在過去的十年中，石墨烯量子點（GQD）是一個新開發的0維石墨烯家族，被認為是一種有前途的光學和電化學傳感材料。 GQD具有出色的熒光特性，高表面積，良好的化學穩定性，顯著的量子限制效應和穩定的光致發光等出色特性，使GQD成為用於選擇性和靈敏地檢測化學物質的新型傳感元件。 GQD已使用各種方法合成了不同的化學和天然前體。從天然來源合成GQD既環保又經濟。但是，由不同碳源合成的GQD在物理性質（如結構，形狀和大小）以及化學性質（如官能團）方面彼此不同，導致它們具有不同的靈敏度和選擇性。本文提出的研究集中在通過使用各種合成技術以及從不同的生物基碳前體中合成摻雜的GQD以及隨後將其作為生物傳感器的應用。
在這項研究中，將合成的GQD與其他材料進一步複合，以增強其在環境和生物醫學傳感領域的應用。這項研究分為以下幾個步驟：首先，對來自不同碳源及其光學傳感器的摻雜GQD進行合成和表徵，其次，使用GQD進行納米複合材料的合成，其中GQD是增強其電化學性能並最終改善其性能的主要材料。測試納米複合材料作為電化學傳感器的功效。
本研究的第一部分涉及磷（P）摻雜的GQD（PGQD），它們是通過微波輔助分別由L-抗壞血酸和磷酸三鈉分別作為碳和磷的前體成功製備的。 PGQD的粒徑約為3.5 ± 0.5 nm，厚度類似於2 - 4 層石墨烯。 PGQD能夠在1 nM – 150 µM的寬範圍內檢測到Ag+，而其檢測限極低，僅為0.02 nM。之後在葡萄糖和巰基琥珀酸分別作為碳和硫源的條件下，通過水熱熱解成功地合成了具有穩定綠色熒光的硫摻雜GQD（S-GQDs），用於快速，靈敏地檢測血紅蛋白（Hb）。所製備的S-GQD顯示出均勻的尺寸分佈，平均粒徑為4.5±0.5 nm。 S-GQD對Hb檢測具有極好的靈敏度，線性檢測範圍為10-1000 nM，在磷酸鹽緩衝溶液（PBS）和人血清（HS）中的檢測下限分別為0.28和0.48 nM。此外，採用新鮮百香果汁和硼酸為原料，通過微波輔助水熱法合成了硼和氮共摻雜的高QY的0維GQD（B，N-GQDs）。然後將平均粒徑為9 ± 1 nm的3 – 6層B，NGQD用於超靈敏和選擇性檢測四環素（TC）。 B，N-GQD在0.04 – 70 µM的線性檢測範圍內顯示出優異的TC測定分析性能，在PBS中檢測限（LOD）為1 nM，在尿液中的檢測限為1.9 nM，在HS中的檢測限為2.2 nM。此外，基於B，N-GQDs的紙納米傳感器對PBS，尿液和HS中0 – 30 µM TC的視覺檢測具有出色的分析性能。此後，在塗有氮摻雜GQD（N-GQDs）的絲網印刷電極（Es）上成功開發了新型超靈敏電化學免疫傳感器。 NGQD易於使用微波合成製造，其NGQD的粒徑範圍為3 – 12 nm。植物血凝素-L（PHA-L）可以通過N-GQDs表面的-COOH和-NH2基團直接附著在N-GQDs上，並作為特異性受體錨定發育中的乳腺癌細胞（MCF-7）。Es|N-GQDs/PHA-L免疫傳感器。 Es | N-GQDs / PHA-L免疫傳感器具有很寬的線性檢測範圍，從PBS中的5-106個細胞mL-1和HS中的20 - 106個細胞mL-1，其LOD極低，分別為1個細胞和2個細胞。此外，這項研究還證明了合成的N-GQDs作為檢測MCF-7的納米熒光生物成像探針的多功能作用，因此在快速、靈敏地診斷和治療乳腺癌方面比現有技術有了重大進展。最後，開發了新型的以3維（3-D）金納米星（AuNS / S-GQDs）納米複合材料修飾的S-GQD，作為電化學免疫傳感器中的活性生物傳感材料。大小為2 – 8 nm的S-GQD被均勻地沉積在大小為35 – 60 nm的AuNS表面。在將AuNS / S-GQDs沉積到絲網印刷電極（Es）上之後，Es || AuNS / S-GQDs / Ab免疫傳感器為PSA檢測提供了穩定，超靈敏且高度選擇性的平台。其檢測範圍很廣，分別在PBS及HS獲得10 fg mL-1至50 ng mL-1和30 fg mL-1至50 ng mL-1的檢測範圍，以及在PBS和HS中的LOD極低，僅為0.29 fg mL-1（9.7 aM）和1.1 fg mL-1。所有開發的用於Ag +，Hb，TC，MCF-7和PSA的納米材料均表現出優異的靈敏度和選擇性，已成功應用於實際樣品中，具有很高的生物傳感探針回收率。
總體而言，獲得的結果證明了基於GQD的納米複合材料作為生物醫學納米傳感、生物成像、藥物輸送和進一步擴展到其他應用的納米材料適用性。

ABSTRACT
During the last decade, Graphene quantum dots (GQDs) are a kind of 0-dimensional (0-D) graphene that has been developed as a material for optical and electrochemical sensing. The properties including remarkable fluorescence and electrical conductivity, a high surface area, good chemical, and photoluminescence stability make GQDs a novel sensing material that has the ability for enhancing sensing performance. GQDs have been synthesized from different chemical as well as natural precursors by using various methods. The synthesis of GQDs from natural sources is eco-friendly and cost effective. However, GQDs synthesized from different carbon sources are different from each other in terms of physical properties such as structure, shape and size as well as chemical properties such as functional groups leading to them possessing individualized sensitivity and selectivity. The study presented in this thesis focused on the synthesis of doped GQDs from different bio-based carbon precursors by using various synthesis techniques and their subsequent application as biosensors.
In this study, synthesized GQDs were further composited with other materials to enhance their application in the field of environmental and biomedical sensing. The study has been divided into the following steps: firstly the synthesis and characterization of doped GQDs obtained from different carbon sources and their optical sensor and secondly, the synthesis of nanocomposites using GQDs, where GQDs is a leading material to enhance its electrochemical properties and ultimately testing the efficacy of the nanocomposite as a electrochemical sensor.
