formulation and evaluation of herbal mouthwash

Shambharkar et al. World Journal of Pharmaceutical Research

www.wjpr.net │ Vol 10, Issue 9, 2021. │ ISO 9001:2015 Certified Journal │

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FORMULATION AND EVALUATION OF HERBAL MOUTHWASH

Shivani B. Shambharkar1* and Vinod M. Thakare

2

1In the Faculty of Science & Technology,

2Head of Quality Assurance (Associate Professor),

Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur.

1. ABSTRACT

The importance of mouth and teeth cleanliness has been recognized

from the earliest days of civilization to the 21st century. Patients and

oral health practitioners are faced with a multitude of mouthwash

products containing many different active and inactive ingredients.

Making informed decisions as to the suitability of a particular product

for a particular patient can be complex task. Although many popular

herbal products have helped to control dental plaque and gingivitis,

they have been used for a short time and only as an adjunct to other

oral hygiene measures such as brushing and flossing. 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 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.

2. INTRODUCTION

Across the world, oral health is becoming a major concern. The world oral health report,

World Journal of Pharmaceutical Research SJIF Impact Factor 8.084

Volume 10, Issue 9, 775-791. Research Article ISSN 2277– 7105

*Corresponding Author

Shivani B. Shambharkar

In the Faculty of Science &

Technology, Rashtrasant

Tukadoji Maharaj Nagpur

University, Nagpur.

Article Received on

28 May 2021,

Revised on 18 June 2021,

Accepted on 08 July 2021

DOI: 10.20959/wjpr20219-20782



776

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 vehical for delivery of chemical 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.

Mouth washes are liquids which contains anti-inflammatory. Anti-microbial and analgesic

action. There are two types of mouthwashes- chemical and herbals Herbal mouthwash

contains a natural ingredients called phytochemicals that contains desired anti-microbial and

anti-inflammatory effects. Herbal mouthwash becomes more popular they work without

alcohol, artificial preservatives, flavours and colors. 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, yavani satva, nagavali, gandhapurataila,

pilu, bibhitaka, ocimum, Echinacea, chameli leaves, etc. some of the herbs that are used in

mouthwashes are clove, which is traditionally used for oral health because of their antiseptic,

antibacterial and antiviral property, peppermint which gives cooling effect to the mouth.

Natural Herbs such as Triphala, Tulsi, Neem, Clove oil, Pudina and many others are used as

single or in combination have been Scientifically Proven to be Safe and Effective Medicine

against Oral Health Problems such as Bleeding Gums, Mouth Ulcers, and Preventing Tooth

Decay without side effects.

Almost all chemical mouthwashes contains 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 chindren. 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.

3. Literature review

1) Rathore et.al (2018):- Worked on antimicrobial effect of mouthwash in patient

undergoing orthodontic treatment and concluded that freshclor mouthwash showed the

maximum potential for the control of pathogenic organism and prevention of gingivitis

and bacterial plaque inhibition.



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2) Renuka et.al. (2017):- Worked on comparision of herbal mouthwashes with

chlorhexidine mouthwash and concluded that besides the disadvantage, chlorhexidine

mouthwash plays effective role during dental treatment on short term usage. Herbal

mouthwash are suitable for maintaining good oral prophylaxis. Many programs have to

be conducted to make them aware about mouthwashes in their oral hygiene.

3) Simiyu N. benjamin et.al (2016):- Worked on knowledge, attitude and use of

mouthwash among dental and medical students of the university of Nairobi. concluded

that in general, dental student have adequate knowledge about mouthwashes. They

practiced the use of mouthwash more than medical students.

4) Amit parashar (2015):- Studied mouthwashs with various different condition which can

vary from malodour to periodontal disease to treatment of secondary infections. The main

objective of the study was to help the oral health care professionals to make the correct

selection of mouthwash while dealing with different condition of oral Cavity. They

concluded that mouthwashes are medicated solutions used for gargling and rinsing the

mouth.

