INTRODUCTION:

Mouthwashes are liquids with pain-relieving, antibacterial, and mitigating properties. Mouthwash is a solution that is most commonly used for its antiperspirant, revitalizing, and cleaning qualities, or to manage plaque.¹Due to their ability to target oral infections, provide immediate pain relief, and have fewer adverse effects, herbal mouthwashes are in high demand.² One serious mouth illness that can impact the teeth is peridontitis. Gum edema, bleeding gums, enamel eruption, cavity development, and the creation of a hollow, black eruption on the tooth surface are all included in dental caries.

Due to poor oral hygiene, childhood and teenage dental caries rates are high in the beginning. Through the jaw bones and the gaps between the fascial planes of the surrounding soft tissue, oral infections propagate from the root of the contaminated tooth.^3,4 Common spices like clove oil, neem, triphala, tulsi, and many more are used either alone or in combination and have been deductively showed to be a safe and effective remedy for oral medical conditions like mouth ulcers, gum disease, and tooth decay prevention without side effects.

Types of mouthwash:

Fig. 1 Type of mouthwash

· Fluoride mouthwash: Mouthwashes with fluoride contain salt, which protects teeth against cavities and pits. Given that fluoride is also included in toothpaste and water, it is appropriate to use caution while using this type of mouthwash because exposure to excessive fluoride levels is harmful to your overall health.^5,6

· Regular mouthwash: A mouthwash that performs the same functions as other mouthwashes but with standard ingredients might also be referred to as normal mouthwash. It's a well-liked option for mouthwash without alcohol as well. Compared to other mouthwash varieties, these ingredients are safer to use.⁶

· Corrective mouthwash: Your breath can be refreshed or bad breath covered up with a mouthwash that doesn't truly affect your overall dental health.^6,7

· Disinfected mouthwash: People with oral infections typically use this mouthwash, which typically contains alcohol, to prevent the growth of germs. For Those who suffer from bad breath or halitosis, this is often also accommodating. In order to help prevent microscopic organisms that cause oral infections and bad breath, this is frequently used in conjunction with the proper cleaning of teeth and flossing.

History:

Ayurveda has the first documented mentions of mouth washing as a gingivitis therapy. Hippocrates advised a solution of salt, alum, and vinegar. Later, in the Greek and Roman eras, mouth washing after mechanical cleansing became popular among the elite classes.^5,8 In order to help fight bad breath, Greek surgeon and physician Pedanius Dioscorides suggested a mixture of "juice and leaves of olives, milk, gum myrrh, pomegranate, vinegar, and wine" between the years of 40 and 90. Mouth wash was originally referenced in Ayurvedic and Chinese medicine about 2700 BC. Patients undergoing chemotherapy can use mouthwash as an effective at-home oral hygiene routine.⁸

Fig. 2 Old mouthwash bottles

Benefits of herbal mouthwash:

· They are less toxic and have very little to no side effects.

· Not all natural mouthwashes include alcohol in addition to sugar.

· Even for the most sensitive mouths, natural mouthwashes are gentle.¹⁰

· Inherently antibacterial are natural mouthwashes.

· No harsh additives are present.

Herbs used as mouth wash:

Clove:

Synonyms: Clove flower, Clove bud

Biological Source: Obtain dried flower bud of Eugenia caryophyllus, family Myrtaceae.^9,10

Geographical Source: Cultivated in Zanzibar, India, Caribbean, sri lanka, clove are also grown in some district of Tamil Nadu state.

Chemical constituents:

About 15% to 16% of volatile oil is found in cloves. 10 to 13 % of tannin, resin, chromone and eugenin. 70% to 75% of the medication eugenol is present in volatile oil. Clove oil ranges in color from colorless to light yellow.¹¹

Uses:

· Used as an carminative, stimulant, flavouring agent.

· Also act as dental analgesic.

· Uesd in formulation of mouthwashes.

· Also used in tobacco products like cigarettes.

· Used as an antiseptic.

