A PROJECT REPORT

ON

Formulation and Evaluation of

Herbal Mouthwas
SUBMITTED

TO

Dr. POOJA

Dr. A.P.J. ABDUL KALAM TECHNICAL UNIVERSITY

TOWARDS PARTIAL FULFILMENT FOR THE DEGREE OF

BACHELOR OF PHARMACY

B. PHARM VII SEMESTER

PRACTICE SCHOOL (BP 706 PS)

SUBMITTED BY SUBMITTED TO

NAME- ASHUTOSH KASHYAP Dr. POOJA

ROLL NO.-2003010500011

SAGAR INSTITUTE OF TECHNOLOGY & MANAGEMENT,

DEPARTMENT OF PHARMACY, BARABANKI – 225001

SESSION-2023-24

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 ACKNOWLWDGEMENT

At the outset of this report I feel great pleasure in rendering my deep sense of
gratitude to my Administrator Director, Director and Head of Department, who
has been an inspirational source of my project report.

I am highly indebted to "Mr. Sunil Jhunjhunwala" (Administrative Director)

for their guidance and constant supervision as well as for providing necessary
information and for his most inspiring guidance and interest which enable me to
complete and present this project report. I am thankful to my Guide Dr. Pooja
(Director). I am also thankful to (Head of Department) Mrs. Sarita Verma
(Associated Professor).

I would like to express my gratitude towards my parents and member of Sagar

Institute of Technology & Management, Department of Pharmacy,
Barabanki (U.P) for their kind co- cooperation and encouragement. My thanks
and appreciation also goes to the people who have willingly helped me out with
their abilities.

Thank you!
Student's Name – Ashutosh Kashyap

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 CERTIFICATE

This is to certify that ASHUTOSH KASHYAP have submitted a Practice

School Project Report on “Formulation and Evaluation of Herbal
Mouthwash " domain, in the partial fulfilment of the requirement for the
Bachelor of Pharmacy to Sagar Institute of Technology& Management,
Department of Pharmacy, Barabanki (Uttar Pradesh) with true and honest
observations. He has completed this project under my supervision and his report
is satisfactory.

Faculty Name – Dr. Pooja

Department–Pharmacy
Institute Name – Sagar Institute of Technology & Management, Department of
Pharmacy, Barabanki (U. P.)

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 DECLARATION

I hereby declare that the information incorporated in the project report is a

bonafide and genuine project report, embodies my own work under guidance and
suggestion and received from my supervisor
Dr. Pooja , Sagar Institute of technology & Management, Department of
Pharmacy , Barabanki (U. P.).

Signature:
Name: Ashutosh Kashyap
Roll No. 2003010500011
Date: -18/12/2023

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 INDEX

S.No CONTENT PAGE . NO

1 Introduction 7

2 Oral disorder 9

3 Types of herbal mouthwash 10

4 Ingredient used in herbal 11

mouthwash
5 Preparation of herbal 15
mouthwash
6 Evaluation 16

7 Result and discussion 18

8 References 23

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 Abstract

Herbal mouthwashes are mouthwashes which are prepared from natural plant
extracts. The use of Herbal mouthwash has grown advantage over chemical
mouthwashes due to their non-irritant and Non-staining properties . The natural
extracts present in these herbal mouthwashes are obtained from various plant
leaves, root, seeds and various tree oils. They have very minimal or no side
effects and they are less harmful. Phytotherapeutic plant extracts and essential
oils are used to create and produce herbal mouthwashes, which contain a variety
of active ingredients such as tannins, and sterols. Herbal mouthwash is used to
promote better oral hygiene. It aids in reducing tooth plaque. It is applicable to
gum diseases. Used to eliminate bacteria in the mouth.

