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博碩士論文 etd-0213108-142314 詳細資訊
Title page for etd-0213108-142314
論文名稱
Title
利用液相層析串聯質譜儀探討孔雀綠及還原型孔雀綠在吳郭魚體內之藥物動力學
Investigation of pharmacokinetics of malachite green and leucomalachite green in Tilapia with liquid chromatography-tandem mass spectrometry
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
140
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2008-01-22
繳交日期
Date of Submission
2008-02-13
關鍵字
Keywords
液相層析串聯質譜儀、藥物動力學、孔雀綠、還原型孔雀綠
malachite green, LC/MS/MS, leucomalachite green, pharmacokinetic
統計
Statistics
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The thesis/dissertation has been browsed 5675 times, has been downloaded 2884 times.
中文摘要
本研究主要目的為以藥物動力學之觀點探討孔雀綠藥浴濃度因子及藥浴時間因子對吳郭魚體內孔雀綠及還原型孔雀綠累積及排除上的差異。本研究以LC-MS/MS作為偵測儀器,對於孔雀綠及還原型孔雀綠的偵測極限為0.51 ppb及0.48 ppb。實驗結果發現,孔雀綠標準品易受高溫影響而出現降解之現象,在分析過程中添加TMPD可以有效的維持孔雀綠標準品之穩定。藥浴水體中之孔雀綠容易附著於藥浴容器上,而造成孔雀綠藥浴時濃度下降。孔雀綠藥浴之時間長短及藥浴濃度高低與體內各部位之孔雀綠及還原型孔雀綠之累積量成現正相關。魚體各部位中,對於外界孔雀綠之累積速率以肝臟最快,累積速率常數為21.62 h-1,而肝臟也是將孔雀綠代謝為還原型孔雀綠之轉換率最主要的部位,還原型孔雀綠之淨累積速率常數為213.67 h-1;對於外界之還原型孔雀綠之累積速率,肝臟也高於其他部位,累積速率常數為28.46 h-1。在水浴時期,對孔雀綠及還原型孔雀綠排除速率最快的部位為鰓,孔雀綠排除速率常數為0.7799 h-1,還原型孔雀綠排除速率常數為0.4658 h-1;脂肪中之還原型孔雀綠在水浴6小時內尚呈現上升之趨勢,累積速率常數為0.1873 h-1∼0.2504 h-1。
Abstract
The purpose of this research is that investigate the effects of time and concentration of exposure for the accumulation and depletion of malachite green and leucomalachite green in tilapia by pharmacokinetics. LC-MS/MS was used as the analytical instrument in this research, and the detection limit of malachite green and leucomalachite green is 0.51 ppb and 0.48 ppb. The results show that malachite green is unstable at high temperature. Addition of TMPD into the standard can stabilize malachite green. The malachite green in exposure water is easy to adhere to the fiberglass tub and cause the decreases of concentration of malachite green in water. The concentrations of malachite green and leucomalachite green in tilapia are positive related to the time and concentration of exposure. In experiments of exposure of malachite green, the highest concentration of malachite green occurs in liver, accumulation rate constant is 21.62 h-1. Liver is also the major organ for transforming malachite green into leucomalachite green, the net leucomalachite green accumulation rate constant is 213.67 h-1. In the period of water bath, Gill is the fastest organ for eliminating malachite green and leucomalachite green, the elimination rate constant is 0.7799 h-1 and 0.4658 h-1; the leucomalachite green concentration in fat is still increase until 6h of water bath.
目次 Table of Contents
謝誌………………………………………………………………………………………………I
中文摘要……………………………………………………………………………………II
英文摘要……………………………………III
目錄……………………………………………………………………………………………V
表目錄……………………………………………………………………………………VIII
圖目錄…………………………………………………………………………………………X

壹、前言...........................................................1
一、孔雀綠及還原型孔雀綠之理化特性…………………………………………1
二、孔雀綠及還原型孔雀綠之用途………………………………………………1
三、孔雀綠及還原型孔雀綠之毒性………………………………………………3
四、孔雀綠及還原型孔雀綠在生物體內的累積………………………………5
五、世界各國對孔雀綠的管制………………………………………………………6
六、檢驗儀器方法比較…………………………………………………………………7
七、研究目的………………………………………………………………………………8

貳、實驗材料及方法……………………………………………………………………10
一、藥品及試劑……………………………………………………………………………10
二、試劑配製……………………………………………………………………………11
三、儀器及器材…………………………………………………………………………13
四、實驗生物……………………………………………………………………………13
五、實驗設計……………………………………………………………………………13
六、萃取步驟……………………………………………………………………………17
七、LC/MS/MS分析測定條件……………………………………………………17

