IOP Conference Series: Earth and Environmental Science

PAPER • OPEN ACCESS Related content

- Optimization and Characterization of
Antibacterial activity of cinnamon ethanol extract Cinnamon Leaves (Cinnamomum
burmannii) Oleoresin
(cinnamomum burmannii) and its application as a L.U. Khasanah, Kawiji, P. Prasetyawan et
al.

mouthwash to inhibit streptococcus growth - The effect of cinnamon bark

(Cinnamomum burmanii) essential oil
microcapsules on vacuumed ground beef
To cite this article: Syahdiana Waty et al 2018 IOP Conf. Ser.: Earth Environ. Sci. 130 012049 quality
I N Brilliana, G J Manuhara, R Utami et al.

- Mycosynthesis of silver nanoparticles

using extract of endophytic fungi,
Penicillium species of Glycosmis
View the article online for updates and enhancements. mauritiana, and its antioxidant,
antimicrobial, anti-inflammatory and
tyrokinase inhibitory activity
M Govindappa, H Farheen, C P
Chandrappa et al.

This content was downloaded from IP address 139.192.255.74 on 14/04/2019 at 16:29

ICBSB IOP Publishing
IOP Conf. Series: Earth and Environmental Science1234567890
130 (2018) 012049 doi:10.1088/1755-1315/130/1/012049

Antibacterial activity of cinnamon ethanol extract

(cinnamomum burmannii) and its application as a mouthwash to
inhibit streptococcus growth

Syahdiana Waty1,2, Dwi Suryanto1*, Yurnaliza1

1
Department of Biology, Faculty of Mathematics and Natural Sciences, University of
Sumatera Utara, Jln. Bioteknologi No. 1, Kampus USU, Medan
2
Medan Health Polytechnique, Ministry of Health, Jln. Jamin Ginting Km. 13.5, Lau Cih,
Medan

Email: dwisuryanto@usu.ac.id

Abstract. Cinnamon bark has been commonly used as spicy and traditional medicine. It contains
several antibacterial compounds such as flavonoids, saponins, and cinnamaldehyde. Several
studies have been done to know the antibacterial effect on bacteria such as Streptococcus in vitro.
This study aimed to examine the antibacterial activity of cinnamon ethanol extract against
Streptococcus and its application as mouthwash to inhibit the bacteria. The cinnamon bark was
macerated followed by extracted in 80% ethanol. Bacterial samples were isolated from dental
plaque of patients visiting dental clinic drg. Syahdiana Waty in Medan, North Sumatra. The
isolates were identified using Vitek 2 compact. Secondary metabolites were detected using
previously described method. Antibacterial assay was done at extract concentration of 6.25%,
12.5%, and 25%. The result showed that alkaloids, flavonoids, saponins, and glycoside were
detected in the extract. Nine bacterial species were identified as Streptococcus mitis, S. sanguinis,
S. salivarius, S. pluranimalium, S. pneumoniae, S. alactolyticus, Kocuria rosea, Kocuria kristinae,
and Spingomonas paucimolis. It showed that the extract of Cinnamon bark significantly inhibited
Streptococcus growth, and it was effective as mouthwash.

Keywords: antibacterial activity, cinnamon bark, mouthwash, Streptococcus

1. Introduction
Cinnamon bark has been used for spicy and traditional medicine.Cinnamon bark and its leaves contain
essential oils, saponins and flavonoids that have been widely used for healing various diseases [1]. The
main compound of the essential oil is cinamaldehyde (60.72%), eugenol (17.62%) and coumarin
(13.39%) which has antibacterial effect [2].
Prevalence of dental and oral diseases in Indonesia tends to increase in which caries is the most
one. The dental caries and abnormalities of the dental begin with the formation of dental plaque [3]. The
formation of plaque in the form of a thin layer attached to the tooth surface and sometimes also found in

Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution
of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI.
Published under licence by IOP Publishing Ltd 1
ICBSB IOP Publishing
IOP Conf. Series: Earth and Environmental Science1234567890
130 (2018) 012049 doi:10.1088/1755-1315/130/1/012049

the gums and tongue is caused by food debris used by pathogenic bacteria in the oral cavity [4].
Pathogenic bacteria present in the oral cavity include Streptococcus mutans, S. viridans,S. pneumoniae,
St. epidermidis, and St. aureus [5].
There are various ways to reduce the accumulation of plaque in the oral cavity, among those are
brushing teeth regularly, rinsing with antiseptic solution, cleaning interdental with dental floss, cleansing
the tongue, chewing gum, and avoiding fermented carbohydrates [6]. The easiest way to remove plaque is
by rinsing. Several studies have demonstrated the effectiveness and usefulness of an antiseptic
mouthwash containing active compounds such as chlorhexidine and essential oils to prevent the formation
of plaque and gingivitis [7].
One of the potential compounds in controlling plaque formation is in cinnamon bark. Puspita et
al. (2013) showed that cinnamon extract has an effect on the growth of S. mutans which is carriogenic
bacteria, but there is no information about the effectiveness of using cinnamon extract as a mouthwash to
reduce dental plaque [2]. Therefore, in this study cinnamon ethanol extract applied as a mouthwash in
inhibiting the growth of these bacteria was conducted.

2. Methods
2.1. Cinnamon bark maceration, extraction, and phytochemical test
Cinnamon bark was macerated followed by extraction in ethanol. A-2 kg of dried cinnamon bark
was immersed in 20 L of distilled ethanol. Maserate was evaporated using a rotary evaporator at 60° C.
The extract was fed into a water bath to remove remaining moisture. The extract was subjected to
phytochemical tests including alkaloids, glycosides, anthraquinone glycosides, saponins, tannins,
flavonoids, and steroids/triterpenoids.

2.2. Patients,plaque samples, and bacterial identification

Forty patients from Dental Clinic of Drg. Syahdiana Waty, Medan were selected based of
Slovin’s equation. Sample inclusion criteria were patients with dental caries of at least one tooth, aged 35-
44 years, in good health and did not previously use antibiotic drug, willing to fulfill informed consent and
willing not to brush their teeth before taking plaque samples. Samples were taken on buccal portion of 16,
26, 36 and 46 and labial of teeth 11 and 31. Samples were stored in containers and immediately taken to
laboratory for bacterial culture. The samples were dissolved in sterilized water and subjected to be
vortexed. Vortexed sample was streak on trypticase yeast cysteine and incubated at 37° C overnight.
Single colony was identified using Vitek 2 compact.

2.3. Antibacterial assay of cinnamon bark extract and prepared mouthwash

Cinnamon bark extract was prepared in concentration of 6.25%, 12.5%, and 25% using dimethyl
sulfoxide as solvent. Bacterial solution was prepared in 0.90% of NaCl and equalised to 0.5 Mc Farland.
The bacterial solution was steak on entire surface of Muller Hinton agar using sterilized cotton bud. Paper
disc immersed in each cinnamon extract concentration was put on the bacterial lawn in petri dish. The
petri dish was incubated at 37° C overnight. Inhibition zone was measured as diameter of clearing zone
around the paper disk subtracted to 0.5 cm. The extract with Minimum Inhibitory Concentration (MIC)
was used as active compound in prepared mouthwash. pH and viscosity of prepared mouthwash were
measured. Application of 10 ml mouthwash was conducted in patients for 30 seconds. The plaque sample

2
ICBSB IOP Publishing
IOP Conf. Series: Earth and Environmental Science1234567890
130 (2018) 012049 doi:10.1088/1755-1315/130/1/012049

and measuring bacterial cell number were conducted as previously described. Number of bacterial cell
was measured as colony growth in total plate count agar.

2.4. Design and data analysis

This study was a pretest-post test one group design.Data was analyzed using Shapiro Wilk,
Kolmogorov-Smirnov, Kruskal-Wallis, Wilcoxon and Man Whitney tests.

