Synthesis of multifunctional au fe3o4 nanoparticles for application in biomedicine

VIETNAM NATIONAL UNIVERSITY, HANOI VIETNAM JAPAN UNIVERSITY - BUI DUC TRI SYNTHESIS OF MULTIFUNCTIONAL Au-Fe3O4 NANOPARTICLES FOR APPLICATION IN BIOMEDICINE MASTER'S THESIS Hanoi, 2018 VIETNAM NATIONAL UNIVERSITY, HANOI VIETNAM JAPAN UNIVERSITY BUI DUC TRI SYNTHESIS OF MULTIFUNCTIONAL Au-Fe3O4 NANOPARTICLES FOR APPLICATION IN BIOMEDICINE MAJOR: NANOTECHNOLOGY SUPERVISOR: Associate Prof Dr NGUYEN HOANG NAM Hanoi, 2018 ACKNOWLEDGEMENT To accomplish this thesis, I have received great support, helpful advices, and guidance from respectful professors, lecturers, researchers, and staff in Vietnam Japan University, Osaka University, and VNU - University of Science At first, I would like to express my gratefulness to my supervisor, Associate Prof Dr Nguyen Hoang Nam, Doctoral student Mr Chu Tien Dung for supplying great researching environment in laboratories, and for giving helpful instructions, guidance, advices and motivation during my research in the laboratory Secondly, I would like to show my gratitude to Prof Tamiya Eiichi, Doctoral student Mr Joyotu Mazumder for providing instructions and knowledge about Electrochemical luminescence, as well as the hospitality during my internship in Osaka University I sincerely thank all professors, staff, and friends in Vietnam Japan University and VNU - University of Science for supplying me the best condition for my research Finally, I am thankful to my family for the support, companionship, and mobilization, which were essential elements for me to finish the thesis Hanoi, 10 June 2018 Author BUI DUC TRI i TABLE OF CONTENTS ACKNOWLEDGEMENT i TABLE OF CONTENTS ii LIST OF FIGURES v LIST OF TABLES vii LIST OF ABBREVIATIONS viii INTRODUCTION CHAPTER LITERATURE REVIEW 1.1 Properties and applications of individual nanoparticles 1.1.1 Magnetite nanoparticles 1.1.2 Gold nanoparticles 1.2 Multifunctional Gold-magnetite nanoparticles 1.2.1 Advantages of multifunctional gold-magnetite nanoparticles 1.2.2 Types of multifunctional gold-magnetite nanoparticles 10 CHAPTER PRINCIPLES OF SYNTHESIZING AND CHARACTERIZING METHODS 13 2.1 Synthesis methods 13 2.1.1 Co-precipitation methods 13 2.1.2 Chemical reduction methods 13 2.1.3 Hydrolysis and condensation methods 15 2.2 Characterizing equipment 17 2.2.1 X-ray Diffractometer 17 2.2.2 Ultraviolet-visible spectroscopy 18 2.2.3 Energy-Dispersive X-ray Spectroscopy 18 ii 2.2.4 Transmission electron microscopy 19 2.2.5 Vibrating Sample Magnetometer 20 2.2.6 Electrochemical luminescence monitoring system 21 CHAPTER EXPERIMENTAL SECTION 23 3.1 Experimental materials 23 3.2 Study the synthesis of gold-magnetite nanoparticles 23 3.2.1 Synthesis of magnetite nanoparticles 23 3.2.2 Synthesis of direct-binding gold-magnetite nanoparticles 24 3.2.3 Synthesis of indirectly-binding gold-magnetite nanoparticles 26 3.3 Characterization 28 3.4 Study the electrochemical luminescence assay of gold-magnetite nanoparticles 28 CHAPTER RESULTS AND DISCUSSION 30 4.1 Study the synthesis of gold-magnetite nanoparticles 30 4.1.1 Crystal structure 30 4.1.2 Size and morphology 33 4.1.3 Localized surface plasmon resonance property 37 4.1.4 Magnetic property 42 4.2 Electrochemical luminescence assay of gold-magnetite nanoparticles 44 4.2.1 Phenomena and mechanism 44 4.2.2 Application in qualitative analyze of gold-magnetite nanoparticles 47 CONCLUSION 49 REFERENCES 50 LIST OF THE AUTHOR’S PUBLICATIONS 58 APPENDIX 59 iii iv LIST OF FIGURES Page Figure 1.1 Crystal structure of Fe3O4 illustrated by VESTA software Figure 1.2 The enhancement of T1 MRI images of a mouse injected with magnetite nanoparticles over time Figure 1.3 Crystal structure of Gold illustrated by VESTA software Figure 1.4 Polarization and oscillations of electron cloud on the surface of gold nanoparticle under the radiation of electromagnetic wave Figure 1.5 Scheme of synthesis of indirectly-binding gold-magnetite nanoparticles with Gold seeds 11 Figure 1.6 Scheme of synthesis of indirectly-binding gold-magnetite nanoparticles by reduction of HAuCl4 12 Figure 2.1 TEM image of gold nanostructured materials synthesized