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 INTRODUCTION
Mouthwash, is an aqueous solution usually used to remove plaque, is held in the
mouth and swished about by the pre-oral muscle to get rid of oral infections. An
active approach is taken by herbal medicine. Since there haven’t been any side
effects related with their use documented to date, this natural herb’s main benefit
is that. Other than this, there is no sugar or alcohol in any herbal mouthwash.
These items pose a problem because the bacteria that cause halitosis and bad
breath like to feed on them and create the by products that lead to halitosis. Using
herbal mouthwash to avoid hazardous elements is therefore a positive start.
Plaque-induced gingivitis, a highly widespread periodontal disease, is commonly
seen in dental practise. By using a number of techniques that raise oral hygiene
standards, plaque accumulation can be prevented and controlled. The mechanical
removal of dental plaque using tooth brushing, dental floss, tooth cleaning sticks,
oral irrigators, and/or professional scaling and polishing are some examples of
this. To control plaque formation, mechanical means alone might not be
sufficient in all cases. Use of antimicrobial mouthwashes in conjunction with
mechanical oral hygiene techniques is strongly advised in such circumstances.
Several well-known herbal products have assisted in controlling. Mouthwashes
are often prescribed in dentistry for prevention and treatment of several oral
conditions. This has now called for a newer age of mouth washes but is the new
age mouth washes at par with the gold standard or even better than them this
study investigates. Herbal medicines, derived from botanical sources, have been
applied in dentistry for a long history to inhibit microorganisms, reduce
inflammation, soothe irritation, and relieve pain It has been recently reported that
a considerable number of herbal mouthwashes have achieved encouraging results
in plaque and gingivitis control Herbal mouthwashes are designed and prepared
with extracts and essential oils from phytotherapeutic plants, containing a
mixture of active agents such as catechins, tannins, and sterols . The mixture of
natural compounds inside the herb- or plant-derived substances usually performs
gentle remedial effects. Compared with the antimicrobial mechanisms by
synthetic chemicals, herbal mouthwashes can have additional anti-inflammatory
and antioxidant properties, which could further benefit gingival health. (2)(3)

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Oral diseas;
1. Dental Caries:
Caries is the most typical oral infection and illness. A persistent, contagious
illness called caries is brought on by bacteria that consume sugar to generate
an acidic environment that erodes teeth. This process causes holes (cavities) in
the tooth’s structure over time. Streptococcus mutants is the main bacterium
implicated, but the disease may be caused by the breakdown of a complex bio
film on the teeth rather than an abundance of one particular species. Saliva and
fluoride are protective elements. Risk elements Caries risk factors are
multifaceted and include both socioeconomic and physical variables. Adult
caries risk factors include: Previous caries High oral bacterial numbers,
especially S mutans .Insufficient use of fluoridated toothpaste and flossing, as
well as insufficient exposure to fluoride Consumption of sugary foods on a
regular basis,(4)(5)

2.Candidiasis:
A candida species infection of the oral mucosa is known as candidiasis.
Candida albicans is the type of candida that affects people most frequently.
Risk elements Species of Candida are typical dweller of the digestive system.
Oral candidal infections are more common among immuno compromised
people, such as the elderly, young children, HIV-positive people, cancer
patients, diabetics, and people with glucose intolerance. People who take
certain treatments, such as chemotherapy, inhaled steroids, broad-spectrum
antibiotics (which alter the body’s usual defensive flora), and antibacterial
therapy are more susceptible. Additionally, dentures may develop a Candida
infection, leaving the surrounding skin erythematous rather than white.

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3.Gingivitis:
An reversible form of gingival inflammation is gingivitis. A gentle form of
periodontal disease, that. There are three classifications: plaque-induced, non-
plaque-induced, and systemic diseases and medication-induced gingivitis.
Poor dental hygiene/plaque formation, primary or secondary tooth emergence,
and dental equipment (braces, dentures) are risk factors for gingivitis.
Crowded teeth or malocclusions, poor dental restorations, uncontrolled
diabetes mellitus, and smoking prevalence When all forms and causes are
taken into account, gingivitis affects up to 50% of youngsters and up to 90%
of adults. The high prevalence of gingivitis during pregnancy is caused by
hormonal changes. Menarche, menstruation, and usage of contraceptives are a
few more hormonal changes in women that can increase the prevalence of
gingivitis.

4. Mouth Ulcer :
Mouth ulcers are small sores that form on gums, lips, inner cheeks or palate
(roof of mouth). They can be triggered by several different factors, including
minor injuries, hormonal changes and emotional stress. Mouth ulcers aren’t
contagious and they go away on their own but there are treatments to help ease
pain and discomfort .