Fig. 3: Picture of Clove

Tulsi:

Synonyms: Holy Basil, Sacred Basil

Biological source:

It Is Obtain from Fresh and Dried Leaves of Ocimum Sanctum Linn, Family Lamiaceae.

Geographical Source:

Australia, Malesia, Asia, and the western Pacific are among its natural tropical and subtropical environments. In all of Southeast Asia's tropical regions, it is extensively grown.¹²

Chemical constituents:

Eugenol, euginal (also known as eugenic acid), urosolic acid, carvacrol, linalool, limatrol, caryophyllene, and methyl carvicol are found in the volatile oil of leaves, while fatty acids and sitosterol are found in the volatile oil of seeds. Sugars are also present in the seed mucilage, and anthocyanins are found in green leaves. Polysaccharides and xylose make up sugars.^12,13

Fig. 4 Picture of Tulsi

Uses:

· Booster of natura immunity.

· Lowers pain (analgesic) and fever (antipyretic).

· Reduces cough, colds, and other respiratory conditions.

· Lowers blood pressure and stress.

· Beneficial to those with diabetes.

Neem:

Synonyms: Indian lilac, Nimba tree, miracle tree

Biological source: Azadirachta indica J. Juss (also known as Melia Indica or M. azadirachta Linn) leaves, either fresh or dried, and seed oil are the components of neem, family Meliaceae.

Geographical Source: According to conventional wisdom, Azadirachta indica is indigenous to Bangladesh, Myanmar, Thailand, and Vietnam in Indochina, as well as the Indian subcontinent.¹⁴

Chemical constituents: Nimbin, nimbanene, and other substances are found in leaves. 6-desacetylnimbinene, amino acids, n-hexacosanol, nimbandiol, and nimbolide, ascorbic acid.

Uses:

· Eliminates Toxins: For a very long time, neem leaves have been used for their purifying qualities.

· Neem is also rich in calcium, which is good for bones.

· It is also iron-rich, has antioxidant qualities.

· Builds immunity, and is good for skin, dental, and hair health.

· It is also anti-fungal and anti-bacterial.¹⁵

Fig. 5: Picture of Neem

METHOD AND MATERIALS:

Collection of plants:

Neem (Azadirachta indica), Tulsi (Ocimum Sanctum Linn), Mentha piperita (Mint), leaf and Clove (Eugenia caryophyllus) buds are collected from the plant species. Saccharine, PEG 40, Glycerol, and alcohol are collected from laboratories of our college.

Extraction process:

Collect the leaf of plant and dried them by shade drying. For extraction different solvent are used.^13,15

Table 1: Active drug and solvent

Sr. No.	Plant Used	Solvent
1	Neem (Azadirachta indica)	Methanol
2	Tulsi (Ocimum Sanctum Linn)	Ethanol
3	Clove (Eugenia caryophyllus)	Water

Collected Herbal extracts were placed in the container after being filtered by using filter paper and stored in adequate space.

Formulation process:

Table 2 List of Ingredients

Sr. No.	Ingredients	Activity	Formula 1	Formula 2
1	Clove	Active drug	1ml	2ml
2	Neem	Active drug	260mg	540mg
3	Tulsi	Active drug	260mg	540mg
4	Mint	Flavoring agent	0.1ml	0.1ml
5	Saccharine	Sweeting agent	50mg	50mg
6	PEG 40	Surfactant	6g	6g
7	Glycerol	Co-surfactant	6.5g	6.5g
8	Methyl paraben	Preservative	2ml	2ml
9	Water	For volume make up	100ml	100ml

· Each ingredient will be measured out in weighing balance.

· The extract will be thoroughly mixed in a mortar and pestle with a small amount of water.

· All other ingredients will be added gradually and Then clove and mint oils will be added drop by drop and mixed carefully and taking care to prevent lump formation. PEG 40 and glycerol will then be added drop by drop and mixed thoroughly water will then be added to make volume and preservative will be added. ^1,2,16

· The product will be packed in an eye-catching, air tight container.