5) P.F. waghmare et.al. (2011):- worked on comparative evaluation of turmeric and

chlorhexidin gluconate mouthwash in prevention of plaque formation and gingivitis : a

clinical and microbiological study. Concluded that chlorhexidine gluconate as well as

effectively used as an adjunct to mechanical plaque control method in prevention of

plaque and gingivitis.

6) B. B. oluremi et.al (2010):- Have studied that the procedure of evaluated for antibacterial

activity of herbal mouthwash with their activity. The cup-plate method was used. The

zones of inhibition produced by the mouthwash against the bacterial isolates were

measured to determined degree of susceptibility. They concluded that preservatives in

mouthwash formulations should be done with caution as this may interfere with

mouthwash activity.



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4. Drug Profile

Biological source

Turmeric consist of dried, as well as fresh rhizomes of plant known as curcuma longa linn

belonging to family zingiberaceae.

Chemical constituents

Turmeric contains 3-6% Polyphenolic compounds, collectively known as Curcuminoids as a

coloring matter. The curcumanoid contains curcumine-I, Curcumine-II, curcumine-III.

Turmeric contains about 5% volatile oil. Their Volatile oil contain sesquiterpenes, alcohol

and ketone and monoterpene Example, zingiberone, turmerone, arturmerone, alcohopl-p-

tolylmethyl, Carbenol, borneole etc. It also contains arabinose, fructose, glucose, and Starch

grains. Others are Cymene, Tumeron, Isdemethoxy curcumin, Demethoxy curcumin, Diaryl

Heptanoids.

Parts used

Rhizomes, tubers.

Uses

Having Anti-plaque, Anti-inflammatory and Anti-micobial property.

Whiten teeth.

Healthy gums.



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Releive the toothach.

Keep your breath fresh.

It is used as stomachic, carminative, aromatic, antiperiodic, blood purifier and stimulant.

It is also an anti-arthritic, anti-inflammatory, anti-lipidemic, carminative and digestant

and anti-fertility.

It is also used in menstrual pain, liver diseases and to produce choleritic and chologogue

action.

Turmeric is used in curry powders, sauces and paper impregnated with its tincture is used

for the borate and boric acid detection.

It is employed as colouring agent for formulation like ointments and creams. Ethanolic

extracts of curcuma longa shows anti-ulcerogenic property. Rhizome extract has anti-

diabetic action. Curcuma is used in conjunctivitis.

A recent study involving mice has shown that turmeric slows the spread of breast cancer

in to lungs and other body parts. Turmeric also enhances the effect of taxol in reducing

metastasis of breast cancer.

The rhizomes is well known for its anti-gastric ulcer and cholagogic properties. It is

prescribed in the therapy of gastric and duodenal ulcer, hepatitis, jaundice, pain in the

extrimities, boils and impetigo. It is also used as a poultice for wounds.

Toxicology

A clastogenic potential of curcuma longa in experimental rats in invivo conditions has been

evaluated. A single acute dose treatment 500 mg/kg body weight could not significantly

induce micronucleated polychromatic erythrocytes but caused considerably higher

chromosomal abberations.

Adverse Reaction and Interaction

Fetal effects are unknown and use during pregnancy. Other than as food spice, is probably

best avoided. Prolonged use has been associated with gastrointestinal upset and may increase

the risk of peptic ulcer. Use in patients with billiary obstructions or gallstones is

contraindicated. There is a potential for interaction with NSAIDs and platelet inhibiting drugs

because of its effects on prostaglandin synthesis.



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TULSI :-

Taxonomical Classification

Kingdom Plantae

Division Magnoliophyta

Class Magnoliopsida

Order Lamiales

Family Labiatae

Genus Ocimum

Species O.Tenuiflorum

Scientific Name Ocimum Sanctum

Vernacular Name

Hindi Tulsi

English Sweet basil

Sanskrit VishwaTulsi

Marathi Tukmaria

Latin Ocimum

basillicum

Biological source

Tulsi consist of the fresh & dried leaves of Ocimum sanctum L. and Ocimum basilicum L.

belonging to family Labiatae.