Types of Mouthwash : Guide to Mouthwash

There are several types of mouthwash which all perform a particular function.
There are fluoride mouthwashes which help to strengthen your teeth,
antiseptic mouthwashes which deal with tooth decay and hide bad breath and

herbal mouthwashes which do not contain alcohol. To re-iterate: the types of

mouthwash available include:
1) Fluoride
2) Cosmetic

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3) Antiseptic
4) Natural (herbal)
5) Total care Herbal Mouthwash
Herbal mouthwashes can be used in addition to different oral hygiene
techniques like flossing and teeth brushing. They can be utilised in supportive
periodontal therapy due to their effective anti-inflammatory and anti-plaque
characteristics, which have been demonstrated. Alcohol, artificial flavours,
colours, or preservatives are not present. Hence Because of the additional
benefits offered by herbal preparations, herbal mouthwashes might be thought
of as a substitute for chemical mouthwashes in maintaining oral hygiene. A
precise diagnosis of the oral condition and in-depth product knowledge are
prerequisites for using mouthwashes. The choice must take into account
elements like the patient’s oral health, disease risk, the effectiveness and
safety of the mouthwash, and the patient’s capacity for practising excellent
oral hygiene.
• Halitosis
• Mucositis
• Periodontal Diseases
• Gum disease
• Xerostomia
• To clean septic sockets
• Vincent’s angina
• To control plaque
• To relieve pain
• To effectively deliver fluoride in order to prevent dental caries
• Reduce inflammation etc.

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1 : Liquorice

Biological Source :
Liquorice consists of subterranean peeled and unpeeled stolons, roots and
subterranean stems of Glycyrrhiza glabra Linn, and other species
of Glycytrhiza,
Fmaily
Leguminosae
Chemical constituents
Licorice extract contains sugars, starch, bitters, resins, essential oils,
tannins, inorganic salts, and low levels of nitrogenous constituents such as
proteins, acids, individual amino and nucleic acids

Fig: 01

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Uses of liquorice

 They will be comforted

 Expectorant
 Sweetening agent
 Flavouring agent
 Antispasmodic
2 Tulsi
Biological source
Tulsi consists of the fresh and dried leaves of Ocimum species like
Ocimum sanctum L. and Ocimum basilicum L. etc.

Family
Lamiaceae family
Chemical constituents
Phytochemical studies have shown that oleanolic acid, ursolic acid ,
rosmarinic acid, eugenol , carvacrol, linalool, and ẞ-caryophyllene are some
of the main chemical constituents of Tulsi

Fig: 02

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 Uses
 Good dental health
 Digestive health
 Good source of Vitamin K
 Promotes respiratory health
3) Clove
Biological source :
Cloves consist of dried flower buds of Eugenia caryophyllus, (Family:
Myrtaceae). It should contain not less than 15% (v/w) of clove oil. acid), resin,
chromone and eugenin.

Fmaily
Myrtaceae family
Chemical constituents
o Volatile oil
o Phenol chiefly eugenol,
o Acetyl eugenol
o Humulen
o α- & β caryophyllene
o tannins.

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 Fig: 03
4 Mentha piperita oil
Products with natural peppermint oil can do wonders for your mouth. The
more you use them, the more benefits you may see. Peppermint oil contains
antioxidants. As a result, it can help lower inflammation levels in your
mouth. If your gums are swollen, you’ll appreciate peppermint oil’s ability
to bring down the puffiness. By catching and reversing inflammation early,
you may be able to keep it from progressing to gum disease. There’s
nothing worse than painful teeth and gums. Fortunately, peppermint oil
may help. The cooling sensation it provides can feel amazing for a sore
mouth. Plus, peppermint oil is touted as a pain reliever throughout the
body, especially for heads and stomachs. You may find that has a numbing
effect on tooth pain as well.

