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FORMULATION AND EVALUATION OF HERBAL MOUTHWASH CONTAINING

NATURAL INGREDIENTS FOR ANTI-MICROBIAL ACTIVITY

Article in World Journal of Pharmacy and Pharmaceutical Sciences · June 2022

DOI: 10.20959/wjpps20226-22344

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 WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
Vijapur et al. World Journal of Pharmacy and Pharmaceutical Sciences
SJIF Impact Factor 7.632

Volume 11, Issue 6, 1820-1828 Research Article ISSN 2278 – 4357

FORMULATION AND EVALUATION OF HERBAL MOUTHWASH

CONTAINING NATURAL INGREDIENTS FOR ANTI-MICROBIAL
ACTIVITY

Praveenkumar V. Vijapur*1, Laxman S. Vijapur1, Pooja Yaragattimath1, Govindraj

Raibagi1, Yogesh Mallapur1 and Bheemareddy Desai1

1
Department of Pharmaceutics, BVVS Hanagal Shri Kumareshwar College of Pharmacy,
Bagalkot, Karnataka, India.

Article Received on
ABSTRACT
10 April 2022, Keeping a decent oral cleanliness supports keeping our general
Revised on 30 April 2022,
Accepted on 20 May 2022 wellbeing in a great shape. The mouth is a fundamental piece of the
DOI: 10.20959/wjpps20226-22344 stomach, and consequently any oral sickness will straightforwardly or
by implication influence the GI plot as well. Neem was used as

*Corresponding Author traditional medicine for several disease including cleaning of oral
Praveenkumar V. Vijapur cavities. Banana peel can act as gentle exfoliator for our teeth. Orange
Department of peels to help whiten teeth. curcumin in turmeric can prevent gingivitis,
Pharmaceutics, BVVS
or gum disease. It helped remove plaque, bacteria, and inflammation
Hanagal Shri Kumareshwar
comparably well with traditional mouthwashes, when used properly.
College of Pharmacy,
Bagalkot, Karnataka, India. Clove have antibacterial and antifungal properties it is widely used in
toothpaste preparations. Cinnamon bark and nutmeg seeds have anti-
inflammatory property. The objective of study is find out the efficacy of these plant extract
against caries inducing microorganisms. further formulating mouthwash which has efficiency
to inhibit the growth of the microorganism.

KEYWORDS: Neem, Banana peel, orange peel, Clove, Cinnamon bark, Nutmeg seeds,
Herbal mouthwash, Antimicrobial.

INTRODUCTION
The world oral health report, 2003, highlighted oral health as an integral and essential
component of general health. Most of the chemical products contains an antiseptic that plays
an important role in controlling plaque accumulation. The vehicle for delivery of chemical

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Vijapur et al. World Journal of Pharmacy and Pharmaceutical Sciences

agents with antiplaque action are toothpaste, mouth washes, spray, irrigators, chewing gums
and varnishes. However, mostly accepted method of delivering the antimicrobial agents after
toothpaste is mouth wash. Mouth wash are an antiseptic solution which is used to reduce the
microbial load in the oral cavity.

Natural mouthwashes may offer significant advantages over the chemical ones. If such
mouthwashes can be formulated which can be easily prepared and used safely by people at
home using natural products, it may leads to improvement in the general dental health of the
population. In this study the various natural ingredients and materials are used. Then the thin
layer chromatography was done to check the quality of materials used. Then the physical
evaluation, ph determination, stability study, thin layer chromatography, antimicrobial study
was studied. By this study it was concluded that the developed herbal mouthwash possess
significant, therapeutically efficacious, suitable vehicle for drug delivery in low cost but
definitely with high potential. Hence there is need for increased usage of herbal preparation
to avoid the adverse effects. This study is an attempt to outline such natural substances,
which may be used as effective mouthwashes.

Almost all chemical mouthwashes contain alcohol and fluoride which is toxic to our body in
overdoses. Hence, most herbal mouthwashes are safe alternative to pregnant women, peoples
with dry mouth, diabetic and to children. The purpose of this study was to determine the
prevalence of mouthwash use and not only the type of mouthwash but quantity of mouthwash
to be taken for use is also important and also this study was performed to evaluate the
efficacy and safety of herbal mouthwash for human medicines.[1]

Various herbal products and their extracts such as Guava, Pomegranate, Neem, Propolis,
Tulsi, Green Tea, Cranberry, Grapefruit etc, have shown significant advantages over the
chemical ones. Natural mouthwashes may offer significant advantages over the chemical
ones. If such mouthwashes can be formulated which can be easily prepared and used safely
by people at home using natural products, it may lead to improvement in the general dental
health of the population.[2]

Herbal mouthwash becomes more popular they work without alcohol, artificial preservatives,
flavor, or colors.[3]

