Karnataka State Council For Science and Technology

KARNATAKA STATE COUNCIL FOR SCIENCE AND
TECHNOLOGY
R.V.COLLEGE OF ENGINEERING, BENGALURU-59

(Autonomous Institution Affiliated to VTU, Belagavi)
DEVELOPMENT OF ECO-FRIENDLY PLANT BASED

MOSQUITO REPELLANTS
PROJECT REPORT
Submitted by
BHARATH.S. 1RV11BT014
KARTHIK.V. 1RV11BT024
SAURAV KUMAR SINGH 1RV11BT047
Under the Guidance of
Dr. Nagashree N Rao Associate Professor and Dr. Ashwani Sharma

Assistant Professor
Dept of Biotechnology
R.V. College of Engineering
in partial fulfilment for the award of degree
of
Bachelor of Engineering
IN
BIOTECHNOLOGY DEPARTMENT
JAN-MAY 2015
i
R.V. COLLEGE OF ENGINEERING, BENGALURU - 560059
DEPARTMENT OF BIOTECHNOLOGY
CERTIFICATE
Certified that the project work titled “DEVELOPMENT OF ECO-FRIENDLY
PLANT BASED MOSQUITO REPELLANTS” is carried out by BHARATH.S
(1RV11BT014), KARTHIK.V (1RV11BT024), SAURAV KUMAR SINGH
(1RV11BT047), who are bonafide students of R.V College of Engineering,
Bengaluru, in partial fulfilment for the award of degree of Bachelor of Engineering in
Biotechnology of the Visvesvaraya Technological University, Belagavi during the
year 2014-15. It is certified that all corrections/suggestions indicated for the internal
Assessment have been incorporated in the report deposited in the departmental
library. The project report has been approved as it satisfies the academic requirements
in respect of project work prescribed by the institution for the said degree.
Guide Head of the Department Principal
External Viva
Name of Examiners Signature with date
ii
R.V.COLLEGE OF ENGINEERING, BENGALURU - 560059
DEPARTMENT OF BIOTECHNOLOGY
DECLARATION
We, BHARATH S (1RV11BT014), KARTHIK V (1RV11BT024), SAURAV
KUMAR SINGH (1RV11BT047), students of eighth semester B.E.,
BIOTECHNOLOGY, hereby declare that the project titled “DEVELOPMENT OF
ECO-FRIENDLY PLANT BASED MOSQUITO REPELLANTS” has been carried
out by us and submitted in partial fulfilment for the award of degree of Bachelor of
Engineering in BIOTECHNOLOGY. We do declare that this work is not carried out
by any other students for the award of degree in any other branch.
Place: Bengaluru Signature
Date: 1. BHARATH S
2. KARTHIK V
3. SAURAV KUMAR SINGH
iii
ACKNOWLEDGEMENT
We would like to thank our project guide Dr. Ashwani Sharma for providing us with
continuous support. He was always there to listen and to give advice, and encouraged
us to perform better every time.
We are thankful to Dr. Nagashree N Rao for providing us with useful inputs in
carrying out our project.
Also we are grateful to Dr. H G Ashok Kumar, Professor and HOD, Dept. of
Biotechnology, RVCE for providing us with the opportunity to work on this project
and also for his guidance without which our project wouldn’t have been successful.
We convey our gratitude to Dr. Satyanarayana B.S., Principal, R.V. College of

Engineering, Bengaluru for the support given to us in many ways.
We are thankful to Dr. N.J.Shetty and Dr. Venkatesha, Mr.Kiran and Mr.Vinay of
Zoology department, Bangalore University for their support in the execution of our
project.
We are thankful to Ms. Lakshmi, Mr.Pradeep, Mr.Harsha and Mr. Armugam T A

for their steady support in carrying out the project.
We convey our gratitude to Karnataka State Council for Science and Technology
(KSCST) for funding our research work.
We also convey heartfelt thanks to our parents for their encouragement and support
throughout the project.
We owe special thanks to all teaching and non-teaching staff of our department and
all our friends who helped us in our work.
iv
ABSTRACT
The mosquito repellents market in India is valued at 16 billion and is growing at a
compounded annual growth rate of 15%. Commercial mosquito coil smoke has
several adverse health effects on human beings as well as other animals such as skin
and eye irritation, etc. In this context, many research works focused to develop herbal
mosquito coils by extracting oils from the plants which can be used as creams and
lotions. But since majority of Indian population uses mosquito coils in their homes
there was a need to prepare an eco-friendly herbal mosquito coil. The objective of our
work was to prepare an eco-friendly mosquito coil completely from organic materials
which are either abundant or waste in our surroundings.
Three Plants namely Citronella grass (Cymbopogon nardus), Eucalyptus tree

(Eucalyptus globulus) and Rosemary plant (Rosmarinus officinalis), Coconut husk as
smouldering agent and Corn cob powder as the binding agent were selected. The coil
was first optimizing for binding strength and harmful gas emission and then the coil
was prepared using plants alone and with combinations using Taguchi’s Design of
Experiments.
The coil prepared was first subjected to physical tests including binding strength,
burning duration, moisture content, harmful gas emission and mosquito repellence test
was carried out using adult female Aedes aegypti mosquitoes. Results of physical
properties testing showed that prepared mosquito coil showed longer duration of
burning and less than half the amounts of harmful gas emission on burning and
prepared mosquito coil had less binding strength compared to the commercial
mosquito coil and repellence testing results showed that prepared mosquito coils take
longer time to repel the mosquitoes compared to the commercial mosquito coil but
our coils only either repelled or caused temporary numbness to the mosquitoes while
commercial coil killed the mosquitoes in a short duration of time. The outcome of the
project is the citronella mosquito coil which has better physical and repellence
properties compared with the commercial mosquito coil. In this direction, further
v
work can be carried out to improve the efficacy of the coil by trying different plant
materials, binder and smouldering agent.
CONTENTS
List of Symbols, Abbreviations and Nomenclature viii
List of Tables ix
List of Figures x
CHAPTER 1
1. INTRODUCTION 2
1.1 Market Survey 2
1.2 Societal Relevance 3
1.3 Problem Statement 3
1.4 Recent work done on mosquito repellents 3
1.4.1 Review on commercial mosquito coils 4
1.4.2 Review of materials 5
1.4.3 Review on processing techniques 6
1.4.4 Unresolved issues 7
1.4.5 Emerging opportunities 7
1.4.6 Research gap 8
1.4.7 Innovativeness in our project 8

1.5 Objectives of the project 8
1.6 Organization of the Report 9
CHAPTER 2
2. BIOLOGICAL MOSQUITO REPELLENT CONCEPTS 12
2.1 Organic Materials 12
2.2 Adult mosquito 13
vi
2.3 Taguchi’s Design of Experiments [DOE] 14
CHAPTER 3
3. METHODOLOGY 16
3.1 Project Methodology 16

3.1.1 Optimization of the Binding and Burning Agents 16
3.1.2 Method to Develop Mosquito Repellent Coil 19
from Individual Plants
3.1.3 Method to Develop Mosquito Repellent Coil with 21
Combination of Three Plants (Citronella, Eucalyptus and Rosemary)
CHAPTER 4
4. BIOLOGICAL MOSQUITO COIL FORMULATION 23
4.1 Development of mosquito coil 23
4.2 Optimization of the Coil 28
4.3 Preparation of Mosquito Coils 31
CHAPTER 5
5. RESULTS AND DISCUSSIONS 38
5.1 Binding strength 38
5.2 Burning duration 38
5.3 Emission testing 39
5.4 Moisture content 40
5.5 Repellence testing 41
CHAPTER 6
6. CONCLUSION AND FUTURE WORK 50
6.1 Conclusion 50
vii
6.2 Scope for future work 50
REFERENCES 52
LIST OF SYMBOLS, ABBREVIATIONS AND NOMENCLATURE

