臺灣博碩士論文加值系統

有機磷農藥被廣泛用於農作物生長，防治病蟲害，其施用後可能經由灌溉或降雨使得這些有毒物質流入河川、湖泊或地下水中，對水體環境生態系造成影響，故本篇研究十種有機硫代磷酸酯類物質對水蚤之急毒性影響。從單一急毒性結果來看，化合物Phoxim對水蚤最具毒性，從結構式來看Phoxim與其他化合物之差異為其帶有苯乙腈官能基，其電荷密度分布不同造成電子傳遞而產生較高毒性。
在QSAR分析方面，疏水性參數log Kow毒性值迴歸相關性佳(r2 = 84.5%)，此結果顯示化合物毒性機制主要貢獻來源為藉由通透細胞膜滲入到細胞體內造成毒性。增加反應性參數只些微提升了方程式r2值，顯示反應性參數並非反應中速率限制步驟。從結果得知有機硫代磷酸酯雖被分類為乙醯膽鹼酯酶抑制有機物，但其對於水蚤毒性機制偏向於麻醉性。
風險評估方面，蒐集文獻數據和本研究急毒性及推估慢毒性數據計算風險商數，大部分RQ＞1，顯示此類化合物具潛在風險。比較急慢毒性數據計算之RQ值，顯示急毒性數據有高估風險的可能。

This study present the toxicity data of ten organothiophosphorus compounds to Daphnia magna using 48-hours acute toxicity test.
Results indicate that Phoxim is the most toxic compound, the reason caused higher toxicity of Phoxim is dependent on its higher charge density due to benzyl cyanide group.
Quantitative structure-activity relationships (QSARs) were established between the EC50 values and various molecular descriptors, and a highly predictive two-variable QSAR models were obtained. According to QSAR models, the toxic mechanism was highly related to hydrophobicity of these compounds, and this result indicated that organothiophosphorus compounds caused toxicity to Daphnia magna mainly due to permeate through membrances, changing the components of the cell membrances. Adding reactive parameters slightly improve the r2 values indicated that the reaction rate of the organothiophosphate with AChE, the conversion of organothiophosphate to organophosphate and the molecule polarity are not rate-limiting steps.
The preliminary environmental risk assessment was conducted following the European Union RQ model. Results point out that most of these compounds exist potential risks to aquatic environment. Risk quotient derived from acute and estimated chronic toxicity data were different, using acute toxicity data may overestimate the risk quotient value.

第一章 前言 1
1.1研究緣起 1
1.2研究目的 2
1.3研究架構 3
第二章 文獻回顧 4
2.1毒性物質-有機磷殺蟲劑之介紹 4
2.1.1有機磷農藥的基本特性 4
2.1.2乙醯膽鹼酯酶於生物體作用機制 4
2.1.3有機硫磷酸殺蟲劑之毒性作用機制 5
2.1.4 有機磷劑代謝 7
2.1.5有機磷農藥於水體環境中之流佈 8
2.1.6 水體中去除方法 9
2.2水蚤急毒性試驗 13
2.2.1 試驗物種簡介 13
2.2.2 水蚤毒性試驗方法 14
2.3定量結構-活性關係 15
2.3.1 QSAR簡介 15
2.3.2 QSAR常用參數 15
2.3.3 QSAR於環境毒理學上的應用 17
2.4文獻中有機磷殺蟲劑QSAR研究 19
2.5水體環境影響評估 21
2.5.1風險評估及風險商數計算 21
2.5.2環境危害評估 23
2.6急慢毒性比值 25
第三張 基本理論 26
3.1毒性物質之濃度反應關係模式 26
3.2 NOEC與CUT-OFF VALUE 27
3.2.1 判定NOEC與LOEC 27
3.2.2 Cut-off value 27
第四章 實驗設備與方法 28
4.1 實驗設備及材料 28
4.2 水蚤毒性試驗 30
4.2.1試驗方法 30
4.2.2試驗步驟 32
4.3藥品配置方式 33
4.4實驗數據處理 34
4.5 QSAR參數計算 40
第五章 結果與討論 41
5.1水蚤急毒性試驗結果 41
5.2 QSAR分析 48
5.2.1單一參數迴歸分析 48
5.2.2 雙參數迴歸分析 53
5.3低影響濃度比較 56
5.3.1 NOEC與EC10之比較 56
5.3.2平均中斷值(cut-off value) 58
5.4環境風險評估 59
5.4.1 風險商數計算 59
5.4.2環境危害性分類 64
5.5試驗物種敏感度比較 66
5.6 急慢毒性比值 70
第六章 結論與建議 71
6.1 結論 71
6.2建議 72
第七章 參考文獻 73
附錄一 單一毒性原始數據 81
附錄二 QSAR 參數計算使用方法 91
附錄三 統計軟體使用方法 96
附錄四 DUNNETT’S TEST計算表使用方法 99
附錄五 水蚤毒性試驗品質管制圖 120

Atkins, P.W.. 1994. Physical chemistry. Oxford:Oxford University Press.