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Vijapur et al. World Journal of Pharmacy and Pharmaceutical Sciences

As it contains natural herbs that have natural cleansing and healing property to teeth and
gums. Many herbal mouthwashes contain herbs with anti-microbial property such as Neem,
Yavanisatva, Nagavalli, Gandha purataila, Pilu, Bibhitaka, Ocimum, Echinacea, Chameli
leaves, etc. Manuka honey, which is rich in flavonoids, increased the glycoprotein production
in the ethanol model of gastric damage. Manuka honey was the most effective antioxidant
and antibacterial honey compared to both acacia honey and wild carrot honey, possibly
because of its high phenol content.[4] Guava (Psidium guajava) as a mouthwash for swollen
gums and ulceration of the mouth and also for bleeding gums.[5] Extracts from neem inhibit
the growth of S. mutans and used in the treatment of periodontitis.[6] Aloe vera mouthwash,
mixture of Terminalia chebula and cinnamon, mixture of Staphysagria, Chamomilla,
Echinacea, Plantago, Ocimum, and cistus extracts, mouthwashes with turmeric, neem and
triphala, mixture of Acacia Arabica, Punica granatum, Chameli leaves, Glycyrrhiza glabra
and neem shows significant reduction of plaque indices and gingival scores comparatively to
chlorhexidine mouthwash and can be a better alternative to chlorhexidine mouthwash.[7-9]

MATERIALS AND METHODS

MATERIALS
Neem leaves, banana peel, orange peel, turmeric root, clove, cinnamon bark and nutmug
seeds were collected from market in Bagalkot Double distilled water was used throughout the
study. Mueller Hilton agar was purchased from HIMEDIA.

METHODOLOGY
Preparations of aqueous extract from different plant sources
The collected plant sources were washed with double distilled water 2-3 times. The washed
plant sources were dried at room temperature in shade. After drying they were crushed and
the powder was stored for further use. 10 grams of orange peel and clove powder were
weighed and were taken in 250 ml conical flask which was containing 100 ml of double
distilled water. The flask was closed with the aluminium foil and flask was kept in dark place
undisturbed for 24 hours. Later the mixture was filtered through muslin cloth and again
filtrate was filtered using Whatmann filter paper grade no.1. The extract was stored in
ambered colour glass container for further use at 4oC. 10 grams of cinnamon, turmeric, neem
and nutmug powder were weighed and were taken in 250 ml Erlenmeyer flask which was
containing 100 ml of 70% ethanol. The flask was closed with the aluminium foil and flask
was kept in dark place undisturbed for 24 hours. Later the mixture was filtered through

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muslin cloth and again filtrate was filtered using Whatmann filter paper grade no.1. The
extract was stored in ambered colour glass container for further use at 4oC. 10 grams of
banana powder was weighed and were taken in 250 ml Erlenmeyer flask which was
containing 100 ml of 70% iso-propyl alcohol. The flask was closed with the aluminium foil
and flask was kept in dark place undisturbed for 24 hours. Later the mixture was filtered
through muslin cloth and again filtrate was filtered using Whatmann filter paper grade no.1.
The extract was stored in ambered colour glass container for further use at 4oC. All the above
obtained extracts were further dried by keeping it in hot air oven for 3 days at 40oC. The
dried powders were stored in closed container and were kept in cool place for further use.

FORMULATION OF HERBAL MOUTHWASH

Required quantity of neem, turmeric, cinnamon, banana peel and nutmug powders were
weighed and mixed with sufficient quantity of 70% ethanol and menthol in a beaker, mixed
properly until all the powders are dissolved in the solvent. Propylene glycol was added to the
above mixture for the complete solubility (solution 1). In another beaker required quantity of
clove and orange peel powders were mixed with required quantity of double distilled water
and to this solution sodium saccharin was added and mixed properly (solution 2). Lastly
beaker containing solution 2 was added to beaker containing solution 1 which was stirred
continuously to get a solution, to this solution sodium benzoate was added as preservative
and sodium lauryl sulfate was added as a foaming agent.

Table No. 01: Composition of herbal mouthwash formulations with different plant
extracts.
F1 F2
Sr.No Ingredients (50 ml)
(gm) (gm)
1 Neem 0.1 0.1
2 Banana peel 0.1 0.1
3 Orange peel 0.25 0.25
4 Clove 0.25 0.25
5 Cinnamon 0.25 0.25
6 Turmeric 0.1 0.1
7 Nutmug -- 0.1
8 Sodium Saccharin 0.1 0.1
9 Menthol 0.1 0.1
10 Propylene glycol 10 10
11 Sodium Lauryl Sulfate 0.5 0.5
12 Sodium Benzoate 0.2 0.2
13 Ethanol 2 2
14 Double distilled water qs qs

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CHARECTERIZATION OF HERBAL MOUTHWASH

