Preparation and Evaluation of Antibacterial Herbal Mouthwash against Oral Pathogens

 

Akshay R. Yadav*, Shrinivas K. Mohite, Chandrakant S. Magdum

Department of Pharmaceutical Chemistry, Rajarambapu College of Pharmacy, Kasegaon,

Maharashtra, India-415404.

 

ABSTRACT:

The present research is aimed to formulate and evaluating herbal mouthwash and evaluating its efficacy against microbial oral cavity load. Due to its physicochemical properties and antimicrobial activity, prepared mouthwash was further evaluated. Malvastrum coromandelianum and Neem extracts and the polyherbal mouth rinse demonstrated good sensitivity to micro-organisms such as E. coli and S. aureus. Natural herbs such as malvastrum coromandelianum, Neem and fennel 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, halitosis, mouth ulcers, and preventing tooth decay. Clove is most commonly applied directly to the gums for toothache, pain control during work, and other dental-related issues. We have developed a mouthwash with some common food materials and herbs and which can replace costly chemicals like alcohol, coloring agents and preservatives making our mouthwash economically more viable than commercial mouthwash. The anti-inflammatory and anti-infectious properties of neem make it a powerful treatment for gum disease. The results of this study indicate that the existence of compounds such as glycosides, terpenoids, flavanoids and saponins that can be responsible for the antibacterial efficacy of extracts and polyherbal mouthwash rinse against these microorganisms that are known to be mostly responsible for oral cavity bacterial infection.

 

 

 

 

INTRODUCTION:

Mouthwash/ rinse is a chemotherapeutic agent used by patients to improve oral hygiene as an effective home care method. Many mouthwash manufacturers say that antiseptic and anti-plaque mouth rinse destroy cavities, gingivitis, and bad breath causing bacterial plaque¹.

 

India is a country with a rich traditional medicine system dating back over three thousand years and possibly herbal combinations must have been recognized since then, but now people have been using modern medicines all along, the significance of the old medicine system has been forgotten². Maintenance of oral hygiene is imperative in preventing the buildup of plaque, a sticky film of bacteria and food that accumulates on teeth³. Oral hygiene measures include mechanical aids such as toothbrushes, interdental cleansers and chemotherapeutic agents such as mouthwashes, dentrifices and chewing gums⁴. Mouthwash (mouth rinses) are solutions or liquid intended to reduce the microbial load in the oral cavity. It may contain alcohol, glycerin, synthetic sweetness, surface active agents, flavoring agents, colouring agents, etc⁵. Many popular herbal products have helped to control dental plaque and gingivitis and they have so far been used as adjunct to other oral hygiene measures such as brushing and flossing. Over 50% of the modern drugs are of natural products origin and as such natural products play an important role in drug development. Malvastrum coromandelianum (L.) Garcke (family Malvaceae), commonly known as false mallow, broom weed, and clock plant⁶. Various parts of this plant are used by numerous tribal populations throughout the world. A common problem among immune compromised, elderly, and chronically ill patients is oral candidiasis. Precipitation of oral candidiasis causes burning sensation and altered taste which further lowers the quality of life⁷. Artificially produced demineralized water, distilled water and also deionized or reverse osmosis-treated water were mainly used in industrial and laboratory applications. Various kinds of mouthwashes have evolved following oral hygienical problems⁸. But apart from this, mouthwashes also serve to refresh breath. Morever, mouthwash also contains some ingredients that serve as digestive aids. Mouthwashes can be chemical or herbal in nature. Mouthwash is a liquid accessory to clean and maintain the health of our teeth for oral hygiene. The mouthwash antibacterial and antimicrobial properties can prevent cavity growth that causes infection, eliminate plaque, combat bad breath, and preserve strong and healthy teeth and gums. Salt treats mouth sores as exosmosis induces many periodontal diseases to improve swollen gum conditions. Herbal drugs have received greater attention in recent times because of their diversity of curing diseases, safety and being well tolerated remedies when compared to the conventional medicines⁹. There are several types of mouthwash available in the market today worldwide. Mouthwash is an aqueous solution which is most often used for its deodorant, refreshing and antiseptic properties or for control of plaque. Maintenance of oral hygiene is imperative in preventing the buildup of plaque, a sticky film of bacteria and food that accumulates on teeth. Oral hygiene measures include mechanical aids such as toothbrushes, interdental cleansers and chemotherapeutic agents such as mouthwashes, dentrifices and chewing gums¹⁰. Mouthwash (mouth rinses) are solutions or liquid intended to reduce the microbial load in the oral cavity. There are several types of mouthwash available in the market today worldwide¹¹. Evaluation of antibacterial efficacy of malvastrum coromandelianum so far has been through in vitro studies. Therefore, this study was designed to assess the clinical efficacy.^10-12

 

MATERIALS AND METHODS:

Collection of plant material:

Malvastrum coromandelianum, fennel, neem and clove were collected and rinsed properly with water to remove contaminants.

