CLOVE OIL-PERIDONTAL DISEASES

1. “FORMULATION AND DEVELOPMENT OF DENTAL GEL CONTAINING CLOVE OIL FOR THE TREATMENT OF
PERIODONTAL DISEASES”
A Dissertation submitted to the
JAWAHARLAL NEHRU TECHNOLOGICAL UNIVERSITY ANANTHAPUR
In partial fulfillment of the requirements for the degree of
BACHELOR OF PHARMACY
Submitted by
SK. SANA BANU (12P21R0050)
K.CHANDANA (12P21R0014)
Y.CHARAN KUMAR (12P21R0015)
P.SATHEESH (12P21R0052)
P.SUBRAMANYAM (12P21R0062)
G.MANIKANTA (12P21R0035)
Under the Guidance of
V.VIJAY KUMAR M.Pharm., (Ph.D)
Associate Professor,
DEPARTMENT OF PHARMACEUTICS.
RAO’S COLLEGE OF PHARMACY
Chemudugunta (P.O.), Venkatachalam (M), Nellore, Andhra Pradesh.
2012 – 2016.

2. ABSTRACT :
Aim The study was aimed to develop and evaluate dental gel containing clove oil as the chief
constituent for the treatment of periodontitis.
Methods It has a wide spectrum of antibacterial activity against a number of periodontal pathogens,
hence it is selected for the treatment of periodontitis. Clove oil gel is formulated by using carbopol 934
as gelling agent, clove oil as medicinal agent, polyethylene glycol co-solvent, methyl paraben and
propyl paraben as preservative and required quantity of distilled water as vehicle.
Results The clove oil was evaluated for physical parameters like acid value, ester value, specific gravity
and refractive index and it shown satisfactory results. The prepared gel was evaluated for various
properties such as antimicrobial activity, pH, spreadability, extrudability, drug content etc. In-Vitro
experiments demonstrated that the formulation F3 is a suitable dosage form for the treatment of
periodontitis. Clove oil showed the zone of inhibition of about 22.05±0.04 mm.
Conclusion On the basis of the result obtained in this present study we conclude that the gel
formulations of clove oil F3 showed good physicochemical properties as well as good drug content
compared to other formulations.
Key words Clove oil, Carbopol 934, Periodontitis, Anti microbial activity.
Ref: Vijaya Kumar Voleti*, Sana Banu Shaik, Chandana Konduru, Sathish Peyam, Charan Kumar
Yaramsetti, Subramanyam Pasala, Shanmuga Pandiyan Pitchaimuthu. Formulation and development of
dental gel containing clove oil for the treatment of human periodontal diseases. JCP. 2016;3(1): 1-7

3. 2nd Indo-Korean Conference,
Sullurpet, 30 th MARCH 2016
Theme: Herbals And Pharmaceuticals: Pivotal Issues & Concerns
Venue: Gokula Krishna College of Pharmacy
(Affiliated to JNTUA, Ananthapuramu, Approved by AICTE & PCI-New Delhi)
SPSR Nellore, A.P., India – 524121
Acceptance of Poster/Oral Presentation
21/03/2016
Dear Delegate
We are pleased to inform you that your paper entitled “Formulation And Devfelopment Of Dental Gel Containing Clove Oil For The
Treatment Of Human Periodontal Diseases ” has been accepted for presentation as “ Poster No:PPCE239” in Poster Session at the
2ndIndo – Korean conference-HPPIC2K16.
As a presenter, you must be a registered delegate for the HPPIC2K16. However only the abstract of those presenters, who are registered by
25 March 2016, will be published in the final abstract book. Selected papers of the seminar might be published in “JOURNAL OF
COMPREHENSIVE PHARMACY”. Those authors, who wish to publish their manuscript in the journal, should submit their full length
papers on or before 25.03.2016.The full length manuscript should be in accordance with the instructions of Journal Of Comprehensive
pharmacy”.
You should have your registration badge at the poster venue in order to present the poster/oral.
For any Queries Contact:
Kindly bring a copy of this letter and photo ID (College ID) for identification at the poster /oral presentation and also have to submit this
letter to the registration committee during conference.
The area provided for poster presentation will be about 1 meter (100cm), width by 1 meter height.
We look forward to meet you at 2ndIndo – Korean conference, Sullurpet.
Kind regards
Chairman, Registration Committee-IKC
Sullurpet, SPSR Nellore.
Dr .Pavan kumar Balagani
Principal, Convenor, HPPIC2K16,
Gokula Krishna college of pharmacy, sullurpet, SPSR Nellore, AP
Email:nchp2k16@gmail.com
Mrs.M.Soujanya For registration: 9494795974
Mr. Deepak kumar For general information: 9652275737
Mr.SK Afsar For scientific session: 98855398761

