Kandarp Dave et al

IJCPS, 2014, Vol.2(3): 666-673

ISSN: 2321-3132

Research Article

International Journal of Chemistry and

Pharmaceutical Sciences
www.pharmaresearchlibrary.com/ijcps

Development and Evaluation of Antibacterial Herbal Toothpaste

containing Eugenia caryophullus, Acacia nilotica and Mimusops elengi
Kandarp Dave*, Lata Panchal, Pragna K Shelat
Laboratory of pharmaceutical Technology, Department of Pharmaceutics, K. B. Institute of
Pharmaceutical Education
Education and Research, Gandhinagar, India
Abstract
The present work dealed with development and evaluation of herbal anti-bacterial
anti bacterial toothpaste containing clove,
babul and borsali as herbal ingredients. Different types of formulations (F1 - F9) were formulated using calcium
carbonate as abrasive and sorbito
sorbitoll solution (70 %) as humectant in varied concentrations based on factorial
design. All the formulations were evaluated for various parameters like dryness, color, appearance, consistency,
wash ability, pH, spread ability
bility and foaming power. In optimized For
Formulation
mulation (F7), the aqueous extracts of
clove (powder of clove buds of Eugenia caryophullus), babul (powder of babul bark of Acacia nilotica) and
borsali (powder of borsali bark of Mimusops elengi) were loaded and herbal toothpaste was prepared. This
Formulation
ation showed good color, appearance, consistency, wash ability, pH, spreadability and foaming capacity.
The formulation showed very good anti-microbial
anti microbial profile during microbial assay. This optimized formulation
was safe to use for dental care.
ve budpowder, Borsali bark powder, Babul bark powder, Microbial assay
Keywords:Clove

Contents
1.
2.
3.
4.
5.
6.

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 666
Experimental . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 667
Results and discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 668
Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . .. . . . . . . . . . . . . 672
Acknowledgement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ... . . . . . . . . . . . . .. . . . . . . 672
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . 672

*Corresponding author
Kandarp Dave
E-mail: kandarpdave007@gmail.com
Manuscript ID: IJCPS1984
Published Online 27 March 2014
2013, IJCPS All Rights Reserved

PAPER-QR CODE

1. Introduction
The anatomic part of tooth consists of crown, root, enamel, dentine and pulp [1]. Many problems associated with the
dental care are bad breath, tooth decay, gum (periodontal) disease, tooth sensitivity, calculus, dental caries and
dental plaque [2]. Plenty of herbal as well as synthetic ingredients are reported to have remarkable beneficial effects
on various dental problems mentioned above. They are Clove, neem, sunthi, mentha, tomar, pippali, aloe vera,
kapoor, babul, borsali. According to the constituent
constituentss they play different role through different mechanisms like
providing freshness, Antibacterial which kill or retard the growth of Streptococcus mutans&Lactobacillus
mutans
acidophilus,the
the main causative organism of dental cavity, by providing protective layer over
ov teeth and/or local
anaesthetic to reduce dental pain.Toothpaste
Toothpaste is a paste or gel dentifrice used with a toothbrush as an accessory to
clean and maintain the aesthetics and health of teeth [3]. Main objectives of the toothpaste are delivering a range of
preventive and therapeutic agents such as fluoride, metal salts and pyrophosphate for calculus inhibition, to reduce
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plaque growth, to treat dentine hypersensitivity and providing oral hygiene. Cleaning of the surface of the teeth is
the primary function of a dentifrice when used with a toothbrush. A dentifrice helps in the removal of food particles,
reduction of superficial plaque or stain, polishing of tooth surface and refreshing mouth breath. Therapeutics &
cosmetic functions may be desired, such as whitening, bleaching, desensitizing, inhibition of plaque formation and
protection against periodontal problems. Dentifrices are disperse systems. They consist of water and water soluble
liquids, oils and both soluble and insoluble solids. As such they are dispersions of solids in a liquid vehicle.
Important characteristic of toothpaste are consistency, abrasiveness, appearance, foaming, taste, stability and safety.
Most commonly ingredients used in toothpaste formulation are Active pharmaceutical ingredients, abrasives,
humectants, detergents, binders, sweeteners, preservatives and antioxidants. List of the ingredients used in
toothpaste are shown in Table 1 [3]. Historically the need and desirability of cleaning the teeth paralleled the
recognition of the necessity to maintain bodily cleanliness. Many of the materials used and recipes suggested,
however, contained materials capable of damaging the teeth and gums. The modern world therefore had a real
social, medical and aesthetic need for well formulated, safe and effective dentifrices. Toothpaste is the daily used
material for cleansing the teeth and to provide freshness to mouth. The rationale behind making herbal toothpaste is
to fight against the bacteria that cause problems regarding to teeth like gum, dental cavity and gingivitis.
The present work deals with development and evaluation of herbal toothpaste. The Plant materials used for the
formulations were aqueous extract of flower bud of clove (Eugenia caryophyllata), powder of bark of babul (Acacia
nilotica) and hydro alcoholic extract of powder of bark of Borsali (Mimusops elengi). The plants have been reported
as good anti-microbial agents in the studies. The present work also dealed with microbial study of these herbal
actives on Gram positive and Gram negative bacteria. Clove has good analgesic, antibacterial and local anaesthetic
activity. It is used in dental pain and teeth hypersensitivity. Babul and Borsali have good antibacterial effect on both,
gram positive and gram negative, spectrum. The plants have been reported as good anti-microbial, local anaesthetic
and anti-bleeding agents in the studies. Natural remedies are more acceptable in the belief that they are safer with
fewer side effects than the synthetic ones.