Evaluation test:

Physical evaluation – The physical parameter evaluated by visual examination like Color, odor and taste.¹⁷

pH determination - A digital pH meter was used to measure the pH of the product herbal mouthwash. The standard buffer was used to calibrate the Ph meter. A pH meter was used determine pH. Dissolve 1ml of mouthwash in 50ml of distilled water. Then put electrode in solution and find pH of mouthwash.¹⁸

Fig. 6 pH Test

Test for microbial growth

1. Prepare the Petri dishes: Get sterile Petri dishes agar medium. Fill the dishes with the agar and let it set.¹⁹

2. Inoculate: Gather bacteria from the mouth or get a standardized culture from a lab. Evenly swab the agar surface to inoculate the bacteria.

Fig. 7 Before mouthwash use

Fig. 8 After mouthwash use

3. Form groups: Assign the Petri plates to various mouthwash samples for testing, with each group serving as a representative sample.²⁰ Add a group that doesn't use mouthwash as a control.

4. Apply mouthwash: Following the manufacturer's instructions, add the recommended quantity of each mouthwash to the corresponding Petri dishes.^21,22

5. Incubate: For 24-48 hours, incubate the Petri dishes at the proper temperature (often about 37°C) to promote bacterial growth.²³

6. Measure inhibition: Track the bacterial growth on each dish following the incubation time. If there are any zones of inhibition surrounding the places where the mouthwash was administered, measure their diameter. More efficacious prevention of microbial growth is indicated by a broader zone of inhibition.²⁴

Stability study - Any pharmaceutical product cannot be fully formulated or manufactured without doing adequate stability tests on the final product. The purpose of this is to ascertain the produced product's chemical and physical stability, which will help assess the product's safety.^25,26 Accelerated stability tests, which involve subjecting the product to elevated temperatures in accordance with ICH rules. Check stability of mouthwash at room temperature for 1 month.²⁵

Homogeneity Test - A clean and dried glass slide was smeared with the herbal mouthwash and covered using glass cover. The appearance was investigated under the light. The mouthwash was also visually tested for homogeneity, aggregates or floccules.²⁶

RESULT:

The finest mouthwash formulation is F2, which has been prepared for a long time and is fresh, effective against germs, and may be used to overcome herbal mouthwashes that contain clove, neem, and tulsi. For extended storage, its ph, color, and odor remain unchanged. There are relatively few adverse effects. Neem and tulsi provide a painless, cooling feeling, fresh breath, and excellent order. Clove and mint plants give the preparation a nice color and help to remove throat infections. They also have good antibacterial properties. Clove oil is useful for dental problems and is found in plants with a good aromatic order.

Fig. 9 Picture of Formulation F1 and F2

Table 3 Evaluations parameter

Sr. No.	Evaluation parameter	Observation	F1 formulation	F2 formulation
1	Physical evaluation	Color	Slightly light Green	Green
		Taste	Bitter	Bitter
		Flavor	Mint	Mint
		State	Biphasic liquid	Biphasic liquid
2.	pH	pH meter	6.99	6.96
3.	Test for microbial Growth	Zone of Inhibition	1.5cm	2cm
4.	Stability test	At room temp.	Flocculation appears	Flocculation appears
5.	Homogeneity		Good	Good

CONCLUSION:

Although they are highly powerful agents, herbs need to be utilized carefully. Active compounds in herbs have the potential to cause undesirable interactions when used with prescription drugs or other treatments. Therefore, if you have any doubts about the suitability of the herb or how it will interact with other treatments, it is advisable to speak with a physician or other health specialist. Evidence of safety and efficacy should underpin the use of herbs in dentistry. The antibacterial properties may be able to eliminate oral pathogens. The results indicate that the F2 formulation's greater active drug concentration than the F1 formulation accounts for its superior efficacy over the former. The F2 formulation is the most efficient against oral infection diseases because it inhibits the upper zone of bacteria.

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Received on 31.08.2024 Revised on 24.09.2024

Accepted on 30.10.2024 Published on 05.12.2024

Available online on December 28, 2024

Research J. Topical and Cosmetic Sci. 2024; 15(2):79-84.

DOI: 10.52711/2321-5844.2024.00014