馬一 中臺灣大學：環境衛生研究所高佩鈴Kao, Pei-LinPei-LinKao2007-11-282018-06-302007-11-282018-06-302007http://ntur.lib.ntu.edu.tw//handle/246246/59792呼氣分析為非侵入性之生物偵測方法，呼出氣體中戊烷被視為氧化壓力指標如同尿液中8-hydroxy-2’-deoxyguanosine (8-OHdG)。抽菸會明顯增加人體氧化壓力，因此本研究以年長抽菸者為對象，觀察抽菸對呼出氣體揮發性有機物和尿液中8-OHdG的影響。 本研究為縱貫式方法，收集受試者（8位抽菸者,8位非抽菸者）呼出氣體和尿液樣本每天早上一次，並紀錄抽菸者每天的抽菸量、上次抽菸的時間、喝酒習慣、服用維他命習慣。另外針對抽菸者採集長時間禁菸下（至少7小時）的呼出氣體。呼氣樣本用採樣袋收集，以複合床熱脫附採樣管來濃縮呼出氣體中揮發性有機物，運用熱脫附儀配合氣相層析質譜儀來進行分析。尿液樣本以酵素免疫吸附分析8-OHdG濃度。統計方面採用混合型模式(Mixed-effect model)校正固定效應和隨機效應後，比較抽菸者和非抽菸者呼氣中揮發性有機物的差異。 結果顯示呼出氣體中揮發性有機物的含量於抽菸後快速上升，並受到年齡和服用維他命習慣的影響。利用模式校正這些干擾因子後，在65個VOCs中有26個物質在抽菸組和非抽菸組有顯著差異。抽菸組長時間禁菸下和非抽菸組的呼氣VOCs比較也有類似結果，並觀察到抽菸者呼氣中pentane含量依然比非抽菸者高。尿液中8-OHdG方面，抽菸組的含量比非抽菸組高出1.5倍，且和呼出氣體中戊烷有相關(r=0.55, p=0.06, n=12)。 呼出氣體VOCs的含量於抽菸後馬上增加並受到年齡和使用抗氧化物情形的影響。呼出氣體中戊烷和尿液中8-OHdG一樣作為氧化壓力的指標，其年長抽菸者含量都比非抽菸者來得高。Breath analysis is a non-invasive biological monitoring method. Exhaled air pentane has been used as a marker of oxidative stress as well as urinary 8-hydroxy-2’-deoxyguanosine (8-OHdG). Because cigarette smoking can significantly increase the oxidative stress, this study used older smokers as the subjects and observed the effect of smoking on exhaled breath volatile organic compounds (VOCs) and 8-OHdG in urine. A longitudinal study of the subjects （8 smokers, 8 non-smokers）was carried out with exhaled breath and urine sampling once every morning and recorded smokers’ daily cigarette consumption, time of the last smoking、alcohol intake and vitamin supplement. In addition, in smokers, exhaled breath was collected after cigarette cessation. Breath VOCs samples in Tedlar bags were preconcentrated with multi-bed sorbent sampling tubes and analyzed by thermal desorption-GC/MS. Urinary 8-OHdG was measured by ELISA. Mixed-effect models were used to investigate the breath VOCs difference between smokers and non-smokers by adjusting fixed effects and random effects. We observed that exhaled breath VOCs increased immediately after smoking and were influenced by age and vitamin consumption. Using model adjusting these factors, 26 of 65 breath VOCs showed significant difference between smokers and non-smokers. Similar results were measured in smokers after cigarette cessation compared to non-smokers. Exhaled breath pentane in older smokers was still higher than non-smokers. In urinary 8-OHdG, smokers had 1.5-fold higher level than non-smokers. There was a correlation between exhaled breath pentane and urinary 8-OHdG in smokers.（r=0.55, p=0.06, n=12）. In conclusion, cigarette smoking caused an immediate increase in exhaled breath VOCs which were also influenced by age and antioxidant supplement. Exhaled breath pentane as well as urinary 8-OHdG were used as a marker of oxidative stress and were increased in older smokers compared to non-smokers.摘要 1 Abstract 2 第一章 緒論 3 1.1 前言 3 1.2 研究目的 4 第二章 文獻探討 5 2.1 呼出氣體之研究 5 2.1.1 呼氣分析研究種類 5 2.1.2 呼氣中VOCs之來源 6 2.1.3呼氣檢測之特性 8 2.1.4呼氣採集方法 8 2.1.5 呼出氣體分析作為肺部疾病之生物指標 9 2.2 氧化壓力指標 10 2.2.1 氧化壓力 10 2.2.2 呼出氣體中氧化壓力指標 11 2.2.3 尿液中氧化壓力指標 11 2.3 抽菸 12 2.3.1 抽菸對氧化壓力的影響 12 2.3.2 抽菸對呼出氣體的影響 13 2.3.3 抽菸對尿液8-OHdG的影響 13 第三章 材料與方法 14 3.1 研究流程 14 3.2 研究設計 14 3.3 研究對象 15 3.4採樣方法 15 3.5 呼氣實驗器材與設備 15 3.5.1藥品、試劑及氣體 15 3.5.2呼氣實驗儀器 17 3.6呼氣樣本前處理 17 3.6.1採樣管之填充 17 3.6.2樣本保存 18 3.6.3採樣管調態(Conditioning) 18 3.6.4採樣管流量校正 18 3.6.5樣本前濃縮步驟 18 3.6.6採樣袋淨化 19 3.7呼氣分析步驟 19 3.7.1熱脫附儀上機前調態 19 3.7.2氣相層析質譜儀 19 3.7.3上機分析 20 3.8儀器分析條件 20 3.8.1熱脫附裝置 20 3.8.2低溫捕集器(Cryo-Focus Module, CFM) 21 3.8.3氣相層析儀 21 3.8.4質譜儀 22 3.9空白樣品分析 22 3.10重複樣品分析 23 3.11內標準品監測 23 3.12破出試驗 23 3.13定性、定量分析 23 3.14 尿液中8-OHdG 25 3.14.1 ELISA Kit 25 3.14.2 Kit內容物 25 3.14.3 分析步驟 25 3.15 統計分析 26 第四章 結果 27 4.1 QA/QC 27 4.1.1 空白試驗 27 4.1.3 破出實驗 27 4.1.2 重覆樣本 27 4.2 定性分析 28 4.3 人口學基本資料 31 4.4 重覆測量試驗 31 4.4.1 呼出氣體VOCs於抽菸者和非抽菸者的情形 31 4.4.2 抽菸影響呼出氣體VOCs的因子 36 4.4.3 呼出氣體VOCs於抽菸組和非抽菸組的差異 40 4.5 抽菸者長時間禁菸下和非抽菸者呼出氣體VOCs的差異 45 4.6 尿液中8-OHdG 52 4.7 氧化壓力指標和抽菸的相關性 56 4.7.1 呼氣中pentane 56 4.7.2 尿液中8-OHdG 56 第五章 討論 63 5.1 影響呼出氣體中VOCs的因子 63 5.2 內生性氣體和外生性氣體 64 5.3 長時間禁菸下抽菸者呼出氣體VOCs情形 65 5.4 尿液中8-OHdG 66 5.5 外生性氣體和氧化壓力指標的相關性 67 5.6 研究限制 69 5.7 結論 70 第六章 參考文獻 71765189 bytesapplication/pdfen-US抽菸呼出氣體揮發性有機物氣相層析儀混合型模式Cigarette smokingExhaled breathVolatile organic compounds(VOCs)GC/MSMixed effect modelthesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/59792/1/ntu-96-R94844010-1.pdf