ISSN : 0975-2366

https://doi.org/10.31838/ijpr/2018.10.03.065
Re se a rc h Art ic le

FORMULATION OF NOVEL HERBAL CREAM AS

MOSQUITO REPELLENT
B. PRABASHEELA 1 *,V. NANDHINI 1 , V. SAKITHYA 2 , NIGARISH ARA 3 AND
R.NAGALAKSHMI 4
1,2,3
Department of Biotechnology, Aarupadai veedu Institute of Technology, Paiyanoor, Kancheepuram district,
Tamil Nadu, India
4
Department of Chemistry, Aarupadai veedu Institute of Technology, Paiyanoor, Kancheepuram district,
Tamil Nadu, India
Email Id: nagalakshmirajan@gmail.com
Received: 01.05.18, Revised: 08.07.18, Accepted: 18.08.18

ABSTRACT
Vector control plays a key role in prevention and control of major vector-borne diseases. Repellents are the first line
of defence against the epidemics of vector-borne diseases. Synthetic mosquito repellent used for control of vectors
cause irreversible damage to ecosystem and are non-degradable in nature. The uses of biologically-active plant
materials with anti-mosquitocidal properties are attracted in recent years, because of their biodegradable nature and
being relatively safer to human and other non-target organism in the environment. Hence the present study aimed to
produce a novel herbal cream as a mosquito repellent. The leaves included for the study are papaya (Carica
papaya), Guava (Psidium guajava), Tawa tawa (Euphorbia hirta) and Neem (Azadirachta indica). The extracts were
prepared by different methods such as Maceration, soxhlet and shaker extraction process and also with different
solvents Ethanol, Ethyl acetate, Acetone. These extracts were qualitatively and quantitatively analysed for active
ingredient. Ethyl acetate (soxhlet) was found to be high concentration of Quercetin. In vitro antioxidant properties
were evaluated by reducing power assay method and ethyl acetate (soxhlet) extract were shown to posses high anti
oxidant activity. This extract was chosen to formulate cream. Piperine extracted from black pepper was used as an
additive in cream formulation. The prepared cream was tried on mosquitoes kept in fabricated mosquito cage. In the
study it was observed that the mosquitoes moved towards the corner of the cage which indicates the repellent
action. The study can be further extended to screen the toxic effect through animal studies to bring it as a potent safe
and eco-friendly mosquito repellent.

Keywords: mosquito repellent, Carica papaya, Psidium guajava, Euphorbia hirta, Azadirachta indica, Quercetin, herbal
cream, Piperine, repellent action, eco-friendly, biodegradable.

INTRODUCTION
Mosquitoes such as Aedes, Anopheles, and Culex combat mosquito nuisance and mosquito borne
are a serious threat to the public health as they are diseases 4. Synthetic chemicalss like N, N-Diethyl-3-
known vectors for various protozoan, viruses, and methylbenzmide (DEET), Dimethyl phthalate (DMP),
bacteria which result in many life threatening Pyrethroids, Octachlorodipropylether (S-2) and
diseases like malaria, filariasis, yellow fever, allethrin, are used in repellent formulations which
Japanese encephalitis, chikungunya, dengue and may lead to their higher exposure to the environment
epidemic polyarthritis in almost all tropical and and hence ,the unacceptable health risks. The
subtropical countries and many other parts of the potential health hazards of repellents are Breathing
world 1. They are major arthropod vectors problem, headache, dizziness, nausea, Eye irritation
responsible to transmit serious pathogenic diseases and skin allergy 5. Allethrin used in the mats
and affect billions of people annually, throughout the increased blood brain barrier (BBB) permeability,
world 2.Mosquitoes are more than 3500 species, suggesting a delayed maturity of BBB and
infecting over 700,000,000 people every year biochemical changes causing health risks, especially
globally and 40,000,000 of the Indian population. at an early age in life (Moya-Quiles et al) 6. DEET is
Mosquitoes alone transmit diseases to more than still the most widely used mosquito repellent. It has
700 million people and over one million deaths are generally been regarded as safe, but toxic effects
reported annually across the globe 3. The use of have been recorded, including encephalopathy in
mosquito repellents is increase day by day due to children, urticaria syndrome, anaphylaxis,
occurrence of mosquito borne diseases. Repellents hypotension and decreased heart rate 7.With an
such as liquid vaporizers, Aerosols, mosquito coils, increasing concern on public safety, a renewed
vapourizing mats are widely used in the country to interest on the use of natural products of plant origin
195| International Journal of Pharmaceutical Research | July- Sept 2018 | Vol 10 | Issue 3
 Nagalaxmi et al / Formulation Of Novel Herbal Cream As Mosquito Repellent

