人體膽固醇的含量與心血管疾病息息相關，當體內的膽固醇超過一定含量時，會在心臟或腦的血管壁上累積，使血液流動遭受阻塞造成病變，所以即時監測膽固醇含量是相當重要的課題。膽固醇與膽固醇氧化酶反應會產生副產物雙氧水，因此本研究利用金屬奈米粒子增加催化雙氧水的能力，製備出高靈敏度的雙氧水感測器，另一方面藉由酵素具良好的專一性，將膽固醇氧化酶修飾於電極，建立具高精確度、高選擇性且實際應用的電流式酵素型膽固醇感測器。
本研究主要分為兩個部分：第一部分為製備金屬奈米粒子，並應用於雙氧水感測器。根據文獻指出，相較於球形奈米粒子，多邊形的奈米粒子具有較多的高活性低配位原子，能提供較多的催化活性位置，故本研究合成二十四面體金奈米粒子 (trioctahedral AuNCs, 24-AuNCs)、多面體金奈米粒子 (multilateral AuNCs, m-AuNCs) 及立方體氧化亞銅奈米粒子 (cubic Cu2O, c-Cu2O)，同時為了進一步提高催化雙氧水的能力，在奈米粒子表面合成奈米鉑微粒，形成以少量鉑合成之核殻式 (core-shell) 奈米金鉑或奈米銅鉑粒子，由掃描式電子顯微鏡 (SEM) 及穿透式電子顯微鏡 (TEM) 觀察奈米粒子的形狀及表面型態，再利用紫外光-可見光光譜 (UV-vis) 及電化學酸處理結果確認鉑還原於奈米金表面，達成核殼式的奈米金鉑結構。最後將合成好之金屬奈米粒子修飾於電極，比較催化雙氧水的能力，其中以核殼式二十四面體金鉑奈米粒子的效果最佳，具有低偵測極限 (10 μM)、線性範圍 (0.01 – 6.0 mM)，及靈敏度 (915 μA/mM cm2, R2: 0.99)。本研究第二部分為將雙氧水感測方法應用於膽固醇感測器，由於膽固醇不溶於水，所以必須使用Triton X-100及isopropanol兩種溶劑，研究中發現Triton X-100不僅具有電化學訊號，且會破壞金屬奈米粒子之修飾層，本研究利用具有良好的生物相容性的甲殼素 (chitosan)，於pH值約為5.5時成膜的特性製備抗干擾層，將酵素層、保護奈米粒子層及抗干擾層透過電沉積的方法，以單一步驟修飾於電極表面，此方法不僅可以節省電極製備時間、增強機械強度且再現性高，透過最佳化電沉積時間，本研究成功阻擋Triton X-100的干擾，製備具有合適線性範圍(0.25 – 5.0 mM)及靈敏度(1.52 μA/mM cm2, R2: 0.97) 的膽固醇生物感測器。

The content of cholesterol in human related closely to the cardiovascular diseases that the monitoring of the cholesterol level is of particular importance. The reaction of cholesterol and cholesterol oxidase (ChOx) is a redox reaction which produces hydrogen peroxide (H2O2) can could be quantified by measuring the electrochemical response. The goal of this research is to establish sensing system for the detection of cholesterol with high selectivity and sensitivity, based on amperometric analyses. Previous studies indicated that the catalysts with polyhedral structure provide high index facets and low coordination atoms showing higher activity than the spherical conformation ones. In order to promote the electrochemical responses, various nanoparticles were synthesized and added on the prepared electrodes for sensing H2O2 and cholesterol.
This study is comprised of two parts: the first part focused on the synthesis of metallic nanoparticles with polyhedral structure. Three types of nanoparticles were synthesized: trioctahedral AuNCs (24-AuNCs), multilateral AuNCs (m-AuNCs), and cubic Cu2O (c-Cu2O). Various analytical methods were employed to verify the physico-chemical and morphology characteristics of the prepared nanoparticles. Moreover, in order to further enhance the catalytic ability for sensing, the metallic nanoparticles with an platinum outer layer was prepared which provides a particular advantage of only utilizing a small amount of platinum and the nanoparticles are termed as core-shelled nanoparticles. The catalytic ability for the different metallic nanoparticles was firstly compared by the sensitivity of sensing H2O2. The results showed that 24-Au@Pt-NCs exhibited the highest sensitivity of 915μA/mMcm2, with a linear range for detecting H2O2 from 0.01 to 6.0 mM (R2: 0.997), and a detection limit of 10μM.
For the second part, the prepared nanoparticles and electrodes were applied for the detection of cholesterol where Triton X-100 and isopropanol were added to increase the solubility of cholesterol. Because the addition of Triton X-100 caused significant interference and the decomposition of nanoparticles, a protection layer on the top of the electrode, applying electroplating to incorporate chitosan and enzymes in a single step procedure, was further added. The combination of the protection layer with nanoparticles showed good mechanical strength for resisting the detachment of enzymes and nanoparticles as well as effectiveness in sample preparation. The resultant cholesterol sensor resists well from the interference of Triton X-100 by the optimized electroplating time and shows suitable linear range of 0.25 - 5.0 mM with a sensitivity of 1.52 μA/mM cm2 (R2: 0.97).

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