DEET - N, N-diethyl-meta-toluamide
DOE - Design of Experiments
BBB - Blood Brain Barrier
EPA - Environmental Protection Agency
CO2 - Carbon dioxide
CO - Carbon monoxide
HC - Hydrocarbons
viii
LIST OF TABLES
Table no. Title Page no.
2.1 DOE method applied for obtaining best combinations 14

4.1 Optimization of coil 29
4.2 Binding strength results for optimization of coil 30
4.3 Emission testing results for optimization of coil 31
4.4 Taguchi’s Design of Experiments 31
5.1 Binding strength analysis results 38
5.2 Burning duration of various coils in comparison to 39
commercial coil
5.3 Emission testing of various coils 39
5.4 Moisture content analysis of various mosquito coils 40
5.5 Negative control results 41
5.6 Mosquito repellence results for commercial Mortein coil 42
5.7 Repellence results for citronella coil 42
5.8 Repellence results for rosemary coil 42
5.9 Repellence results for Eucalyptus coil 43
5.10 Repellence results for coil 4 44
ix
LIST OF FIGURES
Fig. no. Title Page no.
1.1 Graph showing Indian market share of different mosquito repellence 02
products
2.1 Rosemary, Citronella and Eucalyptus plant leaves 12
2.2 Coconut husk and Corn cob 13
3.1 Flowchart for Optimization of Burning and Binding agent 17
3.2 Optimization of Burning and Binding agent 18
3.3 Flowchart for development of mosquito coil based on a single plant 19
3.4 Development of mosquito coil based on a single plant 20
3.5 Flowchart for developing the mosquito repellent coil for
21
combination of plants
4.1 Eucalyptus leaves being ground using mortar and pestle 23
4.2 Powdered leaves being sieved to obtain fine and uniform powder 23
4.3 Coils developed using cow dung which had low binding strength 24
4.4 Corn cob kept in hot air oven at 600 C for 48 hours for drying 24
4.5 Dried binding material corn cob ready to be crushed 25
4.6 Corn cob boiled and mixed with other constituents 26
4.7 Mosquito coil in round shape 27
4.8 Moulds developed from cardboard sheet 27
4.9 Mould being filled with coil paste 27
4.10 Testing Binding strength of the coil 29
4.11 Analysis of smoke for harmful gas emission 30
4.12 Analysis of Burning duration 32
4.13 Aedes aegypti breeding in larva stage 33
4.13 Adult Aedes aegypti Mosquito breeding in cages 33
4.15 Glass box dimensions used for repellence testing 34
4.16 Aedes aegypti adult mosquitoes in mesh cage 34
x
4.17 Transfer of mosquitoes from cage to glass box using aspirator 34
4.18 Testing of mosquito repellence 35
4.19 Testing of repellence for commercial mosquito coil 35
4.20 Testing of repellence for mosquito coil 36
5.1 Emission testing result for commercial coil and Citronella coil 40
5.3 Numbness caused by mosquito coil smoke 43
xi
Development of eco-friendly plant based mosquito repellents
INTRODUCTION
Mosquitos are the irresistible asset of the domestic life which can be useful as well as
deleterious. In the scenario of fast moving domain they are responsible for many
disastrous diseases [1].
1.1 Market survey
Mosquitoes have always been a menace for man as they vector a range of deadly
diseases. Mosquito repellents are available in market as coils, mats, vaporisers,
aerosols, creams and other wide range of products. Coils were the first mosquito
repellents to be introduced into Indian market. In spite of mosquito problem, use of
repellents in India is quite low. It is approximated only 16.4% of the houses in urban
areas and 22.6% in the metros use mosquito repellents. The estimate for rural areas it
is even lower, at 6.9%. Usage of mosquito repellents is expected to rise as people are
becoming more aware of the diseases caused by the mosquitoes [6].
The mosquito repellent market in India is valued at ` 16 billion and is growing at a
compounded annual growth rate of 15%. Major players in the Indian mosquito
repellent industry are Good knight from Godrej Sara Lee Ltd having 20% market
share, Maxo from Jyothi laboratories having 22% market share and Mortein from
Reckitt Benckiser India Ltd having 29% of the market share and the Indian market
share between different types of products is shown in Fig 1.1.
Fig 1.1 Graph showing Indian market share of different mosquito repellence products
2
B.E. Department of Biotechnology, RVCE, Bengaluru.
1.2 Societal Relevance

Mosquito is a small fly and belongs to the family Culicidae. Male mosquitoes depend
on the nectar for food whereas female mosquitoes are ecto parasites which suck on
blood of other animals. Female mosquitoes detect the host by several mechanisms
such as chemical sensors which detect lactic acid present in the sweat of the animals
and the carbon-di-oxide released by the host and, heat sensors which can detect the
bodily produced heat and get attracted to the high humidity around the body of the
host and they are also attracted to dark places and dark clothing. While sucking the
blood of the host mosquitoes can transfer diseases from a patient to a healthy human
being such as yellow fever, dengue and malaria. It is estimated more than one million
deaths occur worldwide due to diseases transmitted by mosquitoes [13].
Mosquito repellents either repel or kill the mosquitoes by either masking the human
scent or blocking the antennae of the mosquito which detects human beings in
proximity or killing the mosquitoes from the chemicals from the repellent ultimately
blocking mosquitoes from biting humans and transmitting diseases [14].
1.3 Problem Statement

Since the commercial mosquito coils have several health effects from the gas they
produce on burning, the problem statement is to identify and test for eco-friendly
biological mosquito repellents as well as binding agents and find the optimized
concentration of the mosquito coil repellence effect of mosquito coil prepared and
compare it with the repellence of the commercially available mosquito coils [8]. Our
mosquito coil prepared is entirely from organic materials which are either waste or
abundant in our surroundings which does not release as much harmful gas as the
commercial ones. This mosquito coil can be prepared easily which will be less
expensive than the commercially available ones [9].
1.4 Recent work done on mosquito repellents
Literature survey includes the review of research works on the materials and methods
of plant based mosquito repellents.
3
1.4.1 Review on commercial mosquito coils
Patel et al [6] investigated the problems regarding the chemicals used in the
commercially available mosquito coils and listed their detrimental effects from the
survey statistics collected. A component named DEET is used in the commercially
available coils for repelling mosquitoes and this component is allergic to many
people. This survey showed the skin irritation at the maximum extent. An article
published in the newspaper ‘The Hindu’ showed the toxicity of commercially
available mosquito coils as shown in Fig 1.2.
Fig 1.2. Health effects of smoke from commercial mosquito coil
A study of Health Effects of Pesticides Used for Mosquito Control showed that a
pesticide used in the commercially available mosquito coils named ‘pyrethrin’ is very
harmful to human beings. It also found that burning of one commercially available
mosquito coil in a closed room is equal to smoking hundred cigarettes, which shows
us that commercially available mosquito coils have severe adverse health effects on
human beings [9].
Pyrethrin amounts to 0.3 to 0.4% of the commercially available mosquito coil weight.
This pyrethrin is an insecticide obtained from the species of the chrysanthemum plant.
The flowers of these plants are dried or oil is extracted using solvents. This is a
natural available insecticide having more adverse effects on human health. So
semisynthetic ones are produced one of them is allethrin. Inhaling this at high levels
cause asthmatic breathing, sneezing, nasal stuffiness, headache, nausea, in
coordination, tremors, convulsions, facial flushing and swelling, and burning and
4
itching sensations. The most adverse health effects are seen in infants who cannot
break the pyrethrin component. Burning of the commercially available mosquito coils
result in release of insecticide along with the smoke which repel the mosquitoes. The
remaining components are organic fillers, binders, dyes, and other additives capable
of smouldering well. Burning of these components produce the gaseous pollutants that
have suspected carcinogenic components [21].
There are two types of pyrethrins:
Type 1 pyrethrin produce reflex hyper excitability and fine tremor.
Type II pyrethrins produce salivation, hyper excitability, choreoathatosis, and

