生化大分子的檢測常需要先製造特異性、生物親和性界面來捕捉目標物質或與目標物種反應。多數電化學式生醫感測器的研究著重於強調其功能性，例如敏感度及特殊性，以反覆摸索的實驗結果作為經驗來設計或修飾電極，然而缺乏一個遵循的準則而導致作法不一，修飾時間及藥品用量過度使用也造成製程上不必要的浪費。因此電化學電極界面量測的基本科學逐漸受到重視，其中衰減全反射式表面增強紅外吸收光譜(ATR-SEIRAS)具有高靈敏度、金屬表面上僅奈米級範圍的界面訊號檢測、時空分辨性、非破壞性檢測生物分子等優點而具有發展的吸引力，並且可應用在跨技術結合的現場臨場分析。本研究主要針對組胺酸、組織胺修飾於金表面作探討，利用時辨光譜記錄乙醇中的11-巰基十一烷酸(11-MUA)自組裝單分子膜修飾、並在2-嗎啉乙磺酸(MES)、4-羥乙基哌嗪乙磺酸(HEPES)等緩衝溶液記綠1-乙基-(3-二甲基氨基丙基)碳醯二亞胺(EDC)及N-羥基硫代琥珀醯亞胺(NHSS)活化及與胺形成醯胺鍵接合的過程，分析與討論其在金電極表面的特徵光譜並作為探討修飾層的依據。
11-MUA自組裝單分子膜修飾僅需數分鐘內即可完成化學性吸附，但碳氫骨架的規則排列可提高膜的覆蓋性與主導該步驟的修飾的反應時間，透過羧酸根(COO)的非對稱伸縮振動的特徵波峰可用來評估單分子膜的排列時間作為評估修飾時間的依據。EDC及NHSS活化的反應過程中，NHSS活化酯的特徵波峰可作為評估活化步驟的修飾，本研究也對EDC酯與NHSS酯的特徵波峰作區別。最後對組胺酸及組織胺修飾的特徵光譜進行探討。

With the development of biotechnology, the detection of macromolecules needs to fabricate a bio-affinity interface to capture or react with the target species. It should be noted that the fabrication of these modified layer often depended on the experience of researchers, and the characteristic of these biosensors were still trial and error. A novel technique is desired to investigate the structure and adsorbed orientation of these layers. ATR-SEIRAS has the advantages of high sensitivity, nondestructive test for biological molecules, nanoscale range of interface signals detection and high spatio-temporal resolution. In this study, The study is interested in the SAMs modification on gold surface. It made time-resolved SEIRAS to investigate the modification of 11-MUA SAMs in ethanol, the activation of EDC and NHSS in MES buffer, and covalent attachment of amide bonds in HEPES buffer, which can be clearly discussed to provide fundamental information for further biosensor application. The chemical absorption of 11-MUA SAMs completed within a few minutes. However, the regular arrangement of the hydrocarbon backbone is related to coverage, which lead to the modified time. The peak of the asymmetric stretch of COO can be used to evaluate the modified time is around 4 hours. The activation time is also done by the stretch of C=O of NHSS activate ester, which for 10 mM EDC and 2 mM NHS is about 50 minutes. Finally, The modification of histidine and histamine are also be investigated.

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