3. Results and Discussion

3.1. Cinnamon bark maceration, extraction, and phytochemical test
Maceration and extraction of 20 kg of the bark yielded 17.5 L of the brown macerate and 670
gram reddish brown extract, respectively. Phytochemical test showed that cinnamon bark ethanol extract
contains secondary metabolite compounds such as alkaloid, saponin, flavonoid, and glycoside groups.
Puspita et al (2013) showed that cinnamon bark extracts contained transinamaldehid, polyphenols,
flavonoids, saponins, and tannins [2]. Awang et al (2013) reported that cinnamon bark essential oil
contained cinnamaldehyde which is potential as an antimicrobial compound [8]. Alkaloid is one of the
most organic compounds found in nature. It has prominent antimicrobial activity and has been widely
used in antimicrobial treatment [9]. Saponin is a compound that has a working mechanism as antibacterial
with its lipophilic properties capable of damaging cell membranes [10]. Flavonoid compounds disrupt
bacteria by destroying the cytoplasmic membrane and causing leakage of important metabolites that
inactivate bacterial enzyme systems [11].

3.2. Isolation of plaque bacteria

There were nine bacterial species found in the plaque samples, in which Streptococcus was the
most common bacteria. There were six species of Streptococcus including S. mitis, S. sanguinis, S.
salivarius, S. alactolyticus, S. pneumoniae, and S. pluranimalium. The other three bacterial species were
Kocuria rosea, K. kristinae, and Sphingomonas paucimolis (Table 1).

Table 1. Frequency of Streptococcus and other bacterial species from dental plaque

No. Bacteria species Number of samples Frequency (%)

1. S. mitis 12 30
2. S. sanguinis 8 20
3. S. salivarius 4 10
4. S. alactolyticus 3 7.5
5. S. pneumoniae 1 2.5
6. S. pluranimalium 3 7.5
7. K. rosea 2 5
8. K. kristinae 6 15
9. S. paucimolis 1 2.5
Total 40 100

3
ICBSB IOP Publishing
IOP Conf. Series: Earth and Environmental Science1234567890
130 (2018) 012049 doi:10.1088/1755-1315/130/1/012049

Streptococcus mitis, S. sanguinis and S. salivarius were the most common bacteria in dental
plaque. Streptococcus mitis is one of the bacteria of the mutans streptococci group (MSG). This group is
known as cariogenic. It also included other species such as S. mutans and S. sobrinus. This group of
bacteria is known as the pioneer bacteria involved in plaque formation and initiates dental caries [12].
Sterptococus sanguinis is one of species of Viridans Group Streptococci (VGS) which is most
commonly found in dental plaque in the oral cavity. S. sanguinis and MSG have a strong association as
oral streptococci and the most commonly found in dental plaque [12]. Streptoccous salivarius together
with S. sanguinis, S. mitis,and S. gordonii known asthefirst colonizer the tooth surface, in contact with
salivary glycoproteins on tooth surfaces via specific polymer capsules such as glucan and fructan.
Furthermore, these bacteria play an important role in the process of dental caries together with other Oral
Streptococci [13]. Streptoccous pluranimalium was a species of VGS that is rarely encountered in dental
plaque. Dhotre et al. (2016) found three strains of Streptococcus that were very rare and unusual in dental
plaque subgingival plaque such asS. pluralimalium, S. thoraltensis and S. hyointestinalis [14].
Bacteria K. rosea and K. kristinae are a gram-positive bacteria belonging to the family
Micrococcus. Sabtie et al (2015) found 16 species of bacteria in dental supragingival plaque from 50
patients, in which two were K. rosea and K. kristinae [15]. The normal habitat of the Kocuria is on the
skin but can also be found in the urinary tract in patients with urinary tract infections. This non-
pathogenic bacteria arehuman normal flora. However, many studies showed that these bacteria might
cause many infections of the urinary and gastrointestinal tract [16]. Many diseases in the gastrointestinal
tract, urinary, and even other vital organ diseases are believed to be associated with the oral cavity
diseases.