2. Materials and Methods

Materials
Clove bud powder, babul bark powder and borsali bark powder were purchased from the ayurvedic shop of
Ahmedabad. Calcium carbonate, glycerine, sorbitol solution (70%), carboxy methyl cellulose, sodium lauryl
sulphate, methyl paraben and sodium saccharine were issued from the laboratory of pharmaceutical technology,
KBIPER, Gandhinagar. During the microbial assay autoclave (electroquip, Ahmedabad) & hot air oven (Nova
instrumentation pvt ltd, Ahmedabad) were used for maintaining sterile condition and culture of nutrient agar bored
with bacteria were incubated using incubator(ambassador). Distilled water was used throughout the study.
Methods
Preparation of Herbal Extract
The net quantity of 5 g,7.5 g and 1 g of clove bud powder, babul bark powder and borsali bark powder were
transferred in a 250 ml conical flask respectively. To that distilled water was added to make volume 100 ml and
allowed to completely moisten. Refluxed on the water bath at temperature not exceed 600C for at least 1 hr. Allowed
to cool at room temperature. Filtered and evaporated to dryness on waterbath.Then dry extract was stored in air tight
zip-bag for further use. 1%, 7.5%, 1% aqueous extract of dried clove, babul and borsali were selected as
antibacterial, as it is reported to have antibacterial and analgesic activity at this concentration [15-17].
General procedure for preparation of Toothpaste Base
For designing of the experimental work, 32 factorial designs were used. Factors selected were conc. of Abrasive and
concentration of Humectant and levels chosen were low (-1), medium (0) and high (+1) and were shown in Table 2.
Different formulations of the toothpaste base were developed according to the Table 3. All the powder materials
were passed through 60# sieve. Accurate quantity of Carboxy methyl cellulose was weighed and transferred into
clean Mortar Pestle. 1/4th quantity of distilled water was added with continuous trituration till gel was formed.
Glycerin and preservatives were added to the Gel. Sorbitol, Sodium Saccharine and remaining quantity of water
were taken in a separate beaker. Above solution was added to the Gel. Mixed well. Calcium carbonate was added
slowly to above mixture. Mixed well. Dried extract of actives prepared previously was incorporated after proper
hydration in the boiling water and mixed well with the base with continuous trituration(for the evaluation of the
base, the actives were replaced with same quantity of water). SLS was dissolved in minimum quantity of water and
added to above mixture. The product was weighed and packed in air-tight container.
Evaluation of Formulations
Preliminary Test
Drying tendency
General Evaluation of tooth paste was done on the basis of drying tendency of the prepared bases (F1 F9) at room
temperature for a week and selected batches were evaluated for further evaluation parameters.
Then following tests were performed for the selected batches of toothpaste base.
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Organoleptic Characters
Selected batches were characterised on the basis of organoleptic characters like appearance, colour, texture, after
taste and extrudability.
Physicochemical Parameters
Determination of grittiness
The paste was extruded about 15 to 20 mm length from collapsible tube of each sample on a butter paper. Then all
the samples were tested by pressing it along its entire length by a finger for the presence of hard and sharp edged
abrasive particles.
Determination of pH
The net quantity of 5 gm of sample was accurately weighed and placed in a 150 ml beaker. To this 45 ml of freshly
boiled and cooled water was added at 27C. It was stirred well to make a thorough suspension. The pH was
determined within 5 minutes by using pH meter (Sartorius CP124S, Swisser instrument, India).
Determination of foaming power
About 5gm of sample was accurately weighed and placed in a 100ml glass beaker. To this10ml of water was added
and the beaker was covered with a watch glass and allowed to stand for 30 minutes. This operation was carried out
to disperse the toothpaste in water. The contents of the beaker were stirred with a glass rod and the slurry was
transferred to a 250ml graduated measuring cylinder, during this transfer ensured that no foam was produced and no
lump paste went into the measuring cylinder. The residue left in the beaker was transferred with further portion of 56 ml of water to the cylinder. The content of cylinder was adjusted to 50ml by adding sufficient water and the
content has to be maintained at 30C. Stirred the contents of the cylinder with a glass rod to ensure a uniform
suspension. As soon as the temperature of the content reached 30C, the cylinder was stoppered and 12 complete
shakes were given to it. The cylinder was allowed to stand for 5 minutes and the volume of foam with water (V1)
and water only (V2) was noted for all samples.
Determination of foaming power:
Foaming power = V1 V2
V1 - Volume in ml of foam with water.
V2 - Volume in ml of water only.
Performance Evaluation
Organoleptic characters
After incorporation of herbal actives final batch wascharacterised on the basis of organoleptic characters like
appearance, colour, texture, after taste and extrudability [18].
Microbial study
The antibacterial activities of different formulations were determined by modified agar well diffusion method. In
this method, nutrient agar plates were seeded with 0.2 ml of 24 h broth culture of S.aureus. The agar plates were
allowed to solidify. A sterile 8 mm borer was used to cut wells of equidistance in each of plates. 0.5 ml of
formulations or herbal extract was introduced into the well. The plates were incubated at 370C for 24 hours. The
antibacterial activity was evaluated by measuring the zones of inhibition (in mm) [19].