because of its effectiveness, environment friendly,

biodegradability and inexpensiveness. A large Materials And Methods
number of plant extracts have been reported to Collection of plant leaves
posse s mosquitocidal activities against mosquito Papaya leaves, Tawa Tawa leaves, Guava leaves
vectors and very beneficial in repelling mosquitoes 8. and Neem leaves were collected from kadapakkam,
Phytochemicals are the bioactive compounds Tamil Nadu, India.
(larvicides, repellents, insect growth regulators, Procedure for preparation of leaves for extraction
antifeedants, ovicides, oviposition deterrents) found Collected leaves were washed with distilled water
in herb and serve as mosquito repellent (Mathivanan and shade dried for 18 days. Dried samples were
et al., 2010) 9. Extracts of plant leaves such as Neem ground in to powder using mortar and pestle.
(Azadirachta indica), papaya (carica papaya) ,Guava Extract Preparation
(Psidium guava), tawa tawa (euphorbia hirta) have Papaya leaf, Tawa Tawa leaf, Guava leaf and Neem
been studied as possible mosquito repellents. Carica leaf powder were taken in ratio (2:1:1:1) for
papaya leaves extract contains antioxidants, phenolic extraction. The powdered plant samples (25 g/250
compounds, chlorogenic acid, quercitin, antiseptic, ml) were extracted successively with Ethanol, Ethyl
antimicrobial, or diuretic, to control parasites, reduce Acetate, Acetone using Solvent Extraction, Shaker
inflammation. The extract has a bitter substance Extraction, Soxhlet Extraction in order to extract the
karpain an insecticide that is not favoured by polar and non polar compounds.
mosquitoes 10. Euphorbia hirta leaves is well known EXTRACTION
for its medicinal properties and used as antiviral and Solvent Extraction: 25 g of dried samples were
anti-inflammatory properties. It posses Larvicidal and dissolved in 250 ml of solvent. Mixtures were kept in
pupicidal activity against malaria vector the dark at room temperature in sterilized conical
(Panneerselvam K et al., 2016)11. Psidium guajava flask and wrapped with aluminium foil to avoid
leaf extract reported as bio insecticide potential and evaporation and exposure to sunlight 16.
contains a number of beneficial substances, Shaker Extraction: 25 g of dried samples were
including antioxidants , flavonoids and rich source of soaked in 250 ml of solvent in 500 ml sterile conical
vitamins , dietary fibers and minerals. It reported that flask and mixed thoroughly. Mixtures were covered
extract posses significant larvicidal activity and also with cotton and wrapped with aluminum foil.
act as effective insecticide to control mosquitoes Mixtures were kept at 60 RPM in orbital shaker for 24
(Satria dharma setiawan et al ., 2014) 12. hours at room temperature 17.
Azadirachta indica, is known for its medicinal, Soxhlet Extraction: 25 g of dried samples were
cosmetic, pesticidal and insecticidal properties which weighed and were successively extracted with 250 ml
act as anti-viral and anti-bacterial, along with being of solvent, in a Soxhlet apparatus at room
a powerful immuno-stimulant. Neem extract have the temperature for 18 hrs 18.
ability to kill Aedes pupae at relatively low PRELIMINARY PHYTOCHEMICAL SCREENING
concentrations and act as an alternative to the use of The crude extracts of leaves were subjected to
synthetic pesticides for control of mosquitoes (Umar preliminary phytochemical screening for possible
et al., 2006) 13. Piperine nigrum act as a potential presence of bioactive compounds. The following
larvicide for the control of mosquito vector species. tests were performed on the extracts to detect various
Alkaloid Piperine from pepper have repellent actions phytoconstituents present in them.
against Anopheles malaria vector (Anopheles Test for tannins
arabiensis, An. colic, An. gambiae, An. Potassium dichromate test: To a 1ml of extract in a
quadriannulatus and An. funestus 14. Plant essential test tube and few drops of 5% ferric chloride was
oils are potential natural repellents that are expected added and observed for the brown colour or dark
to replace synthetic compounds. The use of topical green colour 19.
insect repellents is one of the strongest Test for Steroid
recommendations by health authorities (Med J Aust. Salkowski reaction: 1 ml of extract was shaken with
2015). The use of cream is much safer method to chloroform in a test tube; few drops of acetic
fight mosquitoes than mosquito coils and sprays 15. anhydride was added and boiled in a water bath
Hence the present study has selected four different and rapidly cooled in iced water. Conc. Sulphuric
plant leaves such as papaya, Guava, tawa tawa and acid (2mL) was added alongside of the test tube.
Neem leaves to be used as an ideal ecofriendly Formation of yellow colour and turning the upper
approach for the control of mosquitoes. This study layer to green showed the presence of steroids 20.
reports a novel approach to develop a mosquito Test for Protein
cream formulation and to determine mosquito Xanthoprotein test: To 1 ml of extract, a few drops of
repellent activities of the cream g to obtain safe and Conc. Nitric acid was added. The yellow colour
efficient mosquito repellent by combinations of indicates the presence of protein 20.
plant leaves extract. Test for Terpenoid
Liebermann-Burchard test: To a 1ml of extract, 2 ml
chloroform of was added. 3 ml conc. sulphuric acid

