Novel Formulation and Evaluation of Poly Herbal

Mosquito Repellent
Deepak Kumar Gupta (  Deepak_gupta20072008@yahoo.com )
Indira Gandhi National Tribal University
Revathi Gupta
Dr. A.P.J. Abdul Kalam University, Indore ,India
Akhilesh Tiwari
Indira Gandhi National Tribal University

Article

Keywords: Mosquito repellent gel, mosquito repellent candle, Essential oil

Posted Date: September 26th, 2022

DOI: https://doi.org/10.21203/rs.3.rs-1900478/v2

License:   This work is licensed under a Creative Commons Attribution 4.0 International License.
Read Full License

Page 1/20
Abstract
Aromatic plants produce chemicals that they use to resist phytophagous attacks Mosquito with various
pathways, such as repellents, protective eating, metabolites and development Controllers etc. Now,
present-day conditions result in the burgeoning need for natural mosquito repellents that would have
cheap, reliable, non-toxic, environmentally safe, and biodegradable attributes. Driven by the global need to
move forward, mosquito repellent candle and Mosquito repellent gel formulation based on carbopol 934
was prepared from the Origanum Majorana, Tanacetum Cinerarifolium, Juniperus Communis, Scented
Geranium, Thuja Occidentalis L., Tecoma Stans (L.), Artenmisia Arborescent, And Calamintha Acinos and
They are further tested for their size, pH, viscosity, spread ability, extrudability, swelling index and
accelerated tests. The prepared candle was tested for flammability and was burnt Performance with
respect to burning time and consequently its efficacy Spotting operation. Candle in flammability check
was performed to Checks its consistent laboratory combustibility. Basic Therefore, on performance, the
time taken to light the candle and its Causal symptoms such as pain, coughing has been reported and
recorded in identified mosquitoes vulnerable at night and in the field above. Throughout many developed
countries, where most residents have little exposure to mosquito net, high-cost mosquito repellent
creams, and miscellaneous physical approaches, this formulation of gel may be an efficient, affordable,
and easily accessible way to deter mosquito-borne diseases such as malaria, dengue, etc. in the lower
parts of society.

Background
Popular insects include mosquitoes, flies, bugs, ticks, butterflies, moths, bees, wasps and so on and are
notorious for their unpleasant, itchy sting. The two major classes of insects, that is, insecta and acariens,
are primarily interested in vector-borne disease transmission. [1] Mosquitoes are typically the best known
of the biting insects and potentially the most studied of all arthropods of economic importance. In
addition, mosquito-borne diseases such as malaria, chikungunya, yellow fever, Rift Valley fever, dengue
fever, and filariasis Japanese encephalitis in many parts of the world produce significant morbidity and
mortality in humans and livestock. [2] Most insects are known to be major pests attacking humans and
domestic animals in order to spread various diseases and some of these diseases may also lead to
death. [3] Throughout India the most prevalent diseases transmitted by mosquitoes are malaria,
chikungunya, dengue, encephalitis and filaria. Any 60 species of Anopheles mosquitoes spread the
deadly disease, Malaria. According to one World Health Organization (WHO) report in 1990, there were
approximately 270 million cases of malaria and approximately one million deaths annually. Lymphatic
filariasis is another disease which is spread by insects. It is largely caused by the Culex pipiens common
house mosquito, and is estimated to be between 100 and 250 million a year. Over the past few decades,
the number of individuals at risk from this urban disease has increased. The Aedes aegypti is widely
spread throughout the globe 's tropical, subtropical, and temperate zones, and is one of the largest
domestic mosquito species that are both human pests and vectors of disease agents. [4]

Page 2/20
Classification of Mosquito Repellents-The mosquito repellents currently available can be widely divided
into the following categories:(Figure:1)

Physical methods of Mosquito Repellents-The physical methods of repelling mosquitoes act as a

protective barrier to mosquito bites. Activities related to mosquito repellent physical methods include
emptying the stagnant water into rain gutters, old tyres, buckets, plastic covers, etc. It is highly desirable
to change the water in bird baths, fountains, pools, rain barrels etc. on a regular basis at least once a
week so that mosquitoes can't find the right place to lay their eggs. It is also recommended that you cover
yourself with long sleeved clothing especially during the time of dawn and dusk. One must repair
windows and door screens to further prevent mosquitoes from entering the home. This category includes
mosquito nets, coils, and other repellents available on the market. Mosquito netting is considered to be a
better measure of safety than coils and other repellents particularly during sleep. Such nets serve as a
defensive shield to keep humans from being attacked by mosquitoes and other insects. [5]

