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研究生: 雷秀玲
論文名稱: 飛行時間式二次離子質譜術於大腸桿菌訊號增強、防油紙中防油成分鑑定及早期胃癌診斷之研究
Time-of-flight secondary ion mass spectrometry study of enhancing Escherichia coli signals, identifying greaseproof agents in greaseproof papers and diagnosing early gastric cancer
指導教授: 凌永健
口試委員: 黃賢達
林立元
傅明仁
李正中
學位類別: 博士
Doctor
系所名稱: 理學院 - 化學系
Department of Chemistry
論文出版年: 2013
畢業學年度: 101
語文別: 英文
論文頁數: 117
中文關鍵詞: 飛行時間式二次離子質譜術氧化鋅奈米粒子大腸桿菌全氟多氟胃癌代謝體學
外文關鍵詞: Time-of-flight secondary ion mass spectrometry
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  • 飛行時間式二次離子質譜術使用於工業材料與生醫研究是一個已確立的技術,其所提供的化學資訊如元素和分子離子相當豐富,簡化和解釋二次離子質譜數據對於未來的應用是重要的。基於這個概念,我們提出以飛行時間式二次離子質譜術於大腸桿菌訊號增強、防油紙中防油成分鑑定及早期胃癌診斷之研究。
    穩定的膠體奈米粒子如金和銀奈米粒子已經研究在使用於增強二次離子訊號。半導體氧化鋅奈米粒子具有化學和熱穩定之獨特性與寬的能帶(3.37 電子伏特),促使我們探討使用氧化鋅奈米粒子增強模式生物:大腸桿菌的二次離子訊號的可行性。實驗過程為首先將經過冷凍乾燥後的大腸桿菌菌落上頭沉積氧化鋅奈米粒子,製備完成的試片接著進行飛行時間式二次離子質譜分析(使用鎵一次離子源),我們評估增強的訊號為質荷比688 (PE C32:1)和質荷比719 (PG C32:1),皆是來自於脂質膜。初步結果顯示氧化鋅奈米粒子沉積在大腸桿菌可大約增強3倍訊號。
    全氟和多氟化合物(簡稱PFCs)是人造化學品並且廣泛用於工業與消費性產品,包括防油紙。由於缺乏了解PFCs組成、毒性和裂解產物,因此需要開發新的分析方法來鑑定PFCs。在此,我們提出使用飛行時間式二次離子質譜術用於快速和便利鑑定防油紙中的PFCs。特徵的碳氟類離子和含磷酸基的碎片離子可使用飛行時間式二次離子質譜(使用鎵一次離子源)做偵測且質荷比可高達800。飛行時間式二次離子質譜圖有助於推論可能的防油劑分子結構為14:2/10:2 S-diPAPs。PFCs厚度藉由使用角度解析X光光電子光譜儀估計小於1奈米,並且從CF3/CF2的強度比值證明CF3官能基優先垂直於並接近防油紙表面。總結這部分研究,飛行時間式二次離子質譜術提供幾乎無樣品製備與快速鑑定PFCs的優點,對於監控新穎的PFCs和其降解產物是重要的分析技術。
    代謝體學包含廣泛、同時和有系統性的鑑定生物系統中的小分子內生性代謝物。飛行時間式二次離子質譜成像術有能力偵測胃癌組織中的有機物和無機物分布,具有偵測早期胃癌的潛力,能增加成功處理的機會。在此,我們收集病人的胃組織樣品,其病理分期從早期到晚期為:IA期(病人1),II期(病人2)和IIIA期(病人3和病人4)。飛行時間式二次離子質譜分析(使用25 kV鎵離子,1pA脈衝電流)首先得到500 × 500 微米平方面積的離子影像,再將胃黏膜層經過選區後的影像轉換成離子強度即可做比較。主成分分析結果顯示可區別癌與正常組織。在癌組織統計出低訊號強度的鐵、磷脂醯膽鹼、磷脂醯乙醇胺、尿酸、硫和碳氮與高訊號強度的丙酸、蘋果酸、檸檬酸和棕櫚酸。本研究進一步探討胃的成癌生理機制發現無機鐵可能與其他9種有機成分無關。


    Page Chapter 1 Introduction 1.1 Time-of-flight secondary ion mass spectrometry 1-1 1.1.1 Principal 1-1 1.1.2 Resolution 1-1 1.1.3 Modes of operation 1-3 1.1.4 Instrumentation 1-4 1.2 Environmental applications of ToF-SIMS 1-4 1.3 Biomedical applications of ToF-SIMS 1-5 1.3.1 Sample preparation 1-5 1.3.2 Data processing 1-6 1.4 References 1-7 1.5 Figures 1-10 Chapter 2 ZnO nanoparticles enhancing secondary ion signals of Escherichia coli analyzed by time-of-flight secondary ion mass spectrometry 1.1 Introduction 2-1 2.1 Experimental section 2-2 2.2.1 Reagents 2-2 2.2.2 Synthesis and characterization of ZnO nanoparticles 2-2 2.2.3 E. coli sample preparation 2-3 2.2.4 ToF-SIMS analysis 