Barr, D.B. and Needham, L.L.. 2002. Analytical methods for biological monitoring of exposure to pesticides: a review. Journal of Chromatography B 778:5-29.

Berijani, S., Assadi, Y., Anbia, M., Hosseini, M.R.M, Aghaee, E., 2006. Dispersive liquid–liquid microextraction combined with gas chromatography-flame photometric detection Very simple, rapid and sensitive method for the determination of organophosphorus pesticides in water. Journal of Chromatography A 1123:1-9.

Cabras, P., 2003. Pesticides:Toxicology and Residues in Food. Food Safety: Contaminants and Toxins:91-124.

Cardozo, M.G., Iimura, Y., Sugimoto, H., Yamanishi, Y., Hopfinger A.J., 1992. QSAR Analyses of the Substituted Indanone and Benzylpiperidine Rings of a Series of Indanone-Benzylpiperidine Inhibitors of Acetylcholinesterase. Journal of Medical Chemistry 35:584-589.

Castillo, L.E., Martínez, E., Ruepert, C., Savage, C., Gilek, M., Pinnock, M., Solis, E., 2006. Water quality and macroinvertebrate community response following pesticide applications in a banana plantation, Limon, Costa Rica. Science of the Total Environment 367:418–432.

Commission of the European Communities, 1996. Technical Guidance Documents in Support of The Commission Directive 93/67 EEC on risk assessment for new notified substances and the commission regulation (EC) 1488/94 on risk assessment for existing substances, Italy.

Cronin, M.T.D. and Dearden, J.C., 1995. QSAR in Toxicology. 1. Prediction of Aquatic Toxicity. Quantitative Structure-Activity Relationships14:1–7.

Daneshvar, N., Aber, S., Khani, A., Rasoulifard, M.H., 2007. Investigation of adsorption kinetics and isotherms of imidacloprid as a pollutant from aqueous solution by adsorption onto industrial granular activated carbon. International Journal of Food, Agricalture and Environmen 5:425–429.

Eddleston, M., Szinicz, L., Eyer, P., Buckley, N.. 2002, Oximes in acute organ phosphorus pesticide poisoning: a systematic review of clinical trials. Quarterly Journal of Medicine 95:275-283.

EU, 2003. Technical Guidance Document on Risk Assessment in Support of
Commission Directive 93/67/EEC on Risk Assessment to New Notified Substances and Commission Regulation (EC) No 1488/94 on Risk Assessment for existing Substances and Directive 98/8/EC of the European Parliament and the Council concerning the placing of biocidal products on the market. Luxembourg (LU): Office for Official Publications of the European Communities.

European Community Legislation, 1967. General Classification and Labelling Requirements for Dangerous Substances and Preparations Directive 67/548/EEC Annex VI.

Fukuto, T.R., 1990. Mechanism of Action of Organophosphorus and Carbamate Insecticides. Environmental Health Perspectives 87:245-254.

Hatzios, K.K., 1991. Biotransformation of herbicides in higher plants. Metabolism of herbicides in higher plants:141-185.

Hermans, J., Bruijn, J., Paulty, J., Seinen, W., 1987. QSAR Studies for fish toxicity data of organophosphorus compounds and other classes of reactive organic compounds. QSAR in Environmental Toxicology-II:135-152.
Hind, A.R., Bhargava, S.K., Grocott, S.C., 1999. The surface chemistry of Bayer process solids: a review. Colloids and surfaces A 146:359–374.

Hoeckstra, J.A. and Ewijk, P.H.V., 1993. Alternatives for the no-obderved-effect level. Environmental Toxicology and Chemistry 12:187-194.

Hofman, J.A.M.H., Beerendonk, E.F., Flolmer, H.C., Kruithof, J.C., 1997. Removal of pesticides and other micro pollutants with cellulose-acetate, polyamide and ultra-low pressure reverse osmosis membranes. Desalination 113:209–214.

Jacoby, N.B., 1980. Enzymatic Basis of Detoxication. New York:Academic Press.

Karelson, M. and Lobanov, V.S., 1996. Quantum-Chemical Descriptors in QSAR/QSPR Studies. Chemical Reviews 96:1027-1043.

Kasagami, T., Miyamoto, T., Yamamoto, I., 2002. Activated transformations of organophosphorus insecticides in the case of non-AChE inhibitory oxons. Pest Management Science 58:1107-1117.

Kazemi, M., Tahmasbi, A.M., Valizadeh, R., Naserian, A.A., Soni, A., 2012. Organophosphate pesticides: A general review. Agricultural Science Research Journals 2(9):512- 522.

