1

“Formulation & Evaluation of Mosquito repellent cream using Cinnamon oil

extract”

A Dissertation submitted to the Dr. Babasaheb Ambedkar Technological University, Lonere, Raigad for
partial fulfillment of

the Degree of

Bachelor of PHARMACY

Subject: Project Work

Submitted by

Student Name
Mr. Prathmesh Rakesh Hanje

Research Guide
Mrs. Priyanka G. Kale

Abhinav Education Society’s, College of Pharmacy (B.Pharm),

Narhe, Pune-411 041.
June, 2024
 2

CERTIFICATE
This is to certify that the research work presented in this dissertation entitled,

“FORMULATION AND EVALUATION OF MOSQUITO REPELLENT CREAM ”

for the partial fulfillment of the Degree of

Bachelor of Pharmacy
(Project Work)

has been carried out by PRATHMESH RAKESH HANJE at AES, College of Pharmacy (B.Pharm),
Narhe, Pune-41, of the Dr. Babasaheb Ambedkar Technological University, Lonere, Raigad
under the guidance of PRIYANKA G. KALE.

Date: Dr. T A Tuse

Place: Pune Principal,
AES, College of Pharmacy (B.Pharm)
Narhe, Pune- 41
 3

CERTIFICATE

This is to certify that the research work presented in the dissertation entitled,

“Formulation & Evaluation of Mosquito repellent cream using Cinnamon

oil extract”

By

Mr. Prathmesh rakesh hanje

for the partial fulfillment of the degree of Bachelor of Pharmacy (Practice School/ Project
Work) at AES, College of Pharmacy (B.Pharm), Narhe, Pune-41, of the Dr. Babasaheb
Ambedkar Technological University, has been carried out under my guidance and
supervision. This dissertation is now ready for examination.

Research Guide
Mrs. Priyanka G. Kale
Prof. Of
AESCOP, Narhe, Pune-41

Date-
Place- Pune
 4

STATEMENT BY THE CANDIDATE

I, the undersigned wish to state that the work embodied in the dissertation entitled,

“Formulation & Evaluation of Mosquito repellent cream

using Cinnamon oil extract” forms my own contribution to the

research/Review carried out under the guidance of Mrs. Priyanka G. Kale at Abhinav

Education Society’s, College of Pharmacy (B.Pharm), Narhe, Pune- 411041. This work

has not been submitted for any other degree of this or any other university. Whenever

references have been made to previous works carried out by others, it has been clearly

indicated as such and included in the reference.

Date: Name: Prathmesh R. Hanje

Year: Final year B-Pharm

PRN: 2063813823032
 5

SR.NO CONTENT Page No.

1 List of Figures 6

2 List of Tables 6

3 Abstract 8

4 Chapter no 1- Introduction 7

5 Chapter no 2- Aim and Objective 11

6 Chapter no 3- Literature review 13

7 Chapter no 4- Excipients 17

8 Chapter no 5- Experimental Work 20

9 Chapter no 6- Phytochemical 23
Screening

10 Chapter no 7- Formulation tables 26

11 Chapter no 8- Evaluation Parameters 29

12 Chapter no 9 Conclusion 33

13 Chapter no 10- Reference 35

 6

Sr.no Name of table Page no.

1 Excipient 18
2 Excipient and there Suppliers. 19
3 Equipment use in Preparation. 19
4 Alkaloid Test 24
5 Flavonoid Test 24
6 Saponin Test 25
7 Phenolic Test 25
8 Steroid Test 25
9 Formulation Table 27
10 Evaluation Parameter 30
List of table

List of figure

Sr.no Name of figure Page no.