4. Introduction
Herbal gels:
• Medicinal plants have been used as traditional treatments for numerous
human diseases for thousands of years and in many parts of the world.
In rural areas of the developing countries, they continue to be used as the
primary source of medicine. About 80% of the people in developing
countries use traditional medicines for their health care [1]
Benefits of herbal drugs:
• Herbal drugs have long era of use and better patient tolerance as well as
public acceptance.
• Herbal drugs acts as a renewable source, which is our only hope for
sustainable supplies of cheaper medicines for the worlds growing
population.
• The cultivation and processing of medicinal herbs and herbal products is
environment-friendly.
• Throughout the world, herbal medicine has provided many of the most
useful and vast variety of drugs to the modern medical science[2].

5. Pharmaceutical Gels
• Defintion:
Gels are semisolid organic or inorganic colloids rich in liquid,
consisting of hydrated threads or granules of the dispersed phase
intimately associated with the dispersion medium
• Some gelling agents (carbomers) require a "neutralizer" or a pH
adjusting chemical to create the gel after the gelling agent has been
wetted in the dispersing medium[3].
• Gelling agents are used concentrations of 0.5% to 10%, depending
on the agent.
• It is easier to add the active drug before the gel is formed if the
drug doesn't interfere with the gel formation[4].
• Only Carbopol® 934P, methylcellulose, hydroxypropyl
methylcellulose, and sodium carboxymethyl cellulose are
recommended for oral administration.

6. Periodontal diseases
• Periodontal disease is one of the most important concerns for dentists and
patients[5]. It is recognized as a major public health problem throughout the
world and is the most common cause of tooth loss in
• The periodontium is the specialized tissues that both surround and support
the teeth, maintaining them in the maxillary and mandibular bones. A
variety of triggering factors like bacterial causes, dyscrasias, avitaminosis
etc cause inflamed gums leading to gingivitis.
• In the United States 50% of adults have gingivitis affecting at least 3-4
teeth; two-thirds of the population has sub gingival calculus, and about a
one-third have periodontitis [6] . Periodontal treatment aims to cure inflamed
tissue, reduce the number of pathogenic bacteria and eliminate the diseased
pockets.
• Mechanical therapy, chemotherapy and systemic administration of
antibiotics are some of the clinical methods being utilized currently[7]

7. S.NO. Author Names Abstract References
1. Reenu Yadav Formulation and characterization of antimicrobial oral gel
from some herbal extracts for treatment of periodontal
diseases.
IJPPR. 2016, 5(2)
2. R.Bhramaramba Formulation and Evaluation of herbal gel containing
Terminalia chebula Retz., leaves extract.
(SAJP). 2015, 4(3): 172-176.
3. Biresh Sarkar Formulation and Evaluation of herbal gel containing
extract of Cedrus deodara.
(IJPCS). 2015,4(1)
4. Singh Rampal Formulation, optimizatioon and evaluation of aceclofenac
transdermal gel
(JPSI). 2015,4(5)
5. T. Muthu Lakshmi Formulation and evaluation of herbal gel containing
dalbergia sissoo roxb. bark extract.
(JPRCP).4(1):53-57
6. Dr Rohit Jain Herbs in periodontology – local drug delivery (WJPR).2014,3(2):1831-1840
7. Varsha B. Bagade Study of antimicrobial activity of herbal formulation (IJPLS).2013, 4(11)
8. Deepak p pawar Formulation and evaluation of herbal gel containing
lantana camara leaves extract.
(AJPCR).2013,6(3)
9. CharuM. Marya Investigation of In vitro inhibitory effect of clove essential
oil and its two active principles on tooth decalcification by
applying juice.
(IJD).2012
10. L. Nuñez Investigation of microbicide activity of clove essential oil
(Eugenia caryophyllata) .
(BJM).2012,43(4)
11. Ganesh Misal Formulation and evaluation of Poly herbal gel. (IJNPR).2012,3(4)
12. Manisha singh Formulation and evaluation Herbal Gel Containing
Ethanolic Extract of Ipomoea Fistulosa.
(IJSR). 2012, 3(7)
13. Ilhami Gu¨ lc¸ Investigation of antioxidant activity of clove oil – A
powerful antioxidant source.
(AJP). 2012, 5: 489-499
14. Euge´ nia Pinto Antifungal activity of the clove essential oil from Syzygium
aromaticum on Candida, Aspergillus and dermatophyte
species.
(JMM). 2009, 1454-1462
Literature Review

