The Antibacterial Effect from Combining Cinnamon, Patchouli and

Coriander Essential Oils

Windri Handayani

, Retno Yunilawati

2,3

, Cuk Imawan

Departemen Biologi, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas

Indonesia, Kampus Depok, Indonesia 16424

Badan Penelitian dan Pengembangan Industri, Kementerian Perindustrian

Departemen Fisika, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas

Indonesia, Kampus Depok, Indonesia 16424

Keywords: Essential Oils, Cinnamon, Patchouli, Coriander, vapour test, antimicrobial

Abstract: Essential oils are dynamic organic liquids that work in synergy with each other. In general, essential oils work

better when mixed with other essential oils. Every essential oil has many compounds and known for their

benefits like healing properties and aromatic compound. When mixing essential oils between one oil and

another, they can compensate for their strengths and weaknesses of each other. In this research, Cinnamon,

Patchouli and Coriander essential oil combined to strengthen their antimicrobial activities. Variation was done

by combining between 3 types of essential oils at different combination ratios 1:1 and 1:2 (v/v). Furthermore,

the oil was tested on Gram-positive bacteria Staphylococcus aureus and Gram-negative bacteria Escherichia

coli using the paper disk method and vapour test. From the results obtained it is known the strength of the

antibacterial activity when the oil is in direct contact with microorganisms and the strength of volatile

compounds of essential oils in inhibiting antimicrobial activity. The essential oils were also characterized

using Gas Chromatography Mass Spectroscopy (GC-MS) to determine the levels and presence of compounds

suspected of having antimicrobial activity. The results show the weakest antimicrobial activity EO

combination were when using patchouli and coriander. Meanwhile, the strongest when testing paper disks

and the vapour test is a combination of cinnamon and patchouli, cinnamon and coriander, cinnamon, patchouli

and coriander.

1 INTRODUCTION

Nowadays, the development of active packaging by

utilizing natural ingredients are quite engaging

studies (Calo, et.al., 2015). Aromatic plants and their

extracts have the potential to be applied in the field of

food safety and food preservation. A part of

phytochemical compounds there are group of

essential oils that generally contain a very complex

mixture of several types of aromatic phytochemical

compounds, which had potential to be develop in this

application (Chen et.al. 2018; Marques et.al., 2019).

Essential oils are dynamic organic liquids that

work together in synergy. In general, essential oils

work better when they mixed with other essential oils.

Each essential oil contains many compounds and is

known for its benefits, such as healing properties and

aromatic compounds. When mixing essential oils

between one oil and another, they can compensate for

each other's strengths and weaknesses. Antagonism

activity observed when the effects of one or both

compounds are less when they are applied together

than when applied individually. Synergism observed

when the effects of the combined substances are

higher than the sum of the individual effects

(Davidson and Parish, 1989; Park et.al., 2018).

Certain phytochemical compounds derived from

plants are known to have a role in inhibiting the

growth and survival of microorganism (Guedes et.al.,

2018; Merino et.al., 2019). However, besides their

benefits, the used of essential oil as a food

preservative have some disadvantages because of the

strong smell that will affect the aroma and food

flavour. Therefore, it is necessary to choose the type,

composition, and dosage of a constant essential oil.

Handayani, W., Yunilawati, R. and Imawan, C.

The Antibacterial Effect from Combining Cinnamon, Patchouli and Coriander Essential Oils.

DOI: 10.5220/0009968601530158

In Proceedings of the 2nd International Conference of Essential Oils (ICEO 2019), pages 153-158

ISBN: 978-989-758-456-5

153

Developing effective synergistic EO

combinations could be an approach for improving

their antimicrobial efficacies and food application

potentials. More works are needed in this area as

studies to date have reported inconsistent results in

EO combinations (synergistic, additive and

antagonistic) (Park et.al., 2018). In this research, 3

types of EO from Cinnamon, Patchouli, and

Coriander tested for their antimicrobial activity

during direct contact and in vapour phase. We tested

the EO composition both single and combination

against Gram-positive bacteria and Gram-negative

bacteria to study their synergistic bioactivity for their

potential as active label packaging mixture

application.

2 MATERIALS & METHODS

2.1 Materials

Essential oil Cinnamon (Ci) (Cinnamomum

burmanii), Coriander (Co) (Coriandrum sativum) and

Patchouli (Co) (Pogostemon cablin) used in this

experiment obtained from a local essential oils

company in Indonesia Nusaroma. The bacteria used

in this work were Escherichia coli NBRC 3301 strain

and Staphylococcus aureus NBRC 100910 collection

from UICC CoE IBR-GS, FMIPA UI represented

Gram-negative bacteria and Gram-positive bacteria.

The Muller Hinton Agar (Difco) was used for the

culture of the bacterial medium.