3. Results and Discussion

The optimization of the toothpaste bases was done on the basis of drying tendency. Table No.5 showed drying
tendency of batches F1 - F9. Among these batches F1, F4, F7, F8, F9 were selected for the further evaluation.
Characterization of the selected batches was done on the basis of different organoleptic characters like color,
appearance, pourabilityand surface and after taste feelings. The data are shown in Table 6. Formulation F1, F4, F8
and F9 shown good appearance but have chalky feeling therefore formulation F7 was met the criteria of acceptable
formulation.
Evaluation of selected and marketed formulation was done on the basis of grittiness, pH and foaming power
parameters. Those were shown in Table 7 comparative study indicates high foaming capacity of marketed product.
Formulation F7 has highest foam capacity among prepared batches. So optimized formulation was F7. Herbal
extracts of active ingredients were evaluated for different parameters like pH, specific gravity and color as shown in
Table 8.
Microbial study of herbal extracts and herbal toothpaste was evaluated by determination of zone of inhibition in
nutrient agar plates seeded with broth culture of S.aureus and E. coli. Table 9 shown zone of inhibition of reported
concentration of herbal extract on S.aureus (Gm +ve) &E.coli (Gm-ve) bacterial species. All the extracts and
product showed readily inhibition. Figure 1(A-E) showed antimicrobial activity on S.aureus (Gm+ve) and Figure2
(A-E) showed antimicrobial activity on E.coli (Gm ve).
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Sl no

Ingredients

API

Abrasive

Humectants

Water

Foaming agent

Binder

Flavors

Sweetener

Color

10

Preservatives

Table 1. List of the ingredients used in toothpaste [3]

Concentration
Materials Used
(% w/w)
Clove, Neem, Sunthi, Mentha, Tomar, Pippali, Aloe vera,
Up to 5
Kapoor
Precipitated Calciun Carbonate
Calcium Phosphate
Dicalcium Phosphate dihydrate
Anhydrous dicalcium phosphate
20-40
Calcium pyrophosphate
Sodium metaphosphate
Ammonium phosphate dibasic
Hydrated alumina, Silica
Glycerin, Poplyethylene glycol
20-40
Propylene glycol
Sorbitol solution (70%)
20-40
Distilled water
Sodium Lauryl Sulphate
Sodium Lauryl Sarcosinate
Sodium Lauryl Sulphoacetate
1-2
Magnesium Lauryl Sulphate
Monoglyceride
Dioctyl-Na Sulphosuccinate
Natural gum : Gum tragacanth, Acasia,Carrageenans
Cellulose derivatives :CMC, MC, Hydroxyehylcelullulose
Up to 2
Starch ether
Synth. Resins : Ehtylenoxide polymer
Carbopol (carboxy vinyl polymer)
Up to 2
Spearmint, peppermint, wintergreen, cinnamon mint
Saccharin solution (0.05-0.3%)
Up to 2
Chloroform, Sodium cyclamate, sorbitol
<1
Titanium dioxide
Methyl parahydroxy benzoate (0.15%)
Propyl parahydroxy benzoate (0.02%)
0.25 1.0
Sodium benzoate, Trclosan
Methyl paraben, Propyl paraben