Kolpin, D.W., Barbash, J.E., Gilliom, R.J., 2000. Pesticides in Ground Water of the United States, 1992–1996. Ground Water 38:858-863.

Liang, P., Guo, L., Liu, Y., Liu, S., Zhang, T.Z., 2005. Application of liquid-phase microextraction for the determination of phoxim in water samples by high performance liquid chromatography with diode array detector. Microchemical Journal 80:19-23.

Lin, X.Y., Yu, R.L., Hu, G.R., 2009. Acute toxicities and QSAR of substituted phenols to photobacterium phosphoreum at different pH. IEEE:1-4.
Lipton, J., Galbraith, H., Burger, J., Wartenberg, D., 1993. A paradigm for ecological risk assessment. Environmental Management 17(1):1–5.

Lu, G.H., Yuan, X., Zhao, Y.H., 2001. QSAR study on the toxicity of substituted benzenes to the algae (Scenedesmus obliquus). Chemosphere: 437-440.


Malato, S., Blanco, J., Richter, C., Milow, B., Maldonado, M.I., 1999. Solar photocatalytic mineralization of commercial pesticides methamidophos. Chemophere 38:1145–1156.

Ma, L., Wang, T., Liu, X., 2011. A study on quantitative structure activity relationship between organic phosphorus pesticides and marine microalgae. Water Resource and Environmental Protection 3:2025-2028.

McFarland, J.W., 1970. Parabolic relationship between drug potency and hydrophobicity. Journal of Medical Chemistry 13:1092-1196.

Miliadis, G.E., Malatou, P.T., 1997. Monitoring of the pesticide levels in natural waters of Greece. Bulletin of Environmental Contamination and Toxicology 59:917–923.

Moore, D.R.J. and Caux, P.Y., 1997. Estimating low toxic effects. Environmental toxicology and chemistry 16: 794-801.

OECD, 2004. OECD Guidelines for the Testing of Chemicals. Test No. 202: Daphnia magna Reproduction Test.

OECD, 2004. OECD Guidelines for the Testing of Chemicals. Test No. 202: Daphnia sp. Acute Immobilisation Test.

Palma, G., Sa´nchez, A., Olave, Y., Encina, F., Palma, R., Barra, R., 2004. Pesticide levels in surface waters in an agricultural–forestry basin in Southern Chile. Chemosphere 57:763-770.

Papadopoulou-Mourkidou, E., 2002. Quality of surface waters of Macedonia-Thrace, Northern Greece, Quality control program, Final Report (in Greek). Ministry of Agriculture, Thessaloniki, Greece.

Pasha, F.A., Srivastava, H.K., Singh, P.P., 2005. QSAR Study of Estrogens With the Help of PM3-Based Descriptors. International Journal of Quantum Chemistry 104:87-100.

PNUMA/IPCS, 1999. Evaluation of chemical risks. Produced by the Program of the Nations United for the Ambient, the Organization the International of the Work and the Worldwide Organization of the Health, within the Framework of the Inter-Organization for Programme the Sound Management of Chemical, 234 p.

Qu, C.S., Chen, W., Bi, J., Huang, L., Li, F.Y., 2010. Ecological risk assessment of pesticide residues in Taihu Lake wetland, China. Ecological modelling 222:287-292.

Russom, C.L., Bradbury, S.P., Broderius, S.J., Hammermeister, D.E., Drummond, R.A., 1997. Predicting modes of toxic action from chemical structure: Acute toxicity in the fathead minnow (Pimephales promelas). Environmental Toxicology and Chemistry 16:948-967.

Sanchez-Bayo, F., 2006. Comparative acute toxicity of organic pollutants and reference values for crustaceans. I. Branchiopoda, Copepoda and Ostracoda. Environmental Pollution 139:385-420.

Schultz, T.W., Sinks, G.D., Bearden, A.P., 1998. QSAR in aquatic toxicology:A mechanism of action approach comparing toxic potency to Pimephales promelas, Tetrahymena pyriformis, and Vibrio fischeri. London: Taylor &; Francis.

Schultz, T.W., Wilke, T.S., Bryant, S.E., Hosein, L.M., 1991. QSARs for selected aliphatic and aromatic amines. Science of The Total Environment 109-110:581-587.

Senior, S.A., Madbouly, M.D., Massry, A.M.E., 2011. QSTR of the toxicity of some organophosphorus compounds by using the quantum chemical and topological descriptors. Chemosphere 85:7-12.

Seward, J.R. and Schultz T.W., 1999. QSAR analyses of the toxicity of aliphatic carboxylic acids and salts to Tetrahymena pyriformis. SAR and QSAR in Environmental Research 10.6:557-567.

Seward, J.R., Hamblen, E.L., Schultz, T.W., 2002. Regression
comparisons of Tetrahymena pyriformis and Poecilia reticulata toxicity.
Chemosphere. 47, 93-101.

