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系統識別號 U0026-2201201318514800
論文名稱(中文) 拜香燃煙中多環芳香烴與重金屬污染物之生物毒性研究
論文名稱(英文) A study on the Cytotoxicity of Polycyclic Aromatic Hydrocarbons and Heavy Metal Species from Incense Smoke
校院名稱 成功大學
系所名稱(中) 環境工程學系碩博士班
系所名稱(英) Department of Environmental Engineering
學年度 101
學期 1
出版年 102
研究生(中文) 黃泓鈞
研究生(英文) Hung-Chun Huang
學號 P56991197
學位類別 碩士
語文別 中文
論文頁數 74頁
口試委員 指導教授-林達昌
口試委員-李文智
口試委員-王應然
口試委員-陳瑞仁
中文關鍵字   多環芳香烴  重金屬  懸浮微粒  細胞毒性 
英文關鍵字 incense  PAHs (polycyclic aromatic hydrocarbons)  heavy metals  particulates  cytotoxicity 
學科別分類
中文摘要 華人地區常以燒香(拜拜)來表達對神明與祖先的敬重,但拜香燃燒時會產生大量的懸浮微粒,這些微粒表面會吸附許多有機及無機的化學物質,其中有機目標污染物以多環芳香烴為主,無機部分則為重金屬,而這些化學物質會刺激人體肺部系統,進而可能演變成癌症,根據研究指出人們一天約有80%的時間待在室內,且國人在家亦有燒香之習慣,故拜香燃煙儼然成為台灣室內新興空氣污染源。
本研究利用燃燒室模擬台灣室內環境,以了解拜香於室內空間燃燒之微粒特性,並將所收集到之微粒以傳統萃取方法萃取微粒中多環芳香烴 (PAHs)成分及重金屬成分,且本研究考慮到人類暴露於空氣污染物,首當其衝之器官為肺部呼吸器官,故本研究以人類支氣管上皮細胞株(BEAS-2B cell line)為目標細胞進行毒性測試,包括MTT assay及Comet assay,希冀以化學分析及毒性測試結果,解析拜香微粒潛在毒性特徵。
實驗結果顯示,新山香、沉香、料香懸浮微粒排放係數依序為41.3、19.02、16.05 mg/g-incense,新山香相較於其它兩種香種約高出兩倍以上。而拜香微粒中重金屬部分,以對人體較無害之K、Na、Ca等金屬元素為主,其金屬萃取物對細胞並無毒性反應,但拜香微粒中PAHs部分卻有明顯毒性反應(細胞存活率平均為50.33±15.2%),而各香種固相PAHs排放係數以沉香最高,其次為新山香、最低者為料香,其值依序為8.90±1.11、8.17±0.78 、4.51±0.43 µg/g-incense,本研究進一步探討PAHs之毒性當量與細胞存活率之關係,發現兩者呈現高度負相關性(r= -0.87),此外基因毒性測試結果顯示,拜香微粒中PAHs部分會對細胞造成DNA damage,此實驗結果亦與細胞存活率測試結果相符合。
綜合以上結論吾人可推知拜香微粒生物毒性的主要貢獻來源可能來自於PAHs,因此未來在拜香燃煙固相汙染物的議題上,建議應多關注PAHs之排放量。
英文摘要 Worshipping ancestors and gods by burning incense sticks has been one of the important traditions in many Asian nations. However, burning incense can produce a great amount air particulates. In addition, the surface of the particulates adsorbes many organic (e.g., polycyclic aromatic hydrocarbons, PAHs) and inorganic chemicals, (e.g., heavy metals).These adsorbed chemicals, once enter the human respiratory system, can stimulate human lungs, and possibly induce cancer. According research, people spend approximate 80% of day in the indoors space, and many Chinese burn incenses daily, so incenses smoke has long time been an indoor pollution in Tawan.
Our research use a combustion chamber to simulate the indoor environment. After collecting the combustion products, a traditional extract method was used to extract PAHs and heavy metals of incense particulate, and to investigate the toxicological characteristics of incense particulates. Our Considering that the first target organ is the lungs when human are exposed to air pollution, we utilized normal human bronchial epithelial cell (the BEAS-2B cell line) as the target cell for biological toxicity tests, along with the MTT assay and the Comet assay. Hope chemical analysis result and biological toxicity result can help us to understand incense particulates toxicity characteristic.