Mechanical methods of Mosquito Repellents-

It was well known that yellow light attracted fewer mosquitoes than white lights and this aspect can be
used as a mechanical device for mosquito repellent. Other approaches that fall under the mechanical
methods of mosquito repellents include the Electric Mosquito Zapper, Mosquito magnets, etc. An Electric
Mosquito Zapper operates by using ultraviolet light to trap mosquitoes and then kills them as they come
into contact with their lethal electric charge dose [6]

Chemical methods of Mosquito Repellents-

Chemical repellents are the most effective means of personal insect-seeking protection and help deter
and control insect-borne disease outbreaks such as measles, Lyme disease, Dengue fever, Bubonic
Plague, West Nile fever, etc. [7]

This can be further classified into two groups, based on the source of the chemical repellents:

Natural Mosquito Repellents-

Natural resources with proven insecticidal or repellent capacity have played a significant role in
interrupting vector-borne disease transmission at both human and community level. Human beings and
mosquitoes have existed in close contact from the dawn of human society and the natural resources
were used to shield ourselves from mosquito bite and vector borne diseases. These herbal mosquito
repellents have been developed and promoted since ancient times, and have led to the discovery of
personal protection methods [8]

Synthetic Mosquito Repellents-

While plant-derived compounds have long been used as repellents for mosquitoes, they still suffer from
certain disadvantages. In contrast with the synthetic repellents, plant related mosquito repellents have
Page 3/20
been tested fairly poorly. In comparison, the potency of these herbal repellents appears to last for shorter
duration and to fade away easily. Thus, these compounds have short term protection and need regular re-
application. Some drawbacks include heavy odor, skin irritation and possible health risks because they
were not commonly tested for toxicity. Most significantly, the cost of these repellents is often prohibitive.
Extensive research began to find a synthetic replacement for natural mosquito repellents due to the
limitations associated with the use of natural plant resources as mosquito repellents, such as limited
repellence period and high cost. [9]

There are many other natural and synthetic substances that have the ability to repel insects, but in the EU
and the US, only four primary substances—DEET, Icaridin, Citrioldiol, and IR3535—are recognised for use
as insect repellents. Each of them works in a somewhat different way and has somewhat different
qualities, but they all produce an odour that repels insects, which is how they all operate. Some important
common compounds in insect repellent are mention in figure no.2

THE OBJECTIVE OF THE STUDY- Seeing that most of the mosquito repellent products and devices
available on the market have reported adverse human effects, the aim of this analysis is to develop
effective mosquito repellent products based on plants.

Methods
Plant selection was based on the study of the various literature and its availability as mosquito repellents
with essential oils, experimental data, and folkloric application.

Origanum Majorana, Tanacetum Cinerarifolium, Juniperus Communis, Scented Geranium, Thuja

Occidentalis L., Tecoma Stans (L.), Artenmisia Arborescent, And Calamintha Acinos were Purchased from
a reliable source.

COLLECTION OF PLANT MATERIAL-

The plant material i.e., Thuja occidentalis, Origanum majorana, Juniperus communis Tanacetum
collected from natural habitat at udainagar (22.5327° N, 76.2233° E, 370 m altitude India) of Malwa
region Madhya Pradesh, India and cinerarifolium, Scented Geranium, Tecoma stans, Artenmisia
arborescent, Acinos arvensis (Lam.) were collected from chitrakoot (25.1788° N, 80.8655° E, 137 m
altitude India) area in India on October 2017. The use of plants in the present study complied with the
institutional guidelines of Dr. A.P.J. Abdul Kalam University, Indore (22.7964° N, 75.9316° E, 557 m
altitude, India) and as per the guidelines of Indian Biological Diversity Act 2002 which permits bonafide
Indians to access biological resources for scientific research. [10] The collected plant material is identified
morphologically, microscopically and compared with standard Pharmacopeial monograph and the
sample of plant was identified & authenticated by Dr. S.N. Dwivedi, Prof. & Head, Department of Botany,
Janta PG College, APS University, Rewa (M.P.) INDIA. and assigned with associated voucher specimen
numbers. (Table 0.1) They underwent a 45 °C air drying process to maintain a steady weight. The aerial

Page 4/20
components were pulverised after drying, labelled and stored in airtight glass at 20 °C until analysis. Then
extraction of essential oils was done for the present study.