Shimabukuro, R.H., 1985. Detoxification of herbicides. Pages 215–240 in S. O. Duke, ed. Weed Physiology. Volume 2. Boca Raton, FL: CRC Press.

Slater, D. and Jones, H., 1999. Environmental risk assessment and the environment agency. Journal of Hazardous Materials 65 (1–2):77–91.

Sparling, D.W. and Fellers, G., 2007. Comparative toxicity of chlorpyrifos, diazinon, malation and their oxon derivatives to larval Rana boylii. Environmental Pollution 147:535-539.

Sun, K., Liu, W., Liu, L., Wang, N., Duan, S., 2013. Ecological risks assessment of organophosphorus pesticides on bloom of Microcystis wesenbergii. International Biodeterioration &; Biodegradation:98-105.

Tankiewicz, M., Fenik, J., Biziuk, M., 2010. Determination of organophosphorus and organonitrogen pesticides in water samples. Trends in Analytical Chemistry 29:1050-1063.

Tsuda, T., Nakamura, T., Inoue, A., Tanaka, K., 2009. Pesticides in Water, Fish and Shellfish from Littoral Area of Lake Biwa. Bulletin of environmental contamination and toxicology 82:716–721.

U.S. Environmental Protection Agency, ECOTOX Database.
(http://www.epa.gov/ecotox/).

U.S. Environmental Protection Agency, 1992. Framework for Ecological Risk Assessment. U.S. Environmental Protection Agency, Washington, DC, EPA/630/R-92/001.

U.S. Environmental Protection Agency, 1998. Guidelines for Ecological Risk Assessment. U.S. Environmental Protection Agency, Washington, DC, EPA/630/R-95/002F.

U.S. Environmental Protection Agency, 2002. Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms. U.S. Environmental Protection Agency, Washington, DC.


U.S. Environmental Protection Agency, 2004. Overview of the ecological risk assessment process in the office of pesticide programs, US Environmental
Protection Agency. Office of Prevention, Pesticides and Toxic Substances Office of pesticides Programs. Washington, DC, 92 p.

U.S. Environmental Protection Agency, 2002. Short-term methods for estimating the chronic toxicity of effluents and receiving waters to freshwater organisms. U.S. Environmental Protection Agency. Washington, DC.


U.S. Environmental Protection Agency, 1991. Technical support document for water quality-based toxics control. EPA 505/2-90-001. Washington, DC.

Verhaar, H.J.M., van Leeuwen, C.J., Hermens, J.L.M., 1992. Classifying environmental pollutants. Chemosphere 25:471-491.

Vryzasa, Z., Vassilioub, G., Alexoudisb, C., Papadopoulou-Mourkidoua, E., 2009. Spatial and temporal distribution of pesticide residues in surface waters in northeastern Greece. Water Research 43:1-10.

Wan, M.T., Szeto, S., Price, P., 1994. Organophosphorus insecticide residues in farm ditches of the Lower Fraser Valley of British Columbia. Journal of Environmental Science and Health B 29:917-949.

Wang, S., Zhao, P., Min, G., Fang, G., 2007. Multi-residue determination of pesticides in water using multi-walled carbon nanotubes solid-phase extraction and gas chromatography–mass spectrometry. Journal of Chromatography A 1165: 166-171.

Watts, J.R., Dilly, S.E., 1996. Evaluation of iron catalysts for the Fenton-like remediation of diesel-contaminated soils, Journal of Hazardous materials 51:209-224.

Worthing, C.R. and Walker, S.B., 1987. The Pesticide Manual: a world compendium. British Crop Protection Council.

Zertal, A., Jacquet, M., Lavedrine, B., Sehili, T., 2005. Photodegradation of chlorinated pesticides dispersed on sand. Chemosphere 58:1431–1437.

Zhao, Y.T., Cronin, M.T., Dearden, J.C., 1998. Quantitative structure-
activity relationships of chemicals acting by non-polar narcosis-theoretical
considerations. Quant. Struct. Act. Relat. 17, 131-138
Zvinavashe, E., Du, T., Griff , T., Berg, H.H.J.V.D, Soffers, A.E.M.F., Vervoort, J., Murk, A.J., Rietjens, I.M.C.M., 2009. Quantitative structure-activity relationship modeling of the toxicity of organothiophosphate pesticides to Daphnia magna and Cyprinus carpio. Chemosphere 75:1531-1538.

中華民國環保署。民國100年。生物急毒性檢測方法－水蚤靜水式法。NIEA 2011；NIEA B901.13B。

行政院農業委員會動植物防疫檢疫局。2005-2009年國內成品農藥銷售統計表。 ( http://pesticide.baphiq.gov.tw/web/Insecticides_MenuItem9_2S.aspx?no=31).