摘要 傳統在複雜蛋白質的分離以及定序的分析技術中有著儀器設備昂貴，分析時間過長，分離條件限制較多，分離產物不純等等問題，而近年來質譜技術的進步，可改善這個問題。 質譜儀是偵測質荷比(mass to charge ration, m/z)來決定分析物分子量的技術，相較於其他的質譜分析有著基質(matrix)干擾與分析物帶有多電荷等問題，本實驗發展新一代高靈敏奈米層狀矽基材 (nanofilament Silicon , nSi)質譜分析晶片來改善這些問題。進一步本研究利用表面改質之方式將nSi質譜分析晶片分別處理成疏水性(hydrophobic)、親水性(hydrophilic)、正電性(cation exchange)、負電性(anion exchange)、固定化金屬親和性 (IMAC) 等不同表面特性的質譜分析晶片。如此 nSi 質譜分析晶片表面能對分析物做純化與分離，進而增加質譜分析之選擇性 (selectivity) 以及靈敏度 (sensitivity)。 從實驗中，質譜的分析數據以及表面的化學鑑定來看，本研究所建構之nSi質譜分析晶片： 1. 隨著蝕刻秒數之增加，奈米結構的表面變較粗糙，使得質譜訊號隨之提升，但卻有一定之極限。 2. 不同表面特性之晶片在經過化學分析電子儀(ESCA)分析後，得到其相對應之元素訊號(如:在疏水性晶片上所得到之碳訊號)比之未經過化學改質之晶片所得到之訊號都有增加。 3. 不同特性之胜肽在其相對應的表面所得之質譜測試訊號，比之未經過化學改質之晶片都有明顯之提升。 Abstract Some problems exist in the conventional separation of complex protein mixtures and the sequence analysis technique. These problems include expensive equipment, extremely long analysis time, restrictive separation conditions, and impure separation results. Employing advanced mass spectrometry techniques improves these issues. Mass spectrometry is a technology that determines the molecular weight of the analyte by detecting its mass-to-charge ratio. Compared to other mass spectrometry methods that has matrix interference and analyte with multiple charges. To develop the new nSi mass-analytical chip will improve these problems. This study utilizes surface modification to process the chip surface and forms different nSi mass-analytical chip, such as hydrophobic、hydrophilic、cation exchange、anion exchange、and metal affinity (IMAC). This approach enables analyte purification and separation, increasing the selectivity and sensitivity of mass spectrometric analysis. According to mass spectrometry (MS) analytic data and chemical surface identification in the experiment, the nSi mass-analytical chip established in this study was characterized as the following: 1. As the etching time increased, the nano-structured surface of the mass spectrometric analysis chips became coarser. MS signal intensity subsequently increased, but remained under a specific level. 2. After the analysis of ESCA (electron spectroscopy for chemical analysis), chips with different surface properties obtained more corresponding element signals (e.g., a carbon signal received on a hydrophobic chip) than chips without chemical modification. 3. Chips after chemical modification with different properties peptides could get higher signal intensity than chips without chemical modification.