The first part of this study deals with Phosphorus (P) doped GQDs (PGQDs) that were successfully prepared from L-ascorbic acid and trisodium phosphate as carbon and P precursors, respectively, via microwave-assisted. The PGQDs had an approximate particle size of 3.5 ± 0.5 nm with a thickness resembling 2 – 4 layers of graphene. The PGQDs were capable to detecting Ag+ at a wide range of 1 nM – 150 µM with an extremely low limit of detection (LOD) equivalent to 0.02 nM. Next, sulfur-doped GQDs (S-GQDs) with stable green fluorescence were successfully synthesized by hydrothermal method in the presence of glucose and mercaptosuccinic acid as the carbon and sulfur sources, respectively, for the rapid and sensitive detection of hemoglobin (Hb). The as-prepared S-GQDs showed uniform size distribution with a mean particle size of 4.5 ± 0.5 nm. The S-GQDs exhibited excellent sensitivity for the detection of Hb and a linear detection range of 10 - 1000 nM with a low LOD of 0.28 and 0.48 nM in phosphate buffer solution (PBS) and human serum (HS), respectively. Furthermore, boron and nitrogen co-doped GQDs (B,N-GQDs) with high QY were synthesized via microwave-assisted hydrothermal method using fresh passion fruit juice and boric acid as the beginning materials. The 3 - 6 layered B,N-GQDs with a mean particle size of 9 ± 1 nm were then applied for the ultra-sensitive and selective for tetracycline (TC) detection. The B,N-GQDs showed excellent toward TC determination in linear detection range of 0.04 - 70 µM and with LOD of 1 nM in PBS, 1.9 nM in urine and 2.2 nM in HS were obtained. In addition, B,N-GQDs based paper nanosensor exhibited excellent analytical performance for visual detection of 0 - 30 µM TC in PBS, urine, and HS. Thereafter, a novel ultra-sensitive electrochemical immunosensor was successfully developed on screen-printed electrodes (Es) coated with nitrogen doped GQDs (N-GQDs). N-GQDs were facile fabricated using a microwave process with N-GQDs obtained in a particle range of 3 - 12 nm. Phytohemagglutinin-L (PHA-L) could directly attach on N-GQDs via -COOH and -NH2 groups on the surface of N-GQDs and acted as a specific receptor to anchor breast cancer cell (MCF-7) in the developed Es|N-GQDs/PHA-L immunosensor. Es|N-GQDs/PHA-L immunosensor exhibited a wide linear detection range from 5 - 106 cells mL-1 in PBS and 20 - 106 cells mL-1 in HS with an extremely low LOD of 1 cell and 2 cells, respectively. Moreover, this study also demonstrated the multifunctional role of the synthesized N-GQDs as a nanofluorescent bioimaging probe to detect MCF-7 and thus presented a significant advance over available technologies for the rapid and sensitive diagnosis and treatment of breast cancer. Finally, novel S-GQDs decorated with 3-dimensional (3-D) gold nanostar (AuNS/S-GQDs) nanocomposite was developed as the active biosensing materials in electrochemical immunosensor. S-GQDs with a size in the range of 2 – 8 nm were homogenously deposited onto the surface of AuNS with a size of 35 – 60 nm. After deposition of AuNS/S-GQDs onto screen-printed electrode (Es), the Es||AuNS/S-GQDs/Ab immunosensor provided a stable, ultra-sensitive, and highly selective platform for PSA detection. Wide detection ranges from 10 fg mL-1 - 50 ng mL-1 and 30 fg mL-1 - 50 ng mL-1 with extremely low LOD of 0.29 fg mL-1 (9.7 aM) and 1.1 fg mL-1 in PBS and HS, respectively, we obtained. All of the developed nanomaterial for Ag+, Hb, TC, MCF-7, and PSA exhibited excellent sensitivity and selectivity are successfully applied in real samples with great recovery for sensing probe.