seizures.
Permethrin is a type of pyrethrin, EPA classified permethrin as possible human

carcinogen based on the tests conducted on mice which developed tumours at high
levels. It was also proved that permethrin causes chromosomal aberrations in
laboratory animals.
The mosquito repellents present in the commercially available ones can affect the
nervous system also, one of the studies tell that exposure to mosquito repellents
present in the commercially available mosquito coils lead to delay in the maturation
of the BBB[Blood Brain Barrier] system. One of the case studies tell us that high
exposure to pyrethrin coils lead to the fatal death due to the congenital abnormalities
during 3rd to 8th week of pregnancy.
1.4.2 Review of materials
Many researchers have used different plant repellents, binders, smouldering agents
according to the availability of those components in their locality. The researches [1-
9] tell that people have used almost all kind of plant repellents in various forms such
as oils, coils, creams etc. Nandini Rani et al [1] made use of the citronella as their
mosquito repellent along with the natural binders neem powder, potato starch, corn
starch, coconut shell powder and cow dung. They powdered the above said
components mixed them and made into cakes and dried them. Mandavgane et al [15]
made use of cow dung, tulsi, neem and pallenthrin. They optimized the concentration
of every component and tried to bring out the best repellent cake out of various
5
combinations with the above said materials. Richard J Beever used the extracted oil of
eucalyptus along with the synthetic insecticide DEET. Das et al [4] extracted oils
from Zanthoxylum limonella (fruits), Citrus aurantifolia (leaf) and petroleum ether
extract of Z.Limonella (fruits) through steam distillation and added with coconut oil
and mustard oil separately. Then they tested for the repellency. Duangkamon et al [5]
investigated the repellence activity of the plants garlic (Allium sativum), clove
(Syzygium aromaticum), lemon grass (Cybopogon citratus), citronella grass
(Cymbopogon nardus), peppermint (Mentha piperita), eucalyptus (Eucalyptus
globulus), orange (Citrus sinensis) and sweet basil (Ocimum basilicum) and their
combinations against the mosquitoes Aedes aegypti and Anopheles dirus Peyton &
Harrion under laboratory conditions by extracting the oils from them. After going
through the several papers we checked the availability of the repellent plants in our
locality and selected citronella, eucalyptus and rosemary.
1.4.3 Review on processing techniques
Nandini Rani et al [1] collected all the components required for the development of
the repellent cakes like citronella, neem powder, coconut husk, cow dung and ground
them into fine powder mixed thoroughly and added into moulds to give them the
shape and to this cakes they added the coconut oil. In the reference Effiom et al [3]
and Das NG et al [4] have extracted the oils from the plant repellents using Clevenger
apparatus and steam distillation respectively. They applied the extracted oil to their
skins and checked its repellence. Duangkamon et al [11] extracted essential oils and
formulated for the repellence. In this paper they have extracted oils from different
parts of different plants. Some of the plants they used and extracted oils are:
 Eucalyptus- Oil from leaf

 Peppermint- Oil from leaves
 Garlic- Oil from stem
 Orange- Oil from fruit
 Citronella grass- Oil from stem
 Lemon grass- Oil from stemf
 Clove- Oil from flower
 Sweet basil- Oil from leaves
6
They used Clevenger apparatus to extract the oil. The extracted oil was applied it on
the arms and subjected the free arms to the mosquito cage for three minutes. The arms
were kept till two mosquitoes bite. If no mosquito had bitten then only the arms are
kept for three minutes. The time taken to bite by the two mosquitoes are recorded and
effectiveness of each coil is analysed. The test was done on hundred human
volunteers. It was done against Aedes mosquitoes. Bhoopendra et al [8] explored the
MEDLINE search on the pertinent information about the plants, its components for
their toxicity. The MEDLINE has a database on the toxicity of almost all the plants.
Mshelia et al [9] tested the cognitive effect of smoke of mosquito coil on the mice.
They exposed different mice for the different set of time to the smoke released from
the mosquito coil and effect was noted and toxicity was compared. Egunyomi et al
[10] tested repellence by making four guinea pigs stand in the cages with their hairs
scraped off from their skin. Then ten disease free female mosquitoes were left into the
room. Then the readiness of the mosquitoes to bite the pigs were noted and compared.
1.4.4 Unresolved issues
Das et al [4] and Duangkamon et al [5] have extracted oils which should be used
immediately otherwise the extracted oil gets evaporated. It is clearly said in the
reference [6] too. The component used in the coils to repel mosquitoes that is DEET
should be used cautiously with the children. High concentration of the DEET might
even cause cancer [6] so it should be avoided with the herbal repellent. Some of the
people even worked on the development of eco-friendly mosquito repellents, but
exposure to the smoke causes skin irritation, eye irritation etc.
1.4.5 Emerging opportunities
If there is a possible way to develop an eco-friendly herbal mosquito coil which is

more effective than the commercially available one then it contributes much towards
environment and reduced diseases that is caused by the chemicals used in the
commercially available ones. Due to the use of commercially available coils the
mosquitoes are eradicated which affect the food chain. So in case if the herbal eco-
friendly coil is used it will only repel the mosquitoes and does not kill them.
7
1.4.6 Research gap
Many people have tried making the herbal eco-friendly mosquito repellents in the
form of oils. They are volatile in nature and should be used immediately and the
methods used to develop them is costly. So we worked to develop the eco-friendly
herbal mosquito repellent in the form of coils and not in the form of oils so that it can
be used anytime. The methods used are simple and the materials are cheap, some
materials are waste things so it is cost effective too.
1.4.7 Innovativeness in our project
Most of the papers referred have extracted the oils and formulated them to creams and
lotions. The methods we used are very simple and do not need the expensive
equipment. The corn cob is used as the binding agent. This when boiled with the
water increases its binding strength and make the coil harder and stable. The
smouldering agent used was coconut husk powder which is waste and easily available
and helps the coil to burn continuously. The method used to test for the mosquito
repellence was easier and simple which included the development of the glass
container which mimics a room of the house .
Review of the literature [1-50] indicate that most of the researchers have worked to
develop the mosquito coil which is eco-friendly and effective than the commercially
available ones so as to replace the commercially available one which helps the
environment, mankind and even mosquitoes. But failed to do in a cost effective way.
And more over their development of mosquito coil involved complex procedures and
required expensive apparatus and equipment. Some have developed the coils but are
more fragile and do not burn continuously so this made us to develop the eco-friendly
plant based mosquito repellents.
1.5 Objectives of the project

