進階搜尋


 
系統識別號 U0026-0812200911255563
論文名稱(中文) 藉由減低基質干擾前處理與強陰離子交換樹脂方法分析尿中有機磷代謝物
論文名稱(英文) Method development in diminishing matrix interference during sample preparation incorporated with strong anion exchange technique to determine urinary diakylphosphates
校院名稱 成功大學
系所名稱(中) 環境醫學研究所
系所名稱(英) Institute of Environmental and Occupational Health
學年度 93
學期 1
出版年 94
研究生(中文) 謝昌原
研究生(英文) Chang-Yuan Hsieh
學號 S7691107
學位類別 碩士
語文別 中文
論文頁數 64頁
口試委員 口試委員-桂椿雄
口試委員-許憲呈
指導教授-林維炤
指導教授-張火炎
中文關鍵字 線上衍生化  固相萃取法  去除  干擾  磷酸根    回收率  有機磷農藥  烷基磷酸化合物  強陰離子交換樹脂 
英文關鍵字 solid phase extraction  strong anion exchange  recovery  chloride  phosphates  interference  diminish  in-vial derivatization  alkyl phosphates  organophosphorous pesticides 
學科別分類
中文摘要   由於有機磷農藥易分解、藥效快等特性,廣泛使用於農作物之病蟲害防治。而有機磷農藥的種類繁多,但一般皆以磷酸酯類化合物為主要的結構式,目前對於有機磷農藥之生物偵測是以量測尿中烷基磷酸類化合物(dialkylphosphates, DAPs)的濃度為主。由過去的文獻發現,以往對於萃取尿液中DAPs的前處理方法中,大多以溶劑萃取法為主,其不但消耗大量的有機溶劑,且費時又耗力,因此有研究者擬利用薄膜型之強陰離子交換樹脂(Strong Anion Exchange, SAX)進行DAPs之萃取。但以SAX萃取水中有機磷代謝物之回收率可達76~96%;不過在尿中DETP、DMTP、DEDTP、DMDTP的回收率分別僅有30~58%、18~37%、48~84%、53~77%,推測原因可能是尿中陰離子基質干擾所致。因此本研究將一方面針對尿中陰離子之氯離子及磷酸根離子進行探討;另一方面發展一套有效降低尿中競爭性陰離子且快速有效的測定尿中DAPs濃度之分析方法。本研究以氯化鈉溶液(0.001~10000µg/mL)及磷酸鈉溶液(0.0025~2500µg/mL)評估兩種離子對DAPs的影響,並且以硝酸銀及氫氧化鈉所合成之氫氧化銀(AgOH)來降低尿中陰離子的干擾,尤其是氯離子。由研究結果中發現,尿中磷酸根離子對以SAX前處理萃取尿中DAPs之方法所造成的影響不大(DETP、DEDTP、DMTP、DMDTP之回收率介於74~94%),僅對DEP產生較大的影響(回收率約為14%);而氯離子的干擾對於整個萃取方法則佔比較重要的角色,當氯離子濃度約等於人體尿中氯離子濃度時,DEP、DETP及DMTP的回收率僅約為0%、6%及8%,因此如何有效的降低尿中氯離子的含量即為一重要的課題。而由AgOH降低氯離子干擾的結果發現,當添加量為3倍氯化鈉濃度時,回收率達一最佳的效果,其DEP、DETP及DMTP的回收率分別可提升至14%、98%及106%,但也使得DEDTP及DMDTP的回收率下降至12%及16%。因此為了正確且有效的評估人體尿中DAPs的濃度,建議的實驗流程分成兩部分,一部分以添加AgOH的步驟評估DEP、DETP及DMTP的濃度,另一部分則不添加AgOH以量測DEDTP與DMDTP,方可對尿中DAPs進行測定。

