CHITOSAN BASED ACETONE SENSOR FOR DETECTING

LOW CONCENTRATION ACETONE

by

TULUS IKHSAN NASUTION

Thesis submitted in fulfilment of the requirements

for the degree of

Doctor of Philosophy

July 2013
 Author’s declaration

I hereby declare that I have conducted, completed the research work and written the

dissertation entitled “Chitosan based acetone sensor for detecting low concentration

acetone”. I also declare that it has not been previously submitted for the award of any

degree or diploma or other similar title of this or any other examining body or

university.

Signature of candidate:

Name of candidate: Tulus Ikhsan Nasution

Date :

Witness by:

Signature of witness (main supervisor):

Name : Professor Hj. Zainal Arifin b. Hj. Ahmad

Date :

ii
 ACKNOWLEDGMENTS

In the Name of Allah, the Most Gracious and the Most Merciful...

All praise is for Allah alone, giving me his blessing and the strength to complete my

Ph.D research project entitled: “Chitosan based acetone sensor for detecting low

concentration acetone”.

I would like to take this ample opportunity to express my greatest thankfulness to my

respected supervisor, Prof. Dr. Hj. Zainal Arifin Ahmad who provided me a good

chance to conduct my research under his invaluable guidance, as well as motivating

and supporting me right which enabled me to realize my ideas into a invention.

Moreover, I would like to give my gratefulness to my co-supervisors, Dr. Sabar

Derita Hutagalung and Dr. Khairel Rafezi Ahmad for their support and endurance in

helping and guiding me to complete this research.

My grateful thanks also go to Head of University of Sumatera Utara (USU) who has

provided funding endorsement for me to finish my PhD. My great appreciates also

go to all staffs in School of Materials and Mineral Resources Engineering, Universiti

Sains Malaysia (USM) which has provided a great and comfortable environment for

me to carry out my research.

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Special thank to my beloved wife, Dr. Irwana Nainggolan who has given endless

support, prayers and steadfast motivation whenever I felt challenged in completing

this project. Last but not least, my heartiest gratitude to my family for their

unconditional love, motivation and support that have been a great support either

directly or indirectly in the completion of this study.

iv
 Dedication

This thesis is dedicated to my wife, two my sons, my daughter, my mother in law and

my sisters.

v
 List of contents

Author’s declaration ....................................................................................................ii

Acknowledgement ......................................................................................................iii

Dedication ....................................................................................................................v

List of contents ............................................................................................................vi

List of tables .................................................................................................................x

List of figures ............................................................................................................xiii

List of symbols ………………………………………………………………….....xxi

List of abbreviations.................................................................................................xxii

Abstrak....................................................................................................................xxiii

Abstract....................................................................................................................xxv

CHAPTER 1………………………………………………………………………….1

1.1 Research background…………………………………………………………...1

1.2 Problem statements…………………………………………………………..…4
1.3 Research objectives…………………………………………………………….7
1.4 Project approach………………………………………………………………..7

CHAPTER 2………………………………………………………………………….9

2.1 Chitin…………………………………………………………………………..9

2.2 Chitosan……………………………………………………………………….10

2.2.1 Properties of chitosan…………………………………………………11

2.2.2 Application of chitosan film…………………………………………..12

2.2.3 Methods for chitosan film fabrication………………………………...13

vi
2.3 Amine…………………………………………………………………………17

2.4 Hydroxyl………………………………………………………………………20

2.5 Acetone………………………………………………………………………..22

2.6 Acetone in human breath…………………………………………….………..24

2.7 Conventional methods to determine acetone concentration…………..………25

2.7.1 Colorimeter……………………………………………...…………….26

2.7.2 Gas chromatography (GC)………………………………………….…28

2.8 Acetone sensor materials……………………………………………………...33

2.8.1 Acetone sensor material based on polypyrrole (PPy)………………...35

2.8.2 Acetone sensing properties of

polypyrrole/poly(methylmethacrylate) blend sensor……………..….38

2.8.3 Acetone sensing properties of ferrocenecarboxymethylated

polymer sensor……………………………………………….……….39

2.8.4 Acetone sensing properties of ZnO-based elements………………..…43

2.8.5 Acetone sensing properties of screen printed WO3 thick films……….44

2.8.6 Acetone sensing properties of coralloid SnO2

nanostructures-based sensor…………………………………………..51

2.8.7 Acetone sensing properties of nanoparticles TiO2 thin film sensor…..55

2.8.8 Acetone sensing properties of nano-sized La2O3 sensor………...……59

2.8.9 Acetone sensing properties of LaFeO3 thin film sensor……………....60

2.8.10 Acetone sensing properties of La1−xPbxFeO3 perovskite system….….62

2.9 Diabetes mellitus……………………………………………………………...64

2.10 Deposition techniques………………………………………………………...66

vii
 2.10.1 Sputter deposition……………………………………………………..67
2.10.2 Thermal evaporation in vacuum………...…………………………….68

2.10.3 Electron beam physical vapor deposition………………………..……69

2.10.4 Chemical vapor deposition…………………………………………....71

2.10.5 Sol-gel technology…………………………………………………….72

2.10.6 Electrochemical deposition…………..……………………………….73

2.11 Summary………………………………………………………………………75

CHAPTER 3……………………………………………………………………...…76

3.1 Experimental plan………………………………….……………..…...76

3.2 Material and equipment……………….............................................................77
3.3 Chitosan solution preparation...........................................................................78
3.4 Mask design.......................................................................................................80
3.5 Process of aluminum patterning on silicon wafer.............................................83
3.6 Electrochemical deposition...............................................................................84
3.7 Annealing process.............................................................................................86
3.8 Electrical testing................................................................................................87
3.8.1 Sample selection for electrical testing...................................................87
3.8.2 Electrical testing procedure...................................................................88
3.8.3 Data collection.......................................................................................89
3.8.4 Data interpretation.................................................................................89
3.9 Moicrostructure characterizations.....................................................................94

CHAPTER 4...............................................................................................................96

4.1 Electron microscope image...............................................................................96

4.2 Fourier transform-infrared spectroscopy (FT-IR) results.................................97
4.3 SEM of chitosan film sensors.........................................................................101
4.3.1 SEM analysis for chitosan films with variation of
deposition time....................................................................................102
4.3.2 SEM analysis for chitosan films fabricated from

viii
 various deposition voltage...................................................................106
4.4 Surface analysis by atomic force microscopy (AFM).....................................107
4.4.1 AFM analysis for chitosan films with variation
of chitosan solution concentration.......................................................108
4.4.2 AFM analysis for chitosan films with
variation of annealing temperature......................................................112
4.5 Electrical testing of chitosan film sensor fabricated

from different concentration............................................................................113

4.6 Electrical testing for of chitosan film sensor fabricated

from different deposition time.........................................................................123
4.7 Electrical testing for of CBAS fabricated from
different deposition voltage............................................................................132
4.8 Electrical testing for of CBAS fabricated from
different annealing temperatures.....................................................................139
4.9 Electrical response of CBAS to acetone vapour-contaminated
air as a function of time...................................................................................148

