Efficacy of Horseradish (Moringa Oleifera As An Alternative Mosquito Repellent

Efficacy of Horseradish (Moringa oleifera) as an alternative Mosquito Repellent
Chapter 1
INTRODUCTION
Background of the Study
A mosquito repellent doesn't actually kill mosquitoes. Repellent work by
making people less attractive to mosquito, so they're less likely to bite you. Today
mosquito repellent is very useful and it can be man - made.
Horseradish or also known as Moringa Oleifera is a root vegetable known
for its punget taste and odor. Horseradish (Moringa oleifera) not only helps ward
off diseases, but it makes an excellent mosquito repellent. Horseradish (Moringa
oleifera) Oil can last for as long as 1 - 2 years past of the expiration date of its jar.
Horseradish (Moringa oleifera) is beneficial to human health. The leaves of the
horseradish have 7 times more vitamin C than oranges and 5 times more
potassium than bananas. It also has calcium, protein, iron and amino acids,
which help your body heal and build muscle. It also packed with antioxidants
substances that protect cell from damage and may boast your immune system.
Moringa Oleifera is the scientific name for Horseradish and it is also called
as “miracle tree” because it has a strong reputation of curing many disease and
the most widely cultivated plant species worldwide. The phytochemicals derived
from the Horseradish (Moringa oliefera) can act as larvicides, insect growth
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regulators, repellents and also have a very deterrent activity observed by many
scientist and researchers. In the study of Asian Pacific Journal of Tropical
Biomedicine, the outcome acquired after the treatment of Horseradish (Moringa
Oleifera) for malarial vector were encouraging.it also yields leaves where
government health researchers found extracts for the formulation of an effective
anti-mosquito repellant. It is not only against dengue, in fact, but against all
mosquito-borne diseases like malaria and filariasis especially in areas of the
country where incidence is high. It also serves as a personal care product used
in preventing mosquito bites that cause itchiness, inflammation and discomfort to
individuals. The repellent uses a washable, oil – based formulation and it's
biodegrable since, it is derived from advantage against other well-known
repellents which are chemical based in formulation. The Horseradish (Moringa
oleifra) leaves roots, fruits, as well as its seeds, have been found beneficial to
human use and needs. Studies conducted by big institutions such as the
Department of Science and Technology and by the Philippine Council for Health
Research and Development (PCHRD) shows that the Horseradish (Moringa
oleifera) plant has components that are potent in repelling mosquitos. Infact,
Horseradish (Moringa oleifera) serves a lot of purposes to human health as
shown by the list created by the Bureau of Plant Industry: Medicinal value of
Horseradish (Moringa oleifera) the methanolic extract of Horseradish (Moringa
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oleifera) leaves was evaluated for repellent effect against mosquito. Repellent
effect of treatment was assessed at different concentration (25, 50, 75, and 100
ml) after 5mins, 10mins, 15mins, of exposure against 45 mosquitoes in a cage.
All extract were evaluated in different cages.Percentage repellency (PR) was
determined for each extract following the procedure of WHO. The results
indicated that the significant effect of the treatments and the effectiveness
increased with extended exposure interval and enhanced dose rate. There was a
statically significant difference between the 2 test groups (Control, and
Horseradish (Moringa oleifera) as determined by a one-way Analysis of Variance
(ANOVA) (F (22, 1) = 20921.216, P = 0.000. Since the p-value 0.000 < 0.05, a
Turkey Post-hoc test revealed that at 95% confidence level, Horseradish
(Moringa oleifera) yielded a (highest) mean value of percentage repellency from
those Control (which also yielded different mean values). The LC50 value
recorded for Horseradish (Moringa oleifera) is 18.2ml respectively. The test plant
extracts were screened for phytochemicals such as alkaloids, flavonoids,
saponins, tannins, steroids, anthraquinones and terpenes. The findings of the
present study clearly revealed that methanol extracts of Horseradish (Moringa
oleifera) leaves are good repellent agents for the control of dengue, however the
efficacy depends on dose rates and the exposure interval. Horseradish (Moringa
oleifera) contains properties that are similar to other vegetables, it has a rich
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source alkaloid, and Bioactive compounds. These components are responsible in
producing mosquito repellent. The Horseradish (Moringa oleifera) is often
considered as important famine food because of its high resistance to drought
and arid conditions owing to their tuberous roots (Padayachee Baijnath 2012).
Almost each and every part of Horseradish (Moringa oleifera) tree is useful for
medicinal, functional food preparations, nutraceuticals, water purification, and
biodiesel production; including roots, leaves, flowers, green pods, and seeds
(Saini 2015). Making your own organic and natural mosquito repellent is really
quite simply and much safer for your entire family than store bought repellent that
are loaded with harmful and expensive chemicals )This Horseradish (Moringa
oleifera) repellent could help in saving the lives of upcoming future in preventing
and lessening the case of dengue that could harmfully affect the public health by
promoting more research studies to scientist about this miracle plant .It can be
the answer for our problem with mosquitoes.
Statement of the Problem
This study aimed to investigate the Efficacy of Horseradish (Moringa oleifera)
as an alternative mosquito repellent .Specifically, it sought to answer the
following questions.
1. What are the phytochemical present in Horseradish (Moringa oleifera) that can
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be an alternative ingredient in making mosquito repellent?
2. Can Horseradish (Moringa oleifera) be an effective mosquito repellent?
3. Is there significant difference among treatments of Horseradish (Moringa
oleifera) in killing mosquito?
4. Is there significant difference between the best Treatment of Horseradish
(Moringa oleifera) as mosquito repellent compared to commercial one?
5. Is there a significant difference between the Treatment of Horseradish
(Moringa oleifera) as mosquito repellent as compared to commercial one in terms
of cost?
Statement of Hypotheses
Null Hypotheses
This study was directed by following hypotheses.
1. There is no phytochemical present in Horseradish (Moringa oleifera) that can
be an alternative ingredient in making mosquito repellent.
2. Horseradish (Moringa oleifera) is not effective mosquito repellent.
3. There is no significant difference among treatments of Horseradish (Moringa
oleifera) in killing mosquito.
4. There is no significant difference between the best Treatment of Horseradish
(Moringa oleifera) as mosquito repellent compared to commercial one.
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5. There is no significant difference between the Treatment of Horseradish
of cost.
Alternative Hypotheses
1. Horseradish (Moringa oleifera) contains various phytochemical such as
alkaloids, saponins, tannins, and Flavonoids, that can be alternative
ingredients in making mosquito repellent.
2. Horseradish (Moringa oleifera) is effective mosquito repellent.
3. There is a significant difference among treatments of Horseradish
(Moringa oleifera) in killing mosquito.
4. There is a significant difference between the best Treatment of
Horseradish (Moringa oleifera) as mosquito repellent compared to
commercial one.
5. There is a significant difference between the Treatment of Horseradish
(Moringa oleifera) as mosquito repellent as compared to commercial one
in terms of cost.
Objectives of the Study
The main objectives of the study aimed to determine the efficacy of
Horseradish (Moringa oleifera) as an alternative Mosquito Repellent.

