用於食品安全應用的柔性等離子體紙SERS基材的製備和優化

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研究生：	Menbere Leul Mekonnen Menbere Leul Mekonnen
論文名稱：	用於食品安全應用的柔性等離子體紙SERS基材的製備和優化 Fabrication and Optimization of Flexible Plasmonic-Paper SERS Substrate for Food Safety Applications
指導教授：	黃炳照 Bing Joe Hwang
口試委員:	周澤川 Tse Chuan Chou 今榮東洋子 Toyoko Imae 何國川 Kuo-Chuan Ho 王迪彥 Diyan Wang 蘇威年 Wein Nein Su
學位類別：	博士 Doctor
系所名稱：	工程學院 - 化學工程系 Department of Chemical Engineering
論文出版年：	2018
畢業學年度：	107
語文別：	英文
論文頁數：	144
中文關鍵詞：	SERS 、SHINERS 、Ag@SiO2 nanocubes 、功能性的Ag殼分離的納米粒子(Ag-SHINs) 、Fe-TiO2 Nanosheet 、分析物定位、敏感度、靈活性基質、濾紙、電介質修飾的紙、三聚氰胺阿斯巴甜、噻苯達唑
外文關鍵詞：	Functionalized-Ag-shell isolated nanoparticles (Ag-SHINs), , , ,, Fe-TiO2 Nanosheet, analyte localization, filter paper,, dielectric modified paper, aspartame, thiabendazole
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摘要
將SERS從實驗室內走向現實應用端，需要成本低及品質好的基材，如食品安全。柔性的基板由於易於製造且便於用在非平面的樣品上，因此可以擴展到SERS的應用上，並且處進對食品及其他商品的檢驗。本論文探討了柔性等離子試紙SERS基板的製備和優化，以及他們在食品安全中的應用。

在第一種方法中，使用多元醇方法合成sharp-edged Ag NCs，然後使用改進的Stober方式以超薄之二氧化矽殼層塗佈它。透過抽氣過濾所得之Ag SHINs組裝在微型紙平台上。這使得此便宜感測平台具有合理分配及奈米立體之粒子間距，與傳統的剛性材料相比，紙的纖維結構可以促進奈米立方體的組合，透過等離子體耦合可提高SERS活性。超薄二氧化矽外殼可維持Ag NCs的穩定性，並使基板能夠以SHINERS模式運行。用於評估三聚氰胺檢測分析，評估顯示線性度高達1 mg/L，檢測極限為60 µg/L。對於液態牛奶的檢驗極限為0.62 mg/ L，可檢測值低於實際樣品，表示對實驗樣品具有足夠的靈敏度。
第二種方法顯示了Ag SHINs 在等離子體紙基質上增強SERS表現的功能。這個功能能在等離子體基質上避免在感應體積內的分析局限與不足，這是經常威脅到SERS敏感度的問題。通常納米等離子體基質是通過連接影響SERS活性的主分子來發揮作用的。因此，我們提出了一種3D氨基修飾,有一層極薄的矽殼的Ag@SiO2 nanocubes 來偵測一個平滑SERS平台內的小分子。氨基酸基團的嵌入是將極薄的矽層用不同的氨基矽烷來進行矽烷化，可以在不犧牲Ag nanocube 的膠狀與化學穩定性的前提下提高它的敏感度。通過短鏈氨基矽烷來作用的Ag-SHINs 有著更好的SERS活性。氨基作用的方法使SERS信號提高了3倍，把無標籤的阿斯巴甜的偵測值降低至71 µg/mL ，其LOD值比沒有氨基作用的Ag-SHINs低了1.5倍。在測試有標籤的軟飲料及阿斯巴甜和葡萄糖混合物時，在基質的訊號選擇性及敏感度上有了很明顯的提高。SERS效能的提高主要歸因於氫鍵能誘導在感測體積內分析的定位。除此之外，小型化的紙平台和3D纖維結構也增強了熱點的密度與樣品收集效率。這個基質也可以擴展應用至其它有羧基和氨基酸的小分子，取代複雜的有生物標籤的偵測方法。
最後一個方法展現了由電介質NS修飾的等離子體紙基質的製造方法與其在食品安全上的應用。等離子體奈米顆粒與電介質平台間的相互作用會產生獨特的光學行為使等離子體/SERS作為一個基質的表現型更高。在本篇文章中，電介質修飾的等離子體紙SERS基質由把Ag@SiO2 nanocubes 放在Fe-TiO2 NS修飾的紙上來合成。Fe-TiO2 NS對可見光的反應能顯著地改變紙的光學性質，並作為Ag NC的電介質襯底。因此，從電介質膜反射回來的入射光與來組Ag NCs 的散射光耦合，導致空間電磁場的增強，從而改善SERS的表現。本文製備的電介質修飾的等離子體紙基質的敏感度降至1 pM R6G，而平均EF為1.49×107 。這個值比未修飾的等離子體紙要好很多，突出了電介質NS的耦合效應。這個基質展現了卓越的分析表現，測試噻苯咪唑的LOD值低至19 µg/L，比未修飾的等離子體紙的敏感度要高4倍多。在直接噴灑噻苯咪唑的蘋果上擦拭后進行測試，可識別的Raman訊號降至15 ppb, 證明了本基質在食品安全上的應用是可靠的。