196| International Journal of Pharmaceutical Research | July- Sept 2018 | Vol 10 | Issue 3
 Nagalaxmi et al / Formulation Of Novel Herbal Cream As Mosquito Repellent

was carefully added to the layer, and observed the (2mL) and few drops of ferric chloride was added,
brown colour 21. followed byH2SO4 (1mL) along the side of the test
Test for reducing sugar tube. The formation of brown ring gives positive
Fehling s test: To 1 ml of extract in a test tube , 10 ml indication for cardiac glycerides 28.
de-ionized water and then few drops of Fehling Test for Catechin
solution (Fehling A and B) were added and heated at To 1 ml of extract, a few drops of Ehrlich reagent
100ºC in water bath. Brick red precipitate showed a was added. The pink colour indicates the presence of
positive result 22. Catechin 28.
Test for alkaloids Test for saponins
Wagner s test : 1 ml of extract was taken in test tubes Froth test: The extract was diluted with 20 ml of
and 2- 3 drops of Wagner's reagent was added. distilled water and it was agitated in a graduated
Reddish brown colour precipitation indicates the cylinder for 15 minutes. The formation of froth
presence of alkaloids 23. showed the presence of saponins 29.
Test for Coumerin THIN LAYER CHROMATOGRAPHY
1 ml of extract was dissolved in methanol and Thin Layer Chromatography developed for the best
alcoholic potassium hydroxide. The appearance of separation of the phytoconstituents present in the
yellow colour which decolorizes while adding conc. extract. The solvent system selected for the best
Hydrochloric acid shows presence of Coumerin 23. results of TLC was Toluene - Ethyl acetate - Formic
Test for Anthraquinone acid (5: 4: 2) for extracts. TLC chamber was
Borntrager s test: To 1 ml of extract 5% Sulphuric saturated for 20 minutes. Standard Quercetin (0.1
acid (1mL) was added. The solution was boiled in a µg spot 1 from stock solution) applied, aluminium-
water bath and then shaken with equal volume of packed TLC plates coated with silica. Plant samples
chloroform and kept to stand for 5 min. Then lower were spotted in triplicate. Plates were developed at
layer of chloroform was shaken with half of its room temperature in TLC chamber. After the TLC run
volume with dilute ammonia. The formation of rose and compounds present in the extracts were
pink to red colour of the ammonical layer gives identified visually. Then, Rf value was calculated as
indication of Anthraquinone 23. the ratio of the distance travelled by solute to the
Test for Flavanoid distance travelled by the solvent [30]
Alkaline reagent test: To 1 ml of the extract, a few QUANTITATIVE ESTIMATION OF QUERCETIN
drops of dilute sodium hydroxide were added. An The concentration of Quercetin present in extracts
intense yellow colour was produced, which become was determined by the method of Method of Singh
colourless on addition of a few drops of dilute acid Upendra et al., 2009 31. Aliquots of working stock
indicates the presence of Flavanoid 24. solutions of Quercetin were prepared with developed
Phenolic compounds solvent to get concentration in range of 0.5-2.5
Ferric chloride test: The extract was dissolved in mg/ml. The absorbance of resulting solution was
alcohol and 1 drop of neutral ferric chloride was measured at λ max 256.30 nm. 1ml of extract was
added. The intense black color indicates the dissolved in 10mL of methanol to get 0.1mg/ml
presence of phenolic compound 25. solution. Absorbance of the test solution was
Test for phlobatannins measured at 257nm against blank.
Take 1ml extract in a test tube , 10ml distilled water INVITRO ANTIOXIDANT STUDY
was added and boiled at 100ºC with few drops of Antioxidant activity of the extracts were analysed by
1%Hydrochloric acid. Deposition of red precipitation Reducing power assay method. The reducing ability
gives positive result 25. of extracts was determined by the method of Yildrim
Detection of Oils and Resins et al., (2001) 32.Various standard concentrations
Filter paper test: Extract was applied on filter paper. and test sample were prepared .It was mixed with 1
It develops a transparent appearance on the filter ml of 0.2 mol/L sodium phosphate buffer (pH 6.6)
paper. It indicates that the presence of oils and resins and 1 ml of 1% potassium ferric cyanide. The
25
. mixture was incubated in water bath at 50 °C for 20
Test for Quinone min. Then 1 ml of 10% of trichloro acetic acid was
Sulphuric acid test: Extracts was dissolved in alcohol added and centrifuged at 3 000 r/min for 10 min.
and treated with sulphuric acid. The colour The supernatant (2 ml) was mixed with 2 ml of
developed indicates the presence of Quinone 26. distilled water and 500 µL of 1% ferric chloride. The
Test for Amino acids absorbance was read at 700 nm. Then absorbance
Lead acetate test: To 5 ml of extract, few drops of was compared with standard values.
40% Sodium Hydroxide and 10% Leadacetate PIPERINE EXTRACTION
solution were added and boiled for 10 mins. Black 10 gm of black pepper powder extracted with 100ml
precipitate of lead sulfate was formed 27. of 95% ethanol in Soxhlet extractor for 2 hours. The
Test for Cardiac glycerides solution was filtered and concentrated on the water
Keller-Killiani test: 1 ml of Extract was shaken with bath at 60oC. To 10mL of a 10% solution of KOH in
distilled water (5 ml). To this, glacial acetic acid 95% ethanol contained in a 100ml flask added the
197| International Journal of Pharmaceutical Research | July- Sept 2018 | Vol 10 | Issue 3
 Nagalaxmi et al / Formulation Of Novel Herbal Cream As Mosquito Repellent