8. The study was aimed to develop and
evaluate dental gel containing clove oil
as the chief constituent for the
treatment of periodontitis.
The objectives of the research
work under taken are as follows:
1. To perform clove oil
characterization.
2. To formulate dental gel of
clove oil using gelling agents
and other ingredients.

9. Plant Profile
Synonym:
• Caryophyllum; clove flower; clove bud;
launge
Biological source:
• Cloves consists of dried flower buds of
Eugenia caryophyllus (Myrtaceae). It should
contain not less than 15% (v/v) of clove oil[8].
Chemical constituents:
• 15-20% of volatile oil; 10-13% of tannin
(gallotannic acid), chromone and eugenin [9].
• The volatile oil contains eugenol (about 70 to
90%), eugenol acetate, methylamylketone,
caryophyllenes and small quantities of ester
and alcohols [10].
Uses:
• Dental analgesic, carminative,
• Stimulant, flavouring agent, an aromatic and
antiseptic

10. Taxonomical classification of clove plant (Syzygium
aromaticum)
Taxonomic Hierarchy
Kingdom Plantae - plantes, planta, vegetal, plants
Subkingdom Viridiplantae
Infrakingdom Streptophyta – land plants
Superdivision Embryophyta
Division Tracheophyta – vascular plants, tracheophytes
Subdivision Spermatophytina – spermatophytes, seed plants,
phanerogames
Class Magnoliopsida
Superorder Rosanae
Order Myrtales
Family Myrtaceae – myrtles, myrtacees
Genus Syzigium P.Br.ex Gaertn
Species Syzygium aromaticum (L) Merr. & L.M.Perry – clove

11. Excipient profile
Carbopol[2]
Nonproprietary Names:
• BP: Carbomers
• PhEur: Carbomers
• USP-NF: Carbomer
Structural Formula:
PEG[2]
Use Concentration (%)
Emulsifying agent 0.1-0.5
Gelling agent 0.5-2.0
Suspending agent 0.5-1.0
Tablet binder 0.75-3.0
Nonproprietary Names:
BP: Macrogols
JP: Macrogol 400
Macrogol 1500
Macrogol 4000
Macrogol 6000
Macrogol 20000
PhEur: Macrogols
USP-NF: Polyethylene Glycol
Structural Formula:
Uses:
Ointment base; plasticizer; solvent; suppository
base; tablet and capsule lubricant.

12. S.NO MATERIALS SUPPLIERS FUNCTION
1. Clove oil S.D.Fine chemicals Pvt. Ltd,
Mumbai
Active pharmaceutical
ingredients
2. Carbopol934 Roquette, Mumbai Gelling agent
3. Polyethylene glycol Isp Chemical Inc, Hyderabad Cosolvent
4. Glycerin Bayer Chemicals, Pune,
Maharashtra
Drug solubiliser
5. Methyl paraben Merck Index chemicals,
Hyderabad
Preservative
6. Propyl paraben Merck Index chemicals,
Hyderabad
Preservative
7. Aspartame Nutrasweet, Vadodara, Gujarat Sweetening agent
8. Distilled water Bayer Chemicals, Pune,
Maharashtra
Vehicle
Materials and Methods

13. Physico chemical characteristics of
Clove oil
Acid value
Acid value = potassium hydroxide consumed × 5.611 5.611
Weight (g) of the sample
Saponification value:
• Saponification value = mg of KOH consumed by 1 g clove oil
• Weight of KOH = Normality of KOH × Equivalent weight× volume of KOH in litres
• Volume of KOH consumed by 1 g of oil = [blank- test]
Ester value
• Ester value = Saponification value – Acid value
Solubility:
• Clove oil is freely soluble in ethanol
Colour:
• The colour of the formulation was checked out against a white background.
Odour:
• The odour of the gels was checked by mixing the gel in water and taking the smell.