Table 0.1 Main Group and family of the studied plant species-

Species Group Family Voucher Specimen Number

Origanum Majorana Linn. Angiosperms Lamiaceae J/Bot./OML-236

Tanacetum cinerariifolium Sch. Bip Angiosperms Asteraceae J/Bot./TCF-237

Juniperus communis Linn. Gymnosperms Cupressaceae J/Bot./JCLB-238

Pelargonium graveolens L'Hér. Angiosperms Geraniaceae J/Bot./PGFST-239

Thuja occidentalis L. Gymnosperms Cupressaceae J/Bot.TOL-240

Tecoma stans (L.) Juss. Ex Kunth. Angiosperms Bignoniaceae J/Bot./ TSLF-241

Artemisia arborescens Linn. Angiosperms Asteraceae J/Bot./AAL-242

Acinos arvensis (Lam.) Angiosperms Lamiaceae J/Bot./AAWh-243

Preparation of mosquito repellent hydro gel with carbopol base-

The gel was prepared by combining the constituents in Table 1 taking into consideration the experimental
results obtained using arm-in-cage method and the appropriate quantities of the extracts and essential
oils. The overall active ingredients in the gel are 12% (V / V per cent). Distilled water was applied to the
mix until the final amount was 80.00 ml, and the mechanical stirrer was used to blend all the ingredients
for one hour. Eventually, the mixture was poured into plastic containers, and the gel could be set.
Formulation of polyherbal gel preparation mention in Figure no 3. [11]

Table-1 The Constituents in the Mosquito Repellent Gel.

Page 5/20
 Constituents F1 F2 F3

Occidentalis L Essential oil 2ml 2ml 2 ml

Artenmisia Arborescent Essential oil 0.6 ml 0.6 ml 0.6 ml

Origanum Majorana Essential oil 2 ml 1 ml 1 ml

Calamintha Acinos Essential oil 1.4 ml 1.4 ml 2 ml

Tecoma Stans Essential oil 2.4 ml 3.4 ml 3 ml

Juniperus Communis Essential oil 1.2 ml 1 ml 1 ml

Scented Geranium Essential oil 2.4 ml 2.6 ml 2.6 ml

Tanecetum cinerarifolium Essential oil 0.8 ml 0.8 ml 0.8 ml

Hexane 1.2 ml 1.2 ml 1.2 ml

Ethanol 1 ml 1 ml 1 ml

Carbapol 934 1 gm 1 gm 1 gm

Propylene glycol 4 ml 4 ml 4 ml

Methylene paraben 0.1 gm 0.1 gm 0.1 gm

Tween 80 3 ml 3 ml 3 ml

Triethanol amine Few drops Few drops Few drops

Distilled water 58.7ml 58.7ml 58.7ml

Preparation of polyherbal mosquito repellent candle-

The candle was composed as the hydrocarbon bases of a mixture of hard paraffin and stearic acid.these
constituents is mention in table no.2. The recipe for the insect repellent candle is as follows: When
burned, the candle was heated by heating (700C) heavy paraffin and stearic acid. When temperature
dropped to 550C± 600C the all-essential oils were added. To get the correct diameter, the wick was
plunged repeatedly into liquid wax. The thread was put in the middle of the formed candle. It was inserted
into a mold in the shape of a cup, and was then filled with wax. The solidified candle was taken off after
cooling. Also 3 different candles have been prepared for the current investigation. Formulation of
polyherbal mosquito repellent candle mention in figure no.4.

Table-2 The Constituents in the Mosquito Repellent candle.