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Specifically, it will aimed:
1. To determine what are the phytochemical present in Horseradish (Moringa
oleifera) that can be an alternative ingredient in making mosquito repellent.
2. To determine the efficacy of Horseradish (Moringa oliefera) as mosquito
repellent.
3. To determine the significant difference among treatments of Horseradish
(Moringa oleifera) in killing mosquito.
4. To determine the significant difference between the best Treatment of
Horseradish (Moringa oleifera) as mosquito repellent compared to commercial
one.
5. To determine the significant difference between the Treatment of Horseradish
of cost.
Significance of the Study
This study aimed to determine the efficacy of Horseradish (Moringa
oleifera) as Alternative Mosquito repellent. This research will be beneficial to the
following:
To the community, this study will provide information's that is beneficial to
all the community members on how to turn Horseradish (Moringa oleifera) as an

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alternative mosquito repellent.
To the Department of Science and Technology, this study will encourage
them to develop another alternative mosquito repellent.
To the Doctors and pharmacies, they can create and introduce a product
and tell how effective the product is and they can be paid with high salary.
To the Economy, because this study will create an opportunity for the
agricultural sector of our economy.
To The farmer, they will plant Horseradish (Moringa oliefera) and take care
of it for a long period of time then after doing their job they will get paid.
To the Future researchers, this will help them to attain more knowledge and
information about the Horseradish (Moringa oleifera) leaves extract and the result
of this study will be great help and will serve as a reference for the future
researchers.
Scope and Limitations of the Study
This study will focused to investigate the efficacy of Horseradish (Moringa
oleifera) as an alternative mosquito repellent. It was conducted from November
to March 2021 at Alaminos City National High School and at Researcher House,
Virgen Milagrosa Foundation (VMUF), college of pharmarcy- Laboratory in San
Carlos City, Pangasinan.
The Horseradish (Moringa oleifera) leaves was the only plant part used in
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conducting this study. The leaves will be gathered in the researcher's backyard
where it is abundant.
Six (6) treatment in three replicates was prepared in this study. These
included Treatment zero (T0) as the negative control, Treatement one (T1) 25%
Horseradish (Moringa oleifera) leaves extract solution (consisted of 25 ml
Horseradish (Moringa oleifera) leaves extract solution and 75% ml commercial
repellent). Treatment two (T2) 50% Horseradish (Moringa oleifera) leaves
extract solution (consisted of 50 % ml Horseradish (Moringa oleifera) leaves
solution and 50 ml commercial repellent), Treatment three (T3) 75%
Horseradish (Moinga oleifera) leaves extract solution and 25 ml commercial
repellent), Treatment four (T4) 100 % Horseradish (Moringa oleifera) leaves
extract solution (consisted pure Horseradish Moringa oleifera leaves extract
solution) and Treatment five (T5) as the positive control Commercial repellent
(DEET).
Two hundred seventy (270) mosquito was used as experimental animals and
were acquired at a frankie’s shop San Carlos City, Pangasinan. They were
placed in the metal cage with six divisions. They were brought to the VMUF –
collage of pharmacy laboratory during the experiment.
As the summary of the review of the literature review the researcher
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conclude that Horseradish (Moringa oleifera) grow widely in many tropical and
subtropical countries. Based from the information collected it was confirmed that
Horseradish (Moringa oleifera) leaves extract exhibit the greatest Biochemical
activity that is responsible for producing mosquito repellent. Horseradish
(Moringa oleifera) leaf extract indicate a high degree of safety. No adverse effects
were reported in associated with human studies.
Definition of Terms
Analysis of Variance. Refers to the statistical tool used to determine if there
is a significant difference among treatment.
DEET. In this study, it is referring to the commercial product used as
mosquito repellent.
Horseradish. It is a plant native to northern India and most widely cultivated
plant species worldwide. The leaves extract of this plant was used as an
alternative mosquito repellent.
Mosquito. It is considered as one of the life-threatening species. They are
carriers of certain diseases some of which can deadly and they also transmit