2.2 Methods

2.2.1 Experimental Design

In this study, we tried to improve the antibacterial

activity from 3 different essential oil (EO) which

were cinnamon, patchouli, and coriander by

combining in certain ratio. The combination of 2

essential oil with ratio 1:2 and 1:1 (v/v), respectively

(Table 1). The essential oil used without further

dilution.

Table 1: The ratio combination from 3 kinds of essential

oil in this research

Cinnamon Patchouli Coriande

Cinnamon 1:1 ; 1:2 1:1 ; 1:2

Patchouli 1:2 1:1 ; 1:2

Coriande

1:2 1:2

2.2.2 EO Gas Chromatography

Characteristic

Characterization and analysis from the EO

based on their ratio combination were using GC/MS

and performed using Gas Chromatograph (GC)

Agilent 6890 series with capillary column HP-5MS,

30 m x 0.25 mm id x 0.25 µm film thickness. Helium

gas (65 kPa) was used as the carrier gas at constant

pressure, and an injection volume of 1 μL was

employed (split ratio of 25:1). The oven temperature

was programmed from 60-240° C, with an increase of

3° C/min until it reaches 250° C. Components were

identified based on a comparison of relative retention

time and mass spectrum following the same method

used in Handayani (2019).

2.2.3 Direct Contact Agar Diffusion Test

Paper disc diffusion method used to determine the

antimicrobial activities by direct contact with the EO.

This test using type strain of Staphylococcus aureus

NBRC 100910 and Escherichia coli NBRC 3301.

The Muller Hinton Agar medium was prepared by

pouring 10 ml of molten media into sterile Petri plates

(d=90 mm) and allowed to solidify for 5 minutes.

After that, in a tube, 10 μl of bacteria culture 10-6

CFU/mL added with 10 ml of medium and mixed

gently with the inoculate before poured on the top of

molten media before and allowed to dry for 5

minutes. The negative control (sterile distilled

water), positive control (Tetracycline 7 μg/mL), the

essential oil then loaded on 6 mm disc, whereas the

volume for each disc was 10 µl. The loaded disc

placed on the surface of the medium then incubates at

32o C for 24 hours. After the end of incubation, a

clear zone formed around the disc measured. Each

experiment done in triplicate.

2.2.4 Vapour Phase Antibacterial Test

The antimicrobial activities from volatile compound

from the EO were tested in vapour phase agar

diffusion test. The vapour phase method follows the

method used by Wang (2016) and Zaika (1988). We

used the same bacteria and medium for preparation

the paper disk diffusion assay. The EO loaded on to 6

mm disc and put under the paper disk cover and

incubate incubated in reverse position.

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3 RESULTS & DISCUSSION

3.1 GC/MS Characterization

GC/MS characterization was carried out to determine

the compound content from each EO. Figure 1 shows

the results of the chromatogram from 4 type of EO.

The GC/MS showed that the types of compounds in

each EO are relatively different and varied (Table 2).

In Ci there are 2 major compounds detected, they are

Cinnamaldehyde and Copaene. Meanwhile, the

compounds contained in Po are α-Guaiene, Azulene,

and 4-Aromadendrene. Furthermore, the Co

containing Benzene, 1-methyl-2-(1-methylethyl) and

3-Carene. The abundance of major compounds also

known from the chromatogram results and match

with the know EO compounds from Cinnamon (Yang

et al., 2019), Patchouli (van Beek & Joulain, 2018)

and Coriander (Laribi et al., 2015). Furthermore, the

EO combined to see its bioactivity in inhibiting

bacterial growth both during direct contact and

through vapor phase from the aromatic compounds.

Figure 1: GC/MS chromatogram profile from Patchouli, Coriander, Cinnamon, and combination from Patchouli and

Coriander EO

Figure 2. shows the antimicrobial activity of

each type of EO using Patchouli (Po), Cinnamon (Ci),

and Coriander (Co) in a single mixture. These results

show there are formation of a clear zone from all the

treatments. Among the three types of EO, Ci had the

highest inhibitory activity against both E. coli and S.

aureus, followed by Co. Meanwhile, Po had no

activity against E. coli only has activity against S.

aureus. This shows that there was an activity that

selective to gram-positive or negative bacteria.

Figure 2: Antimicrobial activity from each Single EO.

The Antibacterial Effect from Combining Cinnamon, Patchouli and Coriander Essential Oils

155

Table 2: GC/MS analysis from Cinnamon, Patchouli and

Coriander EO.

3.2 Antibacterial with Direct Contact Assay

Furthermore, the results of combined EO mixture

tested for their antimicrobial activity. The results

obtained can be seen in Figure 3 which shows the

antimicrobial activity from Po/Co. The results

obtained indicate an increase in antimicrobial activity

than single EO mixture. Antimicrobial activity in

Po/Co combination (2: 1) has the best activity. While

the ratio of 1:1 has relatively the same antimicrobial

activity with the single mixture, and the Po/Co ratio

(1: 2) has the weakest antimicrobial activity.