1 MOSQUITO REPELLENT MARKETED PREPARATION 12
2 CINNAMON PLANT 15
3 CINNAMON BARK 15
4 CLEVENGR APPRATUS 22
5 PHYTOCHEMICAL TESTS 25
6 PREPARATION OF CREAM 28
7 PREPARATION OF CREAM 1 28
8 PREPARATION OF CREAM 2 28
9 pH METER 30
10 VISCOMETER 30
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Chapter No.1
 8

 Abstract:
The potential of Cinnamon essential oil as a natural insecticides and mosquito repellent
was studied. Cinnamon cassiabark was extracted using hydrodistillation. All extraction
was carried out for 6 hours at 1 atm. The yield of cinnamon oil was obtained 1.82 %
yields of cinnamon essential oil extracted when using hydrodistillation method. The
results indicated that 9 major volatile compound were presence such as alcohols,
aldehydes, alkenes, carboxylic acids, ether, ester and ketone in the cinnamon essential
oil extracted by hydrodistillation. Trans-cinnamaldehyde was found to be the major
volatile compound with the highest percentage of 86.67 %. The repellency and
insecticidal activity of cinnamon oil by hydrodistillation method obtained was directly
exposed to specimen i.e., mosquito. The repellency and insecticidal activity of
cinnamon essential oil was compared through different concentration of fabricated
repellency paper. As a conclusion, solvent extraction shows an effective method on
cinnamon essential oil extraction with positive insecticidal and repellent activity on
mosquito. The method employed to prepare this cream was very simple. Quality of
formulated product was assessed by different evaluation parameters. The formulated
cream showed good consistency, spreadability, homogenicity, pH.

 KEYWORDS: Cinnamon oil extract, Phytochemical screening, Evaluation,

Preparation of mosquito repellent cream, conclusion.
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 INTRODUCTION:
Essential oils are volatile natural complex secondary metabolites which has a strong
odor and have a generally lower density than water. Approximately, 3,000 esential oils
are known out of which 300 that have been commercialized for cosmetics,
pharmaceuticals, perfume industries and pesticidal potential. Cinnamon is a spice
collected from the bark of several trees from the genus Cinnamomum and Lauracea
family that is used in sweet and savoury foods. According to Vangalapati, Cinnamon
Cassia (C. Cassia) has the strong and spicy-sweet flavour. Vietnamese cassia (Saigon
cinnamon, Cinnamomum loureiroii) and Chinese cassia (Cinnamomum aromaticum) are
the sweetest and strongest varieties. As stated by Janick and Jules, (2011), the name
cinnamon comes through the Greek kinnámōmon from Phoenician. In Sinhala, Sri
Lanka, cinnamon is known as kurundu which was recorded in English in the
17thcentury as Korunda (Knox and Robert, 2008). In Indonesia, where cinnamon is
cultivated in Java and Sumatra, it is called kayu manis ("sweet wood") and sometimes
cassia vera, the "real" cassia (Samat and Bell, 2009)

 Definition:
Repellent is defined by Fried et al. (2007) as substances that cause insect to turn away.
Repellent have been used to prevent insects or specifically in this study, the mosquito
from harming or annoying human daily life as they always swarm over host’s blood.
Mosquito repellent is used to prevent the mosquito from invading the houses and biting
the humans and children’s. Chemicals and poisonous substances are effective, but can
bring harmful in the kitchen, around children and pets. One of the example of
carcinogenic chemical that have been used which either repell or kill the mosquito is
N,N-diethyl-3-methylbenzamide (DEET). Therefore, an alternative natural repellent
from cinnamon oil have been developed to replace the carcinorgenic chemical repellent.
In this study, the extraction of the cinnamon essential oil was carried out by
hydrodistillation in a Clavenger-type apparatus in accordance with the method of Wang
et al. (2009) with slight modification and using Soxhlet extraction in accordance with
the method of Mustafa and Hilal (2004) using three different solvents which are
dichloromethane (DCM), hexane and petroleum ether. The cinnamon essential oil was
analyzed using gas chromatography–mass spectrometry (GC-MS) to determine the
chemical compound especially the cinnamaldehyde. The cinnamon essential oil was
also used in this study to test for the repellency and insecticidal activity of the mosquito.
 10

 Benefits:
An mosquito repellent will help protect you from mosquitoes that spread
malaria and other diseases, such as dengue, chikungunya, and Yellow fever.
You can use an insect repellent on your skin and clothes to keep away
(repel) insects. Your doctor may also prescribe you a medication to prevent
malaria (antimalarial drug).
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Chapter No.2
 12

(Figure 1 )
( MOSQUITO REPELLENT MARKETED REPELLENT )

 Aim: To study formulation and evaluation of mosquito repellent cream using

cinnamon oil.