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malaria and dengue fever.
Mosquito repellent. It is a substance applied from the skin. It evaporates on
the skin and work by blocking a mosquito's sense of smell. Preventing it from
finding its target.
Scheffe Test. It refers to the statistical tool that was used if the result of the
ANOVA is significant in order to determine where the variance lies.
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Chapter 2
REVIEW OF RELATED LITERATURE AND STUDIES
Foreign Literature and Studies
Horseradish (Moringa oleifera) is the scientific name for malunggay and it
is also called as "miracle tree" because it has a strong reputation of curing many
disease and the most widely cultivated plant species worldwide. Horseradish
(Moringa oleifera) is the most widely cultivated species of a monogeneric family -
The moringaceae, native to sub- himalayan tracts of India. This rapidly- growing
tree (also known as the horseradish tree, drumstick tree, benzalire tree, mlonge,
mulangay rebeday, or Ben oil tree), was utilized by the ancient romans, greeks
and Egyptians, and it is now widely cultivated and has become naturalized in
many locations in the tropics and sub tropics. Horseradish (Moringa oleifera)
considered as a rich source of bioactive chemicals and they must be an
alternative source of mosquito control agent. The phytochemicals derived from
the Horseradish (Moringa oleifera) can act as larvicide, insect growth regulators,
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repellent and also have a very deterrent activity observed by many scientist and
researchers. Many of plant extracts are subject to risk factors in mosquito control,
but in the study Horseradish (Moringa oleifera) extract showed a good effect as
an alternative mosquito repellent and does not have a toxic effect on the human
beings. Horseradish (Moringa oleifera) has an impressive range of medical uses.
Different parts of this plant such as the leaves, roots, seed, bark, fruit, and
flowers contain a profile of essential purposes.
The methanolic extract of Horseradish (Moringa oleifera) was evaluated
for repellent effect against aedes aegypti mosquito,repellent effect of treatment
was assessed at different concentration (20,25,30,35,and 40 mg/ml) after 5
mins,10 mins, and 15 mins of exposure against 45 aedes aegypt adult
mosquitoes in a cage. All extracts were evaluated in different cages. Percentage
repellency (PR) was determined for each extract following the procedure of
WHO. All the tested extracts showed moderate to good repellent activities,
however the minimum percentage repellency potential was detected in the 25ml
concentration of Horseradish (Moringa oleifera).
The repellent activity of the extracts was performed with human volunteers
and does response method was used to estimate the percentage protection. The
results indicated that the significant effect of the treatments and the effectiveness
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increased with extended exposure interval and enhanced dose rate. There was a
statically significant difference between the 3 test group (Control Horseradish
(Moringa.oleifera and S. Indica) as determined by a one-way analysis of variance
(ANOVA) (F (221) = 20921.216, P= 0.000. Since the p-value 0.000 < 0.05, a
turkey post- hoc test revealed that at 95% confidence level, S.indica yielded a
different mean value of percentage repellency from those of oleifera and the
control (which also yielded different mean values). The LC 50 value recorded for
Moringa oleifera and stachytarpheta indica are 66.0 mg/ml and 18.2 mg/ml
respectively.
The test plant extracts were screened for phytochemicals such as
alkaloids, flavonoids, saponins, tannins, steroids, anthraquinones and terpenes.
The findings of the present study clearly revealed that methanol extract of
Horseradish (Moringa oleifera) and S. Indica leaves are good repellent agents for
the control of aedes aegypt, however the efficacy depends on does rates and the
exposure interval.
Vector-borne diseases, such as malaria, filariasis, dengue and
hemorrhagic fever (DHF), are still major public health problems in the Southeast
Asian countries. Because of their tropical or subtropical climate. Also owing to
poor drainage System, especially during rainy seasons, the presence of many
fish ponds, Irrigation ditches and the rice fields provide abundant mosquito
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breeding places. Malaria and other vector-borne diseases contribute to the major
disease burden in India. Repeated use of synthetic insecticides for mosquito
control has disrupted natural Biological control systems and led to resurgences in
mosquito populations. It has also resulted in the development of resistance,
undesirable effects on Non-target organisms, and fostered environmental and
human health concern that initiates a search for alternative control measures.
Plants are considered as a Rich source of bioactive chemicals and they may be
an alternative source of Mosquito control agents. Plant products have been used
by traditionally human communities in many parts of the world against the
vectors and species of insects. The phytochemicals derived from plant sources
can act as larvicides, insect growth regulators, repellents and ovipositional
attractants and have deterrent activities observed by many researchers.
Repellents have an important place in protecting man from the bites in insect
pests. An effective repellent will be useful in reducing man vector contact and in
interrupting disease transmission. A repellent compound should be toxic, non-
irritating and long lasting. Amides, imides, esters and other polyfunctional
compounds are known to be good repellents. Plants could be an alternative
source for mosquito repellents because they constitute a potential source of
bioactive chemicals and typically are free from harmful effects. Because of this,
much interest has been focused on plant extracts, or plant essential oils as
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potential mosquito repellent agents, and studied the interactive effect of
botanicals (Neem, Pongamia) and Leucas aspera, Bacillus sphaericus against
the larvae of Culex quinquefasciatus.
Horseradish (Moringa oleifera) is the most widely cultivated species of a
monogeneric family-the Moringaceae, native to the sub-Himalayan tracts of India.
This rapidly-growing tree (also known as the horseradish tree, drumstick tree,
benzolive tree, kelor, marango, mlonge, moonga, mulangay, nebeday, saijhan,
sajna or Ben oil tree), was utilized by the ancient Romans, Greeks and
Egyptians, and it is now widely cultivated and has become naturalized in many
locations in the tropics. All parts of the Moringa tree are edible and have long