4.10 Sensing mechanism of CBAS to acetone vapour............................................153

CHAPTER 5.............................................................................................................160

5.1 Conclusion.......................................................................................................160

5.2 Future recommendations.................................................................................162

5.3 Commercialization Potential...........................................................................163

REFERENCES.........................................................................................................164
APPENDICES
APPENDIX A
APPENDIX B

ix
 List of Tables

Tabel 2.1: The properties of acetone (Tang et al., 2008)..........................................23

Table 3.1: Variation of chitosan solution concentration...........................................79

Table 3.2: Variation of deposition voltage................................................................86

Table 3.3: Variation of deposition time....................................................................86

Table 3.4: Variation of Annealing Temperature.......................................................87

Table 4.1: The roughness of chitosan film surface for different

chitosan concentration............................................................................110
Table 4.2: The roughness of chitosan film surface for different

chitosan concentration............................................................................113

Table 4.3: The minimum and maximum values of relative humidity

and ambient temperature of CBASs fabricated from
different chitosan concentrations during operation................................118
Table 4.4: Standard deviation of maximum output voltage for CBASs

fabricated from the different chitosan solution concentration...............120

Table 4.5: The final output voltage values of CBASs fabricated

from the different chitosan solution concentration

as the stabilization and recovery............................................................120

Table 4.6: The difference between final output voltage as the stabilization

and reco-very of the CBASs fabricated from

the different chitosan solution concentration.........................................121

Table 4.7: The minimum and maximum values of relative humidity

and ambient temperature of CBASs fabricated from

x
 different deposition time during testing.................................................127

Table 4.8: Standard deviation of maximum output voltage for CBASs

fabricated from the different deposition times.......................................129

Table 4.9: The final output voltage values of CBASs fabricated

from the different deposition times

as the stabilization and recovery............................................................130

Table 4.10: The difference between final output voltage as

the stabilization and recovery of the CBASs fabricated

from the different deposition time.........................................................130

Table 4.11: The minimum and maximum values of relative humidity

and ambient temperature of CBASs fabricated from

different deposition time during testing.................................................135

Table 4.12: Standard deviation of maximum output voltage for CBASs

fabricated from the different deposition voltages..................................136

Table 4.13: The final output voltage values of CBASs fabricated

from the different deposition time as
the stabilization and recovery................................................................137

Table 4.14. The difference between final output voltage as

the stabilization and recovery of the CBASs

fabricated from the different deposition time........................................137

Table 4.15: The minimum and maximum values of relative humidity

and ambient temperature of CBASs fabricated from

different annealing temperatures during testing....................................144

Table 4.16. Standard deviations of maximum output voltage for CBASs

xi
 fabricated from the different annealing temperatures............................145

Table 4.17: The final output voltage values of CBASs fabricated

from the different annealing temperatures

as the stabilization and recovery............................................................146

Table 4.18: The difference between final output voltage as the stabilization

and recovery of the CBASs fabricated from

the different annealing temperatures......................................................146

xii
 List of Figures

Fig. 2.1: Chemical structure of chitin (Rinaudo, 2006)..............................................9

Fig. 2.2: Chemical structure of chitosan (Caroline et al., 2003)…………………...11

Fig. 2.3: Photolithographically patterned chitosan features.

(a) SEM of matched comb structure. (b) SEM of 2-mm-wide

(b) lines with 4-mm pitch. (c) SEM of serpentine spring structures.
(d) SEM of a sawtooth structure. Note: All patterns are in
a 2-mm-thick chitosan layer (Cheng et al., 2008)………………..….14
Fig. 2.4:. Schematic demonstrating chitosan film electrodeposition

(Yi et al., 2005)……………………………………………………….…..17

Fig. 2.5: Chemical structure of primary amine (McMurry, 1992)…………………18

Fig. 2.6: Chemical structure of (a) secondary amine and

(b) tertiary amine (Lewis, 1923)………………………………………….20

Fig. 2.7: Chemical structure of hydroxyl (Lide, 2005)…………………………….20

Fig. 2.8: Electron cloud surrounding the whole hydroxyl (Lide, 2005)…………...21

Fig. 2.9: Structural formula of the acetone molecule (Cao et al., 2001)…………...22

Fig. 2.10: Acetone as a final product of metabolic processes (Hamilton, 2007)……25

Fig. 2.11: Colorimeter tube for acetone concentration

determination (Harwood and Moody, 2005)……………………………..26

Fig. 2.12: Colorimetric method with ready-to-use DPD reagents, test kits,
colorimeters and spectrophotometers (Sharma, 2002)…………………...27

Fig. 2.13: Paper indicator for thedetermination of acetone

concentration in urine (Joo, 2010)………………………………………..28

Fig. 2.14: (a) Schematic of a GC-MS, (b) schematic of a FID and

xiii
 (c) actual picture of GC-MS followed

by FID (Phillips, 1997)…………………………………………………...30

Fig. 2.15: Schematic of selected ion flow tube mass spectrometry

(SIFT/MS) (Adams et al., 1976)…………………………………………32

Fig. 2.16: Schematic of proton transfer reaction mass spectrometry

(PTRMS) (Taucher et al., 1996)………………………………………….33

Fig. 2.17: The response of PPy to acetone vapor at the concentration of

17 vol.% in N2 at 24oC and at 49% RH (Ruangchuay et al.

2004)…………………………………………………………………..….35

Fig. 2.18: Schematic representation of a sensor testing system of organic

vapors (Bazzaoui et al., 2007)……………………………………………36

Fig. 2.19: Responsive electrical behavior of ferrocenecarboxymethylated

polymer upon injection and removal of mixed

N2/acetone vapor (Hachawee et al., 2008)……………………………….39

Fig. 2.20: Attachment of two small copper wires on specimen

by silver paint (Hachawee et al., 2008)…………………………………..40

Fig. 2.21: The experimental setup used for measuring electrical

conductivity and gas sensitivity of the electroactive

ferrocenecarboxymethylated polymer specimen

(Hachawee et al., 2008)…………………………………………………..42

Fig. 2.22: Relationship between operating temperature and the sensitivity

of screen printed WO3 thick film for 50 ppm

acetone vapor in air (Khadayate et al., 2007)…………………………….45

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Fig. 2.23: The sensitivity versus acetone vapor concentration

of WO3 thick film (Khadayate et al., 2007)……………………………...48

Fig. 2.24: Response of screen printed WO3 thick film operated at 300 oC

to 50 ppm acetone vapor in air (Khadayate et al., 2007)………………...50

Fig. 2.25: Repetitive response of screen printed WO3 thick film upon

exposure to 50 ppm acetone vapor at 300 oC

operating temperature (Khadayate et al., 2007)………………………….51

Fig. 2.26: (a) A gas sensor, (b) external part of the gas-sensor package

and (c) electronic gas sensor system. Ei is the electrical

source of the measure; Eo is the electrical

source of the heating (Fang et al., 2010)…………………………………52

Fig. 2.27: (a) Response of SnO2-based gas sensor to 50 ppm benzaldehyde

(black curve) and acetone (red curve). (b) Linear dependence

relation between response sensitivity and relative

benzaldehyde (black) andacetone (red)