Page 6/20
 Constituents F1 F2 F3

Occidentalis L Essential oil 2ml 2ml 2 ml

Artenmisia Arborescent Essential oil 0.6 ml 0.6 ml 0.6 ml

Origanum Majorana Essential oil 2 ml 1 ml 1 ml

Calamintha Acinos Essential oil 1.4 ml 1.4 ml 2 ml

Tecoma Stans Essential oil 2.4 ml 3.4 ml 3 ml

Juniperus Communis Essential oil 1.2 ml 1 ml 1 ml

Scented Geranium Essential oil 2.4 ml 2.6 ml 2.6 ml

Tanecetum cinerarifolium Essential oil 0.8 ml 0.8 ml 0.8 ml

Stearic acid 15gm 15gm 15gm

Hard paraffin wax QS QS QS

EVALUATION OF THE FORMULATED GEL –

1) Appearance: The prepared gels were inspected visually for clarity, colour and presence of any
impurities.

2) pH: About 5 ± 0.01gm of the gel was added to 45 ml of distilled water and stirred to uniform
suspension. The pH of the solution was determined by using digital pH meter.

3) Viscosity -Viscosity of the gel was determined by using Brookfield viscometer. Viscosity was measured
for the fixed time 60 second for 0.5 rpm. Viscosity determinations were performed at room temperature.

4) Accelerated Stability test

For the span of 90 days, the optimized formulation was subjected to an accelerated stability test (40 °
C±2 ° C temperature; 75% ±5%, relative humidity). The prepared gel formulation was placed in a PVC
container and sealed with a black foil. as assessment of physical appearance, pH, viscosity, spreadability
and extrudability.

5) Public survey- Community volunteer survey carried out to test the health and effectiveness of
formulations at 10 houses and 10 chicken shops in selected Indore locality. The recipe was circulated to
all participants and collected feedbacks. The analysis took 1 month to finish. The results were obtained
and systematically analysed.

6) Appearance - The appearance of the cream was judged by its colour, texture, roughness and its odour.

Page 7/20
 7) Spreadability - The Spreadability was represented in terms of time in seconds by two slides taken to
slip off the jelly, placed in between the slides under a certain load. The less time it takes to split the two
slides, the greater the quality of the Spread. Two sets of standard sized glass slides have been produced.
The herbal cream mixture had been placed on one of the diapers. The other slide was placed on top of
the solution, and the cream was placed between the weight of the two slides on the upper slides and the
cream was thinly squeezed between the two slides to create a thin sheet. Reduced weight, then brushed
off the excess substance adhering to the diapers.

8) Patch Test Approximately 1-3gm of test substance was mounted on a piece of cloth or funnel and
added to the vulnerable part of the head, e.g., head behind ears. The cosmetic to be checked has been
added to 1sq.m. of the skin region. Command patches were added as well. Within 24 hours the repair site
is inspected. The procedure was replicated three times, so there was no response. Since no reaction has
been observed on third order, the individual can be taken as not hypersensitive.it is mention in figure
no.5. [12]

9) Irritancy Test-The prepared 0.5 g gel was added to the usual hairless skin at an area of 6 cm2 and then
wrapped with a semi-occlusive bandage for 1 hr period. After the application date, the bandage was
removed, the applied gel was completely scrapped off and any rashes or related signs were visually
examined at the location. The research was administered over 7-day duration. The results were expressed
with respect to grades. [13]

10) Extrudability- A basic approach to this analysis has been adopted. After the gel was placed in the
bottle, the formulas were poured into the collapsible tubes. The extrudability of the various mosquito
repellent gel formulations was measured by weight in grams needed to extrude a 0.5 cm of mosquito
repellent gel ribbon within 10 seconds. [14]

11) Washability The washability of formulations was tested by rubbing the gel on the skin and then
determining the ease and degree of washing it with purified water and examining the result manually. [15]

12) Swelling index -Taking 2 g of gel in a beaker comprising 10 mL of distilled water determined the
swelling index of the prepared polyherbal gel. The swelled formulation was withdrawn from the beaker
after 1 hour, and placed on a petridish18. The substance was measured again and the index of swelling
was calculated from the formula:

Swelling index (Si) = Wt-Wo /Wo × 100

Where, Wt = weight of swollen at t time; Wo = original weight of gel at zero time.