 Objectives:
1) To prepare the extraction of cinnamon oil.
2) To perform the phytochemical screening of extract of cinnamon.
3) To prepare mosquito repellent cream of extract of cinnamon.
4) To characterize and evaluate the prepared mosquito repellent cream.

 Plan of work:
1) Literature Review of mosquito repellent cream prepared by herbal ingredients.
2) Selection of herbal active ingredient
3) Procurement of Herbal active ingredient and excipient.
4) Extraction of active Ingredient from extract.
5) Phytochemical Screening of extract .
6) Formulation of mosquito repellent Cream by using extract of Cinnamon Bark oil
as an active ingredients by using other excipients.
7) Characterization / Evaluation of mosquito repellent cream.
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Chapter No.3
 14

 Literature Review
1) Author Names:- Khalid Haddi, Leda R.A. Faroni, and Eugenio E. Oliveira

In investigations conducted by Prajapati et al. (2005), the essential oil of C. zeylanicum

proved to be oviposition deterrent and repellent against three mosquito species tested
(Anopheles stephensi, Ae. aegypti, and Culex quinquefasciatus). Yang et al. (2004)
investigated the repellent activity of methanol extracts and steam distillate from 23
aromatic medicinal plant species against female blood-starved Ae. aegypti, and they
found that at a dose of 0.1 mg/cm2, the repellency of extracts of C. cassia bark and C.
camphora steam distillate was comparable to that of deet. The duration of the
effectiveness for extracts from C. cassia bark was comparable to that of deet.

Laboratory studies suggested that cinnamon may be useful as an insect repellant (Cloyd
et al. 2009). Hori (2003) described C. cassia as having repellent activity against the
cigarette beetle Lasioderma serricorne, while in the study of Jumbo et al. (2014),
cinnamon oil exhibited repellent actions against A. obtectus, in accordance with other
investigations, such as the one carried out by Liu et al. (2006), and where they described
a good repellent activity exhibited by essential oil from the seeds of C. camphora
against storage pests S. oryzae and Bruchus rugimanus. In addition, cinnamon essential
oils displayed repellant action against the red bud borer Resseliella oculiperda (Van tol
et al. 2007). Hanifah et al. (2012) reported that C. zeylanicum showed the highest
repellency rate compared with the other plants extracts in a study carried out to evaluate
the repellency of six plant extracts against the larval stage of Leptotrombidium deliense,
the mite vector of scrub typhus.

 Herbal Ingredients and excipient profile:

a) Cinnamon oil
b) Clove oil

Cinnamon oil:- cinnamon oil can kill off mosquito eggs. It can also act as a
repellent against adult mosquitoes, most notably the Asian tiger mosquito.

Clove oil:- Clove Oil. Studies have found that undiluted topical clove oil is active
against mosquitoes. The studies concluded that it's effective for nearly four hours.
However, like thyme oil, clove oil should not be applied undiluted to skin as it can
be absorbed and result in adverse effects.
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 Plant profile:-

Scientific name: Cinnamomum verum

Family: Lauraceae
Genus: Cinnamomum
Kingdom: Plantae
Order: Laurales

 Part used:- Bark, twigs

 Synonyms:- Cinnamon bark; Kalmi-Dalchini, Ceylon cinnamon

 Biological Source: of coppiced trees of Cinnamomum zeylanicum Nees,

belonging to family Lauraceae Cinnamon consists of dried inner bark of shoots

(Figure 2)
( CINNAMON PLANT )