Abstract
The translation of Surface Enhanced Raman Spectroscopy (SERS) from the laboratory to the real world applications such as in food safety demands inexpensive yet quality substrates suitable for point-of-need applications. Flexible substrates due to their ease of fabrication and convenience to sample nonplanar surfaces can expand the application of SERS and facilitate onsite inspection of food and other commodities. In this dissertation, the fabrication and optimization of flexible plasmonic-paper SERS substrates, as well as their application in food safety, are discussed.
In the first approach, sharp-edged Ag NCs are synthesized using the polyol method followed by coating them with ultrathin silica shell using a modified Stober method. The resulting shell isolated Ag NCs (Ag SHINs) are assembled on a miniaturized paper platform by suction filtration. This results in inexpensive sensing platform with reasonable distribution and interparticle spacing of nanocubes. Compared with the conventional rigid substrates, the fibrous structure of the paper promotes the assemblage of nanocubes rendering enhanced SERS activity through plasmon coupling. The ultrathin thin silica shell maintains the stability of the Ag NCs and enables the substrate to operate in SHINERS (Shell isolated Nanoparticle Enhanced Raman Spectroscopy) mode. Assessment of analytical performances of the substrate for melamine detection showed a good linearity up to 1 mg/L with a limit of detection of 60 µg/L, which is below the permissible residue limit signifying adequate sensitivity for real sample analysis with less sample treatment.
The second approach shows the role of functionalization of Ag SHINs in improving their SERS performance in a plasmonic–paper substrate. Functionalization of a plasmonic substrate circumvents inadequate analyte localization within the sensing volume, which often threatens the sensitivity of a SERS method. Often nanoplasmonic substrates are functionalized by direct attachment to the host molecule, which affects their SERS activity. Herein, we report 3D-amino-functionalized Ag@SiO2 nanocubes with ultra-thin silica shell for the detection of small molecules in a flexible SERS platform. The amino groups are introduced by silanizing the ultra-thin silica layer using different aminoslanes which improves sensitivity without sacrificing the colloidal and chemical stability of the core Ag nanocubes. Ag-SHINs functionalized with shorter chain length aminosilanes shows better SERS activity. The amino-functionalization effects a 3-fold SERS signal enhancement enabling label-free aspartame detection down to 71 µg mL-1 which are 1.5 times lower LOD than the non-functionalized Ag-SHINs. Analysis of spiked soft drink, as well as a mixture of aspartame and glucose, shows a robust performance signaling the selectivity and sensitivity of the substrate. The improved SERS performance is attributed to the hydrogen bonding induced localization of the analyte within the sensing volume. Added to this, the miniaturized paper platform with its 3D fibrous structure enhances the hotspot density and sample collection efficiency. This substrate can also be extended to other small molecules with the carboxylic group and amino acids as well, replacing the complicated bio-labeled detection schemes.
The final approach demonstrates the fabrication of a dielectric NS modified plasmonic-paper substrate and its application for food safety. The interaction of plasmonic nanoparticles with a dielectric platform gives rise to unique optical behaviors and this can be maneuvered to improve the plasmonic/SERS performances of a substrate. Herein, dielectric modified plasmonic-paper SERS substrate is developed by assembling Ag@SiO2 nanocubes on Fe-TiO2 NS modified paper. The Fe-TiO2 NS being visible light responsive significantly alters the optical property of the paper and serve as a dielectric underlay for the Ag NCs. Hence, the incident light reflected back from the dielectric film couples with the scattered light from the Ag NCs leading to spatially enhanced electromagnetic field improving the SERS enhancement. The prepared dielectric modified plasmonic-paper substrate is sensitive down to 1 pM R6G with average EF of 1.49×107. This value is superior to unmodified plasmonic-paper highlighting the coupling effect of the dielectric NS. The substrate shows robust analytical performance for thiabendazole detection with LOD down to 19 µg/ L, which is 4-fold more sensitive than unmodified plasmonic paper. Direct swabbing test of thiabendazole sprayed apple fruit shows a discernible Raman signal down to 15 µg/ L indicating the utility of the substrate for point-of-need applications in food safety.