14. Formulation of clove oil gel
Soaking • Soak carbopol 934 in water
Neutralizat
ion
• Neutralize with triethanolamine to pH
6.4
Addition of
preservative
• Addition of methyl and propyl paraben
Additon of
co solvent
and API
• Addition of propylene glycol and clove oil
in another test tube
Addition of
sweetner
• Finally aspartame is added
Stirring
• Stirring is done until a homogenous
product is formed

16. Composition of gel formulations
INGREDIENTS F1 (gm) F2 (gm) F3 (gm) F4 (gm) F5 (gm)
Clove oil (ml) 0.75 0.75 0.75 0.75 0.75
Carbopol (g) 0.3 0.4 0.5 0.6 1
Polyethylene glycol
(ml)
15 15 15 15 15
Glycerine (ml) 5 5 5 5 5
Methylparaben(g) 0.18 0.18 0.18 0.18 0.18
Propylparaben(g) 0.02 0.02 0.02 0.02 0.02
Aspartame (g) 0.4 0.4 0.4 0.4 0.4
Distilled water q.s q.s q.s q.s q.s

17. Evaluation of gel formulation
Appearance :
• All the formulations of clove oil gel were pale yellow in colour.
Consistency:
• The consistency was checked by applying on skin.
Greasiness:
• The greasiness was assisted by the application on to the skin.
Determination of pH:
• The pH of gel was determined using digital pH meter by dipping the glass electrode completely into the gel
system [11].
Determination of viscosity:
• Viscosities of the formulated gels was determined using Brooke field viscometer, spindle no. 7 and spindle
speed 60 rpm at 25◦C was used for gels, the corresponding dial reading on the viscometer was noted[12].
Determination of spreadability:
• Spreadability was determined using following formula,
S=M.L/T
• Where S is the spreadability in grams.cm/sec, M is the mass in grams, T is the time in seconds.
Determination of extrudability:
• It was determined by sing a tube filled with the gel, having a tip of 5mm opening and by measuring the
amount of gel that extruded through the tip when a pressure was applied on the tube was noted down [13].

18. Determination of antimicrobial activity:
Agar cup plate method was used for screening of antimicrobial activity of
clove oil gel. Different concentrations of clove oil gel were placed
aseptically in cups of agar plate which was previously inoculated with
culture[14]. The plates were left at ambient temperature for 30 mins prior
to incubation at 37◦C for 24 hrs. The broad spectrum antibiotic i.e.,
tetracycline was used as positive control for obtaining comparative
results[15]. Plates were observed after 24-48 hrs incubation for the
appearance of the zone of inhibition. Antimicrobial activity was evaluated
by measuring the diameter of zones of inhibition (millimetres) of
microbial growth.

19. Results
S.NO. PARAMETERS CLOVE OIL
PROCURED
CLOVE OIL
STANDARD
1. Colour Pale yellow Pale yellow
2. Odour Aromatic Aromatic
3. Acid value 3.66 3.84
4. Ester value 37.21 38.22
5. Solubility in ethanol Freely soluble Freely soluble
6. Density 1.02 g/ml 1.05g/ml
7. Refractive index 1.492 1.532
Table : 5 Physicochemical characteristics of clove oil

20. 1
2
3
Fig: Antimicrobial activity on S.salivarius
1 – F3 (Gel Formulation)
2 – Clove oil
3 – Tetracycline
5

21. Formulatio
ns
Appearance pH Spreadability
(g-cm/sec)
Extrudabili
ty %
Homogeneity Drug
Content
F1 Pale yellow 6.6 18.20 92.14 Good 95.00
F2 Pale yellow 6.7 18.14 93.15 Good 95.20
F3 Pale yellow 6.7 17.49 94.10 Very good 95.40
F4 Pale yellow 6.6 16.72 90.23 Good 93.62
F5 Pale yellow 6.4 15.59 89.10 Very good 89.80
Table:6 Characteristics of gel formulations

22. 16.5
17
17.5
18
18.5
19
19.5
20
Zone of inhibition
(S.salivarius)
Zone of inhibition
(S.sanguis)
Zone of
inhibition(L.acidophilus)
F3
Fig: 3 Antimicrobial activity on S.salivarius
0
5
10
15
20
25
F1 f2 F3 F4 F5
Zoneofinhibition(mm)
Zone of inhibition(S.salivarius)
Gel formulations
Fig: 4 Zone of inhibition of Streptococcus salivarius
0
5
10
15
20
25
F1 F2 F3 F4 F5
zoneofinhibition(mm)
Zone of inhibition (S.sanguis)
Gel formulations
Fig:5 Zone of inhibition of Streptococcus sanguis
0
5
10
15
20
25
F1 F2 F3 F4 F5
zoneofinhibition(mm)
Zone of inhibition (L.acidophilus)
Gel formulations
Fig: 6 Zone of inhibition of Lactobacillus
acidophilus