with different stabilizers 14 Figure 2.2 The process of synthesis silica by Stober method 15 Figure 2.3 Mechanism of functionalization a material surface with APTES 16 Figure 2.4 The principle of EDX 19 Figure 2.5 Simplified configuration of VSM 21 Figure 2.6 Setup of Electrochemical luminescence monitoring system 21 Figure 3.1 The synthesizing process of direct-binding gold-magnetite nanoparticles by photo-assisting reduction 24 Figure 3.2 Digital photo of experiment setup for the illumination of UV-light 25 Figure 3.3 The synthesizing process of indirectly-binding gold-nanoparticles 26 Figure 4.1 XRD patterns of directly-binding (FA), indirectly-binding (FSA) goldmagnetite nanoparticles, silica-coating magnetite nanoparticles (FS), and bare magnetite nanoparticles (Fe3O4) 30 Figure 4.2 TEM image and size distribution graph of Fe3O4 nanoparticles 33 Figure 4.3 TEM image and size distribution graph of silica coating magnetite nanoparticles (FS) 34 v Figure 4.4 TEM image and size distribution of indirectly-binding gold-magnetite nanoparticles (FSA) 35 Figure 4.5 a) TEM image of directly-binding gold-magnetite nanoparticles (FA) and b) Illustration scheme of Janus structure of the hybrid of two materials 36 Figure 4.6 UV-VIS spectra of Magnetite nanoparticles (Fe3O4), silica-coating magnetite nanoparticles (FS), indirectly-binding (FSA) and directly-binding (FA) gold-magnetite nanoparticles 37 Figure 4.7 UV-VIS spectrum of indirectly-binding gold-magnetite nanoparticles (FSA) with different amounts of trisodium citrate 38 Figure 4.8 UV-VIS spectrum of directly-binding gold-magnetite nanoparticles (FA) with different amounts of trisodium citrate 41 Figure 4.9 The magnetic hysteresis loops of bare magnetite nanoparticles (Fe3O4), directly-binding (FA) and indirectly-binding (FSA) gold-magnetite nanoparticles 43 Figure 4.10 Digital image of directly-binding gold-magnetite nanoparticle (FA) before and after separating from a solution by a permanent magnet 44 Figure 4.11 Electrochemical luminescence signal of the sample with the same amount of Luminol in the absence and presence of gold-magnetite nanoparticles with different mixing time (t) between FA5 100ppm and Tris Buffer pH = 11.89 44 Figure 4.12 a) Linear sweep voltammetry versus applied potential and b) Electrochemical luminescence signal and Linear sweep voltammetry versus monitoring time of solution mixing between Luminol and Tris buffer 45 Figure 4.13 Mechanism of Electrochemical luminescence of Luminol 46 Figure 4.14 Electrochemical luminescence of luminol in reaction mixtures 47 Figure 4.15 The relation of ECL signal and concentration of gold-magnetite nanoparticles at the mixing time with Tris Buffer pH=11.89 of t = 0, t = 10 mins, and t = 20 mins 48 vi LIST OF TABLES Page Table 1.1 Recent research on the synthesis of directly-binding gold-magnetite nanoparticles and their application in biomedicine 10 Table 3.1 Chemicals used in the research 23 Table 3.2 Preparation of “reaction mixture” for ECL assay 29 Table 4.1 Lattice parameters of crystals in Fe3O4 nanoparticles 32 Table 4.2 Lattice parameters of Au crystals in indirectly-binding gold-magnetite nanoparticles 32 Table 4.3 Absorption peaks in UV-VIS spectra of directly-binding gold-magnetite nanoparticle samples synthesizing with different amount of HAuCl4 40 Table 4.4 Element composition of four samples of directly-binding gold-magnetite nanoparticle synthesizing with different amount of HAuCl4 40 Table 4.5 Absorption peaks in UV-VIS spectra of directly-binding gold-magnetite nanoparticle samples 42 vii LIST OF ABBREVIATIONS Abbreviation Description APTES (3-Aminopropyl)triethoxysilane ECL Electrochemical luminescence EDX Energy-Dispersive X-ray spectroscopy FCC Face-centered cubic IR Infrared Spectroscopy LSV Linear sweep voltammetry MRI Magnetic resonance imaging NPs Nanoparticles ROS Reactive oxygen species TEM Transmission Electron Microscopy TEOS Tetraethyl orthosilicate TEOS Tetraethyl orthosilicate VSM Vibrating Sample Magnetometer XRD X-ray diffraction viii between gold-magnetite nanoparticles and Tris Buffer, the luminescence was amplified more significantly with the maximum