Atomic Spectrometry, Vol:12, pp.417∼420(1997).

Hight, S.C.; Capar S.G."Electron Capture Gas-liquid chromatographic determination of methyl mercury in fish and shellfish: Collaborative study" ,J. Assoc. Off. Anal. Chem., Vol:66 , pp.1121∼1128(1983)

Hight, S.C.; Corcoran M.T."Rapid determination of methyl mercury in fish and shellfish: Method development.?,J. Assoc. Off. Anal. Chem., Vol:70 , pp.24∼30(1987)

Moens, L.; Smaele, T.D.; Dams, R." Sensitive, Simultaneous Determination of Organomercury, -lead, and -tin Compounds with Headspace Solid Phase Microextraction Capillary Gas Chromatography Combined with Inductively Coupled Plasma Mass Spectrometry?,Anal. Chem., Vol:68, pp.3859~3866(1997)

Pereiro, I. R.; Wasik, A.; Lobiski.Determination of methylmercury species in fish reference materials by isothermal multicapillary gas chromatography with atomic emission detection after microwave-assisted solubilization and solvent extraction粑ournal of Analytycal Atomic Spectrometry, Vol:13, pp.743~747(1998)

Gui-bin, J. ; Ni, Z.M.; Wang, S.R. ; Han, H.B.Organic Mercury Speciation in Fish by Capillary Gas Chromatography Interfaced with Atomic Absorption Spectrometry?,Fresenius Z Analytical Chemistry, Vol:334, pp.27~30(1989)

Tesng, C. M; Diego A.D.; Martin, F.M. ; Donard O. F. X.淯apid and Quantitative Mocrowave-Assisted Recovery of Methylmercury From Standard Reference Sediments.?,Journal of Analytical Atomic Spectrometry ,Vol:12, pp.629~635(1997)

Tesng, C. M. ; Diego, A.D.; Martin F. M.; Amouroux D. ; Olivier F. X. Donard, 淯apid Determination of Inorganic Mercury and Methylmercury in Biological Reference Materials by Hydride Generation, Cryofocusing, Atomic Absorption Spectrometry After Open Focused Microwave-Assisted Alkaline Digestion?,Journal of Analytical Atomic Spectrometry, Vol:12, pp.743~750(1997)

Cai, Y. ; Monsalud, S.; Furton ,K.G; Jaff., R.; Jones, R. D., "Determination of methylmercury in fish and aqueous samples using solid-phase microextraction followed by gas chromatography-atomic fluorescence spectrometry.", Applied Organometallic Chemistry, Vol:12, pp.565~569(1998)

Lee, Y. H. ; John, M.; Iverfeldt, A., Experiences with the Analytical Procedures for the Determination of Methylmercury in Environmental Samples.?,Applied Organometallic Chemistry, Vol:8, pp.659~664(1994)

Morrison, M. A.; Weber, J. H. Detremination of monomethylmercury cation in sediments by vaccum distillation followed by hydride derivatization and atomic fluorescence spectrometric detection? Applied Organometallic chemistry ,Vol:11, pp.761~769(1997)

Ritsema, R. ; Donard,O. F. X. ,On-line Speciation of Mercury and Methylmercury in Aqueous Samples by Chromatography-Atomic Fluorescence Spectrometry After Hydride Generation? Applied Organometallic Chemistry, Vol:8, pp.571~575(1994)

Eiden, R. ; Falter, R.;Augustin-Castro, B.; Scholer, H.F. Distillation, on-line RP C18 Preconcentration and HPLC-UV-PCO-CVAAS as a new Combination for the Determination of Methylmercury in Sediments and Fish Tissue.? Fres. J. Anal. Chem., Vol:357, pp:439~441(1994)

Falter, R. ; Scholer, H. F. Determination of Mercury Species in Natural Waters at Picogram Level with on-line RP C18 Preconcentration and HPLC-UV-PCO-CVAAS? Fresenius J. Analytical Chemistry, Vol:353, pp:34~38(1995)

Hempel, M. ; Hintelmann, H.; Wilken, R. D.Determination of Organic Mercury Species in Soils by High-performance Liquid Chromatography With Ultraviolet Detection.? Analyst, Vol:117, pp.669-672(1992)

Bisogni,Jr. J.J.; Lawrence A.W."Determination of submicrogram quantities of monomethyl mercury in aquatic samples.", Environmental science & Technology, pp:850∼852(1974)

Morrison, M. A.; and Weber, J. H. Detremination of monomethylmercury cation in sediments by vaccum distillation followed by hydride derivatization and atomic fluorescence spectrometric detection? Applied Organometallic chemistry Vol:11, pp:761~769(1997)

Mackey, E.A; Becker, D.A."Determination of methylmercury in two mussel tissue standard reference materials by pre-irradiation separation and neutron activation analysis", Analyst, Vol:123, pp.779∼783(1998)