3.3. Antibacterial activity

The result of antibacterial test of cinnamon bark ethanol extract showed all concentration of the
extract inhibited the bacterial growth (Figure 1.). Cinnamon bark extract of 6.25%, 12.5% and 25%
showed inhibitory zone of 6.78, 9, and 11.68 mm, respectively (Table 2).

Figure 1. The bacterial inhibitory zone of cinnamon bark ethanol extract to S. mitis (25) and to S.
sanguinis (34)

4
ICBSB IOP Publishing
IOP Conf. Series: Earth and Environmental Science1234567890
130 (2018) 012049 doi:10.1088/1755-1315/130/1/012049

Table 2. Inhibitory zone of cinnamon ethanol extract to bacteria from dental plaque
Concentration of cinnamon extract Diameters of inhibitory zone(mm)
6.25% 6.78
12.5% 9
25% 11.68
Tetracyclin 28.3

Data analysis showed that there was a significant difference in bacterial inhibition zone of the
three concentrations. Furthermore, it was showneach concentration showed significantly different on
inhibitory zone. The similar result showed in Puspita et al. (2013) to inhibit the growth of S.mutans. Al-
Duboni et al. (2013) reported that cinnamon bark extract showed to have more antibacterial activity than
that of ginger extract on S. mutans [17].

3.4. Examination of cinnamon extract mouthwash as antibacterial agent

It was shown that the prepared mouthwash had pH of 6.97 with viscosity of 1.01 cP.
The results of antibacterial activity found that population of bacteria were reduced (Figure 2.). It was
shown that there was a significant difference to bacterial activity of the prepared mouthwash in the mouth
before and after rinsing.

Figure 2. The average population of bacteria before and after rinsing with the mouthwash of Cinnamon
extract, Chlorhexidin and sterile aquades.

From this study it was suggested to use this extract as an alternative in controlling pathogenic oral
cavity bacteria. Many other plants such as starfruit leaves, gambir leaves, siwak, and lemon were reported
to be used in mouthwash against bacteria in dental plaque in vivo. Fajriani and Mahrum (2015) reported

5
ICBSB IOP Publishing
IOP Conf. Series: Earth and Environmental Science1234567890
130 (2018) 012049 doi:10.1088/1755-1315/130/1/012049

that 40% lemon extract showed a significant effect in reducing bacterial number 30 minutes after rinsing
[18].

4. Conclusions
Cinnamon bark ethanol extract contained phytochemical compounds such as alkaloid, flavonoid,
saponin, and glikosid. The extract of each concentration showed antibacterial activity in vitro, and as
mouthwash as well.

Acknowledgment
Many thanks are addressed to Ministry of Health, Republic of Indonesia for providing a
scholarship and research fund.