英文摘要  Organophosphorous pesticides (OPPs), mainly constituting phosphoric ester, have been extensively used in pest control in agricultural activities owing to their ready decomposition and rapid efficacy. Dialkylphosphates (DAPs) concentrations in urine have been considered as biological exposure markers for the exposure to OPPs. Conventional sample preparation of solvent extraction has been blamed for its considerable consumption of organic solvents, time and manpower demanding. Therefore, Strong Anion Exchange (SAX) technique is becoming a promising alternative. The recoveries of DAPs in water using SAX technique were about 76-96%, but those in urine were most less than 50%. The abrupt decrease in recovery for urine samples could be due to the anion matrix effect of urine. The purposes of this study were to identify the target anions, esp., chloride and phosphate, causing matrix effects and to develop a method to diminish matrix interference during sample preparation incorporated with SAX technique to determine urinary DAPs. The water solutions of sodium chloride (ranging from 0.001–10,000 µg/mL) and sodium phosphate (ranging from 0.0025–2,500 µg/mL) were used to evaluate the target anion influencing SAX extraction. Silver hydroxide, developed by the combination of silver nitrate and sodium hydroxide, was used as a matrix retardant. We found phosphate ion cause very mild effects on SAX recoveries (the recoveries for DETP, DEDTP, DMTP, and DMDTP all lay between 74 and 94%) except for DEP (recovery decreased to only 14%). On the other hand, chloride ion revealed a significant effect on DAPs recoveries; i.e., the recoveries for DEP, DETP, and DETP were down to 0%, 6%, and 8%, respectively. When silver hydroxide as matrix retardant was added in the water solution of sodium chloride at the concentration 3-fold of chloride, the recoveries for DEP, DETP, and DMTP could reach their maximum of 14%, 98%, and 106%, respectively, at the cost of the recovery decreases of DEDTP and DMDTP to 12% and 16%, respectively. We concluded that two separate procedures incorporated with SAX technique for the determination of DAPs in urine: silver hydroxide could be added as a matrix retardant for DEP, DETP, and DMTP; and no any matrix retardant required for DEDTP and DMDTP.

論文目次 中文摘要..............................A
Abstract..............................C
誌謝..............................E
目錄..............................F
表目錄..............................J
圖目錄..............................K

第一章序論..............................1
1-1 前言..............................1
1-2 研究動機..............................3
1-3 研究目的..............................3

第二章文獻回顧..............................4
2-1 農藥發展過程..............................4
2-2 有機磷農藥簡介..............................5
2-2.1 有機磷農藥結構及特性..............................5
2-2.2 有機磷農藥毒性..............................5
2-2.3 有機磷農藥代謝..............................6
2-2.4 有機磷農藥生物指標..............................6
2-3 樣本前處理方法..............................7
2-3.1 溶劑萃取法..............................7
2-3.2 固相萃取法..............................8
2-3.3 衍生化試劑..............................8
2-4 強陰離子交換樹脂..............................9
2-5 尿液基質的干擾..............................10

第三章材料與方法..............................11
3-1 實驗材料與設備..............................11
3-1.1 藥品與材料..............................11
3-1.2 儀器與設備..............................11
3-2 研究架構圖..............................12
3-3 實驗方法..............................12
3-3.1 氫氧化銀(AgOH)配製..............................12
3-3.2 分析條件..............................13
3-3.3 實驗裝置示意圖,如圖3-2 所示..............................13
3-3.4 實驗步驟..............................13
3-3.5 實驗室QA/QC..............................14
3-3.5.1 檢量線配置..............................14
3-3.5.2 分析準確度..............................14
3-4 實驗設計..............................14
3-4.1 衍生化條件測試..............................14
3-4.2 酸鹼值(pH)測試..............................14
3-4.3 尿液基質測試..............................15
3-4.3.1 氯離子的測試..............................15
3-4.3.2 磷酸根離子的測試..............................15
3-4.4 氫氧化銀(AgOH)添加比例測試..............................16
3-4.5 空白尿液樣本測試..............................16
3-4.5.1 添加回收率..............................16
3-4.5.2 不同濃度添加回收率..............................16
3-5 真實樣本測試..............................16
3-5.1 職業暴露族群尿液樣本分析..............................16
3-5.2 職業暴露族群添加回收率測試..............................17
3-5.3 樣品的儲存、運送及處理..............................17

第四章結果與討論..............................18
4-1 衍生化條件測試..............................18
4-2 檢量線及空白尿液測試..............................19
4-2.1 檢量線測試..............................19
4-2.2 空白尿液測試..............................19
4-3 酸鹼值(pH)測試..............................20
4-4 尿液基值測試..............................21
4-4.1 氯離子測試..............................21
4-4.2 磷酸根離子測試..............................21
4-5 氫氧化銀(AgOH)添加比例測試..............................22
4-6 空白尿液測試..............................23
4-6.1 添加回收率..............................23
4-6.2 不同濃度添加回收率..............................23
4-7 尿中DAPs 之分析方法的比較..............................25
4-8 職業暴露族群尿液樣本分析..............................26