Our results show that, in terms of particulate emission, Xin Shan incense, Chen incense, and Liao incense are respectively 41.3, 19.02, and 16.05 mg/g-incense. The emission from the Xin Shan incense stick is higher than both Chen incense and Liao incense. In the heavy metal part, all incense particulates mainly contain K, Na, Ca, which are harmless to human. The heavy metals found in the extract did not exhibit toxicity respond in cell tests. However, PAHs extract of incenses particulate clearly showed toxicity. The respond (average of cell viability is 50.33±15.2%), Chen incense is 8.90±1.11 µg/g-incense, Xin Shan incense is 8.17±0.78 µg/g-incense Liao incense is 4.51±0.43 µg/g-incense. Among the three incences Chen incense was the highest, and PAHs toxicity equivalency had a highly negative correlation with the cell viability (r= -0.87). In addition, genotoxicity test results show PAHs in the incenses particulate can cause DNA damage. This result was also consistent with the cell viability test result.
It is therefore concluded that the particulate toxicity of the incense smoke is mainly contributed by PAHs. It is suggested that we should be more concerned about the PAHs emission in the incense smoke.
論文目次 第1章 前言 1
1-1研究動機 1
1-2研究目的 2
第2章 文獻回顧 3
2-1室內空氣微粒毒性與拜香的相關介紹 3
2-1-1室內空氣微粒毒性 3
2-2-2拜香燃煙中PAHs與重金屬 8
2-3PAHs與重金屬之生物毒性 11
2-3-1 PAHs之生物毒性 11
2-3-2 重金屬之生物毒性 15
2-4生物毒性測試方法與相關研究 16
2-4-1體外暴露研究(In vitro study) 16
2-4-2 MTT Assay 17
2-4-3 彗星試驗(Comet Assay) 18
第3章 實驗方法與步驟 19
3-1實驗設計與規劃 19
3-2採樣設計與規劃 21
3-2-1採樣設備 21
3-2-2採樣設計 21
3-2-3 採樣地點 22
3-3採樣前準備 22
3-3-1 採樣濾紙製備/調理 22
3-3-2 濾紙前處理 22
3-3-3 矽膠與無水硫酸鈉之前處理 23
3-4樣品之PAHs萃取、分析及生物毒性樣品製備 23
3-4-1 萃取 23
3-4-2濃縮、淨化、再濃縮 24
3-4-3生物毒性樣品製備 25
3-5 樣品重金屬萃取、分析及生物毒性樣品製備 26
3-5-1微波消化 26
3-5-2感應偶和電漿原子發射光譜儀(ICP-OES)分析 28
3-6生物毒性 29
3-6-1細胞存活率測試(MTT assay) 29
3-6-2 彗星試驗 (Comet Assay) 35
第4章 實驗品質保證與品質控制 39
4-1重金屬分析程序之QA/QC 39
4-1-1 重金屬標準品檢量線之建立 39
4-2多環芳香烴類化合物分析程序之 QA/QC 40
4-2-1 溶劑空白試驗 40
4-2-2 固相PAHs樣本空白試驗 40
4-2-3 採樣之空白試驗 40
4-2-4PAHs標準品檢量線之建立 40
第5章 結果與討論 44
5-1 拜香燃燒之物理特徵 44
5-1-1拜香之基本物理參數 44
5-1-2拜香燃燒產生之微粒與香灰特徵 45
5-2拜香排放固相PAHs及重金屬濃度分析 47
5-2-1拜香排放固相PAHs濃度分析 47
5-2-2拜香排放固相重金屬濃度分析 52
5-3拜香燃煙中PAHs之生物毒性測試 56
5-3-1拜香燃煙中PAHs之細胞存活率測試(MTT assay) 56
5-3-2拜香燃煙中拜香燃煙中PAHs基因毒性測試結果 59
5-4拜香燃煙中拜香燃煙中重金屬之生物毒性測試 61
5-5小結 63
第6章 結論與建議 65
6-1結論 65
6-2建議 66
第7章 參考文獻 67
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