Title of the Manuscript: Formulation and Evaluation of Herbal Mouthwash

NEED OF HERBAL MOUTHWASH:

• • • Preserves oral hygiene by lowering the amount of microorganisms in the mouth and aiding in the removal of food particles.
    • Breath freshening: Offers a rapid and simple solution for halitosis, or bad breath.
    • Decreases tartar and plaque: helps avoid accumulation that causes cavities and gum disease.
    • Prevents and treats gum disease: can lessen gingivitis risk, bleeding, and irritation.
    • Supports dental care in general: enhances brushing and flossing to provide a more thorough dental hygiene regimen.

AIM: Formulation and Evaluation of herbal mouthwash

OBJECTIVE:

1. To develop formulation of herbal mouthwash:

The present results therefore offer a greater use for traditional use of herbal mouth wash.

2. Safety:

Herbal mouthwash was safe and there was neither report of adverse reactions.

3. Effectiveness:

The objective of present work is to formulate and evaluate herbal mouthwash and to evaluate its effectiveness against microbial load of oral cavity.

4. To maintain oral hygiene:

Oral health is important as overall health. Now-a- days people may faces more oral problems like periodontal disease, sore throat, gingivitis, plaque and so on. For maintaining good oral health various formulations are formulated.

5. To Prevention, control and reduction of oral infection:

It can reduce the plaque growth in your mouth, decrease your chances of developing gum disease, and prevent tooth decay.

6. To reduce side effects by promoting herbal use:

The use of herbs in dentistry should be based on evidence of effectiveness and safety. 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.

7. Herbal medicines:

Ayurvedic medicines give a holistic approach toward entire human beings. It can maintain the balance between general and oral health as well as an environment which is in this era necessary for the well-being of humans.

PLAN OF WORK:

• • Literature Survey .
  • Collection of Plant Material .
  • Selection of Excipients for Formulation.
  • Extraction of Herbal Drugs .
  • Development of Formulation .
  • Evaluation of Prepared Formulation.[12]

DRUG PROFILE:

Table No 1: Drug profile

Chemicals, Glasswares And Insruments:

These are obtained from Research-lab fine chemicals industries, Mumbai 400002 (India) Research- lab. , salunke@gmail.com A GMP Certified company chemicals and Glassware used in the experimental work were calibrated in the laboratory the glassware was washed until it was used for experiment .

LIST OF CHEMICAL:

Table No 2: List of chemical

Fig No 1: Chemical Reagent

Instruments:

• Beakers
• Measuring cylinders
• Electronic weighing balance
• Water bath
• Whatman filter paper
• Steam distillation apparatus
• pH meter
• Ostwald viscometer
• Petri dishes

Materials And Method Of Extraction

1. Tulsi (Ocimum sanctum)

The herb (Tulsi) is useful in teeth disorders. You can use its sun-dried and powdered leaves to wash your teeth. Additionally, it can be combined with mustered oil to create a paste that can be used as toothpaste.

• Biological source:            It is the fresh and dried leaves of plants from the Ocimum genus, primarily Ocimum sanctum L. (also known as Ocimum tenuiflorum)
• Kingdom: plantae.
• Order: lamiales .
• Family: lamiaceae.
• Genus: Ocimum Species: sanctum.
• Geographical Source: North-central India and surrounding South Asian regions.
• Parts used: Leaves, Seeds and Roots.[13]
  1. Chemical constituents:
     • Eugenol, nerol, eugenol methyl ether.
     • Caryophyllene, terpinene-4-ol-decyladehyde.
     • Camphor and carvacrol.
     • Essential oils, ascorbic acid, carotene, calcium, phosphorus and insoluble oxalates.
     • It also contains terpenes, mucilage, fixed oil andfatty acids.
     • Volatile Oil-0.8% .[14]

1. 2. Uses of Tulsi plant:
      • In reduction of oral malodour.
      • Anticancer activity of O. sanctum: Tulsi has been shown to possess an excellent anticancer activity.
      • As an antioxidant, it also has the ability to scavenge highly reactive-free radicals.
      • Modulates immunity.
      • As analgesic, antipyretic, and        anti- inflammatory Activity.[15]

Fig No 2: Tulsi leave [16]

1. 1. Preparation of Ocimum sanctum Leaf Extract. (Tulsi)

Dried Opium sanctum leaves

↓

Powdered (20 g)

↓

Mixed with 100 mL distilled water

↓

Kept overnight at room temperature

↓

Filtered (muslin cloth → Whatman filter paper)

↓

Green filtrate obtained

↓ ried

↓

Residue collected for experiment. [17]

Fig No 3: Tulsi Exract

2. Guava (Psidium guajava)

Psidium Guajava is the plant that gave rise to guava. It is a member of the Myrtaceae family. Shape: Depending on the species, guava fruits can be round or oval in shape and range in length from 4 to 12 centimeters (1.6 to 4.7 inches) .[18]

• Biological source: guava is the evergreen tree Psidium guajava L.
• Kingdom: Plantae.
• Order: Myrtales.
• Family: Myrtaceae Genus: Psidium.
• Geographical Source: America, specifically a region extending from Mexico and Central America to northern South America. Historically, it is believed to have originated in the savannas and semi- deciduous forests of South America.
• Species: Psidium guajava. [19]

Fig No 4: Guava with leave [21]