Primary objectives of this project is to identify and test for eco-friendly biological
mosquito repellents as well as binding agents and to optimize the concentration of
herbal product for efficient and eco-friendly mosquito repellents. This is achieved by
trying out different organic materials both for biological repellents as well as binding
agents and optimize the concentration of herbal product for efficient and eco-friendly
8
mosquito repellents is achieved by first optimizing the coil for binding strength and
harmful gas emission and later for repellence towards the mosquitoes.
To accomplish the above mentioned objectives, the method of approach used was to
first try the crude method to prepare the coil by grinding the plant materials, coconut
husk and corn cob, then check for their burning abilities and later on the project was
standardized in which we removed the moisture from all the substances by drying
them up and make them equal in size and boil the corn cob powder with the water to
prepare a thick sticky slurry and mix the slurry with the other components to prepare a
product which is laid out into a individual coils with specific dimensions and drying
them in hot air oven and once they are dried they are checked for their physical
properties such as harmful gas emission, burning duration, smoke density, binding
strength and at last check their repellence on the mosquitoes reared of the species
Aedes aegypti and choose the best coil from the different combinations of the plant
materials made.
1.6 Organization of the Report
The main body of this report is preceded by detailed table of contents, list of
abbreviations, list of tables and list of Figures. This report is organized into six
chapters, followed by the references.
Chapter 1 INTRODUCTION presents the basic information of market survey,

mosquito repellent types, societal relevance, problem statement, literature survey and
objectives of the project.
Chapter 2 THEORY AND CONCEPTS presents the plant materials used in the
project as well as binding material and smouldering agent, mosquito life cycle, their
breeding mechanism and the Taguchi design used in the formulation of different
combinations of the mosquito coil.
Chapter 3 METHODOLOGY illustrates the methodology used for the project. It

also contains flow chart highlighting the project methodology adapted here.
Chapter 4 PREPARATION OF MOSQUITO COIL presents the details of

preparation of plant based mosquito coil which include obtaining the plants, binder
and smouldering agent and using them appropriately to prepare the final product.
9
Chapter 5 RESULTS AND DISCUSSION incorporates results and tables for each
of the combinations of mosquito coils prepared and tested for each of the physical
properties and repellence ability of the mosquito coils.
Chapter 6 CONCLUSION AND SCOPE FOR FUTURE WORK presents the

conclusion drawn from the results and also scope for future work.
10
BIOLOGICAL MOSQUITO REPELLENT CONCEPTS
Mosquitoes are a menace to the society and they are to be controlled to avoid
spreading the diseases. One of the mechanisms to control mosquitoes is to prepare a
repellent in the form of a coil. Chemicals in the smoke repel mosquitoes.
2.1 Organic Materials
Commercially available mosquito coils have many adverse effects on human health so
we choose to develop a eco-friendly mosquito coil to completely reduce the adverse
effects and which shows the repellence towards the mosquitoes as the same extent as
commercial ones. From the literature survey we got the list of plants that show
repellence towards mosquitoes which include Citronella, Catnip, Marigold, Basil,
Lavender, Peppermint, Garlic, Rosemary and Eucalyptus.
From these plants we selected Citronella, Eucalyptus and Rosemary based on their
availability in our surroundings and based on how much work has been carried out on
them and as these were the ones that showed maximum repellence towards
mosquitoes as suggested by the literature survey which are shown in Fig 2.1 [24].
Fig 2.1 Rosemary, Citronella and Eucalyptus plant leaves
Coconut Husk as Smouldering Agent
Coconut husk is a by-product of coconut industry. It is brownish in colour in form of

long hair like structures as shown in Fig 2.2. Constituents of Coconut husk include 26
12
% water soluble substances, 14.25 % pectin and other constituents soluble in boiling
water, 8.5 % hemicellulose, 28.33 % lignin and 22.92 % cellulose [26][27].
Fig 2.2 Coconut husk and Corn cob
Corn Cob Powder as Binding Agent
Corn cob is a thick fleshy part of the corn situated in the middle of corn stalk as
shown in Fig. 2.2. General constituents of corn cob include 31.8 % cellulose, 34.7 %
hemicellulose, 27.9 % lignin and 5.6 % minerals and waxes [28][46].
2.2 Adult mosquito
A mosquito goes through 4 stages in its lifecycle: egg, larva, pupa and adult. Journey
from egg to adult differs within species and is determined by surrounding conditions.
Some types can form the adult mosquito from egg in a span of 5 days whereas for a
tropic conditioned environment generally it could take approximately forty days.
Changes in the adult imago size relies on density of eggs laid in the water and the
amount of food obtained to them in the water surroundings.
Males last for around five to seven days feeding upon plant flower’s nectar and other
origins of sugar. Females feed on the host by sucking in the blood and once they
complete their full meal females will perch and develops the eggs, this procedure
varies with the environmental conditions and takes around 2 to 3 days in tropic
environmental conditions. As the eggs are formed, female deposits the eggs in
suitable environment and continues its search for food.
These repeat until the female dies. In unconditioned environment, adults do not last
for more than 14 days whereas they can last for a month when held in laboratory
environment. Their lifetimes depend on several factors such as ambient environment
13
and their mechanisms to get a blood meal and not get killed by the predators or the
repellents.
2.3 Taguchi’s Design of Experiments [DOE]
A Japanese statistician named Genichi Taguchi developed a method Design of
Experiments method which is applied to different fields in order to increase quality of
manufactured products.
The DOE includes larger understanding of concept of variance than the previously
used R.A. Fischer’s Analysis of Variance. He developed method of Outer arrays in
which he said mainstream sampling is not sufficient as there is no way for finding a
random sample from the succeeding conditions. The ANOVA (Analysis of Variance)
method developed by Fisher concentrated mainly to reduce the impact of nuisance
components to test comparableness of mean treatment results whereas Taguchi’s DOE
includes Variations more centrally.
Outer array follows the Judgmental sampling in which each try out is broadened to an
outer array which would imitate the random surroundings in which the ware would
function. Thus Taguchi’s method could be used for analyzing the combinations of
ratios which are used to prepare mosquito coils from different components such as
plant leaves from each of the plants. This method provides the required number of
combinations which involve all the best variations the method could provide when
tried out on a trial and error methods as shown in Table 2.1.
Table 2.1 DOE method applied for obtaining best combinations
Citronella (g) Rosemary (g) Eucalyptus (g)
COIL 1 0.834 0.834 0.834
COIL 2 0.500 1.000 1.000
COIL 3 0.357 1.070 1.070
COIL 4 1.000 0.500 1.000
COIL 5 0.714 0.714 1.070
COIL 6 0.834 1.248 0.416
COIL 7 1.070 0.357 1.070
COIL 8 1.250 0.834 0.416
COIL 9 0.938 0.938 0.625
The idea behind the inceptions of this assignment of making of mosquito coil from the
biological sources have been paved through the basic concepts of material, analysis
and designing of the experiment.
14
METHODOLOGY
Methodology followed in the research is to prepare a mosquito coil based out of

completely organic materials which are either waste or abundant using simple and
inexpensive techniques.
3.1 Project Methodology
The best methods was induced in order to develop the proper, effective and herbal
mosquito repellents. There were many components used to develop the herbal eco-
friendly mosquito coil. They were mosquito repelling plant leaves, burning agent and
binding agent. The flow chart of the method used in the development of the eco-
friendly mosquito coil is shown below.
3.1.1 Optimization of the Binding and Burning Agents
In order to test the coil for the emission of toxic gases and the repellence a coil has to
be prepared. There are two important factors to be considered for the development of
the eco-friendly mosquito repellents. One is the binding strength and other is the
burning capability. In order to develop the coil burning agent and the binding agents
should be optimized. The flow chart is as shown below and the coil is prepared as
shown in Fig 3.1 and Fig 3.2.
16
The components required to develop the coil were dried and ground
(plant leaves, coconut husk and corn cob)
The fine powder is separated using sieves
To definite amount of plant material different ratios of corn cob and coconut husk
are chosen making equal weight.
The boiled corn cob paste is mixed with other constituents to

form a paste which is put into mould
The paste in the mould is dried in hot air oven at 60o

C for 48 hours
Each dried coil is checked for binding ability and harmful gas
emission and the results are noted
Based on the test results, coil which has high binding ability and low
harmful gas emission is chosen
The chosen coil corn cob and coconut husk ratio is used in
further preparation of coils
Fig3.1 Flowchart for Optimization of Burning and Binding agent
17
Components ground and added to sieves Sieved to separate fine particles
Components are mixed thoroughly Corn cob boiled in microwave oven
The mixture was poured into beakers Filling in the mixture
Testing for binding strength Test for the burning capability
Fig 3.2 Optimization of Burning and Binding agent
18
3.1.2 Method to Develop Mosquito Repellent Coil from Individual Plants
Corn cob powder was added as a binding agent and coconut husk as a smouldering
agent and before adding these were optimized as shown above. The flow chart to
develop the eco-friendly mosquito repellent coil is shown below and the coil is
prepared as shown in Fig 3.3 and Fig 3.4.
. The components required to develop the coil were dried and ground (plant
leaves, coconut husk and corn cob)
The fine powder is separated using sieves
Each plant material is mixed with optimized quantity of boiled corn cob and
coconut husk composition to form a thick paste
The paste prepared is put into mould to give a specific shape