been consumed by humans. In the West, one of the best known uses
of Moringa is to flocculate contaminants and purify drinking water with its
powdered seeds. This tree has in recent times been advocated as an
outstanding indigenous source of highly digestible protein, Ca, Fe, Vitamin C,
and carotenoids suitable for utilization in many of the so-called “developing”
regions of the world where undernourishment is a major concern. In the present
study an attempt was made to evaluate the toxicity of Horseradish (Moringa
oleifera) on malarial vector, Anopheles stephensi (An. stephensi). Horseradish
(Moringa oleifera) Lam. (syn. M. pterygosperma Gaertn.2n = 28) belongs to the
family Moringaceae, commonly known as the ‘drumstick’ or ‘horseradish’ tree. It
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is an affordable and readily available source of major essential nutrients and
nutraceuticals, and it has the potential to eradicate malnutrition (Kunyanga et
al. 2013). The Horseradish (Moringa oleifera) is often considered as important
famine food because of its high resistance to drought and arid conditions owing
to their tuberous roots (Padayachee and Baijnath 2012). Almost each and every
part of Horseradish (Moringa oleifera) tree is useful for medicinal, functional food
preparations, nutraceuticals, water purification, and biodiesel production;
including roots, leaves, flowers, green pods, and seeds (Saini 2015). The
immature pods, flowers, and foliage of this tree are used for culinary purposes in
different parts of the world (Stevens et al. 2013). The foliage of M. oleifera (MO)
has been established as a rich source of phenolics and glucosinolates (Amaglo
et al. 2010), minerals (Saini et al. 2014a), tocopherols (Saini et al. 2014b),
carotenoids (Saini et al. 2014c), polyunsaturated fatty acids (Saini et al. 2014d),
ascorbic acid (Saini 2015), and folate (Saini et al. 2016). Moringa seed oil (yield
30–40 % w/w), also known as “Ben oil” is used for the production of biodiesel,
because of the high content of monounsaturated fatty acids in the form of oleic
acid (C18:1) (Azam et al. 2005; Rashid et al. 2008). Moringa seed oil is a
potential candidate for biodiesel production, as it meets all the main
specifications of the biodiesel standards of US, Germany, and Europe (Mofijur et
al. 2014). Thus, it has great commercial and industrial importance. The low
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molecular weight cationic proteins (MOCP), chitin binding protein isoforms (Mo-
CBP3), lectins, napins, mabinlins, and other seed proteins extracted from MO
seeds are successfully characterized and used in domestic and industrial water
purification, and hardness removal because of the potent antimicrobial and
coagulant properties (Kansal and Kumari 2014) Horseradish (Moringa Oleifera)
as a Potential Repellent for Malarial Vector
Mosquito
Mosquito (a Spanish word for "little fly") is considered as one of the life-
threatening species. They are carriers of certain diseases some of which
can deadly and they also transmit malaria and dengue fever. Mosquitoes are
increasing in population nowadays, they usually live on dirty areas
and dark areas of our home and uncivilized area such as stagnant water and
storage rooms. They are small midge-like flies that constitute the family
"Eulicidae". Females of most species are ectoparasites whose tube-like mouth
parts pierce the host's skin to consume blood. They feed on the blood of various
kinds of hosts. Though, the loss of blood is seldom of any importance to the
victim, the saliva of the mosquito often causes an irritating rash that is a serious
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nuisance. In warm and humid tropical regions, some species are active for the
entire year, but for temperate and cold regions, they hibernate or enter diapause.
As stated earlier, mosquitoes tend to hate lemon's citrus smell. Considering the
fact that it is only concentrated on its smell means candle could be used as a
means of distributing the lemons scent to repel them and to eliminate them, if
possible at once.
Repellent
Repellent are substance that act locally or at a distance, deterring an
Arthropod from flying to, landing on to biting human or animal skin. Usually,
insect repellent work by providing a vapor barrier deterring the arthropod from
coming into contact with the surface and sometimes, applying on the skin for
protection. Mosquito are a serious threat to public health transmitting several
dangerous diseases to over two billion people in the insecticide resistance of this
vector and has become a global problem.
Insecticides residues in the environment, as a result of chemical insecticide
usage, have turned the researcher's attention towards natural products. In that
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past years, the plant kingdom has been of great interest as a potential sources of
insecticidal products. Many species in the plant kingdom synthesize a variety of
secondary metabolites which play a vital role in defense of plants against insects/
mosquitoes. Plants may be alternative source of bioactive compound. Plant
products can be used either as an insecticide for killing larvae or adult
mosquitoes or as repellent for protection against mosquito bites, depending on
the type of activity they possess products of secondary plant metabolism may be
responsible for the chemical communication between plants and insects.
Phytochemicals have been considered as potential natural insecticide s and can
be used for mosquito management in integrated control. Phytochemicals
obtained from plants are usually less environmental harmful than synthetic
chemicals and it has renewed the interest in the research on Phytochemical
compounds considering them as an ecologically safe alternative for synthetic
insecticides. A review of botanical phtychemicals with mosquitocidal potential
published by haalan. Demonstrates from the botanicals containing active
phytochemicals. The aim of this study was to evaluate the repellency activities of
the Moringa oleifera.
Repellents are one way of protecting man from insect's bite. An alternative
repellent should help us lesser our contact with vector borne disease and
interrupting the process of transmission. This Horseradish (Moringa oleifera)
20
repellent could help in saving the lives of upcoming future in preventing and
lessening the case of malaria disease that could harmfully affect the public health
by promoting more research studies to scientist about this miracle plant it can
essentially be the answer for our problem with mosquitoes.
Chapter 3
METHODOLOGY
Conceptual Framework
Input Process Output
1.Preparation of 1. Collection of 1.Horseradish