Mosquito repellent activity-The volunteers' forearm was immediately thoroughly cleaned with soap and
dried up absolutely. The left arm acted as the hand kept inside the mosquito cage. The vector frequency
landed on the forearm over a 30-second period. If the mosquitoes have arrived > 10 then the analysis has
started. The arm was gently removed from the mosquito cage after a time period of 30 seconds. The right

Page 8/20
arm smudged with mosquito repellent gel formulation was consequently joined, and the analysis was
completed at 30, 60, 120, 240, and 480 minutes analogously. It was estimated the number of mosquitoes
that landed and correlated with the positive control (Odomos Naturals Mosquito Repellent Gel). The
analysis was carried out in triplicate format. It is mention in figure no.6.

EVALUATION OF POLY HERBAL MOSQUITO REPELLENT CANDLE-

Flammable test- To research mosquito repelling behaviors the prepared candle was tested for
flammability and was burnt Efficiency with regard to burning time and consequently its efficacy Spotting
operation. Candle inflammability test was performed to Checks its clear laboratory combustibility. Simple
Therefore, on performance, the time taken to light the candle and its Causal symptoms such as pain,
coughing has been reported and recorded in identified mosquitoes vulnerable at night and in the field
above Period like the chicken shops and village houses. [16]

Result
All the three gel formulations produced (F1-F3) had a white, less or more porous, homogeneous smooth
textured appearance with no firm particles or gritty that was observed while interacting between the
fingertips. The translucency of the gel formulations is primarily determined by the concentration of
carbopol 934. A reduced proportion of carbopol 934 results in an improvement in the consistency of the
gel formulations. No skin irritation, edema, rashes, erythema, or any dermatological reaction or specific
inflammation, when applying the gel formulations for 7 days. Additionally, a genius washability attribute
was observed for all formulations produced (Table 3)

Table 3: Physical evaluation of the formulated gel formulations.

Formulation Color Transparency Smoothness Washability Irritability

F1 White Less Translucent Smooth Good No Irritation

F2 White Translucent Smooth Good No irritation

F3 White More Translucent Smooth Good No irritation

The pH of the gel formulations was observed to be within the range of 7.1-7.3 which is below the usual
skin pH level. Viscosity is an essential aspect that affects pharmaceutical properties such as
spreadability, extrudability, bottle derivative quality, etc. The formulations' viscosity falls within the range
44600-52900 cps. The rheological study indicated that the shear stress increases extensively with an
increase in the torque resulting in a reduction in the viscosity of the formulation. The formulations
presented the spreadability within the 14.10-16.73 g.cm/sec range which reflected that a small amount of

Page 9/20
shear would easily spread the gel formulation. A quantitative analysis of spreadability and viscosity
showed that the spreadability decreases dramatically with an increased viscosity of the formulation. The
prepared gel formulations displayed impressive extrudability with a large amount of extruded content (+ +
to + + +). Despite the formulation’s increased viscosity, the extrudability increases alongside and inhibits
fast extrusion from the collapsible pipeline. The swelling index of 110-123 per cent was observed. The
swelling index indicated the matrix existence of the gel formulation which inhibits the drug’s-controlled
release. (Table 4)

Table 4 Evaluation parameters of polyherbal gel formulations.

Formulation pH Spreadability Viscosity Extrudability Swelling index (%)

(g.cm/sec) (cp)

F1 7.4 16.73 52600 ++ 110

F2 7.3 15.55 49900 ++ 115

F3 7.2 14.10 43600 +++ 123

No major differences in pH, viscosity, spreadability, swelling index, extrudability, and physical appearance
were observed when the optimized formulation (F3) was exposed for 90 days at accelerated conditions
(40±2 ° C and 75±5 percent RH). Changes in pH by 0.1 percent, viscosity by 600 cps, swelling index by
9%, spreadability by 0.79 g.cm/sec were found significantly (table 5) Nevertheless, after research, no
improvements in the physical appearance, translucency, and smoothness were observed. Overall, the
formulation remained constant for the 3-month cycle and in tropical and subtropical regions is predicted
to remain in its original form for a longer time.

Table 5: Accelerated study of optimized gel formulation (F3).