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 Fig: 04
5 Glycerol
Glycerin is a humectant, which means it helps to retain moisture. In the
Icase of mouthwash, it can help to keep the oral cavity hydrated and reduce
dryness or soreness of the mouth andthroat. Additionally, glycerin can act
as a lubricant, which can make it easier to brush or floss the teeth. Some
studies have also suggested that glycerin may have mild antibacterial
properties, which could help to reduce plaque and bacteria in the mouth
However, it is important to note that mouthwash is not a substitute for
brushing and flossing, and it should be used in conjunction with good oral
hygiene practices.
Sodium lauryl sulphate:
Sodium lauryl sulphate (SLS) is used in toothpaste and mouth rinses as an
emulsifying and surface cleaning agent. SLS has been implicated in an
increased incidence of oral irritation in subjects predisposed to recurrent
aphthous stomatitis (RAU)

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 Formulation ingredients Table no 1
Ingredients Uses F1 F2
Tulsi Extract Antibacterial, 10 ml 10 ml
Antiseptic
Liquorice Powder Sweetening agent 1.80 ml 2 gm
, Expectorant.
Clove oil Antiseptic , 0.1ml 1 ml
flavouring
Mentha piperita oil Antibacterial 5ml 2 ml
Glycerol Co-surfactants 6.5 ml 6.5 ml
SLS Surfactants 3 gm 1 gm
Alcohol Preservatives 2 ml 1 ml
water Up to 100 ml - -

List of Equipment Table no 02

Sr.No Equipment
1 Measuring cylinder
2 Beaker
3 Mortar pestle
4 Conical flask
5 Funnel
6 Water bath
7 Heating mental
8 Petri dish
9 pH meter
10 Incubator

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Preparation of herbal mouthwash
 Two different extracts of poly herbal mouthwash were developed.
 The mouthwash formula made use of four main herbal ingredients:
Liquorice ,Tulsi and Clove, Peppermint
 Three minor ingredients added which are; Liquorice, Tulsi extract.
 The minor components were used for the preservation and for improving
the taste.
 In order to test the anti-bacterial activity of the mouthwash herbs, different
percentage of the herbal extract were prepared.
 For the formulation, the mouthwash herbal ingredients were ground to
obtain their powder form.
 10 gram of each Tulsi, Liquorice, and clove, peppermint were separately
soaked into 100 mL of distilled water and incubated
 At 37oC for 48 hours. After incubation, the herbal extract were filtered.
 The extract were then boiled separately and left to cool.
EVALUATION
Physical Evaluation:
Physical assessment (colour, odour, and consistency) was performed using
sensory and visual examination and compared with the marketed mouthwash
preparation .
pH:
A digital pen-style pH meter was used to measure the pH of prepared herbal
mouthwash formulations. For the evaluation of pH, the electrode of the pH meter
was immersed into the mouthwash formulations and kept it in the formulations
until the displayed level was stable. Note down the readings from display.
Viscosity:
Using an Ostwald viscometer, the viscosity of mouthwash formulations was
measured. The viscometer was mounted vertical position on a suitable stand.

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Mouthwash was filled in to the viscometer up to mark A. The time was counted
for mouthwash to flow from A to mark B. Viscosity was measured in triplicate.
After Taste:
The taste is strong and remains unchanged over the week except for the ambient
temperature sample. Healthy male and female candidates, were selected to
participate in the evaluation of taste. About 1 mL of herbal mouthwash was given
to selected candidates and ask for the taste.
Foam Height:
1 ml of mouthwash was mixed in 50 ml distilled water. The mixture was poured
into a 100 ml measuring cylinder. Water was added to the volume to make it 20
ml. The mixture received 25 strokes, after which it was kept aside. The height of
the foam above the aqueous volume was observed.
Turbidity:
Turbidity is the term used to describe a fluid’s cloudiness or haziness as a result
of individual particles (suspended solids or liquids) that are typically invisible to
the unaided eye. The units of turbidity from a calibrated Turbidity meter are
called Nephelometry Turbidity Units (NTUs). A formulation with an NTU of
less than 12 (or about 12) is referred to as a clear formulation. Turbidity of all
three formulations of mouthwash was measured by using Nephelometry
(Turbidity meter).
Stability Studies:
The aim of stability studies was to guarantee that the mouthwash formulations
can be used and can continue to have the same properties over time. Prior to
conducting antibacterial testing, stability studies were conducted on various
mouthwash formulations.
Physical stability:
The visual appearance, homogeneity and physical separation of the formulated
mouthwash were measured throughout this test. Three formulations of
mouthwash were then kept at different temperatures, including 12°C and 25°C.