detected optical value after mixing with Tris Buffer 10 mins and 20 mins being approximately 9.4 times and 13.3 times higher than that of the sample in the absence of gold-magnetite nanoparticles, respectively Figure 4.12 a) Linear sweep voltammetry versus applied potential and b) Electrochemical luminescence signal and Linear sweep voltammetry versus monitoring time of solution mixing between Luminol and Tris buffer To study the mechanism of this enhancement, we consider the linear sweep voltammetry (LSV) of the solution of luminol mixing with Tris buffer was investigated The peak at 365 mV in the LSV graph (Figure 4.12a) versus applied potential is contributed by the oxidization of luminol [61] The peak at 250 mV could be considered as the peak of the redox reaction of reactive oxygen species (ROS) with the voltage applied by the working electrode Moreover, the highest peak in ECL plotting, at monitoring time of 6.5s, is corresponding to the peak of 250mV of LSV plotting not at the peak of luminol (Figure 4.12b) Therefore, the redox reaction of reactive oxygen species affects significantly the ECL signal Interestingly, the presence of gold-magnetite nanoparticles and Tris buffer gives rise in the ECL at monitoring time of 6.5s (Figure 4.11), corresponding the peak of the redox reaction of ROS, this could be stated that the redox reaction of ROS was enhanced This could be due to the higher amount of ROS present in the reaction mixture with the presence of FA nanoparticles as compared to that of the reaction 45 mixture without FA nanoparticles Thus, gold-magnetite nanoparticles could catalyze for the ROS generating reactions of Tris buffer solution [50] The role of gold-magnetite nanoparticles and ROS in the electrochemical luminescence of luminol is described in the Figure 4.13 At first, the reaction mixture of Tris Buffer and FA nanoparticles gives rise in the amount of ROS Besides, luminol was oxidized by the electrode (as the oxidant (1)) to be reducing form The luminol reducing form reacts with the product of redox reaction of ROS with the working electrode (as the oxidant (2)) to convert oxidized-luminol to excited-state luminol, which then transfers to ground state and emits luminescence signal [62] Figure 4.13 Mechanism of Electrochemical luminescence of Luminol [62] To study about the enhancement of ECL signal in the presence of goldmagnetite nanoparticle over different mixing time, the ECL signal at the monitoring time of 6.5s of different reaction mixtures described in Table 3.2 were collected and plotted as Figure 4.14 It can be considered again that the electrochemical luminescence signal in the solution of FA nanoparticles and Tris Buffer were 46 significantly enhanced after the 10 and 20 minutes, whereas there is no enhancement in the sample only containing distilled water, Tris buffer, or FA nanoparticles This means that each component itself does not raise in the electrochemical luminescence, and it can be stated that the enhancement of ECL signal in the presence of FA nanoparticles and Tris buffer could be due to the interaction between FA and Tris Buffer In addition, the Fe3O4 nanoparticles in Tris buffer similarly not exhibit this property Therefore, the enhancement could be caused by the property of gold components in FA nanoparticles, which is in agreement with other research [50, 63] Figure 4.14 Electrochemical luminescence of luminol in reaction mixtures 4.2.2 Application in qualitative analyze of gold-magnetite nanoparticles Figure 4.15 shows the dependence of electrochemical luminescence signal of luminol on the concentration of directly-binding Au-Fe3O4 nanoparticles (FA3 sample) The intensity of electrochemical luminescence increases along with the FA nanoparticle concentration, which matches well with linear relation with R2 > 0.95 In addition, the dependence of ECL signal on particle concentration with different 47 periods of reaction time different time point of measuring were investigated Herein, at a longer point in time, the ECL signal tends to more fluctuate with the value of R2 decrease from 0.995 to 0.985 Although the dependence is most linear at the initial