C for 48 hours
Dried coil is checked for physical properties such as burning duration,

binding strength, moisture content and harmful gas emission
Coil is checked for repellence using live Aedes aegypti mosquitoes and
repellence results are noted down
Fig 3.3 Flowchart for development of mosquito coil based on a single plant
19
The components are weighed Corn cob powder is boiled
All the mixture is mixed in a mixer Filling in the mixture to the moulds
Test for binding strength Test for the burning duration
Repellence test Numbness affected mosquito
Fig 3.4 Development of mosquito coil based on a single plant
20
3.1.3 Method to Develop Mosquito Repellent Coil with Combination of Three

Plants (Citronella, Eucalyptus and Rosemary)
Taguchi’s design is used to obtain different combinations of 3 plants in the best way.
The flow chart is as shown below.
Citronella leaf Rosemary leaf Eucalyptus

powder powder leaf powder
Taguchi’s Design of Experiments (DOE) is applied to get best

combinations of the 3 plants
Corn cob is boiled and mixed with coconut husk and

combination of 3 plants from the Taguchi’s DOE
The paste prepared is put into mould to give a specific shape

C for 48 hours
Dried coil is checked for physical properties such as burning duration,

binding strength, moisture content and harmful gas emission
Coil is checked for repellence using live Aedes aegypti mosquitoes and
results are noted down
Fig 3.5 Flowchart for developing the mosquito repellent coil for combination of plants
21
BIOLOGICAL MOSQUITO COIL FORMULATION
This chapter includes the method to optimize the mosquito coil formulation and
method to prepare the mosquito coil from basic constituents using inexpensive
materials and techniques.
4.1 Development of mosquito coil
Leaves of Citronella and Eucalyptus were collected from Danvantri, a nursery in the
Bangalore University and rosemary from the Lalbagh nursery. Corn cob and coconut
husk were obtained from their respective local industries. They were dried in hot air
oven at 105O C for 5 minutes as they lose all the moisture and were crushed to powder
using pestle and mortar as shown in Fig 4.1.
Fig 4.1 Eucalyptus leaves being ground using mortar and pestle
Since the powder was not fine and uniform, mixer was used to grind the dried
material. Ground powder was then sieved through 2 mm mesh sieve to obtain a fine
powder as shown in Fig 4.2.
Fig 4.2 Powdered leaves being sieved to obtain fine and uniform powder.
23
The binding agent, to start with we developed a coil using cow dung as a binding
agent, but the coil had very low binding strength of 15 g as shown in Fig 4.3 although
the burning property was good, we tried changing the binding agent to corn cob. The
corn cob which is a by-product of corn was brought from a corn cob vendor and dried
in hot air oven at 105o C for 5 minutes until all the moisture is lost as shown in Fig
4.4. The dried corn cob as shown in Fig 4.5 was ground to fine powder with the help
of the locally available commercial grinders. Again the powdered corn cob was finely
grounded using a mixer and was sieved through 2 mm sieve to obtain a uniform fine
powder.
Fig 4.3 coils developed using cow dung which had low binding strength
Fig 4.4 Corn cob kept in hot air oven at 600 C for 48 hours for drying
24
Fig 4.5 Dried binding material corn cob ready to be crushed
Corn cob chosen as the binding agent was mixed with plant leaves powder and the
paste was given a cuboidal shape which was dried in a hot air oven at 60O C for 48
hours until most of the moisture is lost. Prepared coil had very low binding strength of
20 g. Then to an approximate amount of corn cob powder, a little of water was added
and the mixture was boiled in microwave for about 3 minutes until the mixture
became sticky and the plant leaves powder was added to the sticky paste as shown in
Fig 4.6 and the wet coil paste was moulded into a cuboidal shape with dimension
6cmX1cmX0.5cm using a mould of cardboard. The wet coil was then dried in the hot
air oven at 60O C for 48 hours until most of the moisture is lost and the coil had good
binding strength of 90 g. When boiled the structural component of glutin gets
structurally modified and the corn cob get sticky.
The coil developed with corn cob was tested for burning ability and the burning of the
coil was not constant and it used to go off very soon and an organic smouldering
agent Coconut husk was tried. Coconut husk is a by-product of coconut industry and
was collected from a coconut industry. Coconut husk collected was ground into fine
particles using mixer and was sieved through 2 mm sieve to obtain a fine and uniform
coconut husk powder. To the above prepared wet mixture, a finite amount of
powdered coconut husk was added and was given a cuboidal shape which was later
dried using hot air oven at 60O C for 48 hours until most of the moisture is lost and the
dried coil was tested for burning property and the coil had a very good burning
property which could be easily ignited and burned continuously.
25
To know the exact amount of the corn cob and coconut husk to be added to the plant
leaves powder so that the coil has good binding strength as well as burn to
continuously, optimization of these components was carried out. To optimize the
concentrations of the corn cob and coconut husk we developed the combination of the
coils in which concentration of the citronella leaves remained constant i.e. 5g and the
concentrations of corn cob and coconut husk vary amounting to total 5.5g of each
combination. The coil with less than 2.625g of corn cob was fragile and the coil with
less than 0.375g of coconut husk was not burning continuously. Out of the
combinations, coil with 2.625g of corn cob and 0.375g of coconut husk showed best
binding strength and burning property and had least harmful gases emission. So the
concentration of the corn cob and coconut husk was optimized.
Fig 4.6 Corn cob boiled and mixed with other constituents
Coils were developed from individual leaves alone and were tested for the binding
strength, burning duration, emission test for the carbon dioxide, carbon monoxide and
hydrocarbons and finally for the repellence towards mosquitoes. Three different coils
developed from three different plants citronella, eucalyptus and rosemary were tested
for the above properties.
Cuboidal shape with dimensions 6cm length, 1 cm width and 0.5 cm height were
given to the coils. Previously we gave them the round shape which restricted them
from burning continuously as shown in Fig 4.7. So we made the moulds from the
cardboard sheets as shown in Fig 4.8 and the mixture of plant leaves, boiled corn cob
and coconut husk was filled in as shown in Fig 4.9 and dried at 60O C for 48 hours
until most of the moisture is lost.
26
Fig 4.7 Mosquito coil in round shape
Fig 4.8 Moulds developed from cardboard sheet
Fig 4.9 Mould being filled with coil paste
27
Once the coils were prepared, they were tested for the following properties:
 Binding strength- Binding strength was assessed to find out the compactness
of the coil by checking the maximum weight the coil can withstand and plays
an important role in the coil as low binding strength leads to easy breakage of
the coil.
 Harmful gases emission- This property was assessed to check for the amount
of Carbon dioxide (CO2), Carbon monoxide (CO), Hydrocarbons (HC) gases
released while burning the coil by using the vehicle emission testing
mechanism and it is important that the harmful gases be as low as possible.
 Burning duration- This property was assessed to check for the duration of
burning of the coil by burning it in the open and more the burning time the
longer the protection by the mosquito coil.
 Moisture content- Moisture content of the coil was analysed to check for the
water content remaining in the coil just before burning them and it is an
important indicator as to check at what moisture content the coil burns.
 Repellence- This property was assessed to check the repellence of mosquitoes
from the gas produced from the burning coil and is the most important
property of any coil as it determines the efficacy of any mosquito coil.
4.2 Optimization of the Coil