第五章結論..............................27

第六章未來展望..............................28

參考文獻..............................29
參考文獻 Adamis Z, Antal A, Fuzesi I, Molnar J, Nagy L, Susan M, “Occupational exposure to organophosphorus insecticides and synthetic pyrethroid”, International Archives of Occupational & Environmental Health, 1985, 56(4):299-305

Anwar WA, “Biomarkers of human exposure to pesticides”, Environmental Health Perspectives, 1997, 105 Suppl 4:801-806

Aprea C, Sciarra G, Lunghini L, “Analytical method for the determination of urinary alkylphosphates in subjects occupationally exposure to organophosphorus pesticides and in the general population”, Journal of Analytical Toxicology, 1996, 20(7):559-563

Aprea C, Strambi M, Novelli MT, Lunghini L, Bozzi N, “Biologic monitoring of exposure to organophosphorus pesticides in 195 Italian children”, Environmental Health Perspectives, 2000, 108(6):521-525

Carson R (著), 李文昭(譯), “寂靜的春天”, 晨星, 1997

Cerklewski FL, Bills ND, “Determination of chloride procedure based upon diffusion of hydrogen chloride”, Analytical Biochemistry, 1985, 151(2):566-570

Chang HY, Wu HC, Lin WC, Wu PL, Kuei CH, “Determination of organophosphorus pesticide metabolites in surface water by use of a strong anion-exchange disk and in-vial derivatization”, CHROMATOGRAPHIA, 2000, 51(9-10):630-633

Cocker J, Mason HJ, Garfitt SJ, Jones K, “Biological monitoring of exposure to organophosphate pesticides”, Toxicology Letters, 2002, 134(1-3):97-103

Fenske RA, Elkner KP, “Multi-route exposure assessment and biological monitoring of urban pesticide applicators during structural control treatments with chlorpyrifos”, Toxicology & Industrial Health, 1990, 6(3-4):349-371

Field JA, Monohan K, “Chlorinated acid herbicides in water by strong anion-exchange disk extraction and in vial elution and derivatization”, Journal of Chromatography A, 1996, 741(1):85-90

Frobe Z, Stengl B, Drevenkar V, Deljac A, “Accumulation of dialkylphosphorus anions from water by ion-exchange on modified XAD-4 resin”, Journal of Chromatographic Science, 1990, 28(5):269-273

Griffin P, Mason H, Heywood K, Cocker J, “Oral and dermal absorption of chlorpyrifos: a human volunteer study”, Occupational & Environmental Medicine, 1999, 56(1):10-13

Hardt J, Angerer J, “Determination of dialkyl phosphates in human urine using gas chromatography-mass spectrometry”, Journal of Analytical Toxicology, 2000, 24(8):678-684

Hebert VR, Hoonhout C, Miller GC, “Use of stable tracer studies to evaluate pesticide photolysis at elevated temperatures”, Journal of Agricultural & Food Chemistry, 2000, 48(5):1916-1921

Herraiz T, Casal V, “Evaluation of solid-phase extraction procedures in peptide analysis”, Journal of Chromatography A, 1995, 708(2):209-221

Heudorf U, Angerer J, “Metabolites of organophosphorous insecticides in urine specimens from inhabitants of a residential area”, Environmental Research, 2001, 86(1):80-87

Heudorf U, Angerer J, Drexler H, “Current internal exposure to pesticides in children and adolescents in Germany: urinary levels of metabolites of pyrethroid and organophosphorus insecticides”, International Archives of Occupational & Environmental Health, 2004, 77(1):67-72

Janice E. Chambers, “Organophosphate---Chemistry, Fate, and Effects”, Academic Press, California, 1992

Kwong TC, "Organophosphate pesticides: biochemistry and clinical toxicology", Therapeutic Drug Monitoring, 2002, 24(1):144-149

Leng G, Kuhn KH, Idel H, “Biological monitoring of pyrethroid metabolites in urine of pest control operators”, Toxicology Letters, 1996, 88(1-3):215-220

Lin WC, Kuei CH, Wu HC, Yang CC, Chang HY, “Method for the determination of dialkyl phosphates in urine by strong anion exchange disk extraction and in-vial derivatization”, Journal of Analytical Toxicology, 2002, 26(3):176-180

