微生物燃料電池是一種新穎的潔淨能源產出方案來因應即將到來的能源危機，與傳統燃料電池主要的差異在於它毋須使用金屬催化劑修飾陽極，而是使用特定微生物來催化有機分子的氧化，進而將得到的電子傳遞給陽極。接著經由電池外部導線而傳遞給陰極槽的氧氣，並與來自陽極並通透過半透膜的氫離子結合產生水的終產物。然而氧氣的還原反應需要很高的過電壓，在動力學上不利於反應進行，因此需要催化劑。傳統上，白金被廣泛的使用，然而它確有價格過於高昂的缺點，會大幅增加電池的生產成本。因此，本研究以探尋較為廉價的氧氣還原反應催化劑與其反應機制為目標，發現使用金屬氧化物修飾陰極具有能催化氧氣還原並增加微生物燃料電池功率的能力，最終決定使用四氧化三鈷。
本研究中尚探討了四氧化三鈷催化氧氣還原的機制與特性，並進一步進行電池操作參數的最佳化，條件為: 使用Co3O4做為催化劑，修飾比例為70%，陽極槽培養液pH值為5，以葡萄糖做為受質，陰極槽使用磷酸緩衝溶液，pH值設定為5，兩極距離為一公分。研究發現50%的四氧化三鈷可達到5%的白金修飾電極之催化效果，在電池功率方面，與使用僅修飾純導電碳膠的陰極之微生物燃料電池相比，電池最大功率密度在最佳化條件下提升9.53倍。本研究也進行了交流阻抗與電池生命週期的分析。

Microbial fuel cell (MFC) is a novel and clean energy source solution to imminent energy crisis. In contrast to conventional fuel cell, microbial fuel cell, instead of using metal catalyst as anode material, takes advantage of microorganisms to catalyze oxidation organic matters. The released electrons were then transferred to oxygen in catholyte through the external conductive wire. Combining with protons permeates from anolyte, thus the final product was water (H2O). However, the reduction reaction’s nature of high overpotential hinders its spontaneity thermodynamically. As a result, utilizing catalyst for oxygen reduction reaction (ORR) is necessary and inevitable. Conventionally, platinum is served as most frequently used catalyst material, but its high cost may limit large scale production of fuel cell. Therefore, the goal of this study is to search for more cost-effective catalyst material and to evaluate the reaction mechanism for oxygen reduction reaction as replacement of platinum. It is found that metal oxides can be potential candidates for acting as catalyst for oxygen reduction reaction; in the meanwhile, enhance power output efficiency of MFC. In the end, Co3O4 is chosen as the final option.
The catalysis mechanism and performance of ORR for Co3O4 is evaluated. In addition, the optimization of operating parameters has been undergone. The optimized conditions are: adopting Co3O4 as cathode catalyst material, modification ratio 70wt.%, anolyte as medium of pH5, glucose used as substrate, pH5 phosphate buffer solution served as catholyte, electrode distance of 1cm. In this study, the effect of modifying 50wt.% of Co3O4 is comparable to 5wt.% of Co3O4. In comparison to the MFC with pure conductive carbon ink modified cathode, the power density is enhanced 9.53 times. The evaluation of electrochemical impedance spectroscopy and life time of MFC is also studied.