Figure 3: Antimicrobial activity from Patchouli (Po):

Coriander (Co) combination.

Meanwhile, Figure 4 and 5 show that the

combination from each ratio shows inhibitory activity

which tends to be the same in all ratios. The strongest

activity dominated went the mixture contained EO

from Cinnamon. Figure 4 shows the comparison of

the clear zone diameter sizes of each treatment in the

direct contact test. These results indicate that in the

single form the EO from Ci has the strongest activity.

While Patchouli only has positive activity on S.

aureus and Coriander has activity on E. coli and S.

aureus so that when combined with Po/Ci or Co the

antibacterial activity increased. The Ci/Po

combination showed a better activity compared to

other EO combinations in this study.

Figure 4: Antimicrobial activity from Cinnamon (Ci):

Coriander (Co) combination with different ratio.

Figure 5: Antimicrobial activity from Cinnamon (Ci):

Patchouli (Po) combination with different ratio.

This is also seen in Figure 6. which shows that a

single EO Ci tends to have the strongest activity

compared to Po and Co, where the diameter of the

clear zone formed can reach 45 mm. The EO from Ci

had almost equal activity to gram negative and

positive. Co tend to have stronger activity for Gram-

negative bacteria than Po, meanwhile the Po tend to

be weakest activity. Meanwhile, Po had stronger

activity against gram positive bacteria than Co. With

the combination from Po/Co (2:1) the activity against

E. coli increasing this result show possibility

compliment action of synergistic activity with the

mixture from those 2 EO by direct contact. Based on

Figure 1, when we combined the EO from Co/Po the

chromatogram detected increasing peak number from

the previous one, which indicate the increasing of

compound number that contained from both Co and

Po EO.

Cinnamon

PK RT Library/ID

1 18.1664 Cinnamaldehyde

2 22.7314 Copaene

Patchouli

PK RT Library/ID

1 22.94 β-Patchoulene

2 24.5992 Caryophyllene

3 25.483 α-Guaiene

4 26.0791 α-Patchoulene

5 28.3137 Azulene

6 34.2794 4-Aromadendrene

Coriander

PK RT Library/ID

1 5.1052 -Terpinene

2 7.8733 Benzene, 1-methyl-2-(1-

methylethyl)

3 10.8416 3-Carene

4 12.5842 Camphore

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Figure 6: The diameter of inhibition zone from each

treatment with direct contact.

3.3 Antibacterial Vapor Test

In addition, beside the antimicrobial test through

direct contact, the vapor phase from the EO were also

tested for its antimicrobial activity. Of the three EOs

used, Ci showed better inhibitory activity on E. coli

than S. aureus (Figure 7). Meanwhile, the EO vapor

test from Co and Po, there were no clear zone

observed from the two test bacteria (Table 3).

Table 3: Antimicrobial activity from vapour phase test of

single EO

Patchouli

(Po)

Cinnamon

(Ci)

Coriander

(Co)

E. coli - +++ -

S. aureus - ++ -

Figure 8. Shows the inhibition zone of the

combination of Ci with other EO did not show an

increasing in the antimicrobial activity. The activity

tends to decrease, which predicted as the result from

the decreasing in the abundance of aromatic

compounds after combined with the ratio treatment

per 10 μL. These results indicate that the vapor from

Ci EO has strong antimicrobial activity. While other

EOs have volatile compounds that do not have

antimicrobial activity. Cinnamaldehyde also known

as cinnamic aldehyde is known to be an aromatic

compound contained in the EO that gives cinnamon

its flavor and odor. Cinnamaldehyde is the major

component comprising 85% in the essential oil and

the purity of cinnamaldehyde in use is high (> 98%).

Both oil and pure cinnamaldehyde are equally

effective in inhibiting the growth of various

microorganisms such as Gram-positive and Gram-

negative (E. coli) bacteria, and fungi including yeasts,

filamentous molds and dermatophytes (Ashakirin,

2017). Other research by Ács et. al (2018) highlight

that Gram-negative strains were more sensitive to EO

vapours.

Figure 7: Clear zone formed from vapour phase test of

Cinnamon EO for antibacterial inhibition.

Figure 8: The inhibition zone from each treatment with

vapour phase against E. coli and S. aureus.