Parts used

Leaves, Seeds and Roots.

Chemical constituents

Volatile Oil-0.8%

i. Eugenol, nerol, eugenol methyl ether.

ii. Caryophyllene, terpinene-4-ol-decyladehyde

iii. Camphor and carvacrol

iv. Essential oils, ascorbic acid, carotene, calcium, phosphorus and insoluble oxalates.

v. It also contains terpenes, mucilage, fixed oil and fatty acids.



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Uses

Tulsi is a small plant, sub-shrub which has multiple uses.ayurveda mentions the importance

of medicinal uses of it.

1. The leaves are quite effective for the ulcer and infections in the mouth. A few leaves

chewed will cure these conditions.

2. The herb is useful in teeth disorders.

3. Its leaves, dried in the sun and powdered, can be used for brushing teeth.

4. It can also be mixed with mustered oil to make a paste and used as toothpaste.

5. These is very good for maintaining dental health counteracting bad breath and for

massaging the gums.

6. It is also used in pyorrhea and other gum, disorders.

7. The anti-inflammatory and anti-infectious properties of tulsi make it a powerful treatment

for gum disease.

8. It is used to treat bronchitis, cough, earache, gastric disorders, hepatic disorders,

inflammation, otitis, otorrhoea, syphilis, malarial fever, cardiac debility, gonorrhea,

palpitation, urino-genital complaints.

Peppermint :-

Taxonomical

Classification

Kingdom Plantae

Order Lamiales

Family Lamiaceae

Genus Mentha

Species M Piperita

Vernacular Name

Hindi Pudina

Tamil Pudhinna

Telugu Pudhina Koora

Marathi Pudina

Latin Mentha piperita

Biological source

It obtained from fresh leaves of Mentha piperita. belonging to family Labiatae.



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Parts used

Leaves.

Chemical constituent

Peppermint oil mainly contains menthol (about 70%). It also contains menthone, menthyl

acetate, and other terpene derivatives like cineole, pinene, isopulegone, camphene, limonene,

zasmone, menthofurone, menthyl isovalerate.

Uses

i. The dried flowering tops are used to prepare beverages like peppermint tea and in the

formulation of liqueurs and bitter.

ii. The current use of peppermint is mainly for colic and irritable bowl syndrome.

iii. Peppermint oil is used as carminatives, aromatic, stimulant and flavouring agent.

iv. It is widely used as an antiseptic in various preparations and in mouth freshners.

v. It is also used as antipruritic and counter irritant over skin and mucous membrane.

vi. Helps in removing the bad breath.

Adverse Effect and Interaction

Peppermint may relax the lower esophageal sphincter and therefore is contraindicated or

patients with reflux diseases. Peppermint should probably not be used in patients with billiary

tract obstruction, cholecystitis or severe liver damage. Peppermint oil has been shown to be a

moderately potent inhibitor of cytochrome P450 3A4, raising the possibility thatis li ke

grapefruit, juice also may inhibit the metabolism of felodipine.



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5. Aim, Objectives and Need of research

5.1:- Aim

To Formulate and Evaluate Herbal Mouthwash.

5.2:- Objective

The experiment will be carried out to achieve the following

Objectives

1. To determine the antimicrobial activity.

2. To evaluate mouthwash for its consistency.

3. To develop a formulation of herbal mouthwash.

5.3:- Need of research

1. Mouthwash act on oral pathogen and also producing less side effects as compared to

synthetic herbal products.

2. Herbal mouthwash are demanded because they instantly relieves the pain.

3. Herbal mouthwash can help to prevent from gingivitis and various infectious diseases.

6. Plan of work

Literature Survey

Collection of Plant Material

Extraction of Herbal Drugs

Selection of Excipients for Formulation

Development of Formulation

Evaluation of Prepared Formulation

7. Research methodology

1) Collection, Procurement and Extraction of tulsi leaves

The dried leaves of ocimum santum are collected and coarsly cut into small pieces. The dried

coarse piece was extracted to obtain a tulsi extract. The successive extraction was carried out

by maceration method. The extract were filtered through muslin cloth and subsequently the

filtrate was pass through filter paper and then packed in close container.