References

[1] Pitojo S, and Zumiati (2016) Tanaman Bumbu dan Pewarna Nabati, Edisi. VIII. CV. Aneka Ilmu,
Semarang. 64-66.
[2] Puspita A, Kholifa M, and Rochmanita, N (2013) Pengaruh Konsentrasi Ekstrak Kulit kayu manis
(Cinnamomum burmanni) dalam Menurunkan Pertumbuhan Streptococcus mutans secara In
Vitro. Fakultas Kedokteran Gigi UMS.
[3] Pratiwi R (2005) Perbedaan Daya Hambat terhadap Streptococcus mutans dari beberapa Pasta Gigi
yang Mengandung Herbal. Majalah Kedokteran Gigi (Dental Journal). 38.
[4] Chandrabhan D, Hemlata R, Renu B, and Pradeep V (2012) Isolation of Dental Caries Bacteria
from Dental Plaque and Effect of Tooth Pastes on Acidogenic Bacteria. Open Journal of
Medical Microbiology. 2. 65-69.
[5] Volk WA, and Wheeler (1993) Mikrobiologi Dasar, Edisi Kelima, Jilid I. Penerbit Erlangga,
Jakarta.
[6] Ladytama, SR, Nurhapsari A, and Baehaqi M (2014) Efektivitas Larutan Ekstrak Jeruk Nipis
(Citrus Aurantifolia) Sebagai Obat kumur Terhadap Penurunan Indeks Plak Pada Remaja Usia
12-15 Tahun- Studi di SMP Nurul Islami, Mijen, Semarang. Odonto Dental Journal. 1 (1).
[7] Endarti, Fauzia, and Zuliana E. (2006) Manfaat Berkumur dengan Larutan Siwak (Salvadora
persica). Majalah Kedokteran Nusantara. 39(4) : 393-401.
[8] Awang AFIB, Susanti D, and Taher M (2013) Antimicrobial Activity and Synergic Effect of
Cinnamomum burmanii’s Essential Oil & its Isolated Compound (Cinnamaldehyde).
International Conference on Chemical, Agricultural and Medical Sciences. 29-30.
[9] Chandukar PN, Tripathi N, Choudary A, and Murab T (2014) Antibacterial Properties of Cinnamon
Stick Oil with Special References to Streptococcus pyogenes and Pseudomonas aeruginosa.
International Journal of Current Microbiology and Apllied Sciences. 3 : 177-178.
[10] Liantari DS (2014) Effect of Wuluh Starfruit Leaf Extract for Streptococcus mutans Growth. J
Majority. 3(7) : 27-33.
[11] Adzakiyah T, Lipoeta I, and Kusuma N (2015) Pengaruh Berkumur dengan Larutan Ekstrak Siwak
(Salvadora persica) Terhadap pH Saliva Rongga Mulut. Jurnal Sains Farmasi dan Klinis. 2(1) :
74-77.
[12] Dziedzic A, Wojtyczka RD, and Kubina R (2015) Inhibition of Oral Streptococci Growth Induced
by the Complementary Action of Berberine Chloride and Antibacterial Compounds. Molecules.
20 : 13705-13724.

6
ICBSB IOP Publishing
IOP Conf. Series: Earth and Environmental Science1234567890
130 (2018) 012049 doi:10.1088/1755-1315/130/1/012049

[13] Maal KB, Majid, Boujari, and Zavareh FA (2010) Identification of Streptococcus salivarius
Bacteriophage Isolated from Persian Gulf as a Potential Agent for Dental Caries Phage
Therapy. African Journal of Microbiology Research. 4(20) : 2127-2132.
[14] Dhotre S, Suryawanshi N, Nagoba B, and Shelkar S (2016) Rare and Unusual Isolates of Viridans
Streptococci From Human Oral Cavity. Indian Journal Pathology and Microbiology. 59(1) :
47-49.
[15] Sabtie HA, Ali IN, and Humudat YR (2015) Microbial Biofilm Formation as a Dental-fouling
Microorganisme Attachment Phenomenon in Oral Cavity Environment. Journal of Life
Sciences. 9 : 40-46.
[16] Ma ESK, Wong CLP, Lai KTW, Chan ECH, Yam WC, and Chan ACW (2005) Kocuria kristinae
Infection Assosiated with Acute Choleocystitis. BioMed Central Infectious Disease. 5(60) : 1-
3.
[17] Al-Duboni G, Osman MT, and Al-Naggar R (2013) Antimicrobial Effect of Aqueous Extract of
Cinnamon and Ginger on Two Oral Pathogens Causing Dental Caries. Research Journal of
Pharmaceutical, Biological and Chemical Sciences. 4(3) : 957-965.
[18] Fajriani and Mahrum (2015) Efectiveness of Lime (Citrus aurantifolia) Extract Solution in
Inhibiting Bacteria Streptococcus mutans Case of Early Childhood Caries. Donnish Journal.
1(4) : 016-020.