Contents
Chapter 1 Preface	7
1.1	Research motives	7
1.2 Development History of Microbial Fuel Cell	9
1.2.1 Designs of Anode Configuration	11
1.2.2 Microorganisms used in MFCs	15
1.2.3 Various Sources of Substrates for Microorganisms in Anode Chamber	22
1.2.4 Designs of Cathode Configuration	26
1.2.5 MFCs for waste and pollution reduction	37
1.2.6 MFCs for Hydrogen Production	48
1.3 Principle of Microbial Fuel Cells	51
1.4 Performance Characterization of Microbial Fuel Cells	53
1.5 Configurations of MFC	54
1.6 Electrochemical Impedance Spectroscopy	57
1.7 Brief Introduction of Cobalt (II, III) Oxide (Co3O4)	59
1.8	Research goals	60
Chapter 2 Experimental Section	62
2.1 Configuration of Microbial Fuel Cell and its Assembly	62
2.2 Instruments	65
2.2.1 Electrochemical Analysis	65
2.2.2 Power Output Analysis	66
2.2.3 Microorganism Incubation	67
2.2.4 Others	67
2.3 Chemicals	68
2.3.1 Chemicals Preparation	69
2.4 Electrodes Preparation for Electrochemical Analyzer	70
2.4.1 Pretreatment of Electrodes	70
2.4.2 The Preparation of Co3O4 – modified Electrode	71
2.5 Experimental Methods	71
2.5.1 The Evaluation of Oxygen Reduction Mechanism	71
2.5.2 The optimization of the MFC Power Output Performance	72
2.5.3 Electrochemical Impedance Spectroscopy	77
2.5.4 Life Time of the MFC	77
2.6 Characterization of Co3O4	78
Chapter 3 Results and Discussion	79
3.1 Design of a Cathode Suitable for the Development of MFC	79
3.2 The Effect of Catholyte pH	91
3.3 The Effect of Catholyte Buffer Solution Type on the Catalysis Behavior of Co3O4	94
3.4 The Evaluation of Catalysis Mechanism	96
3.5 The Anolyte pH Determination	109
3.6 Substrate for Bio-assisted Organic Compound Oxidation Study	114
3.7 The Effect of Electrode distance on the Performance of MFC	116
3.8 The Modification Ratio of Co3O4 on the Cathode	118
3.9 The Effect of Catholyte pH	121
3.10 Compromised pH in Both Chamber	124
3.11 Electrochemical Impedance Spectroscopy of Optimized MFC	126
3.12 Life Time Study	129
3.13 The Optimized Operating Parameters of MFC	130
3.14 Characterization of Co3O4	131
3.15 General Current-Potential Curves at the RDE	133
Chapter 4 Conclusion	136
Appendix	137
Reference	151