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The appearance was then assessed at various temperatures, and the findings were
recorded.
pH stability:
A calibrated pH meter was used to record pH stability. To evaluate the changes
in the pH readings, different mouthwash formulations were kept on the shelf of a
room at 37°C and at 40°C. After that, the result was noted and compared for a
period of six weeks

Pepermint oil Clove oil

RESULTS AND DISCUSSION

pH:

The pH of all three formulations was recorded by digital pen style pH meter
(Figure 3). The results of pH were shown in Table 6. The pH of formulations F-
1, F-2 was found to be 6.2, 5.3 and respectively.

Viscosity

We used an Ostwald viscometer to measure the viscosity of each of the three

mouthwash formulations According to tests, F-1, F-2, viscosities were 9.2, 8.5,
CPS, respectively.

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After Taste

For the evaluation of taste 10 healthy human volunteers were selected. 1 ml of

each formulation of mouthwash was swished or gargled by all volunteers for
about half a minute and then spat out. According to the feedback of all volunteers
the taste of all three formulations on Day 1 and Day 7 was found to be similar.

Foam Height
The height of the foam of three formulation 1 (F-1), formulation 2 (F-2) was
found to be 3 ml, 5 ml respectively.

Turbidity

Turbidity of mouthwash formulations was measured by using Nephelometry. The

turbidity of F-1, F-2 was found to be 004, 003 NTU respectively.

Stability Studies
The result of physical stability studies was shown . Change in color, odour,
physical separation and homogeneity of all two formulated mouthwash was
observed at 12°C and 25°C. According to the results of Stability studies, there
was a slight change in the pH of formulations at 40°C.

DISCUSSION
The present study was designed to formulate and evaluate an herbal mouthwash
and compare it with marketed mouthwash reported that, mouthwash is a
chemical antiseptic commonly used in mouthwashes and dental products. It has
some disadvantages compared to herbal mouthwashes. It is also more likely to
cause dry mouth, mouth ulcers, and other oral irritations than herbal
mouthwashes. Results of this study have shown that the test mouthwash
formulation-2 (F-2) was better than F-1 and F-2. When compared, the evaluation
parameters of the F-2 formulation produced findings that were highly similar to
those of the standard but had less side effects. The formulation of F-2 mouthwash
was found to have a pH of 5.3 to have a pH of 5.4. When the turbidity of the F-2
formulation was compared to that of standard it was found to be identical.

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 CONCLUSION
The formulated herbal mouthwashes can significantly work in long way to help
people in eradicating dental issues such as decay, gingivitis, sensitive teeth, root
infection, bad breath, enamel erosions and many other oral problems.
Furthermore, we can assure that no harmful ingredients are present in mouthwash
formulations. The herbal mouthwash is better than mouthwash as it does not
cause negative side effects, such as staining of the natural teeth, altered taste
perception due to prolonged use and dry mouth. The phytochemical tests confirm
the presence of flavonoids, phenolic compounds, tannins, alkaloids and
terpenoids. It has been medically demonstrated that the natural herbs utilized in
the current formulations may prevent the other problem of oral cavity such as
tooth decay and gum diseases. This herbal mouthwash makes it simple for a
person to rinse their mouth and avoid a variety of oral health problems. The
present study is crucial for developing an affordable, efficient herbal oral health
intervention for low socioeconomic communities. However, since this study was
brief, longer studies with larger sample sizes are necessary.

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 Physical Evaluation Table no 03
S.no Formulation Parameter Observation
1 F1 Colour Florescent Yellow
Odour Pleasant cool mint
Appearance Visual appearance
Texture Liquid
pH 6
2 F2 Colour Dark yellow
Odour Pleasant odour
Appearance Visual appearance
Texture Liquid
pH 5

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