系統識別號 U0026-2407201816200700
論文名稱(中文) 利用非標的代謝體分析方法找尋老鼠尿液中毒物暴露的生物指標
論文名稱(英文) Untargeted metabolomics analysis of toxicant exposed rat urines for biomarker discovery
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 106
學期 2
出版年 107
研究生(中文) 黃凱盈
研究生(英文) Hoi Ieng Wong
學號 s76045016
學位類別 碩士
語文別 英文
論文頁數 52頁
口試委員 指導教授-廖寶琦
中文關鍵字 生物指標  鄰苯二甲酸二丙基庚酯  代謝體學  尿液  正交偏最小平方判別分析 
英文關鍵字 biomarker  DPHP  metabolomics  urine  OPLS-DA 
中文摘要 鄰苯二甲酸二丙基庚酯(DPHP)是聚氯乙烯中帶有毒性的增塑劑,常被用於商業性的產品,例如電纜和鞋子等。本論文主要目的是利用質譜儀和正交偏最小平方判別分析(OPLS-DA)找出人類暴露DPHP後新的生物指標。首先,在第一階段找尋DPHP暴露的生物指標中,分別餵食Wistar大鼠DPHP兩個口服劑量300和1200 mg/kg body weight各15隻,單獨把大鼠放置在代謝籠中收集其24小時後的尿液樣品,利用固相萃取,超高效液相層析儀串聯質譜儀進行尿液樣品分析,而質譜訊號則是利用正交偏最小平方判別分析(OPLS-DA)建構劑量為兩個口服劑量300,1200 mg/kg body weight兩樣本群差異之模型,再進一步通過S-Plot鑑定DPHP暴露的生物指標。在第一階段總共找出31個可能為DPHP暴露的生物指標。在第二階段為了驗證從第一階段中找到DPHP可能暴露的生物指標,會利用大鼠模型進行驗證,分別餵食Wistar大鼠DPHP五個口服劑量0,150, 300,600和1200 mg/kg body weight各3隻。結果顯示在第一階段找尋DPHP暴露的生物指標中全部31個DPHP暴露指標在大鼠模型中都有劑量效應的關係,而這31個DPHP暴露的生物指標中,其中28個生物指標在過往的文獻中並沒有被報導,這些新的DPHP暴露生物指標可以進一步研究其在人體暴露評估中的應用。
英文摘要 Di-(2-propylheptyl) phthalate (DPHP) is a toxic plasticizer in polyvinyl chloride and it is used in commercial applications such as cables and shoes. The aim of this study is to discover biomarkers for human DPHP exposure assessment using mass spectrometry based-metabolomic approach and orthogonal partial least squares discriminant analysis (OPLS-DA). In phase I (exposure biomarker candidate discovery), OPLS-DA was used to model the difference of two urine sample groups collected from rats orally administered with 2 different DPHP doses of 300 and 1200 mg/kg body weight (n=15, respectively). The urine samples were analyzed by solid phase extraction-ultra performance liquid chromatography-high resolution mass spectrometry (SPE-UPLC-HRMS). Identification of biomarker candidates for DPHP exposure was achieved by S-plot. As a result, 31 exposure biomarker candidates were identified. In phase II (exposure biomarker validation), those 31 candidates were validated as exposure biomarkers by examining dose-response relationship revealed by rat urine samples collected from rats orally administered with 5 different DPHP doses 0, 150, 300, 600, and 1200 mg/kg body weight (n=3, respectively). Out of 31 validated biomarkers, 28 biomarkers have not been reported in the previous literature. These novel DPHP exposure biomarkers can be further investigated for their applications in human exposure assessment.
論文目次 Content
摘要 I
Abstract II
Abbreviation VII
Chapter 1 Overview of the literature 1
1-1 Phthalate 1
1-2 Phthalate toxicity 1
1-3 Phthalate exposure pathway 2
1-4 Di-(2-propylheptyl) phthalate (DPHP) 2
1-5 The usage of DPHP 3
1-6 The toxicity of DPHP 4
1-7 The metabolites of DPHP 5
1-8 The strategies of exposure marker searching 7
1-8-1 Orthogonal partial least squares discriminant analysis (OPLS-DA) 7
Chapter 2 Objective 7
Chapter 3 Materials and methods 9
3-1 Research scheme 9
3-2 Research materials and methods 11
3-2-1 DPHP rat urine collection and pretreatment 11
3-2-2 High resolution liquid chromatography mass spectrometry analysis 12
3-3 Structure identification of DPHP exposure biomarkers 15
3-3-1 UPLC- MS/MS analysis 15
3-3-2 Identification of exposure biomarker structure 17
3-4 Data analysis 14
3-4-1 Data processing 14
3-4-2 Exposure biomarker identification using OPLS-DA 15
3-4-3 Exposure biomarker validation using Spearman correlation coefficient 15
Chapter 4 Result & Discussion 18
4-1 DPHP metabolite biomarkers discovery using OPLS-DA 18
4-1-1 OPLS-DA filter efficiency 18
4-2 DPHP exposure marker candidates discovery using OPLS-DA 22
4-3 Validation of DPHP exposure marker candidates 22
4-3-1 31 DPHP exposure biomarkers signal abundance in rat urine 25
4-4 Chemical structure identification of DPHP exposure biomarkers 27
Chapter 5 Conclusions 39
Reference 41

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