concentrations (Fang et al., 2010)………………………………………..54

Fig. 2.28: Typical dynamic response of the MAPLE-deposited TiO2

sensing layer to different concentrations of acetone vapors

at the working temperature at 400oC (Rella et al., 2007)………………...56

Fig. 2.29: Sensor responses (defined as Ivap/Iair) towards 100 ppm of

ethanol and acetone vapors in dry air as a function

of working temperature (Brousse et al., 1996)…………………………...57

Fig. 2.30: Sensor responses towards ethanol and acetone

xv
 vapors as a function of alcohol vapor

concentration (Brousse et al., 1996)……………………………………...58

Fig. 2.31: Selectivity of the La2O3 sensor. Conditions:

wavelength, 490 nm; temperature, 361oC; and

air flow rate, 200mL min−1 (Tang et al., 2008)…………………………..59

Fig. 2.32: The response property of the LaFeO3 thin film

to 80 ppm acetone gas tested at 400oC (Liu et al.,

2006)…………………….………………………………………………..61

Fig. 2.33: The temperature dependence of the sensitivity in

different concentration of acetone gas for sample

La0.68Pb0.32FeO3 (Zhang et al., 2006)……………………………………..62

Fig. 2.34: The concentration dependence of sensitivity at T = 240oC for

sample La0.68Pb0.32FeO3 to acetone gas (Kong, Shen, 1996)…………….63

Fig. 2.35: Schematic of sputter deposition (Drüsedau et al., 2000)…………………67

Fig. 2.36: An scheme of the deposition equipment used in

the laboratory (Kovacik et al., 2012)……………………………………..68

Fig. 2.37: Schematic of an EBPVD system (Lugscheider et al., 1998)……………..69

Fig. 2.38: Schematic of a chemical vapor deposition

method (Kojima et al., 1990)……………………………………………..70

Fig. 2.39: Sol-gel processing routes (Amorphous particles, nanometer

scale, water based, metal alkoxides or salts, molecular scale,

solvent based) (Adraider et al., 2011)……………………………………71

xvi
Fig. 2.40: Schematic of electrochemical deposition method (Kim et al., 2006)…….73

Fig. 3.1: Project flowchart………………………………………………………….76

Fig. 3.2: Preparation of the transparent yellow chitosan solution…………………78

Fig. 3.3: Dimension of the alignment mask………………………………………..80

Fig. 3.4: Aluminum layer mask (mask 1)………………………………………….81

Fig. 3.5: Chitosan film mask (mask 2)……………………………………………..82

Fig. 3.6: Dimension of aluminum pattern………………………………………….83

Fig. 3.7: Fabrication process of chitosan film sensor in layer by layer form……...84

Fig. 3.8: Chitosan solution electrochemical deposition process…………………...85

Fig. 3.9: Experimental setup for the electrical properties

measurement of the sensor……………………………………………….88

Fig. 3.10: An illustration of exposure of (a) wet air and (b) acetone vapour
contaminated wet air to the surface of chitosan layer……………………91

Fig. 3.11: A calibration curve plot showing limit of detection (LOD),

limit of quantification (LOQ), dynamic range,

and limit of linearity (LOL)………………………………………………93

Fig. 3.12: The schematic of AFM using a cantilever

to scan the specimen surface……………………………………………..95

Fig. 4.1: Image of chitosan film in membrane form

deposited onto aluminium layer………………………………………….97
Fig. 4.2: FTIR spectrum of chitosan powder………………………………………98
Fig. 4.3: FTIR spectrum of chitosan solution……………………………………...99

Fig. 4.4: FTIR spectrum of chitosan film………………………………………...101

Fig. 4.5: The SEM images of chitosan film sensors fabricated

xvii
 from the deposition time: (a) 2 minutes, (b) 4 minutes,

(c) 6 minutes, (d) 8 minutes and (e) 10 minutes………………………...103

Fig. 4.6: Electrochemical deposition process…………………………………….104

Fig. 4.7: SEM images of CBASs fabricated from deposition
voltage of (a) 0.5 V and (b) 2.0 V………………………………………107
Fig. 4.8: AFM images of CBAS fabricated from the various chitosan
concentration: (a) 1.00% w/v, (b) 1.25% w/v,
(c) 1.50% w/v, (d) 1.75% w/v and (e) 2.00% w/v………………………109
Fig. 4.9: The values of roughness (Ra) and root mean square (RMS)

of chitosan film sensor as a function of chitosan concentration………...110

Fig. 4.10: Particle size of chitosan film vs chitosan concentration………………...111

Fig. 4.11: AFM images of CBAS sensor fabricated at the annealing

temperatures of (a) 105oC and (b) 110oC……………………………….112

Fig. 4.12: Output voltage as a function of time for CBASs fabricated

from different chitosan concentration…………………………………..114

Fig. 4.13: (a) the change in relative humidity and (b) the change in

ambient temperature during testing for CBASs fabricated

from different chitosan concentration…………………………………..117

Fig. 4.14: Reproducibility of CBAS fabricated from different chitosan

concentrations for five times measurement……………………………..122

Fig. 4.15: Output voltage vs time for CBASs fabricated

from different deposition time…………………………………………..123

Fig. 4.16: (a) the change in relative humidity and (b) the change in

ambient tempera-ture during testing for CBASs

fabricated from different deposition time……………………………….126

xviii
Fig. 4.17: Reproducibility of CBAS fabricated from different

deposition time for five times measurement……………………………131

Fig. 4.18: Output voltage vs time for CBASs fabricated from

different deposition voltage……………………………………………..133

Fig. 4.19: (a) the change in relative humidity and (b) the change in

ambient tempera-ture during testing for CBASs

fabricated from different deposition voltage……………………………134

Fig. 4.20: Reproducibility of CBAS fabricated from different

deposition voltage for five times measurement…………………………138

Fig. 4.21: The life time of CBAS annealed at different temperatures……………..140

Fig. 4.22: Output voltage as a function of time for CBASs fabricated

from different annealing temperatures………………………………….141

Fig. 4.23: (a) the change in relative humidity and (b) the change in

ambient temperature during testing for CBASs

fabricated from different chitosan concentration……………………….142

Fig. 4.24: Reproducibility of CBAS fabricated from different

annealing temperature for five times measurement…………………….147

Fig. 4.25: The electrical response of CBAS to normal air and

acetone vapour-contaminated air as a function of time…………………149

Fig. 4.26: (a) The changes in humidity and (b) ambient temperature

of CBAS as a function of time during testing in

normal air and acetone vapour-contaminated air……………………….151

Fig. 4.27: The electrical response of CBAS to acetone

xix
 vapour as a function of concentration…………………………………..152

Fig. 4.28: The change in output voltage of chitosan film

sensor as a function of time upon exposed

to normal air under 5.0 V extra voltages………………………………..153

Fig. 4.29: Illustrations of the sensing mechanism of chitosan film sensor:

(a) the adsorption of oxygen, (b) the formation of oxygen species,

(c) exposure to water molecules, and (d) the release of electrons............156

Fig. 4.30: Electron cloud surrounding the whole hydroxyl group…………………157

Fig. 4.31: Two polar bonds of water……………………………………………….158

Fig. 4.32: The formation of hydrogen bonds from the reaction

between water molecules and amino groups……………………………159

xx
 PENDERIA ASETON BERASASKAN CITOSAN UNTUK MENGESAN
ASETON KEPEKATAN RENDAH

Abstrak

Penderia aseton berasaskan citosan (chitosan based acetone penderias, CBASs) yang
mampu mengesan kehadiran wap aseton kepekatan rendah telah berjaya dihasilkan
melalui penggabungan teknik fotolitografi dan pemendapan elektrik. CBAS telah
dibuat dalam bentuk filem dalam mana citosan yang digunakan dihasilkan secara
sintesis dari hasil sisa kulit ketam yang dibeli dari Sigma Aldrich. Telah diperolehi
bahawa kumpulan amin dan hidroksil di dalam struktur molekul filem citosan dapat
bertindak balas secara baik dengan molekul-molekul wap aseton yang menyentuh
permukaannya. Ini ditandakan dengan voltan keluaran CBAS yang berbeza untuk
kepekatan aseton yang berbeza. Pengesanan aseton kepekatan rendah telah
dipertimbangkan sebagai satu bidang kajian yang menarik berasaskan laporan
perubatan yang menyatakan kepekatan aseton di dalam napas berbeza dari 0.3 to 0.9
ppm pada orang sihat sehingga melebihi 1.8 ppm untuk pesakit kencing manis. Ini
menjadikan aseton satu penanda kimia yang sesuai untuk pengenalpastian penyakit
kencing manis melalui napas yang dihembuskan. Oleh itu, kajian mengenai ujian
makmal sifat-sifat elektrik CBAS terhadap udara yang dicemari wap aseton dalam
julat 0.1 – 100 ppm telah dilakukan pada suhu bilik (~25-30oC) di dalam udara
normal. Ini bertujuan untuk membuktikan keupayaan yang besar CBAS untuk
digunakan dalam mengenalpasti penyakit kencing manis melalui nafas yang
dihembuskan oleh pesakit terbabit. Hasil kajian telah menunjukkan bahawa tindak
balas elektrik CBAS meningkat dengan sangat pantas, sekitar 10 saat, segera
permukaannya didedahkan samada terhadap udara bersih ataupun udara yang
dicemari wap aseton. Penderia juga telah menunjukkan penyembuhan yang cepat,
iaitu kurang dari 200 saat apabila pendedahan dihilangkan dan bacaan turun ke nilai
yang asal. Sebagai akibat pendedahan selama 5 minit, CBASs dibawah pengaruh
voltan yang dibekalkan sebesar 1.5 volt menghasilkan nilai balasan anggaran 3.9,
12.3, 40.5, 47, 66.4 dan 70.2% apabila CBASs didedahkan terhadap udara yang
dicemari aseton dengan kepekatan masing-masing 0.1, 1, 10, 20, 50 and 100 ppm.
Apabila pengukuran diulangi secara berterusan, CBASs telah menunjukkan
kebolehulangan dengan cukup memuaskan. Selama pengujian, CBASs berfungsi

xxiii
pada keadaan yang mantap, yang ditandakan dengan tidak terdapatnya keadaan naik
turun yang bermakna dalam bacaan. Dengan demikian, pencapaian sifat-sifat elektrik
CBAS telah mencapai syarat-syarat sebagai penderia yang baik. Selanjutnya CBAS
telah menunjukkan kemampuannya untuk mengesan wap aseton dalam kepekatan
sangat rendah yang menjadi bukti kepada keupayaannya untuk digunakan sebagai
penderia penyakit kencing manis yang praktis, ramah persekitaran dan berpencapaian
tinggi di masa hadapan.

xxiv
 CHITOSAN BASED ACETONE SpENSOR FOR DETECTION OF
LOW CONCENTRATION ACETONE

Abstract

Chitosan based acetone sensors (CBASs) which are capable of sensing the presence
of acetone vapor in low concentration have been successfully fabricated including
the combination of photolithograpy and electrodeposition techniques. The CBASs
were fabricated in film form in which the chitosan used was synthesized from waste
product of crab shells which was purchased from Sigma Aldrich. It was found that
the amine and hydroxyl groups in molecular structure of chitosan film can interact
well with the molecules of acetone vapor touching its surface. This interaction was
indicated by different output voltages of CBAS for different concentration of
acetone. The detection of low concentration acetone has been considered as an
attractive area of study. Some medical reports express that the acetone concentration
in the breath varies from 0.3 to 0.9 ppm in healthy people to more than 1.8 ppm for
diabetics. This makes acetone a suitable chemical marker for diabetes diagnosis
through exhaled breath. Therefore, the preliminary study on the laboratory testing of
the CBAS electrical properties to acetone vapor-contaminated air in range of 0.1 –
100 ppm was carried out at room temperature (~25-30oC) in normal air. This aims to
prove the great potential of CBAS as an application in diagnosing diabetes mellitus
through breath exhaled by diabetics. The results showed that electrical response of
CBAS increased very rapidly, in around 10 s, once its surface was exposed to both
wet air and acetone vapor contaminated wet air. The sensors also exhibited quick
recovery i.e., approximately less than 200 s once the exposure was removed and the
reading dropped to its original values. As a result of exposure for 5 min, CBASs
under 1.5 volts in supplied voltage yield maximum response values of approximately
3.9, 12.3, 40.5, 47, 66.4 and 70.2% when they are exposed to 0.1, 1, 10, 20, 50 and
100 ppm acetone vapor contaminated wet air, respectively. When the measurements
were repeated continuously, the CBASs showed remarkably adequate repeatability.
During testing, the CBASs operated in stable condition, indicated by no significant
fluctuation in reading. Thus, the performance of the CBAS electrical properties
achieved the requirements of a good sensor. Moreover, its ability to detect acetone
vapor in very low concentrations, it was thus established that CBAS produced in this

xxv
work has very promising potential to be applied as a practical, environment friendly
and high performance diabetes biosensor in the future.

xxvi
 CHAPTER 1

INTRODUCTION

1.1. Research background

Diabetes mellitus is a chronic disease indicated by elevated glucose (blood sugar)

levels that result from cells in body cannot effectively use available insulin to burn

glucose for energy. Inability in controlling the progress of diabetes can lead to

serious medical complications such as nerve damage, blindness, limb amputation,

kidney failure, heart disease, premature death and gangrene. Nowadays, the disease

is suffered by millions of individuals worldwide. Current method to test blood sugar

level is by using blood sample which is obtained by piercing the skin on the finger.

The blood is then lied onto to a disposable test-strip for chemically reacting. This

method is tedious, invasive and may be painful. This paves the way for improved

methods to detect diabetes, possibly by using other chemical markers exhibited by

diabetics (Turner et al., 2009).