Since Binding strength and Harmful gases emission are two important components of
any mosquito coil, those 2 properties were chosen to optimize the coil by selecting the
best combination of corn cob and the coconut husk with the plant material.
Leaves of citronella plant, corn cob powder and coconut husk are dried in hot air oven
at 105o C for 5 minutes and are grounded using mixer and are sieved through BSS
sieve with 2 mm aperture and with equal weights of leaf powder, different ratios of
corn cob powder and coconut husk powder are taken making up equal weight as
whole as shown in the table. Corn cob powder is heated in microwave oven for 3
minutes until the corn cob takes up the water and becomes sticky, then the corn cob is
heated and mixed with the definite amounts of coconut husk and the plant material
powder as shown in Table 4.1.
28
Table 4.1 Optimization of coil in reference to the various constituents

Citronella Corn cob Coconut Water Water Total
weight (g) weight husk added remained weight
(g) weight (g) initially after after
(g) boiling (g) mixing
(g)
COIL1 2.5 2.875 0.125 40 18.313 23.813
COIL 2 2.5 2.75 0.25 40 16.685 22.185
COIL 3 2.5 2.625 0.375 40 15.715 21.215
COIL 4 2.5 2.5 0.5 40 15.189 20.689
The mixture is then filled up in the mould which has dimensions of 6cm X 1cm X
0.5cm and the coil is dried up in the hot air oven at 600 C for 2 days as it loses most of
the moisture.
The coils are first tested for binding strength by holding one end of the coil with a
clamp and other is subject to weights in increasing order and the breaking point for
the coil is taken to be the binding strength of the coil as shown in Fig 4.10 and the
results are noted down in the table 4.2.
Fig 4.10 Testing Binding strength of the coil
29
Table 4.2 Binding strength results for optimization of coil

Citronella Corn Coconut Total Moisture % Binding
weight cob husk weight present moisture strength
(g) weight weight (g) (g)
(g) (g)
COIL1 2.5 2.875 0.125 5.528 0.028 0.506 75
COIL 2 2.5 2.75 0.25 5.523 0.023 0.416 95
COIL 3 2.5 2.625 0.375 5.532 0.032 0.578 140
COIL 4 2.5 2.5 0.5 5.529 0.029 0.524 55
MORTEIN 200
COIL
Coils are then tested for the harmful gas emission at emission testing centre which
works by measuring the amount of harmful gases such as Carbon dioxide (CO2),
Carbon monoxide (CO) and Hydrocarbons (HC) present in the smoke from the
burning of coil. The coil is lit up inside one end of the pipe and the ends are blocked
to stop the smoke from escaping for 10 seconds and the emission testing device is
inserted into the coil from the other end as shown in the Fig 4.11 and the device notes
down the amount of harmful gases present in the smoke and the reading from the
computer are noted down in the Table 4.3.
Fig 4.11 Analysis of smoke for harmful gas emission
Based on the results of binding strength [Table 4.2] and emission testing [Table 4.3],
coil 3 was selected as the optimum composition of the corn cob and coconut husk to
be used for further testing.
30
Table 4.3 Emission testing results for optimization of coil

Citronella Corn Coconut Carbon Carbon Hydrocarbons
leaves (g) cob (g) husk (g) monoxide dioxide (PPM)
(% Vol) ( % Vol)
COIL 1 2.5 2.875 0.125 0.225 2.07 114
COIL 2 2.5 2.75 0.25 0.279 2.14 132
COIL 3 2.5 2.625 0.375 0.292 2.68 129
COIL 4 2.5 2.5 0.5 0.249 2.17 157
MORTEIN 0.611 3.45 139
COIL
4.3 Preparation of Mosquito Coils

The coils were prepared from each of the plant alone and with different combinations
of each plant from the Taguchi’s design of experiments (DOE) as shown in Table 5.4
into a mixture with corn cob as the binding material and coconut husk as the
smouldering agent as mentioned in the above section.
Table 4.4 Taguchi’s Design of Experiments:

Citronella (g) Rosemary (g) Eucalyptus (g) Total weight (g)
COIL 1 2.5 0 0 2.5
COIL 2 0 2.5 2.5 2.5
COIL 3 0 0 2.5 2.5
COIL 4 0.834 0.834 0.834 2.5
COIL 5 0.500 1.000 1.000 2.5
COIL 6 0.357 1.070 1.070 2.5
COIL 7 1.000 0.500 1.000 2.5
COIL 8 0.714 0.714 1.070 2.5
COIL 9 0.834 1.248 0.416 2.5
COIL 10 1.070 0.357 1.070 2.5
COIL 11 1.250 0.834 0.416 2.5
COIL 12 0.938 0.938 0.625 2.5
Each of the coil is prepared with definite ratio of corncob and coconut husk as it was
optimized in the previous section i.e., 2.625g of corn cob and 0.375 g of coconut husk
31
with the 2.5 g of plant material, making up the total weight to 5.5 g which is
distributed equally into 4 moulds of coil for drying after the addition of water.
4.3.1 Burning duration was checked for each of the coil by lighting the coil at one
end as shown in Fig 4.12 and the time taken by the coil to burn completely are noted
down and the readings are tabled in the following table:
Fig 4.12 Analysis of Burning duration
4.3.2 Moisture content was calculated in order to find out the amount of moisture
present in the coil while burning the coil and this is calculated by taking the difference
in weights of wet coil prepared and the dry coil before burning.
4.3.3 Repellence testing of the coils was analysed in order to check their efficiency
of the coil in repelling the mosquitoes by their smoke which is produced from the
burning of the coil.
Repellence testing of the coil was carried out as follows. A glass box with dimensions
45cm length, 20 cm width and 35 cm height with bottom end open was chosen which
also had a opening on one of the sides with dimensions 5cmX10cm as shown in Fig
4.15. Glass box had a total volume of 31500 cubic centimetres. This glass box can be
compared with the dimensions of a house in which the opening in the glass box wall
referring to the window in the house. The glass box with its open end was kept
downwards so that the only way air and mosquitoes can pass through was the opening
through the wall of the glass.
32
Fig 4.13 Aedes aegypti breeding in larva stage

Aedes aegypti adult females of 4-5 days age were bred in a mesh cage as shown in Fig
4.14 and using the aspirator the mosquitoes were taken out manually and for
repellence testing of each coil 6 mosquitoes were introduced into the inverted glass
box as shown in Fig 4.17 wherein each mosquito had a volume of 5250 cubic
centimetres of volume inside the box and could move freely around and can go
outside through the opening. As soon as the mosquitoes were in the box, mosquito
coil was lit at one of its ends and kept at the centre of the glass box and it obtained
oxygen for burning through the opening in the wall of the box through which the air
freely moved in and out. Based on the movement of the mosquitoes outside the box or
their killing or their numbness inside the box, repellence of the mosquito coil was
tested.
Fig 4.14 Adult Aedes aegypti Mosquito breeding in cages
33
45 cm
20 cm
35 cm
10 cm
5
5 cm
Fig 4.15 Glass box dimensions used for repellence testing
Fig 4.16 Aedes aegypti adult mosquitoes in mesh cage
Fig 4.17 Transfer of mosquitoes from cage to glass box using aspirator
34
Fig 4.18 Testing of mosquito repellence

Negative control was to leave the mosquitoes in the box and note down the time
taken by the mosquitoes to go out of the box. Negative control was done to check the
movement of mosquitoes in the box and out of the box.
Six adult Aedes aegypti mosquitoes were taken out from the cage using aspirator and
were introduced into the box and the time taken by them to go out of the box was
noted up to 10 minutes as shown in Fig 4.18.
Positive control was to check the repellence of a commercially available Mortein
mosquito coil on the mosquitoes. This control was performed to compare the readings
of our mosquito coil with the commercially available mosquito coil. Six adult Aedes
aegypti mosquitoes were taken out from the cage using aspirator and were introduced
into the box and the Mortein mosquito coil was burnt in the box as shown in Fig 4.19
and the readings were noted down.
Fig 4.19 Testing of repellence for commercial mosquito coil
35
Fig 4.20 Testing of repellence for mosquito coil
Repellence testing of mosquito coils