concentrated pepper extract. Heated the resulting 6pm and 9 pm. 20 mosquitoes were placed in the
solution and water was added drop wise. A yellow cage. Prior to testing, volunteer washed hands with
precipitate formed 33. unscented soap. Treatment comprises a repellent
MOSQUITO REPELLENT CREAM FORMULATION applied to the forearm of a volunteer skin surface
Pure petroleum jelly (25 g) was accurately weighed area. The treated forearm is inserted into the cage
into a 50 ml wide-mouth glass bottle and melted in and exposed to mosquitoes. The number of
water bath at temperature of about 50ºC. At this mosquitoes that land and probe the skin in 15
stage, 1 g of extract and 5ml of oil were transferred minutes was observed and recorded. The
into the molten petroleum jelly. Finally, 10 mg of observations are repeated every 1 hour 35.
benzoic acid was added to the molten state. The
mixtures were stirred properly to ensure that the Results And Discussion
sample was uniformly mixed with the molten stage of PRELEMINARY PHYTOCHEMICAL SCREENING
the cream 34. The present study revealed that ethanol, ethyl acetate
CAGE TEST and acetone leaves extracts contained
The cage test is the most common way of testing the cardiacglycerides, Quinone, protein, amino acid,
effectiveness of a mosquito repellent. To evaluate the Flavanoid, phenols, Saponin, steroids, tannin,
effectiveness of repellent activity against mosquito Terpenoid and reducing sugar in all three solvents.
performed preparatory experiments with mosquitoes No source of Coumerin, Phlobatannin, alkaloids, oil
in cage. The mosquitoes used in this experiment were & resins and Catechin found in any of the solvents.
caught using a net while biting humans between THIN LAYER CHROMATOGRAPHY

Standard Ethanolic extract Ethyl acetate extract Acetone extract

Figure l: Thin layer chromatography
The chromatogram developed with Toluene - Ethyl acetate - Formic acid in ratio of 5: 4: .2 as mobile phase
revealed the presence of Quercetin.

Figure 2: Quercetin Estimation

Quercetin Concentration was found to be high in ethyl acetate extracts. Soxhlet extraction was found to be
effective in obtaining crude extracts. Hence, this extract was chosen to formulate cream.

198| International Journal of Pharmaceutical Research | July- Sept 2018 | Vol 10 | Issue 3
 Nagalaxmi et al / Formulation Of Novel Herbal Cream As Mosquito Repellent

Table 1: Phytochemical screening

MACE ETHY ETHYL

SHAKER ETHYL SOXHL
RATIO L ACET ACETONE ACETA ACET
ETHAN ACETA ETETHA
TEST N ACET ONE TE ONE
OL TE NOL
ETHA ATE
NOL
+
Tannin + + + + + + + +
+ +
Steroid + + + + + + +
Protein
+ + + + + + + + +
+
Terpenoid + + + + + + + +
Reducing
+ + + + + + + + +
sugar
-
Alkaloid - - - - - - - -
Coumerin
- - - - - - - - -
Anthraqui
none - - - - - - - - -

Flavanoid
+ + + + + + + + +

Phenol + + + + + + + + +
Phlobatan
nin - - - - - - - - -

Oil &resin
- - - - - - - - -
Quinone
+ + + + + + + + +
Amino
acid + + + + + + + + +

Cardiac
glycerides + + + + + + + + +

Catechin
- - - - - - - - -
Saponin
+ + + + + + + + +

QUANTITATIVE ESTIMATION OF QUERCETIN

Table 2: Quercetin Estimation

S.NO SOLVENT MACERATION SHAKER SOXHLET

1 Ethanol 0.125±0.0241 0.15±0.0029 0.171±0.0031

2 Ethyl Acetate 0.271±0.003 0.247±0.0035 0.29±0.0071

3 Acetone 0.273±0.005 0.172±0.0258 0.19±0.0039

199| International Journal of Pharmaceutical Research | July- Sept 2018 | Vol 10 | Issue 3
 Nagalaxmi et al / Formulation Of Novel Herbal Cream As Mosquito Repellent