Graph Contents
Figure (1) Schematic diagram of the MFC Principle……………………….…..……55
Figure (2) Schematic diagram of the single-chambered MFC……………….………57
Figure (3) Schematic diagram of the Dual-chambered MFC………………….……..58
Figure (4) Schematic diagram of the Tubular Up-flow MFC………………………..58
Figure (5) Schematic of Co3O4 Structure…………………………………….………61
Figure (6) Schematic of Microbial Fuel Cell Assembly…………………….……….68
Figure (7) I-V Curve of the Co3O4 – modified GCE…………………………..……..84
Figure (8) I-V Curve of the CuO – modified GCE……………………………….….84
Figure (9) I-V Curve of the Fe2O3 – modified GCE…………………...........……….85
Figure (10) I-V Curve of the Fe3O4 – modified GCE……………………………..…85
Figure (11) I-V Curve of the NiO2 – modified GCE………………………..……..…86
Figure (12) I-V Curve of the CaTiO3 – modified GCE…………………………....…86
Figure (13) I-V Curve of the LiMn2O4 – modified GCE……………………….……87
Figure (14) I-V Curve of the WO2 – modified GCE…………………………………87
Figure (15) I-V Curve of the Pb3O4 – modified GCE………………………….…….88
Figure (16) Stable I-V Curve of the Fe3O4 – modified GCE………………….…..…89
Figure (17) Stable I-V Curve of the LiMn2O4 – modified GCE…………….……….89
Figure (18) Steady-state currents at 0V - Fe3O4 – modified GCE……………..…….90
Figure (19) LSV - Effect of catholyte pH……………………………..….………….94
Figure (20) Power density – voltage polarization curve - Effect of catholyte pH…...95
Figure (21) LSV - Effect of catholyte buffer kind……………………………………97
Figure (22) Steady-state reduction current vs. each potential - Pure conductive carbon
ink – modified GCE………………………………………………………………….99
Figure (23) Steady-state reduction current vs. each potential - 10 wt.% Co3O4 -
modified GCE………………………………………………………….…………...100
Figure (24) Steady-state reduction current vs. each potential - 30 wt.% Co3O4 -
modified GCE……………………………………………………………….……...100
Figure (25) Steady-state reduction current vs. each potential - 50 wt.% Co3O4 -
modified GCE……………………………………………………………….……...101
Figure (26) Steady-state reduction current vs. each potential - 70 wt.% Co3O4 -
modified GCE……………………………………………………………….……...101
Figure (27) Steady-state reduction current vs. each potential - 50 wt.% platinum
coated carbon powder - modified GCE….……………………………….….……...102
Figure (28) I-t amperometric plot (0.4V) - 50 wt.% Co3O4 – modified
RRDE….………………………………...…………………………………..……...104
Figure (29) I-t amperometric plot (0.3V) - 50 wt.% Co3O4 – modified
RRDE….………………………………...…………………………………..……...104
Figure (30) I-t amperometric plot (0.2V) - 50 wt.% Co3O4 – modified
RRDE….………………………………...…………………………………..…..….105
Figure (31) I-t amperometric plot (0.1V) - 50 wt.% Co3O4 – modified
RRDE….………………………………...…………………………………..…..….105
Figure (32) I-t amperometric plot (0V) - 50 wt.% Co3O4 – modified
RRDE….………………………………...…………………………….…………....106
Figure (33) I-t amperometric plot (-0.1V) - 50 wt.% Co3O4 – modified
RRDE….………………………………...…………………………….……………106
Figure (34) I-t amperometric plot (-0.2V) - 50 wt.% Co3O4 – modified
RRDE….………………………………...…………………………….……….…...107
Figure (35) I-t amperometric plot (-0.3V) - 50 wt.% Co3O4 – modified
RRDE….………………………………...………………………………….……....107
Figure (36) I-t amperometric plot (-0.4V) - 50 wt.% Co3O4 – modified
RRDE….………………………………...………………………………….……....108
Figure (37) I-t amperometric plot (-0.5V) - 50 wt.% Co3O4 – modified
RRDE….………………………………...…………………………………….…....108
Figure (38) I-t amperometric plot (-0.6V) - 50 wt.% Co3O4 – modified
RRDE….………………………………...……………………………………….....109
Figure (39) Signals of oxygen vs. applied potential plot - 50 wt.% Co3O4 – modified
RRDE….……….……………...……………………………………………....…....109
Figure (40) Calibration plot of H2O2 - 50 wt.% Co3O4 – modified
RRDE….……….……………...………………………………...………….……....110
Figure (41) Proposed ORR mechanism on Cobalt (II, III) oxide modified
electrode…………………………………………………………………………….110
Figure (42) LSV - Effect of culture medium pH on the performance of
anodes….……….…………………...…………………………...…………….…....113
Figure (43) LSV - Effect of culture medium pH on the performance of
anodes….……….…………………...…………………………...………….……....114
Figure (44) Power density – voltage polarization curve - Effect of culture medium
pH….……….…….………...…………………………...………………...…...…....114
Figure (45) LSV - Effect of various subsrates on the performance of the
anode….……….…………………...…………..………………...………….……....117
Figure (46) Power density – voltage polarization curve - Effect of electrode distance
on the performance of the microbial fuel cell…………………………………...….119
Figure (47) LSV - Effect of modification ratio…………………………….……….121
Figure (48) Power density – voltage polarization curve - Effect of modification
ratio….……….…….………...…………………...……………...………….……....122
Figure (49) Modification ratio vs. maximum power density……………………….122
Figure (50) LSV - Effect of catholyte pH…………………………………………..124
Figure (51) Power density – voltage polarization curve - Effect of catholyte
pH…………………………………………………………………………………...125
Figure (52) Plot of pH vs.time…………………………………………..………….127
Figure (53) Power density – voltage polarization curve - Effect of both chambers with
the same pH……………………………………..………………………….…….…128
Figure (54) EIS data of MFC performed as Nyquist plot………………….………..130
Figure (55) Equivalent circuit for two-chambered MFC…………………….……..130
Figure (56) The life time of MFC…………………………………….…………….131
Figure (57) The XRD spectrum of Co3O4…………………………………………..133
Figure (58) FE-SEM images of Co3O4……………………………………………...133
Figure (59) Variation of 1/i with ω-1/2 at 0V………………………………………...136
Figure (60) Variation of i with ω at 0V……………………………………………...136

Table Contents
Table (1) Electrochemical parameters used in MFCs study………………………….56
Table (2) The open circuit potential of each metal oxide – modified
GCE………………………………………………………..………….……………...90
Table (3) The performances evaluation of various metal oxides as cathodic
modifier………………………………………………………………………….…...92
Table (4) The open circuit potential of 50 wt.% Co3O4 – modified GCE in each kind
of buffer solution……………………………………………………………….…….94
Table (5) The open circuit potential of anodes incubated with various pH
medium…………………………………………………………………….…...…...112
Table (6) The open circuit potential of MFC with various electrode
distances.....................................................................................................................116
Table (7) The open circuit potential of GCE coated with various modifier………...120
Table (8) The open circuit potential of cathode modified with 50 wt.%
Co3O4……………………………………………………………………………......122
Table (9) Various parts of internal resistances of MFC……………………….…….127
Table (10) Optimized operating parameters
table………………………………………………………………………………....129
Table (11) Rate constants corresponding to each applied potential………………...137

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