(Figure 3) CINNAMON BARK

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 Uses
1. It is used as antioxidant, anti-inflammatory, antidiabetic, antimicrobial, anticancer,
lipid-lowering, and cardiovascular-disease-lowering compound,
2. Cinnamon has also been reported to have activities against neurological disorders,
such as Parkinson's and Alzheimer's diseases.
3. Cinnamon is used as mosquito repellent.
4. It is also used as insect repellent.
5. Cure for neuro generative diseases.
6. Reduce your risk of cancer.
7. Remedy for toothache.
8. Helps to obtained youthful skin.
9. Enhancing the digestive functions.
10. Lowering blood sugar
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Chapter No.4
 18

 Excipients used in mosquito repellent cream:-

Excipients Structure Melting Uses
Point(℃)
Stearic acid Ideal emulsifying
69.3 agent,It govern
consistency of the
cream, Effective
stabilizer, thickener
and softener.
Glycerin As a 23humectant
290 which helped the
cream from drying
out. Soften and
protect the hair and
prevent chaps.
Triethanolamine Emulsifying agent,
335.4 provide cream with
lesser luster. Serve as
PH adjuster.

Cetyl alcohol Prevent cream from

49.3 separating into oil
and liquid.

Water For stability to cream

0 because hard water
leads to formation of
magnesium causing
inversion of
emulsion.
Preservatives Methyl Inhibit growth of
(Methyl paraben, Paraben- microorganisms.
Propyl paraben 125- 128°c
ratio (4:1 ) Propyl
Methyl paraben Propyl paraben
Paraben
95-98°c
Perfume ( Rose Imparts odor to
Oil) 60-70 preparation.

Table No1. excipient used in preparation of vanishing cream containing extract of

cinnamon oil
 19

 Materials used:-
Excipients and their suppliers and models.
Table No.2excipients and their suppliers and model

Name Of Excipient Name of suppliers

Stearic acid SD lab chemical centre, Mumbai

Glycerin SD lab chemical centre, Mumbai

Triethanolamine Research lab fine chem industries, Islampur.

Cetyl alcohol SD lab chemical centre, Mumbai

Methyl , Propyl Paraben SD lab chemical centre, Mumbai

 Equipment’s used and Name of Equipment’s:-

Table No. 3: Equipment used in preparation of mosquito repellent cream

PURPOSE NAME OF EQUIPMENTS

Extraction clevenger apparatus
Viscosity Brookfield Viscometer
Spreadability Spreadability apparatus
Type of emulsion Optical microscope
 20

Chapter No.5
 21

 Experimental work:-

 Selection of herbal material:-

Selection of cinnamon oil which contains active ingredients.

 Method :
 Material and Chemicals :
The Bark of Cinnamon cassia sp used in this study was purchased from a local
supermarket. All solvents and chemical used such as hexane, dichloromethane,
petroleum ether and anhydrous sodium sulphate were analytical grade.

 Sample Preparation :
1. Fresh bark sample was washed and air-dried, and then the dried sample was ground
into powder.
2. Then, the sample was stored in a drying oven for further analyses.
3. In order to compare the extraction yield of cinnamon esential oil obtained, different
extraction methods were used.
4. Extraction of cinnamon oil was carried out using hydrodistillation and Soxhlet
extraction with hexane, dicholoromethane and petroleum ether as the solvents.

 Extraction of cinnamon oil using Hydrodistillation :

1. The cinnamon essential oil was extracted by using hydrodistilation with a Clevenger
type apparatus.
2. About 30 gm of precisely weighed of cinnamon bark powder has been put into 500
mL distillation flask with four boiling chip and 300 mL of distilled water was
poured into the flask.
3. They were immersed for 1 hr, followed by heating at 100 ºC for 6 hrs.
4. Then, the distillate was transferred to a 250 mL conical flask.
 22

5. Then, the volatile compounds was extracted from the water phase three times using
dichloromethane and dehydrated over anhydrous sodium sulphate for 30 min and
filtered through a mid-speed filter paper.
6. The cinnamon oil obtained was concentrated and stored at 4 ºC for further analyses.

(Figure 4)
CLEVENGER APPARATUS

 Phytochemical screening:
 Phytochemical screening was performed as per the methods and test of earlier

researchers. Procedure and result for these tests in following Table No.