The breath of diabetics is known to be of bad odor, with a fruity scent. This fruity

scent, or the smell of decayed apples, to some, is a direct result of acetone presence

in the breath (Likhodii et al., 2002). In breath of healthy individuals, the acetone

concentration is in range of 0.3 to 0.9 ppm (Deng et al., 2004). However, in the

exhaled air of diabetic patients, the acetone concentration is reported to be more

than 1.8 ppm (Sahay et al., 2005). Such significant difference makes acetone a

suitable breath marker in detecting diabetes (Turner et al., 2009; Likhodii et al.,

2002; Deng et al., 2004). Furthermore, Liu et al. (2006) found that acetone vapor

1
produced from lypolysis in human body can easily carried by the breath into

alveolus walls and finally, it mixes with alveolus air. Hence, the amount of acetone

vapor in the breath exhaled by diabetics must be comparable to the amount of

acetone vapor in the blood. This further increases the attractiveness of using

acetone levels in the breath as a reliable diabetes marker. As only the patient’s

breath is needed for analysis, a breathalyzer-like apparatus is all that is needed to

diagnose diabetes. The only obstacle now in ensuring a trouble free, painless and

steadfast diabetic detection is a reliable acetone sensor.

Acetone sensors have been considered as an attractive area of study due to its

promising application in the diagnosis of diabetes mellitus. However, fabricating

reliable acetone sensors in breathalyzers to obtain accurate data, in the aid of

diabetes diagnosis is far from trivial. Amongst others, reliability under different

working conditions (i.e., different levels of vapor in air and in breath, different

temperatures etc.), sensitivity (i.e., detection of acetone at various levels of

concentrations), fabrication techniques and cost remain as issues. Therefore, the

main objective of this work focuses on the fabrication of a sensor to detect acetone

in human breath. In this study, chitosan-based detectors, or chitosan-based acetone

sensors (CBAS) are proposed based on a few arguments, as follows.

Firstly, chitosan is proven to be able to detect very low concentrations of acetone

vapor. It is conceded that detecting acetone in breath may be more complicated. In

order to prove that the CBAS have the required reliability in detecting breath

acetone in further work, laboratory tests of the electrical properties of chitosan film

sensors must be first conducted. This will be presented later in this thesis. Secondly,

2
CBAS seems to offer advantages, compared to semiconductor acetone sensors

(SAS). Metal oxides such as ZnO, SnO2, TiO2 and WO3 which are usually used to

make SAS, in single phases or in the form of compounds or composites (Anno et

al., 1995; Khadayate et al., 2007; Rella et al., 2007; Patil and Patil, 2007), operate at

high temperatures (usually above 200oC). CBAS proposed in this work can operate

at room temperature and wet environments. This eliminates the need for high power

consumption and complicated designs which require a heating element. Thirdly,

chitosan which is used as a sensitive layer in this sensor design is a natural bio-

polymer, produced by treating seafood waste (shrimps, crabs, lobsters, krills, etc)

(Majeti and Kumar, 2000; Prashanth and Tharanathan, 2007, Nasution et al., 2013;

Pillai et al. 2009). Hence, it is a reproducible resource which does not harm the

environment once they are discarded.

All these advantages suggest that CBAS have a high competitive value in the

commercial market, especially for breath acetone detection application. To date,

other than our published work (Nasution et al., 2013), no reports on the utilization

of chitosan for acetone sensing were found, especially for diabetes analysis

purposes. This is maybe due to its swelling property, which apparently makes it

difficult to obtain reliable measurements.

In this study, the CBASs were fabricated in film forms using an electrochemical

deposition method because the large amine groups presented in chitosan having

~6.5 in pKa value have enabled the chitosan soluble in aqueous acidic media (Pillai

et al. 2009; Gerard et al., 2002). The CBAS was characterized using Fourier

Transform Infrared (FTIR) spectrometer to identify the existence of amine and

3
hydroxyl in CBAS structure. Another characterization used was Scanning Electron

Microscopy (SEM) to observe surface morphology of CBAS. Atomic Force

Microscopy (AFM) was used for measuring the surface roughness and particle size

of CBAS. For laboratory test scale, chitosan film properties including sensitivity,

response, stability, recovery, repeatability, life-time, recovery and selectivity were

tested by measuring the changes in output voltage of CBAS, both in exposure to

wet air and acetone vapor-contaminated air. The electrical testing results were

related to the results of molecular structure characterization to determine electrical

properties of CBAS and in turns, to explain the sensing mechanism of CBAS.

1.2. Problem statements

Commercial semiconducting materials such as TiO2, SnO2, WO3, ZnO and La2O3

have been well known as good sensing materials for acetone vapor measurement.

Although, they have been established as an excellent sensing material for detecting

acetone vapour, but they are not obtained from reproducible resources that make

their cost to be continuously more and more expensive and in turns, uncompetitive in

satisfying the market demand for low cost sensors. Moreover, most of them operate

at high operating temperatures (generally above 200oC). This requires an extra heat

element which results in more complicated design. All those problems have led them

to low fabrication cost (Sahay, 2005; Anno et al., 1995; Khadayate et al., 2007;

Wang et al., 2008; Fang et al., 2010; Pinna et al., 2004; Vuong et al., 2004 and 2005;

Brousse and Schleich, 1996; Rella et al., 2007; Tang et al. 2008).

4
Beside the sensors based on metal oxides, the sensors based on conductive polymers

(CPs) such as polypyrrole, polyaniline and polythiophene have been used for

measuring acetone vapor at room temperature. Over other metal oxides, those CPs

have the other advantages including lighter weight, lower price, non-corrosive and

easy to prepare. However, they are synthetic polymers which are human-made

polymers and therefore, they are mostly non-biodegradable (Ruangchuay et al., 2003

and 2004; Nigorikawa et al., 1995; Bazzaoui et al., 2007; Hachawee et al., 2008).

Chitosan which is a natural bio-polymer appears very promising as an alternative

acetone sensing material to replace the metal oxides and traditional CPs in an effort

to overcome the mentioned problems. This is due to its production comes from the

waste treatment of the crustacean shells such as crabs, shrimps, krills and lobsters, so

that it constitutes a reproducible resource and hence, biodegradable and low cost for

production. The chitosan is also not harmful to the environment when they have

become discarded objects and thus, enables us to develop the environmentally

friendly biosensor (Caroline et al., 2003; Majeti and Kumar, 2000; Prashanth and

Tharanathan, 2007, Nasution et al., 2013; Pillai et al. 2009; Rinaudo, 2006; Luo et

al., 2005; Yang et al.; Spagna et al., 2007).

The other advantage is regarding the active amino group (-NH2) in chitosan. Due to

this chitosan films can be fabricated by a simple electrochemical method (Pillai et al.