Physical evaluation
Physical parameter such as color, odour, taste and consistency were examined by physical
examination.[1]

pH determination
The pH of the herbal mouthwash formulations was determined by pH meter by taking
mouthwash in beaker and pH of the formulations were recorded.[10]

Determination the Viscosity

Ostwald viscometer was cleaned by the water and ethanol and dried. Mouth wash was placed
in viscometer and pull it by pipette until the small bulb is full. Let the liquid to flow through
the capillary tube with run time when the liquid reaches the mark shown on the viscometer
and then stopped time when the liquid reaches the bottom mark and viscosity was recorded
by the below formula.[11]

η1 is viscosity of liquid 1.
η2 is viscosity of water 0.891 poise.
t1 flow time of liquid 1.
t2 flow time of water.
d1 density of liquid 1.
d2 density of water 0.997 g/cm3.

Anti-microbial activity study

Antibacterial activity of the herbal mouthwash was determined using a well diffusion
method. Standard strains of Escherichia coli ATCC25922 and Staphylococcus aureus
ATCC29213 were used and a disk of Amikacin was used as a control or for comparison. The
organisms were inoculated in peptone water and the turbidity obtained was matched with 0.5
McFarland standards. This was inoculated as a lawn culture on Muller Hinton agar. A sterile
filter paper disk impregnated -with the herbal mouthwash and a commercially available
amikacin disk were placed on it. Both the plates were incubated at 37℃ for 24 hours. After
24 hours, the plates were examined for the zone of inhibition which was measured in
millimeters.[9]

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RESULTS
Physical Parameters
Physical parameter such as color, odor, taste and consistency were examined by physical
examination.

pH Determination
The pH of the F1 formulation was found to be 6.1 and F2 was found to be 6.3 as the oral
cavity is having pH around 6.4, this pH range of the formulation is suitable for oral disorder.

Viscosity determination by Ostwald viscometer

Viscosity determination was done by using Ostwald viscometer. The viscosity of F1
formulation was found to be 1.425cps and relative viscosity was found to be 1.70. The
viscosity of F2 formulation was found to be 1.25cps and relative viscosity was found to be
1.46.

Anti-microbial activity
In this work the formulated mouth washes have been investigated for antimicrobial activity
against E. coli and S. Aureus (Fig-1,2) the result showed that the formulated herbal mouth
washes exhibited antimicrobial activity. While the F1 formulation shows less antimicrobial
activity than F2 formulation shown in (Table No. 02). The formulated mouth washes are only
effective against gram positive organism as compared to gram negative organism.

Table No. 02: Antimicrobial activity of F1 and F2

Sl No. Organisms F1 (mm) F2 (mm)
1 Staphylococcus aureus ATCC29213 18 20
2 Escherichia coli ATCC25922 No Zone No Zone

Fig No:- 01 Antimicrobial activity of Staphylococcus aureus.

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Fig No:- 02 Antimicrobial activity of E. Col.

DISCUSSION
The current investigation focused on the formulation of mouth wash comprising of different
plant extract from clove, neem, banana peel, turmeric, orange peel, cinnamon and nutmeg
which was effective against Staphylococcus aureus and E.coli, a common pathogenic bacteria
causing dental caries. The study is important since it focuses on developing herbal product
which can be detrimental to pathogen growth, without having any adverse effect on human
health and hence can be better alternative than the commercially available chemical solutions
and formulations.

The pH of prepared herbal mouthwash was measured by using pH paper. The pH of the F1
formulation was found to be 6.1 and F2 was found to be 6.3 as the oral cavity is having an
acidic ph around 5.5, this ph range of the formulation is suitable for oral disorder.

Viscosity determination was done by using Ostwald viscometer. The viscosity of F1

formulation was found to be 1.425cps and relative viscosity was found to be 1.70. The
viscosity of F2 formulation was found to be 1.25cps and relative viscosity was found to be
1.46.

In this work the formulated mouth washes have been investigated for antimicrobial activity
against E. coli and S. Aureus. the result showed that the formulated herbal moth washes
exhibited antimicrobial activity. While the F1 formulation shows less antimicrobial activity
than F2 formulation.

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Vijapur et al. World Journal of Pharmacy and Pharmaceutical Sciences

CONCLUSIONS
Herbal mouthwash formulations can be prepared by using different plant sources with
varying concentrations. The pH of the herbal mouthwash formulations was in the range of 6.1
to 6.3 which lies in the normal pH range of the oral cavity. All the herbal mouthwash
formulations showed good viscosity and were in the standard range. Antimicrobial activity of
both the formulation was done by well diffusion method and found that the prepared
formulation was effective against gram positive microorganism as compared to gram
negative microorganism.

ACKNOWLEDGEMENT
The authors would like to thank HSK, college of pharmacy Bagalkot for providing all the
needed facilities to perform the study.

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