2-3 2.3 Results and discussion 2-3 2.3.1 ToF-SIMS surface mass spectra 2-3 2.3.2 Enhancement mechanism 2-4 2.4 Conclusions 2-5 2.5 References 2-6 2.6 Figures 2-7 Chapter 3 Rapid identification of trimethyl and triethyl amines using sulphonic acidic ionic liquids: A time-of-flight secondary ion mass spectrometry study of fragmentation reactions 3.1 Introduction 3-1 3.2 Experimental section 3-3 3.2.1 General 3-3 3.2.2 Synthesis 3-3 3.2.3 Sample preparation 3-4 3.2.4 Time-of-flight secondary ion mass spectrometry analysis 3-4 3.3 Results and discussion 3-5 3.3.1 ToF-SIMS spectra of [TMBSA][HSO4] 3-5 3.3.2 ToF-SIMS spectra of [TEBSA][HSO4] 3-7 3.4 Conclusions 3-8 3.5 References 3-10 3.6 Figures 3-13 Chapter 4 Rapid identification of per- and polyfluorinated compounds in greaseproof papers by time-of-flight secondary ion mass spectrometry and related surface analysis methods 4.1 Introduction 4-1 4.2 Experimental section 4-3 4.2.1 Chemicals and reagents 4-4 4.2.2 Sample collection and preparation 4-4 4.2.3 Surface and in-depth analysis by ToF-SIMS 4-4 4.2.4 Quantitative analysis by LC-MS/MS 4-5 4.2.5 Surface and in-depth analysis by XPS and ARXPS 4-6 4.2.6 Suafce roughness analysis by optical microscope 4-6 4.2.7 Water contact angle analysis by optical contact angle meter 4-6 4.2.8 Vibrational spectroscopy analysis by ATR FTIR 4-6 4.3 Results and discussion 4-7 4.3.1 ToF-SIMS spectra 4-7 4.3.2 Thickness and orientation of PFCs 4-10 4.3.3 Thickness and orientation of PFCs 4-11 4.4 Conclusions 4-12 4.5 References 4-14 4.6 Tables 4-17 4.7 Figures 4-19 4.8 Supplementary Materials 4-27 Chapter 5 Early detection of gastric cancer using time-of-flight secondary ion mass spectrometry metabolomics imaging 5.1 Introduction 5-1 5.2 Experimental section 5-3 5.2.1 Sample preparation 5-3 5.2.2 ToF-SIMS imaging analysis 5-3 5.2.3 Data preprocessing 5-4 5.2.3 Principal component analysis and descriptive statistics 5-4 5.3 Results and discussion 5-4 5.3.1 Histopathology of human gastric tissues 5-5 5.3.2 ToF-SIMS imaging and ROI mass spectra analysis 5-5 5.3.3 PCA results 5-5 5.3.4 Descriptive statistics and biological mechanism 5-6 5.3.5 The relation of metabolites and inorganic Fe 5-8 5.4 Conclusions 5-9 5.5 References 5-10 5.6 Tables 5-12 5.7 Figures 5-15 5.8 Supplementary Materials 5-21 Chapter 6 Summary and Perspective 6-1

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