1. 1. Chemical constituents:
      • Phenolic compounds: These are powerful antioxidants that have health-promoting properties.
      • Flavonoids: Such as quercetin, avicularin, apigenin, guaijaverin, kaempferol, and myricetin, which have been studied for their anticancer, antidiabetic, and antioxidant activities.
      • Tannins: Known for their antimicrobial properties.
      • Terpenoids: Which can produce relaxation effects.
      • Saponins: Compounds with potential health benefits.
      • Vitamins: Including ascorbic acid (vitamin C).
   2. Uses of guava leaves:
      • Antibacterial properties.
      • Anti-inflammatory effects, Reduces dental plaque, Relief from toothache.
   3. Extraction process of guava leaves:

Take 25 gm of leaf powder → into a beaker

↓

Add 200 ml of sterile double distilled water → to the beaker

↓

Heat the mixture → in a water bath

↓

Reduce the volume → till it's less than 50 ml

↓

Filter the liquid

↓

By using Whatman's filter paper

Fig No 5: Guava Extract

3. Peppermint oil:

Peppermint oil appears to be safe when taken orally (by mouth) or applied topically in the doses commonly used. Peppermint oil has been safely used in many clinical trials. Possible side effects of peppermint oil taken orally include heartburn, nausea, abdominal pain, and dry mouth. [23]

• Biological Source: Peppermint consists of the leaves and flowering tops of the plant Mentha piperita L.
• Kingdom: Plantae (Plants).
• Order: Lamiales.
• Family: Lamiaceae (also known as the mint family).
• Geographical Source: it originated in Europe and the Middle East, but the majority of the world's peppermint leaves are now harvested in India (Uttar Pradesh), the United States (Pacific Northwest), and Morocco.
• Species: Mentha piperita L.

1. 1. Chemical constituents
      • Essential oil including menthol, menthone, menthofuran.
      • 1,8-cineole, and menthyl acetate. [24]
      • 1,8-cineole, limonene.
      • Beta-pinene and beta- caryophyllene.[25]
   2. Uses of Peppermint Oil:
      • Antioxidant.
      • Anti-inflammatory.
      • Anti-bacterial properties.[26]

Fig No 6: Peppermint leaves [27]

1. 1. Extraction process of peppermint leave:

Weigh mint leaves (60 g)

↓

Place in distillation flask and add sufficient water

↓

Heat to generate steam (steam distillation process)

↓

Volatile oil vaporizes with steam

↓

Condense vapours (oil + water) using condenser

↓

Collect distillate

↓

Separate oil using separating funnel

↓

Collect and weigh peppermint oil [28]

Fig No 7: Peppermint Extract

Methodology:

1. Maceration method / Aqueous extraction (hot)

Used for Tulsi leaves (boiling leaf powder in water and filtering).

2. Hot Aqueous Exraction

Used for Guava leaves (boiling leaf powder in water and filtering).

3. Steam distillation method

Used for Peppermint oil extraction.

4. Simple mixing / Solution Method

Used for final formulation of herbal mouthwash (mixing extracts, excipients, preservative, & vehicle.

Formulation Of Herbal Mouthwash:

Table No 3: Formulation of herbal mouthwash

METHODS OF PREPARATION:

Wash all laboratory apparatus and

accurately weigh all ingredients

Collect fresh guava leaves

Dry guava leaves for 10 days at room temperature

Grind dried leaves to obtain coarse powder

Store powder in airtight container

(cool, dry place; away from sunlight)

Boil guava leaf powder at 90 °C for 15–20 minutes & Filter extract using filter paper

Beaker I: Add ethanol and peppermint oil and mix

Beaker II: Add glycerol, propylene  glycol, Tulsi extract, tween-80, warm  water, and guava extract; dilute

Add sodium benzoate & Add FDA-approved green colorant

Make up final volume with purified water

Transfer prepared mouthwash into tightly closed container. [29]

Evaluation Test Of Herbal Mouthwash:

• Physical parameters: such as color and odor, were assessed visually.
• PH: A digital pH meter was used to measure the prepared herbal mouthwash's . A standard buffer solution was used to calibrate the pH meter. One milliliter of mouthwash was weighed, dissolved in fifty milliliters of purified water, and its pH was then measured. [30]
• Determination of Antibacterial Property of Mouthwash:
  1. Agar media was prepared then the formulated mouthwash was inoculated on the plates agar media by steak plate method and controlled is prepared by mouthwash. The plates were placed in the incubator and are incubated 37⁰C for 24 hours. After the incubation period the plates were taken out and the Microbial growth were checked and compared with the control.
  2. The antibacterial activities were evaluated by measuring the zones of inhibition (in mm).[31]
• Viscosity: A Ostwald viscometer was used to measure the mouthwash's viscosity .The table provides a description of the viscosity results for each created formulation. [32]
• Foam test: The product's ability to foam was assessed by adding a small amount of water to a measuring cylinder, noting the starting volume, then shaking the cylinder 10 times. The final foam volume was recorded. [33]

Result And Discussion:

1. 1. Physical Parameter:

Table No 4: Physical parameter of Herbal Mouthwash

2. pH Stability analysis:.

Table No 5: pH of Herbal Mouthwash

Fig No 8: pH Analysis of Mouthwash

3. Antibacterial activity:

Table No 6: Antibacterial Activity of Mouthwash

1. Before keeping in incubator

                   (B) After incubator

Fig no 9: Microbial Activity