Materials Needed Materials Needed (Moringa oleifera) as
an alternative
(Horseradish 2.Making the mosquito repellent.
(Moringa oleifera) Horseradish (Moringa
leaves, Mosquito, oleifera) extract.
Blender, container,
Spray bottle,
Laboratory Gown 3.Testing and data
and Mask.) gathering
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Flowchart of the Procedure
Collection of Materials Preparation of Horseradish

Horseradish (Moringa (Moringa oleifera) leaves
oleifera) leave, Mosquito, (Collecting Horseradish
Laboratory Gown and Mask.) (Moringa oleifera) Leaves
Exact)
Preparation of
Treatment
(T0 Placebo, T1- 25%
Horseradish (Moringa
oleifera) leaves Extract, T2 Preparation of Treatement
- 50% Horseradish for Phytochemical Analysis
(Moringa oleifera) Leaves
Extract, T3 -75% Moringa
oelifera Extract, T4 – 100%
moringa oleifera Leaves
Extract, T5 – Positive
Control)
Pre- treatment Evaluation, Data Gathering

Application of
Treatment,Post- treatment
Evaluation 22
Statistical Analysis
(ANOVA, T- test, Cost
Analysis)
I. Research Design
This study used Complete Randomized Design to solve the problems
Stated. The method was used to obtain most of the data. The researcher
conducted the study with six (6) treatments in three (3) replicates: Treatment 0
(T0) as the negative control, 25 % Horseradish (Moringa oleifera) leaves extract
solution (consisted of 25 ml Horseradish (Moringa oleifera) leaves extract
solution and 75 ml commercial repellent)was labeled as Treatment 1 (T1); 50%
Horseradish (Moringa oleifera) leaves extract solution and 50 ml commercial
repellent ) was labeled as Treatment 2 (T2); 75% Horseradish (Moringa
oleifera) leaves extract solution (consisted of 75 ml Horseradish (Moringa
oleifera) leaves extract solution and 25 ml commercial repellent ) was labeled as
Treatment 3 (T3); 100% Horseradish (Moringa oleifera) extract solution
(consisted pure Horseradish (Moringa oleifera)leaves extract solution) was
labeled as Treatment 4 (T4) and the positive control Commercial repellent
( DEET) was labeled as Treatment 5 (T5). Two hundred seventy mosquitoes
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were used as experimental subjects. They were randomly chosen and group into
six. Forthy - five mosquito in each group. Group a Served as the control group
took treatment 0. Group B – F serve as the experimental groups.
The methods used in the study included 1) Collection of materials 2)
Preparation of Horseradish (Moringa oleifera) extract 3) Preparation for
Phytochemical Analysis 4) Preparation of treatments 5) Pre-treatment Evaluation,
Application of Treatments, Post-treatment Evaluation 6) Data Gathering 7)
Statistical Analysis. This method of research design was also be used in the
presentations and interpretations of results.
The Horseradish (Moringa oleifera) extract sample was brought to the
Virgen Milagrosa University Foundation (VMUF), College of Pharmacy, San
Carlos City, Pangasinan for Phytochemical Screening.
The results were presented in the forms of tables and graphs.
Photographs were also presented to support the results.
II. Preparation and Gathering of Materials
A.Collection of Materials
The young and fresh Horseradish (Moringa oleifera) leaves was gathered
from the premises of the researcher where it is abundant. The glass container,
spray bottle and blender came from the researcher’s residence. The other
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materials that were used like laboratory gown, Mosquito cage, gloves and masks
were supplied in the Laboratory Room of the Alaminos City National High School,
Alaminos City, Pangasinan and Virgen Milagrosa University Foundation (VMUF) -
College of Pharmacy laboratory, San Carlos City, Pangasinan.
B. Cleaning and Washing of leaves
The collected leaves were washed using the basin. Then, the damaged
Horseradish (Moringa oleifera) Leaves were removed and the fresh and young
leaves were selected. Then, the selected young Horseradish (Moringa oleifera)
Leaves were washed thoroughly and were drained separately. The leaves that
will be collected will be cut / blender, weight and undergo the ethanolic extraction.
C. Materials/Instruments
The following materials and apparatus were used in this study:
Glass Jars It refers to the jars where the treatment was placed for laboratory
testing.
Electrical Blender. It refers to the blender that was used by the researcher
during experimentation procedure.
Mosquito cage. It refers to the cage that was used by the researcher during
experimentation procedure.
Spray bottle. It refers to the bottle where the treatment was placed for laboratory
25
testing
Gloves. It refers to the covering of the hands that was used by the researcher to
avoid contact with chemicals during the preparation of the treatments.
Laboratory Gown. It refers to the cloth that the researcher worn during the
experimentation process.
Mask. It refers to the protective gear of cloth that was used by the researcher to
avoid inhalation of odor of harmful chemicals.
D. Preparation of Horseradish (Moringa oleifera) leaves extract samples for
Testing
The gathered extract was placed in sterilized bottles. It was filtered to
remove the sediments and other solid materials in the oil. It was kept in the
refrigerator ready for testing.
E. Safety Precautions
Safety precautions and prescriptions given by the project adviser were
observed. The whole process of experimentation/procedures were reviewed to
Refrain from untoward incidents or circumstances. Hair nets, gloves, and
laboratory gowns were used to avoid contamination to the samples.
26
III. Phytochemicals screening of test plant
The phytochemical in investigation of Horseradish (Moringa oleifera) was
carried out using standard method described by SAFOROWA. The plant extract
was screened for alkaloids, flavonoids, Tannis, and saponins.
IV. Preparation of Treatments
The amount of Horseradish (Moringa oleifera) leaves were prepared with
the following treatment in three replicates.
Table I A: Horseradish (Moringa oleifera) leaves extract per treatment
(Mosquito repellent)
TREATMENT AMOUNT OF MORINGA AMOUNT OF
OLEIFERA LEAVES COMMERCIAL
EXTRACT REPELLENT DEET

