fabrication of antibacterial composite from bacterial cellulose and betel leaves

26 Pages·2011·0.6 MB·English

THE FABRICATION OF ANTIBACTERIAL COMPOSITE FROM
BACTERIAL CELLULOSE AND BETEL LEAVES

NOOR IZYANNISA’ BINTI SARJUNI

A thesis submitted in fulfillment
of the requirements for the award of the degree of
Bachelor of Chemical Engineering (Biotechnology)

Faculty of Chemical & Natural Resources Engineering
Universiti Malaysia Pahang

DECEMBER 2010
ABSTRACT

Wound healing was a complicated process that needs to be take care from
infection and inflammation. Infection that occurs on wound will slow down the
healing process. Therefore the objectives of this research were to produce
antibacterial biocomposite from bacterial cellulose with betel leaves. The films were
prepared by mixing the bacteria cellulose, betel leaves and gelatin in four different
samples that contain from 0% to 70% composition of betel leaves extract. These
biocomposite films were characterized by using Universal Testing Machine, Fourier
Transform Infrared (FTIR) Spectroscopy, Scanning Electron Microscope (SEM),
Gas Pycnometer, Antibacterial Effect Testing, water absorption testing and
Biodegradable Testing. The biocomposite film that contains 70% Betel Leaves
extract displayed better antibacterial properties, biodegradability and optimum water
absorption for wound healing process. The film showed high effectiveness
antibacterial activity towards gram positive bacteria rather than gram negative
bacteria where the inhibition area was 0.20 cm2 and the effectiveness activity was
100%. There was about 343.87 mg/L of phenolic content in this sample. The film
also showed the highest rate water absorption value which was 1.812 x 10-12 cm2/s
and the film completely degraded in 24 days. The addition of betel leaves extracts
and bacteria cellulose can be a promising source to produce biocomposite film that
contains antibacterial effect which was suitable for wound healing and also can
shorten the wound healing process.
ABSTRAK

Penyembuhan luka adalah satu proses yang rumit yang memerlukan
penjagaan yang rapi daripada sebarang jangkitan dan radang. Jangkitan yang berlaku
pada luka akan melambatkan proses penyembuhan.Oleh itu, objektif kajian ini
adalah untuk menghasilkan antibakteria biokomposit daripada selulosa bakteria dan
daun sireh. Biokomposit filem dihasilkan dengan campuran selulosa bakteria, daun
sireh dan gelatin dalam 4 sampel yang berbeza dengan mengandungi 0% hingga 70%
kandungan perahan daun sireh. Bio-komposit filem ini di kenal pasti dengan
menggunakan, Fourier Transform Infrared (FTIR) Spectroscopy, Mikroskop
Elektron Imbasan, Gas Piknometer, ujian antibakteria, ujian serapan air dan ujian
biodegradasi. Biokomposit yang dihasilkan daripada selulosa bakteria dan 70%
kandungan perahan daun sireh menunjukkan sifat antibakteria dan biodegradasi yang
baik serta mempunyai kadar penyerapan air yang optimum untuk penyembuhan luka.
Filem ini menunjukkan tahap keberkesanan antibakteria yang tinggi kepada Gram
positif dan Gram negative bakteria dimana kawasan yang terbantut adalah 0.20 cm2
dan keberkesanan aktiviti antibakteria adalah 100 % dengan kehadiran 343.87 mg/L
kandungan fenolik. Filem ini turut menunjukkan kadar serapan air yang tinggi
sebanyak 1.812 x 10-12 cm2/s dan boleh terurai sepenuhnya dalam masa 24 hari.
Campuran daun sireh dan selulosa bakteria akan menjanjikan penghasilan bio-
komposit yang mempunyai kesan antibakteria serta sesuai bagi merawat luka dalam
masa yang singkat.
TABLE OF CONTENT