Loewenherz C, Fenske RA, Simcox NJ, Bellamy G, Kalman D, “Biological monitoring of organophosphorus pesticide exposure among children of agricultural workers in central Washington State”, Environmental Health Perspectives, 1997, 105(12):1344-1353

Lu FC (著), 劉宗榮(譯), “BASIC TOXICOLOGY”, 藝軒, 1994

Moate TF, Lu C, Fenske RA, Hahne RM, Kalman DA, “Improved cleanup and determination of dialkyl phosphates in the urine of children exposed to organophosphorus insecticides”, Journal of Analytical Toxicology, 1999, 23(4):230-236

Monohan K, Tinsley IJ, Shepherd SF, Field JA, “Quantitative-determination of the acidic metabolites of dacthal in ground water by strong anion-exchange solid-phase extraction”, Journal of chromatographic and food chemistry, 1995, 43(9):2418-2423

Moretto A, “Experimental and clinical toxicology of anticholinesterase agents”, Toxicology Letters, 1998, 102-103:509-513

Nutley BP, Cocker J, “Biological monitoring of workers occupationally exposure to organophosphorus pesticides”, Pesticide Science, 1993, 38:315-322

Quistad GB, Casida JE, "Sensitivity of blood-clotting factors and digestive enzymes to inhibition by organophosphorus pesticides", Journal of Biochemical & Molecular Toxicology, 2000, 14(1):51-56

Richter ED, Rosenvald Z, Kaspi L, Levy S, Gruener N, “Sequential cholinesterase tests and symptoms for monitoring organophosphate absorption in field workers and in persons exposed to pesticide spray drift”, Toxicology Letters, 1986, 33(1-3):25-35

Simcox NJ, Camp J, Kalman D, Stebbins A, Bellamy G, Lee IC, Fenske R, “Farmworker exposure to organophosphorus pesticide residues during apple thinning in central Washington State”, American Industrial Hygiene Association Journal, 1999, 60(6):752-761

Sultatos LG, “Mammalian toxicology of organophosphorus pesticides”, Journal of Toxicology & Environmental Health, 1994, 43(3):271-289

US EPA, “Pesticide Assessment Guidelines, Subdivision U-Applicator Exposure Monitoring”, U.S. EPA, Washington, D.C., 1987

Vale JA, “Toxicokinetic and toxicodynamic aspects of organophosphorus (OP) insecticide poisoning”, Toxicology Letters, 1998, 102-103:649-652

Vander AJ, Sherman JH, Luciano DS, “The Kidneys and Regulation of Water and Inorganic Ions”, in Vander AJ, Sherman JH, Luciano DS, (ed.) “Human Physiology”, fifth ed., McGraw-Hill Publishing Company, New York, USA, 1990, 471-511

Vasilic Z, Stengl B, Drevenkar V, “Dimethylphosphorus metabolites in serum and urine of persons poisoned by malathion or thiometon”, Chemico-Biological Interactions, 1999, 119-120:479-487

Willems JL, De Bisschop HC, Verstraete AG, Declerck C, Christiaens Y, Vanscheeuwyck P, Buylaert WA, Vogelaers D, Colardyn F, “Chloinesterase reactivation in organophosphorus poisoned patients depends on the plasma on centrations of the oxime pralidoxime methylsulphate and of the organophosphate”, Archives of Toxicology, 1993, 67(2):79-84

吳鴻奇, 利用薄膜型陰離子交換樹脂分析水中及尿液樣品中微量有機磷農藥代謝指標物之研究, 成功大學化學研究所碩士班碩士論文, 1998

國科會, 永續台灣評量系統-經濟壓力組, 2001, 於2004,9,20瀏覽
(http://www.law.ntu.edu.tw/sustain/SDIWEB/frameset5.htm)

農業雜誌, 農藥對土壤及地下水之污染探討, 2002, 於2004,9,20瀏覽
(http://www.egabio.com/admin/newshow.asp?pno=17)

蕭志杰, 尿液中有機磷農藥代謝指標物分析方法改善之研究, 成功大學化學研究所碩士班碩士論文, 2001
論文全文使用權限
  • 同意授權校內瀏覽/列印電子全文服務,於2005-02-04起公開。
  • 同意授權校外瀏覽/列印電子全文服務,於2005-02-04起公開。


  • 如您有疑問,請聯絡圖書館
    聯絡電話:(06)2757575#65773
    聯絡E-mail:etds@email.ncku.edu.tw