T0 Negative Control (Placebo)
T1 (25%) 25 ml 75 ml
T2 (50%) 50 ml 50 ml
T3 (75%) 75 ml 25 ml
T4 (100%) 100 ml 0 ml
T5 Commercial repellent (DEET)
V. Pre-Treatment Evaluation
The repellency study followed the method of World Health Organization
27
(WHO). The test concentration used for the repellent activity were 25ml, 50ml,
75ml,100ml. The volunteer had no contact with lotions, perfume, oils, or soaps on
the day of the assay. Each test concentration of Horseradish (Moringa oleifera)
were tropically applied to the arm of the volunteers (from palm to elbow) using
cotton wool. An arm without any test concentration applied on it served as
control. The treated arm was then inserted into mosquito cage for one full minute.
Each test concentration was repeated three times to get a more accurate mean.
The number of mosquitoes that landed on the treatment arm were recorded and
the shaken off before they imbize any blood. The same procedure was repeated
for all the test concentrations of different cases. The percentage repellency was
calculated in the following formula.
% repellency = Tc-Tt × 100
Where Tc=mean number of mosquitoes that landed on the control arm.
Tt= mean number of mosquitoes that landed on the treated arm.
VI. Application of Treatments
The result of the repellent activities of Horseradish (Moringa oleifera) is
presented on table 1&Figure 1. The result showed that Horseradish (Moringa

28
oleifera) leaf extract recorded 66.66% repellency at 25 ml concentration and
71.42% repellency at 50 ml, 80.95% repellency at 75 concentrations and 90.47%
repellency at 100ml.
The highest percentage repellency occurred at the highest concentration
(100ml) of Horseradish (Moringa oleifera) the lowest concentration (25ml) of
Horseradish (Moringa oleifera). The control (with number extract) should no
repellency against the mosquitoes. The LC50 values of the extract of
Horseradish (Moringa oleifera) is 18.2 ml respectively.
Table 1: Percentage Repellency of Horseradish (Moringa oleifera) leaves
extract.
Concentration (ml) Number of mosquitoes that landed on the treatment arm Mean number that landed Mean number that did not landed
29
P l a n t Repellency%
Time interval in minutes
5mins 10mins 15mins
C o n t r o l
(number extract ) 0 19 18 17 18 2 7
Horseradish
( M o r i n ga o l ei f e r a ) 25 15 14 13 14 3 1 66.66
5 0 12 12 12 12 3 3 71.42
7 5 8 8 8 8 3 7 80.95
100 5 4 3 4 41 90.47
Number of tested mosquitoes = Two Hundred Seventy (270) Mosquito. They
were randomly chosen and group into six. Forthy five (45) mosquito in each
Concentration in ml.
Estimated Marginal Means of Percentage repellency (%)
100.00
80.00
60.00
40.00
20.00
0.00
0 25 50 75 100
Concentration ml
30
Figure 1: Percentage of doe’s responses of Horseradish (Moringa oleifera)
VII. Statistical analysis
The mean percentage protection data where subjected to Profit analysis for
calculation of LC50. Data were analyzed using SPSS software. Microsoft excel
was uses for calculating means and standard deviations and performing of
analysis of variance (ANOVA). Results with P< 0.05 were considered to be
statistically significant.
1. Analysis of Variance was used to determine if there are significant
differences among the treatment and Scheffe Post Hoc Test determine
where the variation exists
ANOVA TABLE
Source of dF Sum of Mean of F- Value

Computed Tabular
Variation Square Square
Between
Groups (k-1)
Within Groups
(n-1)
Total
2. T-test was utilized to determine if there was a significant difference
31
between The Horseradish (Moringa oleifera) leaves extract and the
Commercial repellent (DEET).
T-test Table
Treatment Repellency Level of Tabular Computed Interpretati

Significance Value on
Treatment 4
Treatment 5
3. Cost Analysis was used to compare the Horseradish (Moringa oleifera)
with the commercial repellent DEET as mosquito repellent.
VIII. Level of Significance
The significant difference among six treatments was tested at a = 0.05 level of
significance.
VIIII. Disposal of Material
After 2 weeks of conducting the in the study, all the materials will
immediately have decontaminated in the autoclave for thirty minutes at one
hundred and twenty degrees Celsius. After gathering the data on the seventh
32
day. The decontamination materials will be disposed and cleaned immediately
using muriatic acid. After the in vitro study, all the Horseradish (Moringa oleifera)
extract remains to be buried in the ground for at least 1 foot. Table 1 shows the
phytochemicals screening of the extract revealed the presence of alkaloids,
tannins, flavonoids, and saponins in Horseradish (Moringa oleifera) as an
alternative ingredient in making mosquito repellent. anthraquinones, terpenes
and steroids were absent in oils from Horseradish (Moringa oleifera)
Chapter 4
RESULTS AND DISCUSSIONS
This portion present data gathered in table form, as well as the result and
discussion of the statistical interpretation of data. These are result in hypothetical
data of efficacy of Horseradish (Moringa oleifera) as Alternative Mosquito
Repellent.
Table 1. Phytochemicals in oils extract from Horseradish (Moringa oleifera)

leaves
Phytochemicals Horseradish (Moringa oleifera)

Alkaloids +
33
Tannins +
Flavonoids +
Saponins +
Anthraquinones -
Terpenes -
Steroid -
+ indicate present; - indicate absent