CHAPTER TITLE PAGE

DECLARATION ii
DEDICATION iii
ACKNOWLEDGEMENTS iv
ABSTRACT v
ABSTRAK vi
TABLE OF CONTENT vii
LIST OF FIGURES x
LIST OF TABLES xi
LIST OF SYMBOL/ABBREVIATIONS xii
LIST OF APPENDICES xiii

1 INTRODUCTION
1.1 Background of Study 1
1.2 Problem Statement 3
1.3 Research Objectives 3
1.4 Scopes of Study 4
1.5 Significance of Study 4

2 LITERATURE REVIEW
2.1 Bacterial Cellulose 5
2.2 Acetobacter Xylinum 6
2.3 Betel Leaves 7
2.4 Biocomposite 7
2.5 Wound 9
2.6 Fourier Transform Infrared Spectroscopy 10
(FTIR)
2.7 Scanning Electron Microscopy (SEM) 11
2.8 Gas Pycnometer 12

3 METHODOLOGY
3.1 Introduction 13
3.2 Material 14
3.3 Experimental Procedure 14
3.3.1 Preparation of Bacteria Cellulose 15
3.3.2 Preparation of Betel Leaves Extract 15
3.3.3 Preparation of Biocomposite Film 16
3.3.4 Film Characterization 16
3.3.4.1 Antibacterial Testing 16
3.3.4.2 Phenolic Content Analysis 17
3.3.4.3 Biodegradable Testing 18
3.3.4.4 Fourier Transform Infrared 18
Spectroscopy (FTIR)
3.3.4.5 Scanning Electron Microscopy 18
(SEM)
3.3.4.6 Water Absorption Testing 18
3.3.4.7 Density Analysis 19

4 RESULT AND DISCUSSION
4.1 Antibacterial Testing 21
4.1.1 Disc Diffusion Assay 21
4.1.2 Diffusion and Spread Plate Technique 24
4.2 Phenolic Content Analysis 25
4.3 Fourier Transform Infrared Spectroscopy (FTIR) 26
4.4 Scanning Electron Microscopy (SEM) 27
4.5 Biodegradable Testing 28
4.6 Water Absorption Testing 29
4.7 Density Analysis 31

5 CONCLUSION AND RECOMMENDATIONS
5.1 Conclusion 32
5.2 Recommendations 33

REFERENCES 34
APPENDICES 39
LIST OF FIGURES

FIGURE NO. TITLE PAGE

3.1 Experimental Procedures for fabrication of 14
antibacterial composite film

4.1 The inhibition zone of film sample against 22
S.aureus

4.2 The inhibition zone of film sample against 23
E.coli

4.3 The graph of percentage EAA against S.aureus 24
and E.coli

4.4 FT-IR spectra 26

4.5 Scanning Electron micrograph 27

4.6 Plot percentage weight loss for each sample 28

4.7 Plot of the water uptake ratio, M/M versus 30
t m
time, t 1/2
LIST OF TABLES

TABLE NO. TITLE PAGE

4.1 Zone of inhibition area for S.Aureus 22

4.2 Zone of inhibition area for E.coli 22

4.3 Percentage of EAA against S.aureus and 24
E.coli

4.4 The phenolic concentration for each sample 25

4.5 Weigh of sample every 6 day 28

4.6 The maximum water uptake, M and diffusion 30
m
coefficient, D

4.7 The experimental and true density value 31
LIST OF SYMBOLS/ ABBREVIATIONS

0 - Degree
% - Percent
C - Celcius
g - gram
cm - centimeter
Kg - Kilogram
L - Liter
mL - mililiter
min - minute
s - second
BC - Bacterial Cellulose
BL - Betel Leaves
EAA - Effectiveness Antibacterial Activity
CPU - Number of colony forming unit
LIST OF APPENDICES

TABLE NO. TITLE PAGE

A Data of standard calibration curve for 39
phenolic content analysis

B Data of CFU in dilution and spread plate 40
technique

C Data of density analysis by experimental 41

D FTIR spectra for bacterial cellulose 43