臺灣博碩士論文加值系統

基質輔助雷射脫附游離質譜法（MALDI/TOF）是現代生物質譜分析技術中，極為靈敏且重要的偵測方法，使用此法可在短時間內偵測到各式大小分子分析物的訊號，因此已成為現代蛋白體質譜學不可缺少的分析儀器。本研究將以此技術來偵測糞便中的潛血及殘餘血跡中主要蛋白質訊號-hemoglobin，由於此蛋白質溶於水的特性，所以利用微量水來萃取糞便及殘餘血跡中的hemoglobin。研究結果發現其偵測糞便中的潛血極限可到達0.1μg/100mg糞便，而鑑定殘餘血跡的極限則可到達血跡以純水稀釋到106倍時，仍有明顯來自血跡中蛋白質的質譜訊號，這比一般傳統化學試劑的靈敏度高10-100倍，且能避免因環境干擾所產生的偽陽性反應。
此外經過半年後稀釋10倍的血液滴在布上，使用萃取溶劑萃取血跡，以MALDI/TOF分析仍能清楚看見蛋白質的質譜訊號，且各別不同生物物種的血液，也可快速得到鑑別，甚至是混和不同物種的血液，再以MALDI/TOF快速分析下亦可輕易得到各該混和物種的訊號，也可以將血液先以酵素消化後，再結合資料庫比對，進行不同物種血液的鑑定，則其準確度可達百分之百，這顯示MALDI/TOF的確可有效應用在糞便潛血的檢驗及刑案中微量血跡的快速鑑識上。

Abstract
In this paper we demonstrate that matrix-assisted laser desorption ionization/time-of-flight (MALDI-TOF) mass spectrometry can be used to rapidly detect occult blood in human feces. The water-soluble hemoglobins, which are the most predominant proteins found in blood, were used as protein biomarkers for the presence of occult blood. The proteins were best extracted from feces using a small amount of pure water and then separated from the feces through centrifugation. We found that saturated 4-hydroxy-α-cyanocinnamic acid (α-CHC) solution was the best MALDI matrix for hemoglobin analysis; this matrix was mixed with the aqueous solution extracted from the feces at a volumetric ratio of 1:1. Although the MALDI-TOF MS results obtained using real samples were consistent with those provided through conventional chemical analysis, the detection limit of occult blood (0.1–1 μg/mg of feces) in feces when using MALDI MS was approximately 10–100 times lower than that of the conventional approach. In addition, materials that commonly interfere with the conventional chemical assaying of occult blood in feces—such as foods containing animal blood—do not interfere with the detection of the hemoglobin biomarkers during MALDI-TOF MS analysis.

目 錄
論 文 摘 要
目 錄
圖 目 錄
表目錄
壹、緒論
一、前言：
二、質譜法的發展：
三、基質輔助雷射脫附游離質譜法（MALDI)
(1)有效吸收雷射能量
(2)隔絕分析物
(3)提供氫離子給分析物
四、大腸癌之簡介（Introduction of Colorectal Cancer）
五、大腸癌之癌化分期（The TNM Staging of Colorectal Cancer）
六、大腸癌之臨床症狀及診斷 （The clinical staging and diagnosis of Colorectal Cancer）七、血跡型態及血跡鑑定
（Bloodstain pattern and Bloodstain identification）
(1)血跡型態
(2)血跡鑑識方法
(3)螢光顯現法：
(4)DNA鑑定法：
八、血液的組成與功能
九、論文目標
第一部份：以基質輔助雷射脫附游離法檢驗糞便潛血
第二部份：以基質輔助雷射脫附游離法快速鑑定殘餘血跡
貳、實驗
一、儀器裝置
1. 基質輔助雷射脫附游離飛行時間質譜儀 ( MALDI-TOF MS )
2. 迴旋式震盪器
3. 超音波震盪器
4. 高速離心機
二、樣品的製備及分析流程：
參、結果與討論
第一部份：以現代質譜法快速的檢驗糞便潛血
第二部分：以基質輔助雷射脫附游離法進行殘餘血跡的快速鑑定
肆、結論
伍、參考文獻

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