West?, G.Determinnation of Methylmercury Mercury Compounds in Foodstuff,Ⅰ,Methylmercury Compounds in Fish, Identification and Determination?? Chem. Scan. Acta., Vol:20, pp.2131~2137 (1966)

West?, G."Determinnation of Methylmercury Mercury Compounds in Foodstuff,ⅡDetermination of Methylmercury Compounds in Fish, eggs, and liver?? Acta. Chem. Scan., Vol:21, pp.1790~1800(1967)

Horvat, M.; May, K.; Stoeppler, M.; Byrne, A. R."Comparative Studies of Methylmercury Dtermination in Biological and Environmental Samples.? Applied Organometallic Chemistry, Vol:2, pp.515~524(1988)

Horvat, M. ; Byrne, A. R. " A Modified Method for the Determination of Methylmercury by Gas Chromatography.? Talanta , Vol:37, pp.207~212(1990)

Horvat, M.; Bloom, N. S.; Liang, L."Comparison of Distillation with other Current Isolation Methods for the Determination of Methlmercury Compounds in low level Environmental Samples. PART1: Sediments? Analytica Chimica Acta, Vol:281, pp.135-152(1993)

Horvat, M. ; Bloom, N. S.; Liang, L.;Mandic, V.; Padberg, S.; Lee, L.-H.; Hintelmann, H.毧ertification of Methylmercury Compounds Concentration in Marine Sediment Reference Material, IAEA-365.紓ol:8, pp.533~540(1994)

"Mercury(Methyl) in Fish and Shellfish" 1st Action, Method 988.11, AOAC(1990)

Hight, S.C.; Corcoran M.T."Rapid determination of methyl mercury in fish and shellfish: Method development.?J. Assoc. Off. Anal. Chem., Vol:70, pp.24~30(1987)

Newsome, W. H."Determination of Methyl Mercury in Fish and in Cereal Grain Products.? J. Agr. Food Chem., Vol:19, pp.567~569 (1971)

Schafer, M.L.; Peeler J. T.; Hamilton C. H.; Camphell J.E. Method for the Estimation of Methyl Mercury Compounds in Fish.紖.S. Department of Health, Education and Welfare, Division of Microbiology, Food and Drug Administration, Bureau of Foods, (1988)

Gilman, L.B.General Guidelines for Microwave Sample Preparation.? CEM. U.S.A.,?Ch.7(1988)

Kingston, H.M.; Haswell S.T."Microwave-Enhanced Chemistry", ACS. U.S.A., Ch.1(1997)

Skoog, D.A.; Leary, J. J. "Principles of Instrumental Analysis" , 4th ed.(1993)

中國層析公司：氣相層析儀維護訓練使用手冊。

Vazquez, M. J. ; Carro, A. M.; Lorenzo, R. A.; Cela, R.Optimization of Methylmercury Microwave-Assisted Extraction from Aquatic Sediments.? Anal. Chem., pp.221~225(1997)

Rub? E. ; Lorenzo R. A.; Casais. C. ; Carro, A. M. ; Cela R.Evaluation of Capillarry Columns used in the Routine Determination of Methlmercury in Biological and Environmental Materials. J. Chromatogr. , Vol:605, pp.69~80(1992)

CEM Corporation MES-1000 Microwave Solvent Extraction System Operation Manual?1993)

Kanno, A.H.A.; Takabataka E.,"A Method for the Determination of Mercury in Environmental Sample Particularly Sediments", Sanitation Chemistry, Vol:31, pp260~268(1985)

EPA Method 8000B, "Determinative Chromatography Separation", U.S.A.(1995)

EPA Method 8082, "Polychlorinated Biphenyl(PCBs) by Capillary Column Gas Chromatography", U.S.A.(1995)

化學製劑中甲基汞檢測方法－氣相層析儀電子捕捉偵測器法(GC-ECD)，環署檢字第26926號公告，NIEA T901.10T

Eiden, R.; Falter, R.; Augustin-Castro, B.; Scholer, H.F. Distillation , on-line RP C18 preconcentration and HPLC-PCO-CVAAS as a new combination for the determination of methylnercury in sediments and fish tissue?,Fresenius J. Anal. Chem., Vol:357, pp439~441(1997)

Code of Federal Regulations,笈etefinition and Procedures for the Determination of the Method Determination Limit? 40CFR, Part1, Appendix B, US GPO, Washington, D.C., pp.537(1990)

Boumans, P.W.J.M., Detection Limites and Spetral Interence in Atomic Spectrometry? Anal. Chem., Vol.66(1994)

Klmbrough, D.E.; Wakakuwa, J.粈ethod Detection? Environ. Sci. Technol., Vol.27, pp.2692~2699(1993)