23. Discussions
• The procured clove oil was characterized for the following parameters:
 Acid value : 3.66
 Ester value : 37.43
 Saponification value: 41.09
 Density : 1.02gm/ml
 Refractive index: 1.492
• The formulations were developed by using clove oil of same concentration and carbopol 934 at
different concentrations.All the formulations were pale yellow in colour and had characteristic
odour of clove oil. The pH of all formulations ranged from 6.4-6.7, which was well within the
normal pH range of buccal cavity 6-7. The spreadability of the gels was found to be in the range
of 15.59-18.20 g-cm/sec, confirming that these gels may spread smoothly and uniformly. The
formulations were glossy and translucent. The homogeneity and tube extrudability of all
formulations was good.
• The drug content of the formulations was ranged from 89.8%-95.40% Table-6.The formulation F3
was found to have maximum drug content.
• The gel formulations of clove oil F3 showed good physicochemical properties as well as good
drug content compared to other formulations.(Table -5,6). Hence, theses formulations were
further selected for anti microbial studies. The results of anti microbial studies showed that gel
formulation of clove oil F3 showed a maximum zone of inhibition on Streptococcus salivarius.

24. Conclusion
• The clove oil was found to have anti microbial activity
against Streptococcus salivarius, Streptococcus sanguis,
Lactobacillus acidophilus. The formulations developed
from clove showed significant results so it can be further
used commercially to develop dental gels after
conducting clinical trials on human beings. Nevertheless
further research is still needed in order to determine if
they efficiently could substitute the synthetic antibiotics
are used in combinations.

25. References
1. Euge nia Pinto; Lui s Vale-Silva; Carlos Cavaleiro and Li gia Salgueiro. Antifungal activity
of the clove essential oil from Syzygium aromaticum on Candida, Aspergillus and
dermatophyte species. Journal of medical microbiology. 2009: 1454-1462.
2. Author. Handbook of pharmaceutical excipients. Pharmaceutical press. 2009.
3. Chaieb, K. et al. (2007) The Chemical Composition and Biological Activity of Clove
Essential Oil, Eugenia caryophyllata (Syzigium aromaticum L. Myrtaceae): A Short
Review. Phytotherapy Research, 21, pp. 501-506.
4. Gupta A. K., Tandon Neeraj, Dr. Sharma Madhu (2011). Quality Standards of Indian
Medicinalplants, Indian Council of Medical Research, New Delhi, Volume 3
5. Ates D.A. and Erdogrul O.T. (2003) Antimicrobial activities of various medicinal and
commercial plant extracts. Turkish Journal of Biology, 27: 157-162.
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antioxidant and antimicrobial activities of turmeric clear liquid soap for wound treatment of
HIV patients. Journal of Science Education and Technology.27: 569-578.
7. Das K, Dang R, Machale UM, Fatepuri S. Formulation and evaluation of herbal gel
containing Stevia leaves extract, The Pharma Review 2010, 8(44): 112-118.

26. Contd...
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9. Satpathy B, Sahoo M, Sahoo P, Patra SR, Formulation and evaluation of gel containing
essential oils of Piper betle against skin infection pathogens. Int . J. Res Phar. Sci 2011;
2(3): 373-378.
10. Hanoch Julianus Sohilait(2015). Chemical Composition of the Essential Oils in Eugenia
caryophylata, Thunb from Amboina Island, Science Journal of Chemistry ; 3(6): 95
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13. Lobene RR, Kashket S, Soparkar PM, Shloss J, Sabine ZM. The effect of cetylpridinium
chloride on human plaque bacteria and gingivitis. Pharmacol Ther Dent. 1979; 4:33-47
14. Pawar Vinita A*, Bhagat Trupti B, Toshniwal Mitesh R, Mokashi Nitin D, Khandelwal K.R;
Formulation and evaluation of dental gel containing essential oil of coriander against oral
pathogens; Pawar Vinita A et al. Int. Res. J. Pharm. 2013, 4 (10).
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Evaluation of Herbal Gel containing Extracts of Albezia Lebbeck linn; Am. J. PharmTech
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