Statistical analysis was carried out by t test calculator using a statistical package program (SPSS 10.0). Values
were performed in triplicates and represented as mean ±SD. Mean values are significant at (P<0.05).
IN VITRO ANTIOXIDANT STUDY- REDUCING POWER ASSAY METHOD
Table:3

S.NO Concentration (mg/ml) Absorbance (nm)

1 .5 0.062

2 1 0.124

3 1.5 0.199

4 2 0.297

5 2.5 0.34

STANDARD CURVE

0.4
0.35
0.3
0.25
Absorbance (nm)

0.2
0.15
0.1
0.05
0
0 1
Concentration(mg2
) 3

The reducing power of the extracts may serve as a significant indicator of its potential antioxidant activity.
Higher absorbance of the reaction mixture indicates greater reducing power. The results were compared with
that standard antioxidant. Ethyl acetate showed high amount of reductones and hence the antioxidant
property.
Table 4
SOLVENT MACERATION SHAKER SOXHLET
S.NO
0.229 ±0.004
1 Ethanol 0.366±0.0041 0.346±0.3035
Ethyl Acetate 0.291±0.003
2 0.352±0.00294 0.427±0.3055
Acetone
3 0.253±0.002 0.215±0.0036 0.302±0.0416

Statistical analysis was carried out by t test calculator using a statistical package program (SPSS 10.0). Values
were performed in triplicates and represented as mean ±SD. Mean values are significant at (P<0.05).

ADDITIVE (PIPERINE) Medicinal herbs serve as an important source of

Piperine from black pepper was extracted with bioactive molecules for novel therapeutic agents and
ethanol using soxhlet extraction at room have been used widely to treat a variety of vector
temperature. Piperine was isolated and the yield ailments such as malaria, dengue etc., (Dalziel J. M.
was found to be 2.8 g. Hence, this Piperine was 2005)37. Plant extracts have been used to repel or kill
added as additive to the cream. mosquitoes and other domestic pest insects for a
long time before the advent of synthetic chemicals
Discussion (Sukumar et al., 2016) 38. Plants synthesize a wide
Many plant extracts possess repellent properties array of secondary metabolism compounds that are
against various insect pests, and several plant oils generally thought to be involved in plant insect
have been identified as potential botanical sources of interactions .They can act as repellent, antifeedant,
insect repellents (Ansaria M.A et al., 2005)36. or toxic (Wollenweber 1994; Rao 1982)39.These
200| International Journal of Pharmaceutical Research | July- Sept 2018 | Vol 10 | Issue 3
 Nagalaxmi et al / Formulation Of Novel Herbal Cream As Mosquito Repellent