point, the ECL intensity at t = is weaker than that after 10 minutes and 20 minutes Therefore, choosing the reaction time of 10 minutes is the most reasonable, which exhibits more sufficient strong ECL intensity than that at the initial, and more acceptable linear dependence of ECL signal on particle concentration than that of reaction time is 20 minutes Therefore, this method could be applied as a quantitative method to measure the amount of FA nanoparticles Moreover, by modifying the surface of the FA nanoparticles for the attachment of bio-molecule, we can measure the amount of biomolecule by detecting Au-Fe3O4 NPs from ECL signal Figure 4.15 The relation of ECL signal and concentration of gold-magnetite nanoparticles at the mixing time with Tris Buffer pH=11.89 of t = 0, t = 10 mins, and t = 20 mins 48 CONCLUSION In conclusion, directly-binding and indirectly-binding gold-magnetite nanoparticles have been synthesized by the incorporation of co-precipitation and reduction methods Both directly-binding and indirectly-binding gold-magnetite multifunctional nanoparticles exhibit superparamagnetic property for separation purposes with the value of saturation magnetization approximately to 60 emu/g and 31 emu/g, respectively; as well as plasmonic property originated from localized surface plasmon resonance phenomena with tunable absorption peak in the region 500 - 600 nm by modifications in synthesis conditions The two types of nanoparticles were different in structure, where directly-binding gold-magnetite nanoparticles exist in form of Janus structure, indirectly-binding gold-magnetite have the structure as small gold nanoparticles attached on silica-shell magnetitecore nanoparticles In addition, the phenomena and mechanism of the enhancement in electrochemical luminescence signal caused by gold-magnetite multifunctional nanoparticles were examined The electrochemical luminescence signal was significantly enhanced over time in the presence of gold-magnetite multifunctional nanoparticles and Tris Buffer, which is attributed to the catalyst property of gold component present in the nanoparticles In addition, the enhancement of electrochemical luminescence signal is depended on the concentration of multifunctional nanoparticles, which is potential for quantitative analysis of nanoparticles and for further detection of biomolecules conjugated to the nanoparticles 49 REFERENCES [1] D Bobo, K J Robinson, J Islam, K J Thurecht, S R Corrie (2016) 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Iss 10, pp 3324-3329 [63] E Tamiya, Y Inoue, M Saito (2018) "Luminol-based electrochemiluminescent biosensors for highly sensitive medical diagnosis and rapid antioxidant detection" Japanese Journal of Applied Physics Vol 57, Iss 3S2, pp 03EA05 57 LIST OF THE AUTHOR’S PUBLICATIONS [1] Paper title: “ẢNH HƯỞNG CỦA NHÓM AMIN TRÊN BỀ MẶT HẠT NANO Fe3O4 ĐẾN CẤU TRÚC, TÍNH CHẤT CỦA NANO COMPOSIT Fe3O4Ag” Author: Chu Tien Dung, Bui Duc Tri, Tran Thi Hong, Nguyen Hoang Nam Published at the scientific conference: The 10th Vietnam National Conference on Solid State Physics and Material Sciences – SPMS 2017 (Hội nghị Vật lý Chất rắn Khoa học Vật liệu Toàn quốc – SPMS 2017) 58 APPENDIX Appendix 1.1 Identification of gold nanoparticle and magnetite nanoparticles in TEM image of directly-binding gold-magnetite nanoparticles - EDX spectrum and TEM image of FA sample at the region without the presence of “dark objects” - EDX spectrum and TEM image of FA sample at the region with the presence of “dark objects” 59 ... setup for the illumination of UV-light 25 3.2.3 Synthesis of indirectly-binding gold-magnetite nanoparticles Figure 3.3 The synthesizing process of indirectly-binding gold -nanoparticles Indirectly-binding... layer of Silica Figure 1.6 Scheme of synthesis of indirectly-binding gold-magnetite nanoparticles by reduction of HAuCl4 [40] 12 CHAPTER PRINCIPLES OF SYNTHESIZING AND CHARACTERIZING METHODS 2.1 Synthesis. .. studies the ? ?Synthesis of multifunctional Au- Fe3O4 nanoparticles for application in biomedicine? ?? This research aims to synthesize and characterize two types of multifunctional gold-magnetite nanoparticles,