 

Preparation of Extraction:

Shade drying was done for almost a month as to avoid chemical degradation due to sunlight. Grinding of the dried material was done, with the aid of a grinder and converted into coarse powder. Extraction of malvastrum coromandelianum was done by microwave extraction. The crude powder was defatted subjected to extraction with ethyl acetate using a microwave extraction. The excess solvent present was evaporated¹³.

 

Formulation of herbal mouthwash:

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

 

Table 1: Formulation table for herbal mouthwash

Sr. No	Ingredients	Role	quantity
1	Malvastrum coromandelianum extract	Antibacterial	1g
2	Neem extract	Antimicrobial	0.5g
3	Fennel extract	Antibacterial	1g
4	Clove powder	Antibacterial	0.5g
5	Glycerine	Humectant	0.5ml
6	Methyl paraben	Preservative	1g
7	Distilled water	Vehicle	q.s to make 10ml

 

Procedure:

1g of Malvastrum coromadelianum, 0.5g of neem extract, 1g fennel extract, Clove powder were dissolved in 4 ml of water, filtered and made up to 10 ml. 0.5 ml of glycerin was added by continuous shaking, followed by the addition of 1gm of methyl paraben for preservation¹⁴.

 

Antibacterial Studies of the Extracts:

Disc Diffussion Method:

1.     Media Used: agar (For bacteria)

 

2.     Temperature:

Bring agar plates to room temperature before use.

 

3.     Inoculum Preparation:

Using a loop or swab, transfer the colonies to the plates. Visually adjust turbidity with broth to equal.

 

4.     Inoculation of Agar Plate:

Dip a sterile cotton swab into the inoculum and rotate it against the tube wall above the liquid to remove excess inoculum within 15 minutes of adjusting the inoculum to a McFarland 0.5 turbidity standard.· Swab three times the entire surface of the agar plate, rotating plates about 60 degree between the streaks to ensure even distribution. Do not touch petri plate sides and produce aerosols.

 

5.     Addition of extract into Plate:

Take hollow tube of 5 mm diameter, heat it. Press it on above inoculated Agar plate and remove it immediately by making a well in the plate. Likewise, make four well on each plate. Add 10µg/ml, 25µg/ml, 50µg/ml, 100µg/mlof compound into the respective Wells on each plate.

 

6.     Incubation:

Incubate plates within 15 min of compound application. Invert plates, and stack them no more than five high. Incubate for 18 - 24 hr at 37 ºC in incubator.

 

7.     Reading Plates:

Read plates only if the lawn of growth is confluent or nearly confluent.

 

Measure diameter of inhibition zone to nearest whole millimeter by holding the measuring device^15-16.

 

RESULTS AND DISCUSSION:

Phytochemical Study Results:

Mouthwash containing antimicrobial ingredient like malvastrum coromandelianum, neem, funnel and other essential plant extracts have been found to reduce oral pathogens when combined with daily brushing and flossing.

Evaluation of herbal mouthwash:

Colour and odour:

physical parameters like odour and colour were examined by visual examinations.

 

pH:

pH of mouthwash was measured by using pH paper.

 

Test for microbial growth in formulated mouthwash:

The formulated mouthwash was inoculated in the plates of agar media by streak plate methods and a control was prepared. The plates were placed in the incubator and are incubated at 37 ºC for 24 hours. After incubation periods plates were taken out and checked for microbial growth by comparing it with standard.

 

Determination of Antibacterial Activity of the Extracts:

Antibacterial potency against S. aureus and E. coli by the standard method using Ciprofloxacin as a standard drug was carried out.

 

 

Table 2: Antibacterial screening by measuring the zone of inhibition

Sr.No	Concentration	E. coli	S. aureus
1	10 µg/ml	+	++
2	25 µg/ml	+++	-
3	50 µg/ml	++	+++
4	100µg/ml (ciprofloxacin)	+++	++

Below 4mm shows (-) Negative activity.

Between 4mm-6mm shows (+) Slight activity.

Between 6mm-8mm shows (++) Moderate activity.

Between 8mm – 10mm (+++) Higher activity.

 

CONCLUSION:

The present liquid herbal mouthwash can work in long way to help people to get rid of bad breath and many oral disorders. This mouthwash is purely herbal without adding any kind of alcohol and any other additives as other products found in the the market. By performing antibacterial activity, the results also showed antibacterial activity in mouthwash formulations. Phytoconstituents, such as azardichtin, tresvetrol, eugenol and palmitic acid which are the main active ingredients of neem, and clove respectively, were responsible for the antibacterial efficacy of the herbal formulation.

 

ACKNOWLEDGEMENT:

I express my sincere thanks to Vice-principal Prof. Dr. S. K. Mohite for providing me all necessary facilities and valuable guidance extended to me.

 

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Accepted on 01.05.2020      ©Asian Pharma Press All Right Reserved