secondary metabolites are reported to have many Additive and Preservative in the ratio of 25: 1: 5: 1:
biological and therapeutic properties and expected 0.01. The repellent activity of cream was evaluated
to have many medicinal uses. Preliminary by cage method. From the test performed, it was
phytochemical screening is an important initial step observed that the mosquito repellent cream repelled
to find out the phytoconstituents present in the plant the mosquitoes to corner of the cage. This shows that
extract, which further leads to the isolation of active mosquito repellent cream provide substantial
compounds, responsible for many pharmacological protection against mosquitoes. It can be used as
actions. Phytochemicals are non-nutritive plant effective mosquito repellent cream.
chemicals that have protective or disease preventive
properties. The result of Phytochemical screening Conclusion
showed that the plant exhibit cardiacglycerides, Mosquitoes are vectors for humanity s most deadly
Quinone, protein, amino acid, Flavanoid, phenols, illnesses, and they are public enemy number one in
Saponin, steroids, tannin, Terpenoid and reducing the fight against global infectious disease. Plant-
sugar and absence of Coumerin, Phlobatannin, derived repellents usually do not pose hazards of
alkaloids, oil & resins and Catechin. The biological toxicity to humans and domestic animals and are
activity of the plant extract might be due to a variety easily biodegraded. Compared to synthetic
of compounds in this plant including phenolics, compounds, natural products are presumed to be
terpenoids, flavonoids and alkaloids existing in plant, safer for human. . This study has attempted to
these compounds may jointly or independently highlight the plants claimed to be used as mosquito
contribute to produce larvicidal, adult emergence repellent. The present study has shown that leaves of
inhibition, and oviposition deterrent activity against Papaya, Tawa Tawa, Guava, and Neem have
both species of mosquitoes40.Asha S et al., (2005) Mosquito repellent action. Safe and effective
have reported the presence of proteins, fats and oils, mosquito repellent cream was successfully developed
gums and mucilage, alkaloids, flavonoids, using plant leaves extract and repellent action was
terpenoids, tannin, phenol, steroids, glycoside, confirmed by cage test. Further investigations are
saponin, Quinone, protein, in ethanolic and needed to elucidate the efficacy of the mosquito
methanolic extract of Euphorbia hirta leaf41, wang P repellent formulations against a wide range of
G et al., (2008) in the aqueous guava leaf extract42 , mosquito species and also to identify active
Zainab saif salef al-hashemi et al.,( 2016)in Neem compounds responsible for mosquito repellent
leaves extract 43 and .Vaijayanthi R et al., (2013) in activity in order to utilize them if necessary, in
Carica papaya leaves extract 44 . Phytochemicals preparing a commercial product to be used as a
present in plant extracts act as general toxicants both mosquito repellent.
against adult as well as larval stages of mosquitoes
45
.Thin Layer Chromatography was developed for the References
best separation of the phytoconstituents present in 1. Shyamapada Mandal , Repellent activity of
the extract. The solvent system selected for the best eucalyptus and Azadirachta indica seed oil against
results of TLC was Toluene - Ethyl acetate - Formic the filarial mosquito Culex quinquefasciatus say
acid (5: 4: 0.2) for extracts. In this study, five spots (Diptera: Culicidae) in India, Asian Pacific Journal of
were identified in the extract. The Rf values of the Tropical Biomedicine , vol.1, 2011, no.1, pp: S109
extract were reveals the presence of phenolic S112. DOI: 10.1016/S2221-1691(11)60135-4.
2. Idin Zibaee, Hasan Mastari Farahani, Javad Mirarab
compounds and flavonoids in accordance with Asha
Razi and Mohsen Nourzad Moghaddam , Synergistic
verma et al., 2014 30 .The Quercetin was the most
effect of two medicinal plants in cream formulation
active substance providing the evidence of its activity
on repellency of vector mosquito and rove beetle ,
against Plasmodium falciparum parasites and
Journal of Entomology and Zoology Studies, Vol. 4,
demonstrates significant anti-DENV inhibitory Issue 4, 2016, 73-79 E-ISSN: 2320-7078 , P-ISSN:
activities [46]. The presence of Quercetin in the 2349-6800.
extracts was confirmed in the present study by 3. Mohomed AA, Tarek IAAM, Zarrag IA, Larvicidal
running TLC along with Standards. Quantitative and repellent effect of some Tribulus terrestris L.,
determination of Quercetin Concentration was (Zygophyllaceae) extracts against the dengue fever
found to be high in soxhlet Ethyl acetate extracts was mosquito, Aedes aegypti (Diptera: Culicidae) ,
compared with other extract .The Anti oxidant activity Journal of Saudi chemical Society ,vol 20, 2012, 13-
was also found to be high in soxhlet Ethyl acetate 16.
extract. . Hence this study, has selected soxhlet ethyl 4. Sharma Diel F, Detscher M, Schock B and Ennis M,
Acetate extract for repellent cream formulation. Epidemiologic notes and reported seizures
Piperine from black pepper was used as additive in temporally associated with use of DEET insect
cream formulation to enhance the repellent action repellants, New York and Connecticut. MMWR
and the yield was found to be 2.8 g. Scott et al., Morb Mortal Wkly Rep1989, 38:678-80.
(2007) have also isolated Piperine from Piper Lon 5. Solomon B, Sahle F, Geber Mariam T, Asres K and
gum Linn (Piperaceae)47The cream formulation Neubert H , Micro encapsulation of citronella oil
includes petroleum jelly, Essential oil, Extract , form mosquito repellent application: Formulation

201| International Journal of Pharmaceutical Research | July- Sept 2018 | Vol 10 | Issue 3
 Nagalaxmi et al / Formulation Of Novel Herbal Cream As Mosquito Repellent