Overall, the obtained results prove the applicability of GQDs based nanocomposites as the capability nanomaterial for biomedical nanosensing, bioimaging, drug delivery, and further expansion to other applications as well.

OUTLINE
ABSTRACT v
Acknowledgement viii
LIST OF TABLES xv
LIST OF FIGURES xvii
ABBREVIATIONS, UNITS, AND SYMBOLS xxv
Chapter 1. Introduction 1
1.1. Motivation 1
1.2. Overviews of GQDs 4
1.2.1. GQDs structure 5
1.2.2. Dopant and functionalization of GQDs 6
1.2.3. Modification of GQDs base on nanocomposites 8
1.2.4. GQDs properties 8
1.3. Synthesis methods of GQDs 10
1.3.1 Top-down strategy 11
1.3.2. The bottom-up strategy 12
1.4. Precursors synthesize graphene quantum dots 13
1.4.1. Chemical precursors 13
1.4.2. Natural precursors and sensing applications 13
1.5. GQDs and GQDs base on nanocomposites applications 17
1.5.1. GQDs for sensing application 18
1.5.2. Bio-imaging, drug delivery, and cancer therapy 22
1.6. Cytotoxicity 24
1.7. Tagets detection and nanotechnology on nanosensor 25
1.7.1 Heavy metal and antibiotic pollutants 25
1.7.2 Disease biomarkers 26
1.7.3. Detection methods on nanomatrials 29
1.8 Objectives 30
Chapter 2. L-Ascorbic acid derived blue luminescent phosphorus doped graphene quantum dots for rapid detection of silver ions 34
2.1. Introduction 35
2.2. Experimental 38
2.2.1. Chemicals and materials 38
2.2.2. Synthesis of PGQDs 38
2.2.3. Preparing of interference chemicals and Ag+ stock solution 38
2.2.4. Detection of Ag+ by PGQD 39
2.2.5. The applicability of PGQD for in real samples 39
2.2.6. Cytotoxicity assay of pure GQDs, PGQD and AgNP/PGQD 39
2.2.7. Surface characterization 40
2.3. Results and discussion 41
2.3.1. Characterization of PGQD 41
2.3.2. The optical property, selectivity, and the pH effect of PGQD 47
2.3.3. PGQD as a fluorescence nanoprobe for Ag+ detection 51
2.3.4. Fluorescence nanoprobe of PGQD in real samples 54
2.3.5. The mechanism of PGQD nanosensing toward Ag+ 55
2.3.6. The changing morphology after toward Ag+ 58
2.3.7. Cytotoxicity of GQD, PGQD and AgNP/PGQD in cell culture 62
2.4. Conclusions 63
Chapter 3. Sustainable fabrication of green luminescent sulfur-doped GQDs for rapid visual detection of hemoglobin 65
3.1. Introduction 66
3.2. Experimental section 69
3.2.1 Chemicals and materials 69
3.2.2 Synthesis of S-GQDs 69
3.2.3. Characterization techniques 69
3.2.4. Preparation of stock solutions of Hb and S-GQDs 70
3.2.5. Detection of Hb by S-GQDs 71
3.3. Results and discussion 71
3.3.1. Characterization of S-GQDs 71
3.3.2. Optical property of S-GQDs 79
3.3.3. pH effect on the Detection of Hb using S-GQDs 81
3.3.4. Analytical performance of S-GQDs on Hb detection 84
3.3.5. Selectivity of S-GQD toward Hb detection 89
3.3.6. Cytotoxicity of S-GQDs 90
3.4. Conclusions 91
Chapter 4. Ultrasensitive detection of tetracycline using boron and nitrogen co-Doped GQDs from natural carbon source as the paper-based nanosensing probe in difference matrices 93
4.1. Introduction 94
4.2. Experimental section 97
4.2.1. Chemicals and materials 97
4.2.2. Synthesis of B,N-GQDs 97