Duration pH Spreadability Viscosity Extrudability Swelling index (%)

(g.cm/sec) (cp)

0th day 7.1 16.73 44800 +++ 123

90th day 7.0 17.56 42000 +++ 114

The F3 formula demonstrated the maximum 90.45 percent mosquito repellent activity in the 0th hr, and
proceeded to work up to 88.35 percent in the 3rd hr. (Table 6) The other formulations F1 and F2 displayed
Page 10/20
less operation 83.22% to 85.66% after 3 hours, while the standard product (Odomos natural’s mosquito
repellent gel) demonstrated almost 98% efficacy after the 3rd hour lapse.

Table 6: Mosquito screening results of the formulated gel.

Treatment stability Repellency (%)

0hr 1 hr 2 hr 3hr

Positive control 100±1.92 99.68±1.23 97.88±1.86 97.78±1.44

F1 86.27±1.89 84.48±1.26 83.45±1.25 83.22±1.23

F2 88.38±1.49 87.48±1.88 86.17±1.76 85.66±1.45

F3 90.45±1.29 89.88±1.65 88.74±1.92 88.35±1.88

In patch test Within 24 hours the repair site is inspected. The procedure was performed three times, since
there was no response. Since no reaction on third application has been detected, the individual can be
taken as not hypersensitive. (Table 7)

Table 7 Mosquito repellent gel patch test

Patch test (Hypersensitivity)

Formulation 1st 24 hr 2nd 24hr 3rd 24 hr

F1 NO REACTION NO REACTION NO REACTION

F2 NO REACTION NO REACTION NO REACTION

F3 NO REACTION NO REACTION NO REACTION

Page 11/20
Flammable test- Flammable test of poly herbal mosquito repellent candles are mention in table no.08.
Each formulated candles were tested in laboratory at uniform normal room condition in confined area
where mosquitoes’ population were more, by lightning and compared with the same size (3 inches)
aromatic herbal candle and time noted for the flammability, burning efficiency with respect to burning
time and eventually its effective repellent activity. The result revealed more effectiveness and efficacy
than the marketed candle (Aura Decor Anti Mosquito & Bug Repellent Jar Candle). Owing to the mixture
of very active essential oils such as Origanum Majorana, Tanacetum Cinerarifolium, Juniperus
Communis, Scented Geranium, Thuja Occidentalis L., Tecoma Stans (L.), Artenmisia Arborescent, And
Calamintha Acinos, the higher burning time was found within the laboratory. These all-essential oils are
more effectiveness for mosquitoes.

Table-8 Flammable test of poly herbal mosquito repellent candle

Flammability Application Burning Effective mosquito repellency

time time
(Min.)

Marketed mosquito repellent 7:00pm 60 Lower and efficient mosquito

aromatic candle and density till 1.25 hrs.
5:00am

Prepared poly herbal F1 7:00pm 90 There were no mosquitoes around

mosquito repellent candle and for a min of 3 hours.
5:00am

F2 7:00pm 90 There were no mosquitoes around

and for a min of 2.30 hours.
5:00am

F3 7:00pm 90 There were no mosquitoes around

and for a min of 2.30 hours.
5:00am

The F1 formula demonstrated the maximum mosquito’s repellency compare to the F2, F3 and marketed
polyherbal mosquito repellent candle.

PUBLIC SURVEY OF FINAL PRODUCT-

All the formulations were tested on the basis of the laboratory performance for human sample inhabited
areas where mosquitoes and households are Density of flies became higher. 10 Separate areas where 10
were selected Specific houses and 10 shops for chicken were chosen based on the People's permission
and as per experimental process It was executed for a month. The findings showed according to the
survey Significant success rate, with no side effects. The result was Tabulated in Table 9 where best
formulation was revealed was a candle and gel.

Table-9 Human survey for efficacy of formulated products (n=10 houses, 10 chicken shops)

Density of insects was measured by house index method.