Six adult Aedes aegypti mosquitoes were taken out from the cage using aspirator and
were introduced into the box and each of the coils prepared were burnt as shown in
the Fig 4.20 and the effect of smoke on the mosquitoes were noted down and the
repellence was checked as shown in the Fig 4.22.
36
RESULTS AND DISCUSSION
This chapter includes the results and discussion of physical properties testing and
repellence testing of the prepared coil and the inferences referred from the results of
physical and repellence testing of prepared coils.
5.1 Binding strength: The coils are tested for binding strength by holding one end
of the coil with a clamp and other is subjected to weights in increasing order and the
breaking point for the coil is taken to be the binding strength of the coil. Binding
strength is checked for each of the coil and the results are as follows in Table 5.1.
Table 5.1 Binding strength analysis results

Citronella (g) Rosemary (g) Eucalyptus (g) Binding strength (g)
COIL 1 2.5 0 0 140
COIL 2 0 2.5 0 107
COIL 3 0 0 2.5 70
COIL 4 0.834 0.834 0.834 95
COIL 5 0.500 1.000 1.000 87
COIL 6 0.357 1.070 1.070 77
COIL 7 1.000 0.500 1.000 100
COIL 8 0.714 0.714 1.070 115
COIL 9 0.834 1.248 0.416 105
COIL 10 1.070 0.357 1.070 97
COIL 11 1.250 0.834 0.416 110
COIL 12 0.938 0.938 0.625 107
5.2 Burning duration was checked for each of the coil by lighting the coil at one
end and the time taken by the coil to burn completely are noted down and the readings
are tabled in the following table 5.2.
38
Table 5.2 Burning duration of various coils in comparison to commercial coil

Citronella Rosemary (g) Eucalyptus Burning duration
(g) (g) (minutes)
COIL 1 2.5 0 0 25”26’
COIL 2 0 2.5 0 19”48’
COIL 3 0 0 2.5 23”02’
COIL 4 0.834 0.834 0.834 20”36’
COIL 5 0.500 1.000 1.000 24”41’
COIL 6 0.357 1.070 1.070 22”11’
COIL 7 1.000 0.500 1.000 23”33’
COIL 8 0.714 0.714 1.070 23”51’
COIL 9 0.834 1.248 0.416 20”08’
COIL 10 1.070 0.357 1.070 21”58’
COIL 11 1.250 0.834 0.416 18”34’
COIL 12 0.938 0.938 0.625 18”06’
MORTEIN 20”05’
COIL
From the Table 5.2 it can be inferred that most of the prepared mosquito coils have
longer burning duration compared to the commercial Mortein coil, especially
Citronella coil has the longest burning duration.
5.3 Emission testing was carried for each of the coils as explained in the above
section and the results are tabulated in the Table 5.3.
Table 5.3 Emission testing of various coils
Carbon monoxide Carbon dioxide Hydrocarbons
( % Vol) ( % Vol) (PPM)
COIL 1 (Citronella) 0.225 2.07 114
COIL 2 (Rosemary) 0.248 2.21 120
COIL 3 (Eucalyptus) 0.268 2.02 118
COIL 4 0.238 2.09 112
COIL 5 0.249 2.15 112
COIL 6 0.252 2.13 110
COIL 7 0.248 2.12 111
COIL 8 0.242 2.20 119
COIL 9 0.235 2.09 115
COIL 10 0.258 2.20 117
COIL 11 0.242 2.15 114
COIL 12 0.239 2.18 105
MORTEIN COIL 0.611 3.45 139
39
Fig 5.1 Emission testing result for commercial coil and citronella coil
From the Table 5.3 and fig 5.1 it can be inferred that all of the prepared mosquito
coils have lesser emission of toxic gases than the commercial Mortein coil sometimes
half the emission of commercial mosquito coil which is advantageous to the user.
5.4 Moisture content was calculated in order to find out the amount of moisture
present in the coil while burning the coil and this is calculated by taking the difference
in weights of wet coil prepared and the dry coil before burning. Moisture content
values of each coil are tabulated below in Table 5.4.
Table 5.4 Moisture content analysis of various mosquito coils

Weight of wet Weight of dried Moisture % moisture
coil (g) coil (g) present (g)
COIL 1 24.428 5.529 .029 0.524
COIL 2 21.658 5.526 .026 0.470
COIL 3 22.154 5.530 .030 0.542
COIL 4 23.110 5.525 .025 0.452
COIL 5 24.089 5.527 .027 0.488
COIL 6 22.482 5.529 .029 0.524
COIL 7 20.416 5.531 .031 0.560
COIL 8 23.659 5.518 .018 0.326
COIL 9 22.554 5.529 .029 0.524
COIL 10 25.268 5.520 .020 0.362
COIL 11 21.992 5.524 .024 0.434
COIL 12 24.239 5.527 .027 0.488
40
From Moisture content analysis results it can be inferred that the coil loses all most all
the moisture content upon drying in hot air oven at 60o C for 48 hours and the coils
have least amount of water present just before using them.
5.5 Repellence testing

Negative control was to leave the mosquitoes in the box and note down the time
taken by the mosquitoes to go out of the box. Negative control was done to check the
movement of mosquitoes in the box and out of the box. Six adult Aedes aegypti
mosquitoes were taken out from the cage using aspirator and were introduced into the
box and the time taken by them to go out of the box was noted up to 10 minutes and
the results are tabulated in Table 5.5.
Table 5.5 Negative control results
Mosquito Effect on the mosquito Remarks
(minutes)
1 6”28’ Moved out of the box
2 - Stayed in the box
From the negative control experiment it can be inferred that mosquitoes would not
move out of the box without any external factor repelling them. 5 of the 6 mosquitoes
stayed in the box even after 10 minutes into the experiment.
Positive control was to check the repellence of a commercially available Mortein
mosquito coil on the mosquitoes. This control was performed to compare the readings
of our mosquito coil with the commercially available mosquito coil. Six adult Aedes
aegypti mosuitoes were taken out from the cage using aspirator and were introduced
into the box and the Mortein mosquito coil was burnt in the and the readings were
noted down table 5.6.
From the repellence activity of commercially available coil (mortein) it is inferred that
it has harmful chemicals which kill the mosquitoes in a short period of time and is an
effective mosquito killer.
41
Table 5.6 Mosquito repellence results for commercial Mortein coil

(minutes)
1 1”20’ Mosquito died
Repellence testing of prepared mosquito coils

6 adult Aedes aegypti mosuitoes were taken out from the cage using aspirator and
were introduced into the box and each of the coils prepared were burnt and the effect
of smoke on the mosquitoes were noted and tabulated in the following tables 5.7 for
coil 1 till 5.18 for coil 12.
Citronella coil (COIL 1):
Table 5.7 Repellence results for citronella coil
(minutes)
6 3”19’ Numbness
From the Table 5.7 results it can be inferred that Citronella coil smoke repels
mosquitoes effectively in less time at par with the efficiency of commercial Mortein
coil.
Rosemary coil (COIL 2):
Table 5.8 Repellence results for rosemary coil
(minutes)
42
From Table 5.8 results it can be inferred that mosquitoes move out of the box due to
smoke of the coil and those which cannot move out of the box get numbness and they
get active when the smoke reduces and the Rosemary coil is slow acting.
Eucalyptus coil (COIL 3)