參、結果……………………………………………………………………………………24
一、 實驗方法部分……………………………………………………………………24
1. LC/MS/MS層析圖譜………………………………………………………………24
2. 無抗氧化劑下光照與否對孔雀綠及還原型孔雀綠的影響…………24
3. 無光照下抗氧化劑對孔雀綠及還原型孔雀綠的影響………………27
4. 光照下抗氧化劑對孔雀綠及還原型孔雀綠的影響……………………27
5. 溫度與抗氧化劑對孔雀綠及還原型孔雀綠的影響……………………30
6.檢量線……………………………………………………………………………………32
7.偵測極限…………………………………………………………………………………32
8.-20℃保存五天後,魚體各組織之孔雀綠及還原型孔雀綠的變化.32
二、 孔雀綠及還原型孔雀綠之藥浴濃度……………………………………37
三、 魚體中的孔雀綠及還原型孔雀綠變化…………………………………37
1. 孔雀綠之升降趨勢…………………………………………………………………37
2. 還原型孔雀綠之升降趨勢………………………………………………………38
3. 各器官中孔雀綠及還原型孔雀綠之最高累積濃度……………………51
4. 藥物動力學-累積速率常數……………………………………………………54
5. 藥物動力學-排除數率常數……………………………………………………57

肆、討論……………………………………………………………………………………66
一、分析方法……………………………………………………………………………66
1. 光照、溫度、抗氧化劑對孔雀綠及還原型孔雀綠的影響…………66
2. 冷凍保存期間魚體各部孔雀綠與還原型孔雀綠之變化……………67
3. 分析方法比較…………………………………………………………………………68
二、藥浴濃度……………………………………………………………………………70
三、體內累積……………………………………………………………………………71
1. 孔雀綠及還原型孔雀綠升降趨勢……………………………………………71
2. 各組織間最高濃度之比較..…………………………………………………74
3. 生殖腺發育與累積…………………………………………………………………75
4. 藥浴期之藥物動力學……………………………………………………………77
5. 水浴期之藥物動力學………………………………………………………………80
6. 組織內孔雀綠及還原型孔雀綠降低至偵測極限下所需時間……82
伍、結論……………………………………………………………………………………85
陸、參考文獻……………………………………………………………………………87
柒、附錄……………………………………………………………………………………95
參考文獻 References
Alderman, D. J. and Clifton-Hadley, R. S., 1993, Malachite green: a pharmacokinetic study in rainbow trout, Oncorhynchus mykiss (Walbaum), Joural of Fish Diseases, 16, 297-311.

Alderman, D. J., Clifton-Hadley, R. S., 1988, Malachite green therapy of proliferative kidney disease in rainbow trout: Field trials, The Veterinary Record, 122, 103-106.

Andersen, W. C., Turnipseed, S. B., and Roybal, J. E., 2006, Quantitative and Confirmatory Analyses of Malachite Green and Leucomalachite Green Residues in Fish and Shrimp, Journal of Agricultural and Food Chemistry, 54, 4517-4523.

Bergwerff, A. A., Kuiper1 R. V., Scherpenisse, P., 2004, Persistence of residues of malachite green in juvenile eels (Anguilla anguilla), Aquaculture, 233, 55–63.


Bergwerff, A. A., Scherpenisse, P., 2003, Determination of residues of malachite green in aquatic animals, Journal of Chromatography B, 788 , 351–359.

Bly, J. E., Quinuou, S. M.-A., Lawson, L. A., Clem, L. W., 1996, Therapeutic and prophylactic measures for winter saprolegniosis in channel catfish, Diseases of Aquatic organisms, 24, 25-33.

Crosbie, P. B. B., Munday, B. L., 1999, Environmental factors and chemical agents affecting the growth of the pathogenic marine ciliate Uronema nigricans, Diseases of Aquatic Organisms, 36, 213-219.

Culp, S. J., 2004, NTP Technical Report on the Toxicity Studies of Malachite Green Chloride and Leucomalachite Green administered in Feed to F344/N Rats and B6C3F1 Mice, National Toxicology Program Toxicity Report Series Number 71.
 
Culp, S. J., Blankenship, L. R., Kusewitta, D. F., Doerge, D. R., Mulligan, L. T., Beland, F. A., 1999, Toxicity and metabolism of malachite green and leucomalachite green during short-term feeding to Fischer 344 rats and B6C3F1 mice, Chemico-Biological Interactions, 122, 153–170.

Culp, S.J., Beland, F.A., 1996. Malachite green: a toxicological review. Journal of the American College of Toxicology, 15, 219–238.

Diggles, B. K., 2000, Chemotherapy of the ciliate Trichdina sp. On juvenile turbot (Colistium nudipinnis) with notes on the susceptibility of fish with abnormal pigmentation, Zealand Journal of Marine and Freshwater Research, 34, 645-652.