2009; Gerard et al., 2002). The use of the electrochemical method for sensor

fabrication can overcome the problem of expensive equipments and high energy

consumption like inherited by the physical deposition techniques including vacuum

evaporation, sputtering, electron beam deposition and thermal evaporation. And

when compared to chemical methods such as sol–gel deposition and chemical vapor
5
deposition (CVD), the electrochemical method also provides advantage because

irregularly shaped specimens can be deposited at short time even at low temperature

with a homogeneous or coherent layer. The chemical composition and thickness of

deposited layer can be well controlled through adequate conditions of the

electrodeposition process.

Besides that, because of chitosan contains large groups of amino (–NH2) and

hydroxyl (–OH) in its molecular structure, there are much active sensing sites on the

chitosan film surface to interact with certain analytes. This makes its sensing

mechanism can be done by direct interactions between chitosan surface and analyte.

Thus it does not require the aid of light for the sensing process in which the sensor

surface interacts indirectly with any analyte, but relied on the change in light

intensity exposed to sensor surface. The magnitude of intensity is determined by the

absorptive capacity of the analyte. Although, indirect interactions can avoid the

surface damages of the sensor, however, the sensitivity to contamination such as

grease or dust in sensing path can lower the accuracy in measurement.

Although the chitosan have several advantages as an acetone sensing material, but its

inherent swelling property has become an obstacle to stick strongly the chitosan layer

on the patterned aluminum layer during the electrochemical deposition process.

Therefore, the study on chitosan solution concentration, deposition voltage,

deposition time and annealing temperature has become very important to be done to

obtain most suitable fabrication parameters. The selected fabrication parameters were

used to fabricate chitosan based acetone sensor (CBAS) to detect acetone

concentration in range of 0.1 until 100 ppm.

6
1.3. Research objectives

The research is focusing on a study of the electrical properties to find out the sensing

mechanism of chitosan film sensors. The objectives are:

i. To fabricate chitosan film sensors capable of sensing acetone vapor at

room temperature by using electrochemical deposition method.

ii. To investigate the electrical properties of chitosan film sensors under wet

air and acetone vapor-contaminated wet air exposures.

iii. To propose sensing mechanism of chitosan film to acetone vapor.

1.4. Project approach

This study was divided into three main parts; sensor fabrication, electrical testing and

molecular structure characterizations. Sensor fabrication was carried out at room

temperature using the electrochemical deposition method. The fabrication process

consists of chitosan solution preparation, aluminum patterning, electrochemical

deposition and annealing. In solution preparation, the chitosan solution

concentrations were varied in range of 1% w/v until 2% w/v with the increment of

0.25%. And then in deposition process, the deposition voltage was varied in range of

0.5, 1, 2 and 5 V and deposition time was varied in range of 2 until 10 minutes with

the increment of 2.0 minutes. Finally, the variation was done for the annealing

process in which the annealing temperatures are in range of 85 until 110oC with the

increment of 5oC.

7
The electrical properties of the CBAS fabricated with the various parameters were

tested using a potential method at room temperature (~25-30oC) and in humidity

around ~56-68%. The electrical properties include sensitivity, responsibility,

stability, repeatability, recovery and reproducibility. The measurement results in

graphs were compared each other to obtain the optimum parameters for fabrication.

The electrical testing was carried out under wet air and acetone vapor-contaminated

wet air exposes in which the acetone concentration vaporized are varied between 0.1

and 100 ppm. Each measurement was repeated as many as five times.

The different electrical properties of chitosan film sensors for the variation of

fabrication parameters were related to the results of molecular structure

characterization. The characterization of the sensors was carried out by using Fourier

Transform Infrared (FTIR), Atomic Force Microscopy (AFM) and Scanning Electron

Microscopy (SEM). The information about the relationship between electrical

properties and molecular structure was utilized to propose the sensing mechanism of

chitosan film sensors in detecting the presence of acetone vapor in wet air.

8
 CHAPTER 2

LITERATURE REVIEW

2.1 Chitin

Chitin as shown in Fig. 2.1 is naturally second most abundant biopolymer and can be

presented in the fungal cell wall as well as the exoskeletons of lots of arthropods

such as shrimps, lobster, crabs or squid pens. Chitin is hard to be dissolved and the

only way to make chitin soluble is by decomposing it slowly using a certain enzyme.

Fig. 2.1: Chemical structure of chitin (Rinaudo, 2006).

9
2.2 Chitosan

The deacetylation of chitin has been done to derive chitosan which is a functional

biopolymer with the units of primarily of β (1→4) linked 2-amino-2-deoxy-β-D-

glucopyranose (Caroline et al., 2003). The groups of amine and hydroxyl in its

molecular structure, as shown in Fig. 2.2, can act as the active sites which can

directly interact with certain analytes. Because chitin is available in large number,

the fabrication of chitosan is rather cheap.

Nowadays, the application of chitosan is in wide range such as in area of agriculture

(Pornpienpakdee et al., 2010; Galed et al., 2004), food and beverage (Spagna et al.,

2001; Lee et al., 2012; de Britto et al., 2012), biopharmaceuticals (Fan et al., 1012),

water and waste treatment (Lalov et al., 2000; Miretzky et al., 2009; Bhatnagar and

Sillanpää, 2009), material science (Jayakumar et al., 2010; Ma and Sahai, 2013;

Jayakumar et al., 2010), cosmetics (Onésippe and Lagerge, 2008) and recently in

gene therapy too (Jayakumar et al., 2010). The increasing interest of chitosan

applications since it has some attractive properties such as good biodegradability,

good biocompatibility, excellent ability in filmforming, good adhesion, high

mechanical strength, hydrophilicity, physiological inertness and non-toxicity (Luo et

al., 2005; Yang et al., 2004).

10
 Fig. 2.2: Chemical structure of chitosan (Caroline et al., 2003).

Chitosan application has been popularly recognized in biomedical area (Rinaudo,

2006) because its anti-microbial, anti-viral, anti-fungal, absorption of exude and

wound-healing properties (Jeon et al., 2001; Wu et al., 2004; Liu et al., 2006). The

antibacterial activity of chitosan can be determined by controlling the variation of

molecular concentration and weight (MW) (Liu et al., 2006; Jeon et al., 2001).

Additionally, chitosan has also been applied to remove the scar tissue which

functions as a wound management aid as well as serving as a supporting material for

gene delivery, cell culture, tissue engineering, drug delivery, anti-microbial agents

and adsorption agents (Jayakumar et al., 2005).

2.2.1 Properties of chitosan

The chitosan’s properties are explained by the presence of its primary amine groups

with pKa ~6.5. At pH lower than 6.5, the amines are positively charged and chitosan

11
is soluble. At higher pH, the amines are increasingly deprotonated, and it becomes

insoluble. This pH dependent solubility allows stable chitosan films to be deposited

on device surfaces through several methods. Importantly, the film can be dissolved

and removed by a mildly acidic wash, and the device can be re-used multiple times.

The structure of the film depends on the deposition conditions, the chitosan

molecular weight, and its degree of deacetylation (Pillai et al. 2009; Gerard et al.,

2002).