4.2.3. Surface characterization 98
4.2.4. Detection of TC by B,N-GQDs in PBS, urine, and human serum 98
4.2.5. B,N-GQD based paper strip nanosensor for TC detection in human serum 99
4.2.6. Cytotoxicity assay of B,N-GQDs 99
4.3. Results and discussion 100
4.3.1. Characterization of B,N-GQDs 100
4.3.2. Optical Property of B,N-GQDs 107
4.3.3. Detection of TC by B,N-GQDs 113
4.3.4. Selectivity of B,N-GQDs 117
4.3.5. Possible sensing mechanism for TC detection by B,N-GQDs 119
4.3.6. Cytotoxicity of GQDs based nanomaterials 124
4.4 Conclusions 125
Chapter 5. Early detection of breast cancer using electrochemical immunosensor based on phytohemagglutinin-L specific receptor carried via N doped GQDs functioning as nanocarriers and nanofluorescent bioimaging probes 127
5.1. Introduction 128
5.2. Experimental section 131
5.2.1. Chemicals and materials 131
5.2.2. Synthesis of N-GQDs 132
5.2.3. Preparation of N-GQDs modified on Es and Es|N-GQDs/PHA-L immunosensor 132
5.2.4. Incubation of the Es|N-GQDs/PHA-L immunosensor with MCF-7 133
5.2.5. MCF-7 cell analysis using HS sample 133
5.2.6. Selectivity and stability of Es|N-GQDs/PHA-L immunosensor on sensing performance 133
5.2.7. Electrochemical measurement procedure 134
5.2.8. Cytotoxicity of the N-GQDs and N-GQDs/PHA-L on MCF-7 and MCF-10A cells 134
5.2.9. Bio-imaging MCF-7cell fluorescence and dark-field microscopy 135
5.2.10. Characterization techniques 135
5.3. Results and discussion 137
5.3.1. Characterization 137
5.3.2. Optical and electrochemical properties of N-GQDs 144
5.3.3. Analytical performance and sensitivity of various matrix immunosensor on MCF-7 147
5.3.4. The nanosensing mechanism for selectivity and stability of the Es|N-GQDs/PHA-L immunosensor 154
5.3.5. Cytotoxicity study of Es|N-GQDs/PHA-L and bioimaging of MCF-7 157
5.4. Conclusion 159
Chapter 6. Ultra-sensitive electrochemical immunosensor for attomolar detection of prostate specific antigen with sulfur-doped graphene quantum dot@gold nanostar 161
6.1. Introduction 162
6.2. Experimental section 165
6.2.1. Chemicals and reagents 165
6.2.2. Synthesis of AuNS/S-GQDs nanocomposites 166
6.2.3. Preparation of AuNS/S-GQDs modified electrode 166
6.2.4. Construction of the Es||AuNS/S-GQDs/Ab based immunosensor 167
6.2.5. Detection of PSA in human serum 167
6.2.6. Electrochemical measurement procedure 168
6.2.7. Selectivity of immunosensor 168
6.2.8. Cytotoxicity of the AuNS/S-GQDs nanocomposites 168
6.2.9. Characterization 169
6.3. Result and discussion 169
6.3.1. Characterization of AuNS/S-GQDs nanosensing materials 169
6.3.2. Electrochemical property of the Es||AuNS/S-GQDs immunosensor 177
6.3.3. Sensitivity of the Es||AuNS/S-GQDs immunosensor 179
6.3.4. Nanosensing of PSA in human serum 185
6.3.5. Stability and selectivity of AuNS/S-GQDs based immunosensor 186
6.3.6. Cytotoxicity of AuNS/S-GQDs based immunosensor 189
6.4. Conclusions 189
Chapter 7. Conclusions and future scope of the work 191
7.1. Conclusions 191
7.2. Future scope of the work 197
References 200

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