Page 12/20
Area Gel TS Density SE Candle TS Density SE Remark
DM after DM after
(%) using using
(%) (%)

SouthTukogaj 75 ± 6.00 10 ± None 75 ± 8.00 12 ± None Excellent

0.06 pm 0.03 0.06 pm 0.03
and
6.30
am

Vijay nagar 90 ± 6.00 15 ± None 90 ± 8.00 15 ± None Very

0.5 pm 0.03 0.5 pm 0.12 good
and
6.30
am

Simrol 75± 6.00 17 ± None 75± 7.00 16 ± None Excellent

0.03 pm 0.04 0.03 pm 0.10
and
6.30
am

Datoda 80± 6.00 12 ± None 80± 7.30 16 ± None good

0.13 pm 0.01 0.13 pm 0.11
and
6.30
am

Nehru nagar 85 ± 6.00 10 ± None 85 ± 8.00 12 ± None Very

0.02 pm 0.18 0.02 pm 0.04 good
and
6.30
am

Shivaji nagar 84 ± 6.00 14 None 84 ± 8.00 14 None good

0.05 pm ±0.04 0.05 pm ±0.10
and
6.30
am

Dhar road 70 ± 6.00 15± None 70 ± 7.30 20 ± None Excellent

0.15 pm 0.10 0.15 pm 0.10
and
6.30
am

Rau 72 ± 6.00 20 ± None 72 ± 8.00 22 ± None Very

0.01 pm 0.04 0.01 pm 0.04 good
and
6.30
am

MR 10 70 ± 6.00 15± None 70 ± 7.00 22 ± None Very

0.02 pm 0.08 0.02 pm 0.10 good
and
6.30
am

Page 13/20
 LIG 78 ± 6.00 22 ± None 78 ± 8.00 18 ± None Excellent
0.04 pm 0.12 0.04 pm 0.05
and
6.30
am

Discussion
Many studies have demonstrated that plant essential oils are more effective repellents than plant
extracts. All essential oils, on the other hand, are quite powerful. This makes them unstable, which adds
to their short lifespan.as insect repellents as a result, numerous essentials are required. Oils are not
acceptable for use as a solo mosquito repellent. Repellents. This issue is addressed in the current report.
Prepare two natural mosquito repellents as part of your research.

compositions using essential oils that are very volatile with the use of herbal extracts. The extracts,
essential oils, and two insect repellent formulations caused no skin irritations or rashes on the test
participants' arms.

Clove bud essential oil, on the other hand, caused a burning sensation on the test participants' arms. As a
result, the small burning sensation felt on the arms of the test participants after applying two mosquito
repellent formulations might be attributed to pyrethrum essential oil. Reducing the Clove bud essential oil
to the point where no scorching feeling is felt will enhance the two insect repellent formulas even further.
Additionally, formulations with lower concentrations of active substances must be produced and tested
for effectiveness.

Conclusion
Natural base mosquito repellent was successfully developed in this research study. The drug has been
found to be very effective and safe to use. Owing to its beneficial use in the human body, the prepared
eight separate essential oil-based mosquito repellent gel and candle formulations have proven to be an
effective tool in preventing mosquito-borne diseases. In many developed countries, where most residents
have no access to mosquito net, high-cost mosquito repellent creams, and miscellaneous physical
methods, this formulation of gel and candle can be an efficient, affordable, and easily accessible way to
deter mosquito-borne diseases such as malaria, dengue, etc. in the lower parts of society.

Abbreviations
DM = Density of mosquitoes before; TS= Time of spray; SE= Side effects; TL=Time of
lightening=Resolution per minute: PVC =Polyvinyl chloride: OC=Celsius: D.S-Dark skin, F.S –Fair skin, M.S-
Medium skin: Wt = weight of swollen at t time; Wo = original weight of gel at zero time.

Declarations
Page 14/20
Acknowledgements: We are thankful to Dr. A.P.J. Abdul Kalam University, Indore, (M.P.) India for
supporting this study. We are also thankful to Dr. S.N. Dwivedi, Prof. & Head, Department of Botany, Janta
PG College, APS University, Rewa (M.P.) India for authentication of plants.

Studies involving plants: The plant material i.e., Origanum majorana, Tanacetum cinerarifolium,
Juniperus communis, Scented Geranium, Thuja occidentalis, Tecoma stans ,Artenmisia arborescent and
Acinos arvensis (Lam.) were collected from various study sites of Malwa region of Madhya
Pradesh(India) and were authenticated by Dr. S.N. Dwivedi, Prof. & Head, Department of Botany, Janta PG
College, APS University, Rewa Madhya Pradesh(India). Voucher Specimen No. J/Bot./OML-
236;J/Bot./TCF-237;J/Bot./JCLB-238;J/Bot./PGFST-239;J/Bot.TOL-240;J/Bot./ TSLF-241;J/Bot./AAL-
242 and J/Bot./AAWh-243

Author contributions: DKG conceptualized and drafted the manuscript and also designed the experiment.
AT was a major contributor in editing and revising with help in collection of plant and prepared figures 1–
2. RG substantively reviewed the draft with given idea of Development of the both formulations. All
authors read and approved the final manuscript.