Table 1 shows the phytochemicals screening of the extract revealed the
presence of alkaloids, tannins, flavonoids, and saponins in Horseradish (Moringa
oleifera) as an alternative ingredient in making mosquito repellent.
anthraquinones, terpenes and steroids were absent in oils from Horseradish
(Moringa oleifera).
Table 2. Percentage repellency of efficacy of Horseradish (Moringa oleifera)
leaves extract as an alternative Mosquito Repellent.
Subject Trail Trial Mean Mean Repelle Efficacy
Trial1 2 3 numbe number ncy (%)
r that that
landed didn’t
landed
T1 25% 15 14 13 14 31 66.66 75%
Horseradish %
34
Leaves
T2 50% 12 12 12 12 33 71.42 80%
Horseradish %
Leaves
T3 75% 8 8 8 8 37 80.95 80%
Horseradish %
Leaves
T4 100% 5 4 3 4 41 90.47 85%
Horseradish %
Leaves
T5 4 3 4 3 42 90.50 95%
Commercial %
Repellent
Table 2 shows the efficacy results for the difference treatment of Horseradish
(Moringa oleifera) leaves extract as an alternative Mosquito repellent. Treatment
1 has 66.66 % repellency at 25 ml concentration and its efficacy is 75% and
treatment 2 has 71.42 % repellency at 50 ml and its efficacy is 80%. Treatment 3
has 80.95 % repellency at 75 ml concentration and its efficacy is 80% and
treatment 4 has repellency of 90.47 % at 100 ml concentration and its efficacy is
85% while Treatment 5 which is the commercial repellent has 90.50 % and its
efficacy is 95%. It is stated in the table that the highest percentage repellency
occurred at highest (100 ml) concentration of Horseradish (Moringa oleifera). The

35
LC50 values of the extract of Horseradish (Moringa oleifera) is 18.2 ml
respectively.
Figure 1. Means of Percentage repellency of Horseradish (Moringa oleifera)
extract concentration on Mosquito repellent
90.50
90.47
80.95
71.42
66.66
T1 T2 T3 T4 T5
Estimated Marginal Means of Percentage repellency (%)
Figure 1. Shows the repellent results of the different treatment of Horseradish
(Moringa oleifera) concentration on mosquito repellent. Treatment 1 has mean
percentage repellency of 66.66% and treatment 2 has mean percentage
repellency of 71.42%. Treatment 3 has mean percentage repellency of 80.95%
and treatment 4 has a mean percentage repellency of 90.47 %. While treatment
5 has mean of 90.50%. It is stated from the table that mosquito repellent is
susceptible as the prepared treatment from Horseradish (Moringa oleifera)
leaves extract.
36
Table 3. Result of Analysis of Variance of Horseradish (Moringa oleifera)
Leaves extract concentration on Mosquito repellent.
Groups Count Sum Average Variance

14 4 27 6.75 16.91666667
31 4 153 38.25 16.91666667
Source Sum of Mean of DF F cal- value F crit- P- Value Interpre

of square squares value at tation
variance (ms) (SS) 5% level
Between 1984.5 1984.5 1 117.3103 5.987378 3.6677E H0
groups -05 REJECT
ED
Within 101.5 16.91667 6
groups
Total 2086 7
Table 3 shows the computed F-ratio which is 117.3103 which is greater than
tabular value of 5.9873778 at 0.5 level of significant. Thus null hypothesis is
rejected hence; there is significant difference between the treatments and the
commercial one in terms of their efficacy Horseradish (Moringa oleifera) leaves
extract concentration on Mosquito Repellent.
Table 4. Results of T-test comparing the mosquito repellent effect of
treatment four (100% Horseradish (Moringa oleifera) leaves extract) and
37
treatment 5 (Pure commercial repellent DEET) against Mosquitoes.
Treatment Repellenc Level of Tabula Computed Interpretat

y Significance r Value ion
Treatment 4
90.47 0.05 3.707 -7.68624 H0
Treatment 5 90.50 REJECTE
D
Table 4 shows the computed value which is 7.68624 greater than the tabular
value of 3.707 at 5% level of significant, thus, null hypothesis is rejected.
Therefore, there is a significant difference in efficacy of the prepared treatment 4
(100% Horseradish (Moringa oleifera) leaves extract) and the commercial one
which is pure commercial repellent (DEET).
Table 5. Cost Analysis
Materials Cost
Horseradish (Moringa oleifera) Free
leaves
Plastic Container P 10.00
Labor P 5.00
Total P 15.00
38
The table below shows the cost of the raw materials used in making an
alternative mosquito repellent properties of Horseradish (Moringa oleifera) leaves
extract. Based on the total amount, making an alternative mosquito repellent of
Horseradish (Mornga oleifera) leaves extract is cheaper than buying a
commercial repellent DEET which cost P 500 or more.
Chapter 5
SUMMARY, OF FINDINGS, AND RECOMMENDATIONS
Summary of Findings
Horseradish (Moringa oleifera) leaves were gathered from the researcher
premises. The main purpose of this study was to determine the Efficacy of
Horseradish (Moringa oleifera) as alternative Mosquito Repellent. Based on the

39
study, the researcher found that Horseradish (Moringa oleiera) leaves extract
inhibits growth of Mosquito Repellent. It was found that the treatments of
Horseradish (Moringa oliefera) and the commercial one which is the pure
commercial DEET efficacy as an alternative mosquito repellent. Therefore, the
Researcher concluded that pure Horseradish (Moringa oleiera) leaves extract is
a potential Mosquito Repellent.
Conclusion
Based on the findings of the study, the following conclusions were drawn:
1. Horseradish (Moringa oleifera) has a phytochemical that can be as alternative
ingredient in making mosquito repellent.
2. Horseradish (Moringa oleifera) is effective mosquito repellent.
3. There is a significant difference among treatments of Horseradish (Moringa
oleifera) in killing mosquito.
4. There is a significant difference between the best Treatment of Horseradish
(Moringa oleifera) as mosquito repellent compared to commercial one.
5. There is a significant difference between the Treatment of Horseradish
40
of cost.
Recommendations
For those who like to go deeper into the study, the researcher would like
to recommend the Horseradish (Moringa oleifera) leaves extract will be used to
another study, a possible source of biodesel, use other plan extraction method,
and consider using different statistical tools to support the significant of the study.
The researcher would like to recommend the following:
1. Additional research and studies must be made on other significant uses of
Horseradish (Moringa oleifera) plant.
2. Further studies must be conducted about the composition of Horseradish
(Moringa oleifera)
3. Further studies on alternative plants that contains repellent activity as an
Alternative mosquito repellent.
41
4. Mass production of Horseradish (Moringa oleifera) leaves extract
recommended for the community since they are cheaper and for
producing an alternative mosquito repellent.
REFERENCES
42
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of Mandala district, Madhya Pradesh. Indian Journal of Malariology. 32:2007, 99-
103
Wink, M. Protection and application of phytochemicals from an agricultural

perspective, phytochemistry and agriculture. JAMA. 34:2003? 171-175
Jilan,i G.and Su, H.. Laboratory studies on several plant materials as insect
repellents for protection of cereal grains. Journal of Economic Entomology. 76:
2002, 154-157
Isman, M. Botanical insecticides, Deterrents and Repellents in Modern