and invitro permeation studies, European Journal of Acta Poloniae Pharmaceutica -Drug Research, vol 68
Pharmaceutics and Biopharmaceutics ,vol.80, 2012, (5), 2011, 687-692.
no.1,61 66. 19. Pandey A, Jagtap JV, Patil AA, Joshi RN, Kuchekar BS
6. Moya-Quiles ,Mather TN, DeFoliart GR, Repellency , Formulation and evaluation of antibacterial and
and initial toxicity of Abate and Dursban antifungal activity of a herbal ointment containing
formulations to Aedes triseriatus in oviposition sites Aloe-Vera, Azardirachta indica and Curcuma-longa,
,Mosq News , vol 43, 2014,474 479 .DOI: Journal of Chemical and Pharmaceutical Reaearch,
10.1007/s00436-011-2540-z vol 2 (3) 2010, 182-186.
7. Sharma Diel F, Detscher M, Schock B and Ennis M, 20. Banso A, Adeyemo S, Phytochemical screening and
Epidemiologic notes and reported seizures anti malarial assessment of Abutgilon mauritianum,
temporally associated with use of DEET insect Bacopa monifera and Datura stramonium,
repellants, New York and Connecticut. MMWR Biokemistrics vol 18, 2006, 39-44
Morb Mortal Wkly Rep, 1989; 38:678-80 2012 21. Sivaraj R, Balakrishnan A, Thenmozhi M, Venckatesh
8. Kumuda Sukumar, Michael J Perich , Lewis R R , Preliminary phytochemical analysis of Aegle
Boobar , Botanical derivatives in mosquito control: a marmelos, Ruta graveolens, Opuntia dellini,
review , Journal of American Mosquito Control Euphorbia royleana and Euphorbia antiquorum,
Association , vol 7, 1991 ,2lO-237. MD 21702-5010 International Journal of Pharmaceutical Sciences and
9. Mathivanan N, Muley BP, Khadbadi SS, Banarase NB, Research, vol 2 , 2011,132-136.
Phytochemical Constitutions and Pharmacological 22. Venkatesan D, Karrunakarn CM, Selva Kumar S,
Activities of Medicinal plants - A review, Tropical Palani Swamy PT, Identification of phytochemical
Journal of Pharmaceutical Research, vol 8, 2015, constituents of Aegle marmelos responsible for
455-465. antimicrobial activity against selected pathogenic
10. Rasman, Tiwari, Kumar B, Kaur M, Kaur G and Kaur organisms, Ethnobotanical Leaflet , vol 13, 2009,
H , Phytochemical Screening and extraction: A 1362-1372.
review , Int. Pharm Sci vol1(1), 2007, 98-106. 23. Joshi A, Bhobe M, Saatarkar, A Phytochemical
11. Panneerselvam K, Abdul Rahuman A, investigation of roots Grewia microcos Linn , J,
Gopalakrishnan G, Venkatesan P and Geetha K , chem. Pharma Res 2013, 80-87.
Larvicidal activity of some Euphorbiaceae plant 24. Senthilkumar PK, Reetha D, Screening of
extracts against Aedes aegypti and Culex antimicrobial properties of certain Indian medicinal
quinquefasciatus (Diptera: Culicidae), Parasitology plants, Journal of Phytology, vol 1 (3), 2009, 193-
Research , 2016, 839-836. 198.
12. Satria dharma setiawan, Shirtcliffe P ,Sen S, 25. Brain K.R and Turner T.D , The practical evaluation
Chakarabarty R, Sridhar C, Reddy Y, Biplab D , Free of phytopharmaceuticals, Wright- science technical,
radicals, antioxidants, diseases and phytomedicines: 1st Ed 1975, Bristol Britain., 144.
current status and future prospect, Int J Pharm Sci 26. Justin Y and subashri B , Phytochemical Analysis and
Rev Res, vol 102 , 2014, 91100 invitro antioxidant activity from extract of Bacopa
13. Umar A, Makhaik M, Sofowora EA , Medical plants monneri a multipurpose medicinal plant,
and Traditional Medicine in Africa , John Wiley and International journal of pharma and biosciences,
Sons. Inc, 2006 New York, USA.12. ISSN 2016, 0975-6299
14. Michael Samuel, Uddin N, Siddiqui BS, Naqvi SN, 27. Ejekeme CM, Ezeonu CS, Eboatu AN,
Begum S, Tariq RM , New constituents from the Determination of physical and
dried fruit of Piper nigrum Linn., and their larvicidal phytochemicalconstituents of some some tropical
potential against the dengue vector mosquito Aedes timbers indigenous to Niger Delta area of Nigeria ,
aegypti, Phytochem Lett , vol 6 ,2002, 219 23. European scientific journal 2014 ,vol 10 , 247- 270
15. Med J Aust. Amer A, Mehlhor H , Larvicidal effects 28. Ayoola G. A, Coker H .B, Adesegun S.A, Adepoju
of various essential oils against Aedes, Anopheles bello, A Phytochemical screening and anti oxidant
and Culex larvae , Parasitol Res ,vol 99, 2006, 466- activity of some selected medicinal plants for malaria
472. therapy in south western Nigeria, Trop .J. Pharm
16. Pandey A, Mishra R , Antibacterial properties of Res, vol 7, 2008, 1010-1024.
Aegle marmelos leaves, fruits and peels against 29. Harbone J.B, Phytochemical methods. London.
various pathogen , Journal of Pharmaceutical and Chapman and Hall, Ltd., 1973; 49-188
Biomedical Sciences , vol 13, 2011, 1-6. 2 30. Asha verma, Namrata singh, Arvind kumar,
17. Gohil T, Pathak N, Jivani N, Devmurari V, Patel J , phytochemical investigation and thin layer
Treatment with extracts of Eugenia jambolana seed chromatography of Aspsragus racemosus methanolic
and Aegle marmelos leaf extracts prevents leaf extract, International journal of advanced
hyperglycemia and hyperlipidemia in alloxan induced research in pharmaceutical and biosciences , vol(1) ,
diabetic rats , African Journal of Pharmacy and 2013, 15 18 ISSN: 2277-6222.
Pharmacology vol 4 (5), 2010, 270-275. 31. Singh U, Baldi A, Simultaneous, Estimation of
18. Kothari S, Mishra V, Bharat S, Tonpay S, Quercetin and Silymarin: Method Development and
Antimicrobial activity and phytochemical screening Validation, int. J pharm. & boil. Arch, vol 4(3), 2013,
of serial extracts from leaves of Aegle marmelos , 527- 531.