Table of contents
Contents
摘要    i
Abstract    iii
Acknowledgment    vii
Table of contents    ix
List of figures    xiii
List of tables    xix
List of schemes and equations    xx
Acronyms    xxi
Chapter 1 Introduction    1
1 Background and Rationale    1
2    Objectives of the study    5
3 Structure of the dissertation    5
Chapter 2 Literature Review    7
1 Background of Raman Spectroscopy    7
2 Basic principles of Raman Spectroscopy (RS)    7
3 Surface Enhanced Raman Spectroscopy (SERS)    9
3.1 Overview of Surface Enhanced Raman Spectroscopy    9
3.2 SERS Enhancement Mechanisms    10
3.3 SERS Enhancement Factor    13
4 SERS substrates    15
4.1 Colloidal suspensions    15
4.2 Nanoparticles immobilized on a planar substrate    17
4.3 Flexible substrates    18
5 Ag as a SERS substrate and the reason for choice    23
5.1 Synthesis of Ag nanoparticles    25
5.2 Shape controlled the synthesis of Ag NPs    28
6 Silica coating of Ag NPs    30
7 Principle of shell-isolated nanoparticle-enhanced Raman Spectroscopy    32
8 Application of SERS in food analysis    33
Chapter 3 Experimental    37
1 Materials    37
2 Synthesis of Ag@SiO2 NCs    37
3 Amino functionalization of Ag@SiO2 NCs    38
4 Synthesis of Fe-TiO2 NS    39
5 Plasmonic-paper SERS substrate fabrication    39
6 Characterizations    40
6 Preparation of analyte solutions    40
7 SERS Measurement and spectral analysis    41
Chapter 4 Synthesis and characterization of Ag@SiO2 nanocubes    43
1 Scope of the study    43
2 Results and Discussion    44
2.1 Morphology, structure and optical properties of Ag NCs    44
2.2 Silica coating and optical property changes    47
3 Summary    51
Chapter 5 Flexible plasmonic-paper SERS substrate with improved assemblage of Ag@SiO2 NCs for trace melamine detection    53
1 Scope of the study    53
2 Results and Discussion    56
2.1 Fabrication and characterization of miniaturized plasmonic-paper SERS substrate    56
2.2 SERS performance    58
2.3 Analytical performance for Melamine Detection    62
3 Summary    69
Chapter 6 Functionalized Ag SHINs assembled on paper for sensitive and selective SERS detection of small molecules    71
1 Scope of the study    71
2 Results and Discussion    74
2.1 Characterization of functionalized Ag SHINs    74
2.2 Characterization of the plasmonic-paper substrate    78
2.3 SERS performance for aspartame detection    81
2.4 Effect of interferants    84
3 Summary    88
Chapter 7 Highly sensitive and stable Fe-TiO2 NS modified plasmonic-paper SERS substrate for pesticides detection    89
1 Scope of the study    89
2 Results and Discussion    92
2.1 Characterization of TiO2 NS    92
2.2 Morphology and structure of the dielectric modified plasmonic-paper substrate    92
2.3 SERS performance    96
2.4 Detection of thiabendazole    101
2.5 Analysis of real sample and swabbing test    104
3 Summary    107
Chapter 8 Conclusion and outlook    108
1 Conclusion    108
2 Outlook    110
REFERENCES    112
Curriculum Vitae    141
Appendix    143


                                

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