1. Test for Alkaloids – Mayer’s Test, Hager’s Test, Dragendorff’s

2. Test for flavonoid – Shinoda’s test
3. Test for saponin – Foam
4. Test for tannin - Ferric chloride Test
5. Test for steroid – Salkowski reaction
 23

Chapter No.6
 24

 Phytochemical Screening:

 Test for alkaloids:-

Table No. 4:- ALKALOID TEST

Test Procedure Observation Inference
Mayer’s Test 1 ml of extract +few White ppt Alkaloid present
drops of Mayer’s
reagent(Pot. Mercuric
iodide solution)
=yellowish/White ppt

Hager’s Test 1 ml extract + Hager’s Yellow ppt Alkaloid present

reagent (saturated
picric acid solution) =
Yellow ppt

Dragendorff’s 1 ml extract + Brown ppt Alkaloid present

Test
dragendorff’s reagent (
Pot. Bismuth iodide
solution)=Reddish
brown ppt

Alkaloidal test for was successfully performed.

 Test for flavonoids:

Table No. 5 :- FLAVONOIDS TEST
Test Procedure Observation Inference
Shinoda Test Extract + Fragments Pink color Flavonoids present
of magnesium
ribbons + conc.
HCL
Test for flavonoids for was performed successfully.
 25

 Test for saponins:

Table No. 6 :- SAPONINS TEST
Test Procedure Observation Inference
Foam Test Extract + Distilled Foam produced Saponins present
water= foam
produced
Test for saponin was performed successfully.

 Test for Phenolic compound and Tannins:

Table No. 7 :- PHENOLIC &TANNIN TEST

Test Procedure Observation Inference
Ferric chloride test Extract+ d/w + Deep-blue/black ppt Phenolic compound
ethanol present
+FeCl3=
deep blue/ black ppt
Test for phenolic compounds and Tannins for extract was performed
successfully.
 Test for steroid:
Table No. 8 :- STEROID TEST
Test Procedure Observation Inference
Salkowski reaction Extract+ Greenish yellow Steroid is present
chloroform + Conc. fluorescence
H2SO4
Test for steroid was performed successfully

(FIG NO.5)

PHYTOCHEMICAL TEST
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Chapter No.7
 27

 Formulation table of mosquito repellent cream:

Table No. 9 :-FORMULATION
Ingredients Quantity Taken (100 gm )
Stearic acid 12 gm
Glycerine 6 ml
Triethanolamine 1 ml
Cetyl alcohol 0.5 gm
Water 78.3 ml
Cinnamon oil 2 gm
Clove oil 2gm
Preservatives (Methyl paraben, propyl 0.18 mg, 0.02 mg
paraben)
Perfume q.s

 Procedure:
1) All required ingredients of mosquito repellent cream was properly
weighed and kept separately.
2) Oil soluble material like Stearic acid and cetyl alcohol was added in a
beaker and melted at 60°C.
3) In another beaker, Water soluble material like glycerin , Triethanolamine,
water was added and heated up to 60°C.
4) Preparation of first beaker (Oil soluble) was added in a second beaker
(Water soluble) drop by drop with continuous stirring.
5) After cooling Methyl paraben, propyl paraben, and perfume was added
and mixed them thoroughly to obtain uniform product. Professional finish
was provided.
6) The extract of cinnamon oil and clove oil was added to prepared cream.
 28

(Figure 6) (Figure 7)

(Figure 8)
( FIG. NO. 6, 7 & 8 PRAPERATION OF CREAM )

Mosquito repellent cream containing cinnamon oil was successfully prepared by

using melt emulsification method.
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Chapter No.8
 30

 Evaluation of mosquito repellent cream containing

cinnamon oil extract:

TableNo.10 Evaluation of mosquito repellent cream

Parameters Procedure Result
Physical Color White
Properties Odour
Pleasant
Appearance
Smooth
Washability The cream was applied on the hand and Easily washable
observed under running water.
pH Weighed 10 mg of cream, dissolved in 10 7
ml of d/wand its pH was measured with the
help of digital pH meter.