Funding: This research did not receive any specific grant from funding agencies in the public,
commercial, or not-for profit sectors.

Availability of data and materials: All data generated or analysed during this study are included in this
published article.

Ethics approval and consent to participate: The study including sample collection was conducted
according to India’s Biological Diversity Act 2002 which permits bonafide Indians to use biological
resources for scientific research. [10]

Consent for publication: Yes

Competing interests: The authors declare that they have no competing interests.

References
1. Mimms C. A., Playfair J. H. L., Roitt I. M., Wakelin D., R., Williams In: Clinical Microbiology, Europe
Limited, 1993, Mosby Year Book.
2. Eldridge B. F., Edman J. D., In: Introduction to medical entomology. Medical Entomology: A Textbook
on Public Health and Veterinary Problems Caused by Arthropods, Eds B. F. Eldridge, J. D. Edman
(Kluwer, London), 2004 p 1.
3. Olkowski W., Larval Mosquito management, Common Sense pest Specific Quarterly, 2001, 17, 8.
4. Halstead S. B., Dengue Bull. 2000 24, 77.
5. http://www.mosquito netting.com/mosquito control.html
6. Enayati A. A., Hemingway J., Garner P., Cochrane Database Syst. Rev2007. 18, 34.
Page 15/20
 7. Mishra, A. Singh K. N., Sharma V. P., Indian J Malariol, 1995, 32 (3), 99.
8. ICMR Bulletin, Prospects of using herbal products in the control of mosquito vectors, 2003, 33 (1), 1.
9. Peterson C., Coats J., Pesticide Outlook, 2001 12, 154.
10. Venkataraman K. India's Biodiversity Act 2002 and its role in conservation. Tropical Ecology. 2009
Jun 1;50(1):23.
11. Ranasinghe MS, Arambewela L, Samarasinghe S. Development of Herbal Mosquito Repellent
Formulations. Int J Pharm Sci Res, 2016; 7(9):3643-3648.
12. Vijayalakshmi A., Tripura and Ravichandran A. V.; Development and Evaluation of Anti- Acne
Products from Terminalia arjuna Bark; IJCRGG Vol. 3, No.1, pp 320-327.
13. Bhowmik Debjit, Gopinath Harish, B. Kumar Pragati, Duraive S. l, G Aravind., K. P. Kumar Sampath;
Medicinal Uses of Punica granatum and Its Health Benefits; Journal of Pharmacognosy and
Phytochemistry; 2013, Vol. 1 No. 5.
14. K.R Vinod, Santhosha D, Anbazhagan. S; Formulation and Evaluation of Piperine Creama New Herbal
Dimensional Approach for Vitiligo Patients; Int J Pharm PharmSci, 2011, Vol 3, Suppl 2, 293.
15. Gupta R, Gupta GD. Formulation Development and Evaluation of Anti-inflammatory Potential of
Cordia oblique Topical Gel on Animal Model. Pharmacogn J, 2017; 9(6): s93-s98.
16. Das Kuntal, Vasudeva C. Vasudeva and Dang Raman. Economical novel formulation and evaluation
of herbal oils for mosquito and house fly repellent activities. Annals of Phytomedicine, 2016: 5(2):
91-96.

Figures

Page 16/20
Figure 1

Broad Classification of Mosquito Repellent Methods

Figure 2

Chemistry of insect repellent

Figure 3

Formulation of polyherbal mosquito repellent gel

Page 17/20
Figure 4

Formulation of polyherbal mosquito repellent candle

Page 18/20
Figure 5

Patch test

Page 19/20
Figure 6

The arm-in-cage (AIC) test for measuring the efficacy of topical mosquito repellents Under laboratory
conditions

Supplementary Files
This is a list of supplementary files associated with this preprint. Click to download.

DeepakGuptaPlantAuthentication.pdf

Page 20/20