API- Active Pharmaceutical Ingredient, CMC-Carboxy Methyl Cellulose, MC- Methyl Cellulose, %w/wPercentage weight by weight
Table 2. Level based design of the batches as per factorial design
Factor(2)
Concentration of Abrasive
Concentration of Humectant(Sorbitol solution)
Levels(3)
Low
-1 (20%)
-1 (30%)
Medium
0 (30%)
0 (40%)
High
+1 (40%)
+1 (50%)
Table 3. Developed batches from F1-F9 as per factorial design
Batches

Abrasive

Humectant

F1
F2
F3
F4
F5
F6
F7
F8
F9

-1
0
1
-1
0
1
-1
0
1

-1
-1
-1
0
0
0
1
1
1

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Table 4. Composition of the batches: (all quantity in %w/w)
Quantity taken per 100 gm paste ( in grams)

Ingredients
F1

F2

F3

F4

F5

F6

F7

F8

F9

3.0

3.0

3.0

3.0

3.0

3.0

3.0

3.0

3.0

20.0
5.0

30.0
5.0

40.0
5.0

20.0
5.0

30.0
5.0

40.0
5.0

20.0
5.0

30.0
5.0

40.0
5.0

25.0

25.0

25.0

35.0

35.0

35.0

45.0

45.0

45.0

0.25
0.5
0.25
1.0

0.25
0.5
0.25
1.0

0.25
0.5
0.25
1.0

0.25
0.5
0.25
1.0

0.25
0.5
0.25
1.0

0.25
0.5
0.25
1.0

0.25
0.5
0.25
1.0

0.25
0.5
0.25
1.0

0.25
0.5
0.25
1.0

10.0

10.0

10.0

10.0

10.0

10.0

10.0

10.0

10.0

q.s.

q.s.

q.s.

q.s.

q.s.

q.s.

q.s.

q.s.

q.s.

35.0

25.0

15.0

25.0

15.0

5.0

15.0

5.0

Color
q.s.
q.s.
q.s.
q.s.
+ SLS- Sodium Lauryl Sulphate, q.s. - Quantity Sufficient

q.s.

q.s.

q.s.

q.s.

q.s.

Carboxy methyl
cellulose
Calcium carbonate
Glycerin
Sorbitol solution
(70%)
Sodium Saccharine
Methyl Paraben
Propyl Paraben
SLS
Combined Herbal
extract
Flavor
Purified Water

Table 5. Evaluation of the toothpaste base on the basis of drying tendency

Batches Dryness test
F1
Not dried
F2
Dried
F3
Dried
F4
Not dried
F5
Dried
F6
Dried
F7
Not dried
F8
Not dried
F9
Not dried

Batches
F1
F4
F7
F8
F9

Table 6. Evaluation of the toothpaste base on basis of organoleptic characters.

Organoleptic character
Color
Appearance
Extrudability
Texture
After Taste
White opaque
Creamy
Easy
Smooth
Chalky
White opaque
Paste
Easy
Smooth
Chalky
Slightly
White opaque
Paste
Easy
Smooth
Chalky
White opaque
Paste
Easy
Smooth
Chalky
White opaque
Paste
Easy
Smooth
Chalky

Table 7. Evaluation of the toothpaste bases and marketed formulation on the Basis of different
Evaluation parameter
Batches Grittiness
pH
Foam capacity (ml)
F1
Absent
8.56
39
F4
Absent
8.70
38
F7
Absent
8.50
45
F8
Absent
8.56
40
F9
Absent
8.51
38
Amar
Absent
7.00
52
Vicco
Absent
7.00
52
Manufacture and marketed by: amar
remedies ltd.
Amar
Mfg date: 09/2011
Exp. Date: 08/2013
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Sl no.
1
2
3

Sl.No.

1
2

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Table 8. Characteristic of herbal extracts
Aqueous extract
Color
pH
Specific gravity
Clove
Light brown
6-8
1.019
Babul
Dark brown
7-8
0.899
borsali
Dark brown
7-8
0.911

Table 9. Microbial study of the Herbal extract & Herbal toothpaste.

ZONE OF INHIBITION(mm)
Aqueous
Aqueous
Hydro alcoholic
Microorganism
Combined
extract of
extract of
extract of
Extract
Clove (1%)
Babul (7.5%)
Borsali (1%)
Staphylococcus
1
3
3
2
aureus
Escherichia coli
1
2
3
3

A
)

2
2

C
)

B
)

D
)

Herbal
toothpaste
Formulation

E
)

Fig.1.Microbial Study on S.aureus of (A)aqueous extract of clove, (B)aqueous extract of babul, (C)aqueous extract
of borsali, (D)combined extract and (E)product.