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2) Collection, Procurement & Extraction of nagarmotha powder

Nagarmotha powder was collected from local shop. Accurate quantity of powder was

weighed and mix with desired quantity of alcohol and stirred well and kept it for 24 hours,

the extract was filter and collected in a container.

3) Method of preparation of herbal mouthwash

a. Material used

Tulsi Extract, Cinnamon Oil, Clove Oil, Neem Oil, Turmeric Oil, Peppermint Oil,

Nagarmotha Extract, SLS, PEG 400, Distilled Water.

b. Formulation of herbal mouthwash

Dissolve the Tulsi extract 2.0ml and Nagarmotha extract 1.0ml in a separate container

(no.1) and add distilled water to obtain a clear solution and well shake it.

In a separate container(no.2), add 2 drops of peppermint oil, 2 drops of clove oil, 2 drops

of neem oil,2 drops of cinnamon oil and 1 drop of turmeric oil and mix it properly. Then



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slowly add polyethylene glycol 400 and slowly add SLS, stirr well to obtain a clear

solution (table:1).

The 1st solution is slowly added in the 2

nd mixture, stirr well and slowly add a distilled

water to make a volume upto 100ml and then shake it continuosly and clear liquid has a

fresh mint taste.

The herbal mouthwash was prepared by the formula given in table 1.

Sr. no. Ingredient Function Percentage (%)

1 Neem Oil Antimicrobial 0.2%

2 Cinnamon oil Bactericidal 0.2%

3 Clove oil Analgesic, anti-

inflammatory 0.2%

4 peppermint oil Freshner, antibacterial,

antifungal 0.2%

5 Turmeric oil Antimicrobial, anti-

inflammatory 0.1%

6 Tulsi extract

(ocimum santum)

Anti-inflammatory,

antioxidant 2.0%

7 Nagarmotha extract

(cyperus scariosus) Flavouring agent 1.0%

8 PEG400 Solubilizing agent 16.00%

9 SLS Foaming agent 00.20%

10 Distilled water vehicle q.s.

Evaluation test

1. Physical evaluation:- Physical parameter such as color, odour, taste and consistency

were examined by visual examination.

2. ph determination:- The ph of prepared herbal mouthwash was measured by using digital

ph meter. The ph meter was calibrated using standard buffer solution about 1ml of

mouthwash was weighed and dissolved in 50ml of distilled water and its ph was measure.

3. Test for microbial growth in formulated mouthwash:- The formulated mouthwash was

inoculated in the plate of agar media by streak plate method and a control was prepared.

The plates were placed in the incubator and are incubated at 37⁰c for 24 hours. After the

incubation period plate were taken out and checked for microbial growth by comparing it

with the control.

4. Thin layer chromatography:- Thin Layer Chromatography is a technique used to isolate

non-volatile mixtures. The experiment is conducted on a sheet of aluminium foil, plastic,

or glass which is coated with a thin layer of adsorbent material. The material usually used

is aluminium oxide, cellulose, or silica gel.

https://byjus.com/chemistry/al2o3/



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On completion of the separation, each component appears as spots separated vertically. Each

spot has a retention factor (Rf) expressed as:

Rf = dist. travelled by sample / dist. travelled by solvent

The factors affecting retardation factor are the solvent system, amount of material spotted,

absorbent and temperature. TLC is one of the fastest, least expensive, simplest and easiest

chromatography technique.

A small drop of the mixture is placed on the base line of the thin layer plate, and similar small

spots of the known amino acids are placed alongside it. The plate is then stood in a suitable

solvent and left to develop as before.