4 CONCLUSIONS

The results showed that EO, with the most potent

activity in direct contact using paper disks and vapour

tests, were Ci/Po and Ci/Co. Meanwhile, the weakest

antibacterial activity with EO combination was when

using patchouli and coriander. The combination of

Po/Co improves their strength against both bacteria

than when they were as single EO. The factors that

were affecting EO’s antimicrobial activity in this

study are the abundance and the EO species, which

depend on a specific active compound in each plant

species — combining the essential oil need to be more

selective, to improve antimicrobial activity and

determine the synergistic and antagonistic effect from

the compound.

The Antibacterial Effect from Combining Cinnamon, Patchouli and Coriander Essential Oils

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ACKNOWLEDGEMENTS

We would like to thank The Center of Excellence

Biology Resources-Genome Study (CoE IBR-GS)

FMIPA UI for the facilities and equipment to support

this research.

REFERENCES

Ács, K., V.L. Balázs, B. Kocsis, T. Bencsik1, A.

Böszörményi & G. Horváth, 2018, Antibacterial

activity evaluation of selected essential oils in liquid

and vapour phase on respiratory tract pathogens. BMC

Complementary and Alternative Medicine 18: 227

Ashakirin, S.N., M. Tripathy, U.K. Patil & A.B.A. Majeed,

2017, chemistry and bioactivity of cinnamaldehyde: a

natural molecule of Medicinal importance. IJPSR 8(6):

2333-2340.

Calo, J.R., P.G. Crandall, C.A. O’Bryan & S.C. Ricke,

2015, Essential oils as antimicrobials in food systems e

A review, Food Control 54: 111-119

Chen C., Z. Xu, Y. Ma, J. Liu, Q. Zhang, Z. Tang, K. Fu, F.

Yang & Jing Xie. 2018. Properties, vapour-phase

antimicrobial and antioxidant activities of active poly

(vinyl alcohol) packaging films incorporated with clove

oil. Food Control 88: 105-112

Davidson, P.M. and M.E. Parish, 1989, Method for testing

the efficacy of food antimicrobial, Food Technology

43(1): 148-155.

Guedes, J.P. de Sousa & E.L. de Souza, 2018, Investigation

of damage to Escherichia coli, Listeria monocytogenes

and Salmonella Enteritidis exposed to Mentha arvensis

L. and M. piperita L. essential oils in pineapple and

mango juice by flow cytometry, Food Microbiology 76:

564-571

Handayani, W., Y. Yasman, R. Yunilawati, V. Fauzia &

Cuk Imawan. 2018. Coriandrum sativum L. (Apiaceae)

and Elettaria cardamomum (L.) Maton (Zingiberaceae)

for antioxidant and antimicrobial protection. Journal of

Physics: Conference Series 1317 (1), 012092

Laribi, B. et al., 2015, Coriander (Coriandrum sativum L.)

and its bioactive constituents, Fitoterapia, 103: 9–26.

Marques, C.S., R.P. Grillo, D.G. Bravim, P.V. Pereira,

J.C.O. Villanova, P.F. Pinheiro, J.C.S. Carneiro & P.C.

Bernardes, 2019, Preservation of ready-to-eat salad: A

study with combination of sanitizers, ultrasound, and

essential oil-containing β-cyclodextrin inclusion

complex, LWT - Food Science and Technology 115:

108433

Merino, N., D. Berdejo, R. Bento, H. Salman, M. Lanz, F.

Maggi, S. Sánchez-Gómez, D. García-Gonzalo & R.

Pagán, Antimicrobial efficacy of Thymbra capitata (L.)

Cav. essential oil loaded in self-assembled zein

nanoparticles in combination with heat, Industrial

Crops & Products 133: 98–104

Park, J.B., J.H. Kang & K.B. Song, 2018, Antibacterial

activities of a cinnamon essential oil with

cetylpyridinium chloride emulsion against Escherichia

coli O157:H7 and Salmonella Typhimurium in basil

leaves, Food Sci Biotechnol. 2018 Feb; 27(1): 47–55

van Beek, T.A. & D. Joulain, 2017, The essential oil of

patchouli, Pogostemon cablin: A review, Flavour

Fragr. J. 33:6–51

Wang, T.H., S.M. Hsia, C.H. Wu, S.Y. Ko, M.Y. Chen,

Y.H. Shih, T.M. Shieh, L.C. Chuang & C.Y. Wu. 2016.

Evaluation of the Antibacterial Potential of Liquid and

Vapor Phase Phenolic Essential Oil Compounds against

Oral Microorganisms. PLOS ONE DOI:

10.1371/journal.pone.0163147.

Yang, Y. Feng et al. (2019) ‘Effects of dietary graded levels

of cinnamon essential oil and its combination with

bamboo leaf flavonoid on immune function,

antioxidative ability and intestinal microbiota of

broilers’, Journal of Integrative Agriculture 18(9):

2123–2132.

Zaika, L.L., 1988. Spices and herbs: Their antimicrobial

activity and its determination1. J. Food Safety 9: 97-

118.

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