Agriculture and an increasingly regulated world. Entomology Journal. 55;
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Shaalan, E. A. A review of botanical phytochemicals with mosquitocidal potential.

International Journal of Environment. 31: 2005,1149-1166
Sofowora, A. Medicinal plants and traditional medicine in Africa. Spectrum books

Ltd. Ibadan, Nigerian. 2000, Pp. 289
World Health organization. Vector resistance to pesticides. Fifteenth report of

WHO export committee on vector biology and control. WHO Tech Ser.2009, 818:
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Muhammed,A. and Umair .A. Evaluation of mosquitocidal activity of water extract

of Moringa oleifera seeds against Culex quinquefasciatus in Pakistan. J. Vet.
Borne Dis., 42:2012, 159-161
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(L.)Poit. (Bush tea) leaves from Nigeria. Journal of Essential Oil Resources. 12:
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Efficacy of Horseradish (Moringa Oleifera As An Alternative Mosquito Repellent

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Efficacy of Horseradish (Moringa Oleifera As An Alternative Mosquito Repellent

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Efficacy of Horseradish (Moringa oleifera) as an alternative Mosquito Repellent

Background of the Study

A mosquito repellent doesn't actually kill mosquitoes. Repellent work by

mosquito repellent is very useful and it can be man - made.

Horseradish or also known as Moringa Oleifera is a root vegetable known

off diseases, but it makes an excellent mosquito repellent. Horseradish (Moringa

Horseradish (Moringa oleifera) is beneficial to human health. The leaves of the

the most widely cultivated plant species worldwide. The phytochemicals derived

from the Horseradish (Moringa oliefera) can act as larvicides, insect growth

scientist and researchers. In the study of Asian Pacific Journal of Tropical

Biomedicine, the outcome acquired after the treatment of Horseradish (Moringa

Oleifera) for malarial vector were encouraging.it also yields leaves where

government health researchers found extracts for the formulation of an effective

mosquito-borne diseases like malaria and filariasis especially in areas of the

in preventing mosquito bites that cause itchiness, inflammation and discomfort to

biodegrable since, it is derived from advantage against other well-known

repellents which are chemical based in formulation. The Horseradish (Moringa

Research and Development (PCHRD) shows that the Horseradish (Moringa

Horseradish (Moringa oleifera) serves a lot of purposes to human health as

Horseradish (Moringa oleifera) the methanolic extract of Horseradish (Moringa

ml) after 5mins, 10mins, 15mins, of exposure against 45 mosquitoes in a cage.

All extract were evaluated in different cages.Percentage repellency (PR) was

statically significant difference between the 2 test groups (Control, and

Horseradish (Moringa oleifera) as determined by a one-way Analysis of Variance

Turkey Post-hoc test revealed that at 95% confidence level, Horseradish

(Moringa oleifera) yielded a (highest) mean value of percentage repellency from

extracts were screened for phytochemicals such as alkaloids, flavonoids,

saponins, tannins, steroids, anthraquinones and terpenes. The findings of the

present study clearly revealed that methanol extracts of Horseradish (Moringa

source alkaloid, and Bioactive compounds. These components are responsible in

producing mosquito repellent. The Horseradish (Moringa oleifera) is often

considered as important famine food because of its high resistance to drought

and arid conditions owing to their tuberous roots (Padayachee Baijnath 2012).

medicinal, functional food preparations, nutraceuticals, water purification, and

the answer for our problem with mosquitoes.

Statement of the Problem

This study aimed to investigate the Efficacy of Horseradish (Moringa oleifera)

as an alternative mosquito repellent .Specifically, it sought to answer the

be an alternative ingredient in making mosquito repellent?

2. Can Horseradish (Moringa oleifera) be an effective mosquito repellent?

3. Is there significant difference among treatments of Horseradish (Moringa

oleifera) in killing mosquito?

4. Is there significant difference between the best Treatment of Horseradish

(Moringa oleifera) as mosquito repellent compared to commercial one?

5. Is there a significant difference between the Treatment of Horseradish

(Moringa oleifera) as mosquito repellent as compared to commercial one in terms

This study was directed by following hypotheses.

1. There is no phytochemical present in Horseradish (Moringa oleifera) that can

be an alternative ingredient in making mosquito repellent.

2. Horseradish (Moringa oleifera) is not effective mosquito repellent.

3. There is no significant difference among treatments of Horseradish (Moringa

oleifera) in killing mosquito.

4. There is no significant difference between the best Treatment of Horseradish

(Moringa oleifera) as mosquito repellent compared to commercial one.

5. There is no significant difference between the Treatment of Horseradish

(Moringa oleifera) as mosquito repellent as compared to commercial one in terms

1. Horseradish (Moringa oleifera) contains various phytochemical such as

alkaloids, saponins, tannins, and Flavonoids, that can be alternative

ingredients in making mosquito repellent.

2. Horseradish (Moringa oleifera) is effective mosquito repellent.

3. There is a significant difference among treatments of Horseradish

(Moringa oleifera) in killing mosquito.

4. There is a significant difference between the best Treatment of