Diggles, B. K., 2001, A mycosis of juvenile spiny rock lobster, Jasus edwardsii (Hutton, 1875) caused by Haliphthoros sp., and possible methods of chemical control, Journal of Fish Diseases, 24, 99-110.
 
Fessard, V., Godard, T., Huet, S., Mourot, A. and Poul, J. M., 1999, Mutagenicity of Malachite Green and Leucomalachite Green in in vitro Tests, Journal of Applied Toxicology , 19, 421–430.

Halme, K., Lindfors, E., and Peltonen, K., 2004, Determination of malachite green residues in rainbow trout muscle with liquid chromatography and liquid chromatography coupled with tandem mass spectrometry, Food Additives and Contaminants, 21, 641–648.

Meinertz, J. R., Stehly, G. R., Gingerich, W. H. and Allen, J. L., 1995, Residues of [14C]-malachite green in eggs and fry of rainbow trout, Oncorhynchus mykiss (Walbaum), after treatment of eggs, Journal of Fish Diseases, 18, 239-247.

Meyer, F.P., Jorgensen, T.A., 1983. Teratological and other effects of malachite green on the development of rainbow trout and rabbits. Transactions of the American Fisheries Society, 112 , 818–824.


Mitrowska, K., Posyniak, A., Zmudzki, J., 2005, Determination of malachite green and leucomalachite green in carp muscle by liquid chromatography with visible and fluorescence detection, Journal of Chromatography A, 1089, 187–192.

Plakas, S. M., El Said, K.R., Stehly, G. R., Gingerich, W. H., and Allen, J. L, 1996, Uptake, tissue distribution, and metabolism of malachite green in the channel catfish (Ictalurus punctatus), Canadian Journal of Fisheries and Aquatic Sciences, 53, 1427-1433.

Rushing, L. G., Thompson Jr, H. C.., 1997, Simultaneous determination of malachite green, gentian violet and their leuco metabolites in catfish or trout tissue by high-performance liquid chromatography with visible detection, Journal of Chromatography B, 688, 325-330.

Scherpenisse, P., Bergwerff, A. A., 2005, Determination of residues of malachite green in finfish by liquid chromatography tandem mass spectrometry, Analytica Chimica Acta, 529, 173–177.

Srivastava, S. J., Singh, N. D., Srivastava, A. K. and Sinha, R., 1995, Acute toxicity of malachite green and its effects on certain blood parameters of a catfish, Heteropneustes fossilis, Aquatic Toxicology, 31, 241-247.

Srivastava, S., Sinha, R., Roy D., 2004, Toxicological effects of malachite green, Aquatic Toxicology, 66, 319–329.

Stammati, A., Nebbia, C., Angelis, I. D., Albo, A. G., Carletti, M., Rebecchi, C., Zampaglioni, F., Dacasto, M., 2005, Effects of malachite green (MG) and its major metabolite, leucomalachite green (LMG), in two human cell lines, Toxicology in Vitro, 19, 853-858.

Valle L., D´ıaz, C., Zanocco, A. L., Richter, P., 2005, Determination of the sum of malachite green and leucomalachite green in salmon muscle by liquid chromatography–atmospheric pressure chemical ionisation-mass spectrometry, Journal of Chromatography A, 1067, 101–105.

中國國家食品安全信息中心http://www.fsi.gov.cn/news.view.jsp?sort=305&id=2430

中華人民共和國商務部出口商品技術指南http://kjs.mofcom.gov.cn/table/manyu.pdf

水產品中孔雀石綠和結晶紫殘留量的測定,中華人民共和國國家標準(GB/T 19857-2005)

水產品中孔雀綠及期待謝物檢驗方法之研究,行政院衛生署九十五年度科技發展研究計畫(計畫編號:DOH95-FD-2023)

香港附屬法例-食物內有害物質規例http://www.hklii.hk/hk/legis/ch/reg/132AF/sch1.html

郭素蓮,中央畜產會 http://www.naif.org.tw/livestock/2005109.htm

漁業署民國九十五年漁業年報http://www.fa.gov.tw/chn/statistics_price/year_book/2006c/95tab7.pdf

漁業資訊服務網 http://www.fa.gov.tw/chn/media/text/chinese.txt

漁業資訊服務網-全國養殖漁業會議總結論報告http://www.fa.gov.tw/chn/organization/inshore_fishing/inshore_fishing_conclusion.php

澳門特別行政區政府衛生局疾病預防控制中心環境及食物衛生部新聞稿第045/CDC/2005號http://www.ssm.gov.mo/design/news/Document/CDC200508a.doc
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