Table 5.9 Repellence results for Eucalyptus coil
(minutes)
smoke of the coil and those which cannot move out of the box get numbness and they
get active when the smoke reduces and the Eucalyptus coil is slow acting.
Fig 5.2 Numbness caused by mosquito coil smoke
As shown in the Fig 5.2, prepared mosquito repellent coils only either repel the
mosquitoes or cause numbness to the mosquitoes unlike the commercial Mortein coil
which killed most of the mosquitoes in short amount of time and the numbed
mosquitoes get active when the smoke reduces.
43
COIL 4
Table 5.10 Repellence results for coil 4
(minutes)
smoke of the coil and those which cannot escape out of the box get numbness and
they get active when the smoke reduces and the Coil 1 is slow acting.
COIL 5
(minutes)
smoke of the coil and those which cannot escape out of the box get numbness and
they get active when the smoke reduces and the Coil 5 is slow acting.
COIL 6
(minutes)
44
smoke of the coil and those which cannot escape out of the box get numbness due to
the smoke of the burning coil and they get active when the smoke reduces and the
Coil 6 is very slow acting.
COIL 7
(minutes)
the smoke and they get active when the smoke reduces and the Coil 7 is slow acting.
COIL 8
(minutes)
the smoke produced by the burning coil and they get active when the smoke reduces
and the Coil 8 is slow acting.
45
COIL 9
(minutes)
the smoke and they get active when the smoke reduces and the Coil 9 is very slow
acting.
COIL 10
(minutes)
the smoke and they get active when the smoke reduces and the Coil 10 is relatively
fast acting in comparison with the commercial Mortein mosquito coil.
46
COIL 11
(minutes)
the smoke and they get active when the smoke reduces and the Coil 11 is fast acting
on the mosquitoes.
COIL 12
(minutes)
From the results of Table 5.7 to Table 5.18, it can be inferred that prepared mosquito
coils have repellence to mosquitoes or make them numb which after conditions get
better get active and start flying.
Out of the 12 mosquito coils tested for repellence, coil with citronella plant alone
gives the best results in which all the mosquitoes are repelled within 3 minutes and 19
seconds. Although not fast acting as the commercial Mortein mosquito coil, prepared
plant based mosquito coils show repellence on longer time and also plant based
47
mosquito coils does not kill the mosquitoes as the commercial Mortein coil but either
repel or numb the mosquitoes which is eco-friendly.
An eco-friendly, biodegradable, organic citronella based mosquito coil is prepared
which is entirely organic without involving any expensive instruments and is par with
commercial mosquito coil in repellent efficiency.
48
CONCLUSION AND SCOPE FOR FUTURE WORK

This chapter contains the outcome of the research work and the work which can be
carried in the future using our results.
6.1 Conclusion
The efficiency of the plant based mosquito coil was studied. This was primarily done
in two steps, assessing physical properties and assessing repellence property of the
mosquito coil. First, physical properties such as harmful gas emission, burning
duration, moisture content and binding strength were calculated and repellence for the
coil was tested and later all the results were compared with the commercial mosquito
coil properties. The following conclusions were drawn:
 Three Plants namely Citronella grass (Cymbopogon nardus), Eucalyptus tree
(Eucalyptus globulus) and Rosemary plant (Rosmarinus officinalis) were
selected as they have shown are eco-friendly biological repellence towards
mosquitoes, Coconut husk as smouldering agent and Corn cob powder as the
binding agent were selected.
 Coil from the citronella plant alone gave the best results by repelling all the
mosquitoes from the glass box in 3 minutes 19 seconds compared to the
commercial mosquito coil which killed all the mosquitoes in 3 minutes and 12
seconds, citronella coil has less harmful gas emission compared to Mortein
coil and has longer burning duration i.e. 25 minutes and 26 seconds compared
to the Mortein coil which has 20 minutes 05 seconds burning duration but
only has lower binding strength i.e. 140 g compared to the Mortein mosquito
coil . From the above results we can infer that citronella mosquito coil has
better physical and repellence properties than the commercial Mortein
mosquito coil.
6.2 Scope for Future Work

There are several areas that require further elaborate study. They are:
 Many plants are yet to be explored for their repellence and many organic
materials can be tried for binder and also smouldering agent. Organic
materials which add a pleasant smell can also be tried in preparation of plant
based mosquito coils.
50
 Not much is known about how the mosquitoes are repelled by the repellent
chemicals and this mechanism has to be worked on so that better mosquito
repellents are prepared.
 Essential oils of many plants are yet to be extracted and their repellence can be
checked to improve the present plant based mosquito coils.
 Different binder material and smouldering agent can be tried with the same
plants chosen by us to improve the physical properties such as binding
strength, harmful gas emission, burning duration which will all improve the
quality of the final product.
 Improvisations can be done on the same product by incorporating organic
material which gives a pleasant smell and increases its aesthetic properties
and its acceptability.
51
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KARNATAKA STATE COUNCIL FOR SCIENCE AND

R.V.COLLEGE OF ENGINEERING, BENGALURU-59

DEVELOPMENT OF ECO-FRIENDLY PLANT BASED

Under the Guidance of

Dr. Nagashree N Rao Associate Professor and Dr. Ashwani Sharma

(Autonomous Institution Affiliated to VTU, Belagavi)

Guide Head of the Department Principal

Name of Examiners Signature with date

(Autonomous Institution Affiliated to VTU, Belagavi)

Place: Bengaluru Signature

3. SAURAV KUMAR SINGH

We convey our gratitude to Dr. Satyanarayana B.S., Principal, R.V. College of

We are thankful to Ms. Lakshmi, Mr.Pradeep, Mr.Harsha and Mr. Armugam T A

Three Plants namely Citronella grass (Cymbopogon nardus), Eucalyptus tree

1.1 Market Survey 2

1.2 Societal Relevance 3

1.3 Problem Statement 3

1.4 Recent work done on mosquito repellents 3

1.4.1 Review on commercial mosquito coils 4

1.4.2 Review of materials 5

1.4.3 Review on processing techniques 6

1.4.4 Unresolved issues 7

1.4.5 Emerging opportunities 7

1.4.6 Research gap 8

1.4.7 Innovativeness in our project 8

2.1 Organic Materials 12

2.2 Adult mosquito 13

3.1 Project Methodology 16

3.1.2 Method to Develop Mosquito Repellent Coil 19

from Individual Plants

3.1.3 Method to Develop Mosquito Repellent Coil with 21

Combination of Three Plants (Citronella, Eucalyptus and Rosemary)

4. BIOLOGICAL MOSQUITO COIL FORMULATION 23

4.1 Development of mosquito coil 23

4.2 Optimization of the Coil 28

4.3 Preparation of Mosquito Coils 31

5. RESULTS AND DISCUSSIONS 38

5.1 Binding strength 38

5.2 Burning duration 38

5.3 Emission testing 39

5.4 Moisture content 40

5.5 Repellence testing 41

6. CONCLUSION AND FUTURE WORK 50

LIST OF SYMBOLS, ABBREVIATIONS AND NOMENCLATURE

DOE - Design of Experiments

BBB - Blood Brain Barrier

EPA - Environmental Protection Agency

CO2 - Carbon dioxide

2.1 DOE method applied for obtaining best combinations 14

The mosquito repellent market in India is valued at ` 16 billion and is growing at a

1.2 Societal Relevance

1.3 Problem Statement

1.4 Recent work done on mosquito repellents

1.4.1 Review on commercial mosquito coils

Fig 1.2. Health effects of smoke from commercial mosquito coil

There are two types of pyrethrins:

Type 1 pyrethrin produce reflex hyper excitability and fine tremor.

Type II pyrethrins produce salivation, hyper excitability, choreoathatosis, and

Permethrin is a type of pyrethrin, EPA classified permethrin as possible human

during 3rd to 8th week of pregnancy.

1.4.2 Review of materials

1.4.3 Review on processing techniques