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Fig 2. Microbial Study on E.coli of (A)aqueous extract of babul, (B)aqueous extract of clove, (C)aqueous
extract of borsali, (D)combined extract and (E)product.

4. Conclusion
Herbal formulations have growing demand in the global market. Natural remedies are more acceptable in the belief
that they are safer with fewer side effects than the synthetic ones. It is a very good attempt to establish the herbal
toothpaste containing hydro alcoholic extract of flower bud of clove (Eugenia caryophullus), bark powder of babul
(Acacia nilotica) and bark powder of borsali (Mimusops elengi).The study revealed that the developed single herbal
formulation F7 consisting 1% clove extract, 1% borsali and 7.5% babul was comparatively better than later other
formulation. They can be used for antibacterial activity.

5. Acknowledgement
The assistance of the department of pharmaceutics and pharmaceutical technology, K. B. Institute of pharmaceutical
education and research, Gandhinagar, India is gratefully acknowledged.

6. References
1.
2.
3.
4.
5.
6.
7.
8.
9.

AT Gray et al., Anthonys textbook of Anatomy physiology, 18th ed., Elsevier publication, 932-3.
PP Sharma, Cosmetics: formulation, manufacture, quality control, 7th ed., Pardana publication pvt ltd, 50719.
MR Martin, G David, Pharmaceutical Dosage forms: Disperse system, Inflorma healthcare publication, 2,
423-47.
CK Kokate., AP Purohit. et al., Pharmacognosy, 4th ed., Niraliprakasan, 11.81-94.
AR Pradeep, E Agarwal, SB Naik, Clinical and Microbiological Effects of Commercially Available
Dentifrices Containing Aloe vera: A Randomized Controlled Clinical Trial, 2011, J Periodontol.
AI Kholani, Comparison between the Efficacy of Herbal and Conventional Dentifrices on Established
Gingivitis, 2011, Dent Res J (Isfahan). Spring; 8(2):57-63.
E Herro, SE Jacob, Menthapiperita (peppermint):Dermatitis, 2010, 21(6),327-9.
NC Mohire, AV Yadav, Chitosan-based polyherbaltoothpaste: as novel oral hygiene product, 2010, Indian J
Dent Res., 21(3), 380-4.
W Scherer et al, The ability of an herbal mouth rinse to reduce gingival bleeding, 1998, J Clin Dent.; 9(4):97100.

International Journal of Chemistry and Pharmaceutical Sciences

672

Kandarp Dave et al

IJCPS, 2014, Vol.2(3): 666-673

10. BH Mullally, JA James, WA Coulter, GJ Linden, The efficacy of a herbal-based toothpaste on the control of
plaque and gingivitis,1995, J ClinPeriodontol. Sep, 22(9), 686-9.
11. J Moran, M Addy, R Newcombe, Comparison of an herbaltoothpaste with a fluoride toothpaste on plaque and
gingivitis, 1991, ClinPrev Dent., 13(3),12-5.
12. J George, S Hedge, KS Rajesh, A Kumar, The efficacy of aherbal-based toothpaste in the control of plaque
and gingivitis: a clinico-biochemical study, 2009, Indian J Dent Res., 20(4), 480-2.
13. AN Ellepola, ZU Khan, R Chandy, L Philip, A comparison of the antifungal activity of herbaltoothpastes
against other brands of toothpastes on clinical isolates of Candida albicans and Candida dubliniensis, 2011,
Med PrincPract., 20(2),112-7. Epub 2011 Jan 20.
14. NC Mohire, AV Yadav, Chitosan-based polyherbaltoothpaste: as novel oral hygiene product, 2010,Indian J
Dent Res., 21(3), 380-4.
15. B Joshi et al, physiochemical extraction and antimicrobial property of different plants: Ocimum sanctum
(tulsi), Eugenia caryophyllata (clove), Achyranthusbindentata (Datiwon), andAzadiractaindica (neem), 2011,
Journal of microbiology and antimicrobial, 3(1), 1-7.
16. AB Chaoudhari, Phytochemical screening and evaluation of Acacia nilotica leaves, 2011, IJPRD, 3(5), 1-7.
17. D Shahwar et al, in vitro antibacterial activity of extract of Mimusops elengi against gram positive and gram
negative bacteria, African j of microbiology res,2009, 3(8), 458-62.
18. T Akelesh et al, Evaluation of standards of some selected cosmetic preparations, JPRHS,2 (4), 302-6.
19. Indian pharmacopoeia 2010 volume I, II, III.

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