202| International Journal of Pharmaceutical Research | July- Sept 2018 | Vol 10 | Issue 3
 Nagalaxmi et al / Formulation Of Novel Herbal Cream As Mosquito Repellent

32. Yildirim A, Mani A, Kara AA, Determination of extracts, World Journal of Pharmaceutical Sciences,
antioxidant and antimicrobial activities of Rumex vol 6, 2015, ISSN : 2321-3310.
crispus L. extracts, J Agric Food Chem , 49(8), 42. Fang Wang, Yong-Hong Chen, Yu-Jie Zhang , Gui-
2001, 4083-4089. Fang Deng , Zhi-Fei Zou , An-Na Li , Dong-Ping Xu ,
33. Piperine ext Lee EB, Shin KH, Woo W S , Hua-Bin Li , Chemical Components and Bioactivities
Pharmacological study on Piperine.,Archives of of Psidium guajava International Journal of Food
Pharmacal Research, 7(2), 1984, 127132. Nutrition and Safety, vol 5(2), 2014, 98-114 ISSN:
34. Adeniran O. I and Fabiyi E , A cream formulation of 2165-896X.
an effective mosquito repellent: a topical product 43. Zainab saif salef al-hashemi , Mohamad hamzad
from lemongrass oil (Cymbopogon citratus) Stapf, J. hossain, Phytochemical Analysis of Neem Extract,
Nat. Prod. Plant Resour., 2 (2), 2012, 322-327. Volume 48 - Issue 3 , 2016 , 264-274.
35. Curtis C Lines JD, Ijumba J, Callaghan A, Hill N, 44. Vaijayanthi R, Olagunju JA and Thompson DG,
Kar_ imzad MA, The relative efficacy of repellents Phytochemical investigation of Carica papaya , Afr. J.
against mosquito vectors of disease, Med Vet Biotechnol, vol 8(23), 2009, 6677-6682.
Entomol , vol1 ,1987, 109-1 19. 45. Mathivanan N, Muley BP, Khadbadi SS, Banarase NB,
36. Ansaria M.A , Mittalb M.A , Razdana R.K , Sreeharia Phytochemical Constitutions and Pharmacological
U , Larvicidal and mosquito repellent activities of Activities of Medicinal plants - A review, Tropical
Pine (Pinus longifolia, Family: Pinaceae) oil, J Vect Journal of Pharmaceutical Research, vol 8(5), 2015,
Borne Dis, vol 42, 2005, 95 99. 455-465.
37. Dalziel J. M, Maragathavalli S, Brindha S, Kaviyarasi 46. Deepa Ganesh, Hans- Peter Fuehrer,
N. S, Annadurai B & Gangwar S. K. ,Medicinal plants Peter Starzengrüber, Paul Swoboda, Wasif Ali Khan,
A review , International journal of advanced Johannes A. B. Reismann, Milena S. K. Mueller,
biotechnology research , Vol 2(1) ,2012,138-142. Peter Chiba, Harald Noedl, Antiplasmodial activity
38. Sukumar K, Gopinesh VK Kannabiran K , Larvicidal of flavonol Quercetin and its analogues in
effect of guava leaf against Culex quinquefasciatus Plasmodium falciparum: evidence from clinical
mosquito larvae , African journal of biotechnology, isolates in Bangladesh and standardized parasite
vol 6 (3 ) 2007 , 301-317. clones parasitology research, Volume 110,issue 6 ,
39. Wollenweber E and Harborne J , The avonoids: 2012, 2289 2295.
advances in research , Chapman and Hall, London, 47. Scott I.M, Helson B.V, Strunz G.M, Finlay H,
1994,259 335. Sanchez-Vindas P.E, Poveda L, Lyons D.B, Philogene
40. Isman M.B, Botanical Insecticides, deterrents, and B.J.R, and Arnason J.T , Efficacy of Piper nigrum
repellent in modern agriculture and an increasingly (Piperaceae) extract for control of insect defoliators
regulated world , Ann. Rev. Entomol, vol 51, 2006 of forest and ornamental trees, Journal of
,45-66. Entomology, vol 139(4) ,2007, 513-522.
41. Asha,Thirunavukkarasu, Mohamad Sadiq,
Phytochemical screening of Euphorbia hirta linn leaf

203| International Journal of Pharmaceutical Research | July- Sept 2018 | Vol 10 | Issue 3