Chitosan films immersed in liquid experience strong pH dependent swelling (Wang

et al., 2004). The pH of the solution determines thamount of positive charge on the

chitosan chains, which in turn affects the electrostatic repulsion between the chains

and the volume of the polymer. This swelling behavior can be utilized for actuation

or for controlled drug release (Spinks et al., 2007). Perhaps the most significant

property of chitosan is its ability to be modified with other substances. The amine

groups can be used as sites for covalent attachment of various biomolecules,

including enzymes, DNA, and antibodies. The modification can also be performed

by physical interaction (e.g.surface absor-bance, entrapment) instead of chemical

bonding, especially in the case of negatively charged substances. The chitosan chains

can also be covalently crosslinked with each other instead of with biomolecules. This

crosslinking improves the strength and chemical resistance of the film.

2.2.2 Application of chitosan film

Up to now, the application of chitosan films has reached the biosensing area for

preparation of taste sensors (Odaci et al., 2008), and templates for immobilization of

12
enzymes (Krajewska, 2004). In order to advance the chitosan application, the

nanotechnology has been explored in which chitosan in nano size particles has been

employed as a delivery of insulin and albumin and as bactericide (Finotelli et al.,

2010; Limapornvanich et al., 2009; Chung et al., 2010). It has been verified that

chitosan with the particles in nano size exhibited better performance in preventing

the growth of microorganisms compared to that of the plain/unmodified chitosan.

2.2.3 Methods for chitosan film fabrication

In acetic media such as acetic acid and formic acid, chitosan can be dissolved

completely that enables us to produce it in layer-by layer (LbL) films. The

fabrication of chitosan in thin film form possesses several advantages in improving

its properties, especially regarding the interaction with cholesterol (Qian and Yang,

2006) and other biomolecules (Tkac et al., 2007), and heavy metal ion binding (Wan

Ngah et al., 2011; Juang et al., 2002). Several different methods have been developed

to deposit and pattern chitosan films.

a. Solution casting

The simplest fabrication method consists of coating the surface with chitosan

solution and evaporating the solvent. This leaves behind a solid chitosan film with a

thickness dependent on the solution concentration. The coating procedure can be

performed with a pipette or with a spin coater (if a highly uniform film is needed).

Elevated temperatures can be applied to the substrate to speed up the solvent

evaporation.

13
Solution casting is the predominant method for applying chitosan to macroscale

devices. However, it does not inherently provide the spatial control necessary for use

in bioMEMS. For this reason, several approaches have been demonstrated to pattern

the cast film. These include a containment method in which chitosan droplets are

laterally constrained and a thermolithography method in which areas of the chitosan

film are masked with a heat sensitive thermoresist (Fernandes et al., 2004). More

recently, spin-cast chitosan films were patterned by conventional photolithography

by masking them with photoresist and etching them with oxygen plasma (Fig. 2).

Fig. 2.3: Photolithographically patterned chitosan features. (a) SEM of matched

comb structure. (b) SEM of 2-mm-wide lines with 4-mm pitch. (c) SEM of
serpentine spring structures. (d) SEM of a sawtooth structure. Note: All patterns are
in a 2-mm-thick chitosan layer (Cheng et al., 2008).

14
An interesting extension of the solution casting method is the layer-by-layer

assembly (LBL) technique. The substrate is exposed to alternating washes in the

cationic chitosan solution and a solution of some anionic species. Due to electrostatic

attraction, a multi-level composite film is produced. This method has been used for

the entrapment of enzymes (Wu et al., 2007), nanoparticles (Siqueira et al., 2006),

DNA (Pedano et al., 2004) and cells (Ang et al., 2002).

b. Printing

Both nanoimprinting and microcontact printing have been successfully demonstrated

for patterning chitosan films with nanoscale resolution. These methods make use of a

patterned mold or stamp, which is typically a flexible polymer such as PDMS. For

nanoimprinting, the PDMS mold is placed over a substrate coated with chitosan

solution while applying heat and pressure. After cooling, the PDMS is peeled off,

and molded structures of chitosan remain patterned on the substrate. Park et al. in

2007 demonstrated the use of nanoimprint lithography to create nanowire and

nanodot chitosan structures. Tan et al. in 2004 added plasticizers to the chitosan to

allow for imprinting at reduced temperature and pressure. For microcontact printing,

the chitosan solution is applied to the PDMS stamp surface, which is then contacted

with the substrate. The pattern formed on the substrate corresponds to the geometry

of the raised features on the stamp. Feng et al. in 2004 demonstrated co-patterning of

chitosan and bovine serum albumin on the same substrate using microcontact

printing.

15
Printing methods can easily create arrays of chitosan struc-tures for spatially resolved

functionalization. However, both nanoimprint lithography and microcontact printing

are inherently planar processes and cannot be used to pattern chitosan films on non-

planar surfaces (such as microchannels or side walls). Furthermore, alignment of the

mold or stamp to pre-existing features on the substrate can be difficult.

c. Electrodeposition

A very promising method for chitosan integration in microdevices is

electrodeposition (Wu et al., 2003). It allows for both spatial and temporal control of

the chitosan film location and thickness, and it can be used with a variety of device

geometries. Electrodeposition takes advantage of chitosan’s pH dependent solubility.

At pH lower than 6.5, chitosan is soluble and cationic due to its protonated amino

groups. At higher pH values, chitosan becomes deprotonated and is no longer

soluble. When an anode and cathode are immersed in a chitosan solution and a

voltage is applied, electrochemical reactions lead to a locally high pH adjacent to the

cathode surface. While Fig. 3 illustrates these cathodic reactions as a net

consumption of hydrogen ions, we should note that the mechanistic details are more

complex and likely involve the reduction of oxygen ions as well. Chitosan forms a

thin film over the cathode surface as a result of the high pH there. The rate of the

chitosan deposition is influenced by many factors: the molecular weight of the

chitosan, the pH of the solution, the sizes and separation of the anode and cathode,

and the applied voltage. Susan et al. in 2009 has extensively characterized the spatial

resolution of electrodeposited chitosan films. Recentin situ visualization and

characterization of elec-trodeposited chitosan done by Cheng et al. further proved

16
that the gelation and immobilization of chitosan onto the cathode are due to the

electrochemically generated pH change by OH ions and the density distribution of

deposited chitosan hydrogel is electric field-dependent.

Fig. 2.4:. Schematic demonstrating chitosan film electrodeposition (Yi et al., 2005).

2.3 Amine

Amines is composed of one nitrogen atom functional group which has a lone pair

because it is an inorganic compound ammonia (NH3) derivative. The unshared

electron pair of nitrogen atom in amine structure may bind with a proton (Lewis,

1923) that makes amine acts as a weak base. When an alkyl or aryl group replaces

one, two, or all three of the hydrogens in ammonia, new compounds are formed and

can be classified into primary, secondary and tertiary amines (McMurry, 1992; Lide,

2005). The formation of primary amine occurs when an alkyl replaces one of three

hydrogen atoms in ammonia as shown in Fig. 2.5.

17
 Ammonia Primary amine

Fig. 2.5: Chemical structure of primary amine (McMurry, 1992).