8. RESULTS AND DISCUSSION

Physical parameter such as color, odor, taste and consistency were examined by visual

examination.

The ph of prepared herbal mouthwash was measured by using digital ph meter.

The ph of the formulation was found to be 6.1, as the skin is having an acidic ph around

5.5, this ph range of the formulation is suitable for oral disorder.



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Thin layer chromatography

R. no. Oils Solvent

system

Rf value Tlc plates

1 Clove oil Hexane :

acetone

9 : 1

0.43, 0.63

2 Cinnamon oil Toluene :

ethyl acetate

9.3 : 0.7

0.45, 0.66

3 Peppermint oil Ethyl acetate :

hexane

0.5 : 9.5

0.52

4 Neem oil methylene

chloride :

methanol

9.9 : 0.1

0.16 , 0.42 ,

0.72 , 0.95



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5 Turmeric oil Chloroform :

ethanol :

glacial acetic

acid

9.4 : 0.5 : 0.1

0.67

Stability study were done with open and close container. Here, by subjecting the product

to room temperature for one month.

This mouthwash is a purely herbal prepared without addition of alcohol. The formulation

undertaken stability studies for physical and chemical change (table: 2).

The test of stability study of herbal mouthwash tabale 2.

Sr. no. Temperature (⁰c) Evaluation

parameter

Observation (month) 0 1

1. 3-5⁰c Visual appearance

Phase separation

Homogeneity

Light brown Light brown

Nil Nil

Good Good

2. Room

temperature25⁰c

(RH=60%)

Visual appearance

Phase separation

homogeneity


Nil Nil

Good Good

3. 40⁰c (RH=75%) Visual appearance

Phase separation

homogeneity


Nil Nil

Good Good

The formulation was free from microbes as they have not produced any microbial growth

when they get inoculated in the agar medium.

The antimicrobial activity was evaluated by streak plate method for different concentration of

mouthwash. (Table: 3)

Herbal mouthwash mask bad odor and produce a pleasing flavor. Herbal mouthwash with

therapeutic agents like anti-microbials, however may be effective for some long term control

of bad odor.



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Alcohol consumption as well as alcohol and tobacco use are known risk factor for head and

neck cancers. When used in mouthwashes antimicrobial ingredient like Neem, clove and

other essential plant extracts have been found to reduce plaque and gingivitis.

9. CONCLUSION

The data presented in 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. This study should be completed with more investigations

and studies, to explore the product in long term follow-up and laboratory tests to improve all

the effects and side effects of the new products, since it will be used as medical product.

Herbal mouthwash preparation have potent action and minimal side effects when compared

with that of the other marketed mouthwashes, hence there is need for increased usage of

herbal preparation to avoid the adverse effects. Thus there is a need to creat awareness among

prescibers and public about the use of herbal mouth washes and many other such studies

should be encourage.



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10. ACKNOWLEDGEMENT

I would like to commence this acknowledgement memorandum on a pious note by thanking the

Almighty for making this entire mission a triumphant experience.

I am also deeply gratified by my parents and family members, whose blessings,

ideals & values have been a guiding light on every step of this competitive journey.

I express my most cordial and humble thanks to my eminent guide respected Dr. Vinod M.

Thakare (Associate Professor), Dadasaheb Balpande College of Pharmacy, Nagpur, for his

meticulous guidance, which exonerated me to consummate the small work assigned to me. His

inspirations, profound knowledge rectitude is inexplicable in few words. With small interaction

with his during the course of dissertation work made me advertent and prudent about research

work.

I express my sincere thanks to Dr. Amol G. Warokar (Associate Professor) for his help and

encouragement throughout the course of my study and dissertation. Without his critical

evaluation, this thesis would not have become a reality.

I am sincerely thankful to Mr. Manoj V. Balpande (President) and Dr. Mrs .Ujwala N.

Mahajan (Principal). Dadasaheb Balpande College of Pharmacy, Nagpur, for providing good

infra-structure and adequate facilities.

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