Viscosity Viscosity of cream was determined by 1110cps

Brookfield viscometer

(Figure 9) (Figure 10)

PH METER VISCOMETER
 31

spreadability The spread ability of Spread ability =

formulated cream was = m*l/t
judged by spread ability = 50*12/6
apparatus = 600/6
= 100 gm cm/sec
Uniform and easily
spread

Irritancy test Marked an area (1 No Irritancy were

sq.cm) on left hand occurred.
dorsal surface. The
cream was applied to the
specified area and time
was noted. Irritancy,
erythema, edema, were
checked if any for
regular intervals up to 24
hrs. and
reported.
homogeneity homogeneity tested via Homogenous
visual appearance
 32

Formation Heated the sample cream in Creaming is upward

of creaming suitable test tube for 10 mins hence o/w type emulsion
and observed the result. If
creaming is upward then
emulsion is o/w type. If the
creaming is downward then
the emulsion is w/o type.

Patch test About 1 gm of material to be No any reaction is

tested was applied directly to observed.

the hands on skin. The

cosmetic to be tested was
applied on area of 1 sq.m of
the skin. The site of patch was
inspected after 24 hrs. As
there was no reaction the test
was repeated three times. If
reaction is not observed on
third application, the person
may be taken as
non- hypersensitive.
Smear type The test was conducted after Greasy in nature
the application of cream on
the hair. The smear formed
can be oily or aqueous in
nature.
Evaluation of cinnamon oil containing mosquito repellent cream evaluated successfully.
 33

Chapter No.9
 34

 Conclusion & Discussion:

Discussion: The formulated mosquito repellent cream was evaluated for several
physicochemical tests and the results were shown. The type of smear formed on the skin
was not greasy after the application of creams. The creams were easy to remove after
applications of cream. The cream was easy to remove after application by washing with
water. The formulations were able to produce a uniform distribution of extracts from the
cream. This was confirmed by visual examination and by evaluating the texture of the
skin. Even though there is no change in a colour reaction is observed when it was kept
for a longer time in a room temperature which indicates the stability of the product.

Conclusion: Using cinnamon oil as a mosquito repellent cream shows promising

potential due to its natural properties and effectiveness in repelling mosquitoes. Its
pleasant aroma and non-toxic nature make it a preferable alternative to chemical-based
repellents, especially for those seeking natural and eco-friendly solutions . cinnamon oil
holds promise as a viable option in the ongoing search for effective and sustainable
mosquito repellents.
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Chapter No. 10
 36

Reference:
1. https://bioone.org/journals/journal-of-the-american-mosquito-
control-association/volume-24/issue-1/8756-
971X(2008)24[161:MLAOBM]2.0.CO;2/Mosquito-Larvicidal-
Activity-of-Botanical-Based-Mosquito-Repellents/10.2987/8756-
971X(2008)24[161:MLAOBM]2.0.CO;2.short

2. https://bioone.org/journals/journal-of-the-american-mosquito-
control-association/volume-24/issue-1/8756-
971X(2008)24[161:MLAOBM]2.0.CO;2/Mosquito-Larvicidal-
Activity-of-Botanical-Based-Mosquito-Repellents/10.2987/8756-
971X(2008)24[161:MLAOBM]2.0.CO;2.short

3. https://academic.oup.com/jme/article-abstract/56/5/1346/5506539

4. https://link.springer.com/article/10.1186/s43088-023-00455-9

5. https://www.sciencedirect.com/science/article/pii/
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971X(2008)24[161:MLAOBM]2.0.CO;2/Mosquito-Larvicidal-
Activity-of-Botanical-Based-Mosquito-Repellents/10.2987/8756-
971X(2008)24[161:MLAOBM]2.0.CO;2.short

7. Nur Nasulhah Kasim, SNAS Ismail, ND Masdar, F Ab Hamid, WI Nawawi

International Journal of Scientific and Research Publications 4 (7), 2250-3153,
2014

8. DHADHANG WAHYU KURNIAWAN, VN Agustina, Wibowo GA Sunarto,

MUHAMMAD ZAENURI SYAMSU HIDAYAT
Int J App Pharm 14 (1), 208-212, 2022