In acetone vapor detection, the presence of the primary amine group in chitosan is

very important since it can well react with acetone vapor. It is a nucleophilic species

(negatively charged species) because of donating an electron pair to an electrophilic

species (positively charged species) to bind each other through a chemical reaction

process. For example, OH− and NH3+ are nucleophilic species, because they can

donate a lone pair of electrons. While H+ which is an acid to be an example for

electrophilic species since it requires two electrons to complete its stable form, which

is by accepting a lone pair.

Gilbert N. Lewis in 1923 specifically defined the term acid as follows: An acid

substance is one which can employ an electron lone pair from another molecule in

completing the stable group of one of its own atoms (Lewis, 1923). Then,

International Union of Pure and Applied Chemistry (IUPAC) provided a modern

definition of acid as follows: An acid is an electron-pair acceptor in the form of a

18
molecular entity and hence, it can react with a base to result in an adduct, by sharing

the electron pair furnished by the base (Lewis, 1923).

Special compound such as H2O can be nucleophilic or electrophilic species since it

can donate or accept a pair of electrons, depending upon the chemical reaction. For

another example, in the chemical reaction between Me3B and NH3 to produce

Me3BNH3. It has been known that Me3B serves as a acid and NH3 serves as a base,

and the reaction product, Me3BNH3 becomes adduct. Therefore, the adduct is defined

as a single reaction product of a combination of two or more distinct molecules

wherein the product is considered a distinct molecular species (Nic et al., 2004).

On the other hand, secondary amine is performed when N together with one

hydrogen atom binds with two alkyl substituents as shown in Fig. 2.6 (a). Two

examples of secondary amine are dimethylamine and methylethanolamine. While, in

tertiary amines is formed when the organic substituents replace three hydrogen atoms

(Fig. 2.6 (b)) and for example is trimethylamine. It has been reported that the

physical characteristic of primary and secondary amines is quite affected by

hydrogen bonding (McMurry, 1992; Lide, 2005). Their ability to form hydrogen

bonds reflects their solubility in water wherein the solubility decreases with the

increase in the number of carbon atoms.

19
 (a) (b)

Fig. 2.6: Chemical structure of (a) secondary amine and (b) tertiary amine

(Lewis, 1923).

2.4 Hydroxyl

A hydroxyl is recognized as a substructure of the water molecule because its

chemical functional group is performed from a covalent bond between an oxygen

atom and a hydrogen atom as shown in Fig. 2.7. It has interactive and reactive

properties such as hydrogen bonding and ionizability. There are two forms of

hydroxyl; the first is radical form (called hydroxyl radical) which is neutral in charge

and the second is anion form (OH−), called the hydroxide anion. A single negative

charge resides on the electronegative oxygen of hydroxide anion.

Figure 2.7: Chemical structure of hydroxyl (Lide, 2005).

20
But when the two atoms form a bond, and a hydroxyl group is formed, something

changes. Between the two, there are still an equal number of electrons and protons,

so the group is neutral. However, the electrons tend to hang out more on one side

than the other, so their negativity concentrates a little more on that side. Naturally,

the other side becomes a little more positive.

Fig. 2.8 shows a hydroxyl group is surrounded by electron cloud which tends to

gather towards the oxygen than hydrogen. This has caused the electrical charges

distribute unevenly and naturally, hydrogen side becomes a little more positive.

However, due to the hydroxyl group has still equal concentration of proton and

electron, the group keeps at neutral. This condition is termed as partial charge. The

hydrogen in hydroxyl group is termed as partially positive and the oxygen is termed

partially negative.

Electron cloud

+Q
-Q
O H

Fig. 2.8: Electron cloud surrounding the whole hydroxyl (Lide, 2005).

21
2.5 Acetone

Sometimes many people are not aware have breathed by inhaling the volatile organic

compounds (VOCs) in large amount from the air. The VOC often consists of various

components which may affect the human health in short-or long-term time. Most of

VOCs sources come from paints, dry cleaning compounds, solvents, degreasers,

assorted industrial products and chemical intermediates such as rubber, lubricating

oil and dyestuff (Cao et al., 2001). Acetone is one of volatile organic compounds

with the formula (CH3)2CO as illustrated in Fig. 2.9 and Table 2.1 listed its

properties.

Fig. 2.9: Structural formula of the acetone molecule (Cao et al., 2001).

22
 Tabel 2.1: The properties of acetone (Tang et al., 2008).

Properties

Molecular formula C3H6O

Molar mass 58.08 g mol−1
Appearance Colorless liquid (white snow-like form when solid)
Density 0.7925 g/cm3
Melting point −94.9 °C
Boiling point 56.53 °C
Solubility in water miscible
Acidity (pKa) 24.2
Refractive index (nD) 1.35900 (20 °C)
Viscosity 0.3075 cP

Structure

Molecular shape trigonal planar at C=O

Dipole moment 2.91 D

The acetone is characterized with easy evaporation at room temperature. Therefore, it

is much more employed as an organic solvent for spray-paint and plastic dissolution,

tissues dehydration and paraffin purification. Acetone is very hazardous at a

temperature of 465°C since it can auto-ignites caused by its high flammability. It

often becomes a cause of fire in chemical stores and construction sites when its

concentration in the air is in range of 2.5% for lower explosive limit (LEL) to 12.8%

for upper explosive limit (UEL).

The concentration of acetone in air above 10,000 ppm in air usually evokes a fatigue,

headache and at the worst, narcosis. It can also irritate mucous membranes, eyes and

upper respiratory tract. The other effect, it can cause muscle weakness, difficult

23
breathing and loss of consciousness if inhaling the high concentration acetone (Cao

et al., 2001; Tang et al., 2008). Besides that, the metabolic changes, coma and the

damage of kidney and liver can occur for a great irritation (Tang et al., 2008). The

American Conference of Governmental Industrial Hygienists reported that the

threshold limit value (TLV) of acetone vapour in our living environment is 750 ppm

(Tang et al., 2008). Therefore, it is to be very important for a family to have an

acetone concentration detector.

2.6 Acetone in human breath

Acetone in human breath is a product from the breakdown of fat in liver for energy

in the body, called a lipolysis. The acetone is easily excreted in the breath and pass

through the alveolus, as shown in Fig. 2.10. The medical reports showed that diabetic

patients exhale higher acetone content in their breath, blood and spittle compared to

healthy people due to the insulin in diabetic body unable to function effectively

(Likhodii et al., 2002; Makisimovich et al., 1996). The presence of acetone in breath

ranges from a relatively high 0.5 ppmv for healthy individuals to hundreds of ppmv

for critically ill, ketoacidotic diabetics. If the fat resolution produces a few acetone,

the tissues (especially muscles) can exploit it completely. On the contrary, when the

acetone produced exceeds the exploitation capacity, it is excreted from ketonuria and

released through the exhaled breathe, indicated by the odor of a decaying apple (Ping

et al., 1997; Makisimovich et al., 1996; Bogdan et al., 2003; Gerard et al., 2002;

Fleischer et al., 2002).

24