Journal of Ethnopharmacology 77 (2001) 123– 127

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Short communication
Antibacterial activity of leaf essential oils and their constituents
from Cinnamomum osmophloeum
Shang-Tzen Chang a,*, Pin-Fun Chen a, Shan-Chwen Chang b
a
Department of Forestry, National Taiwan Uni6ersity, No 1 Section 4, Roose6elt, Taipei, Taiwan, ROC
b
Department of Internal Medicine, National Taiwan Uni6ersity, Taipei, Taiwan, ROC

Received 4 June 2000; received in revised form 28 February 2001; accepted 18 May 2001

Abstract

The antibacterial activities of the essential oils from leaves of two Cinnamomum osmophloeum clones (A and B) and their
chemical constituents were investigated in this study. The nine strains of bacteria, including Escherichia coli, Pseudomonas
aeruginosa, Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, methicillin-resistant Staphylococcus aureus
(MRSA), Klebsiella pneumoniae, Salmonella sp., and Vibrio parahemolyticus, were used in the antibacterial tests. Results from the
antibacterial tests demonstrated that the indigenous cinnamon B leaf essential oils had an excellent inhibitory effect. The MICs
(minimum inhibitory concentrations) of the B leaf oil were 500 mg/ml against both K. pneumoniae and Salmonella sp. and 250
mg/ml against the other seven strains of bacteria. Cinnamaldehyde possessed the strongest antibacterial activity compared to the
other constituents of the essential oils. The MICs of cinnamaldehyde against the E. coli, P. aeruginosa, E. faecalis, S. aureus, S.
epidermidis, MRSA, K. pneumoniae, Salmonella sp., and V. parahemolyticus were 500, 1000, 250, 250, 250, 250, 1000, 500, and 250
mg/ml, respectively. These results suggest that C. osmophloeum leaf essential oil and cinnamaldehyde are beneficial to human
health, having the potential to be used for medical purposes and to be utilized as anti-bacterial additives in making paper
products. © 2001 Elsevier Science Ireland Ltd. All rights reserved.

Keywords: Antibacterial activity; Cinnamaldehyde; Cinnamomum osmophloeum; Essential oil; Leaf

1. Introduction food additives. It has also been demonstrated that

Plant-derived essential oils have long been used as
flavoring agents in food and beverages and, due to the
presence of antimicrobial compounds, they have poten-
tial as natural agents for food preservation (Helander et
al., 1998). Cinnamon oil is commonly used in the food
industry because of its special aroma. In addition, its
antimicrobial activity has also attracted great attention
from many researchers. Cinnamomum cassia bark oil is
generally used in food and beverages and is very valu-
able in commerce. The main constituents of C. cassia
bark oil are cinnamaldehyde and coumarin (Hu et al.,
1985). These two compounds have also been used as
* Corresponding author. Tel.: + 886-2-23630231x3196; fax: +886-
2-23654520.
E-mail address: peter@ms.cc.ntu.edu.tw (S.-T. Chang).
Cinnamomum zeylanicum oil has an inhibitory effect
against meat spoilage organisms (Ouattara et al., 1997).
Singh and his co-workers (Singh et al., 1995) have
demonstrated that C. zeylanicum bark oil has fungitoxic
properties against fungi involved in respiratory tract
mycoses, such as Aspergillus niger, A. fumigatus, A.
nidulans, and A. fla6us. In addition, C. zeylanicum oil
has been tested for maize kernel protection against A.
fla6us (Montes-Belmont and Carvajal, 1998). The re-
sults have revealed that it can effectively inhibit the
growth of A. fla6us and has no phytotoxic effect on
germination and corn growth. The important chemical
constituents of C. zeylanicum oil are cinnamaldehyde
and eugenol (Ross, 1976). These two compounds have
also been demonstrated to have inhibitory properties
against A. fla6us (Montes-Belmont and Carvajal, 1998).

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124 S.-T. Chang et al. / Journal of Ethnopharmacology 77 (2001) 123–127
Cinnamomum osmophloeum Kaneh. (Lauraceae) is an
endemic tree in Taiwan. It has been of interest to
researchers because the chemical constituents of its leaf
essential oils were similar to those of C. cassia bark oil.
In order to explore the potential usefulness of the leaf
essential oils of C. osmophloeum, it is important to
know their chemical constituents. Therefore, the chemi-
cal constituents of leaf essential oils from various C.
osmophloeum clones found in different regions of Tai-
wan have been studied by many researchers. Hu and his
coworkers analyzed the composition of the essential oils
of C. osmophloeum leaves collected randomly from 21
provenances in central, southern, and eastern parts of
Taiwan. It was found that cinnamaldehyde was the
major constituent in the leaf essential oils of some C.
osmophloeum clones, and that eugenol was present in
the leaf essential oils of the other C. osmophloeum
clones. In addition, based upon the chemical composi-
tion of the different leaf essential oils, they classified C.
osmophloeum into nine types such as cassia type, cin-
namaldehyde type, coumarin type, linalool type,
eugenol type, camphor type, 4-terpinenol type, linalool-
terpinenol type, and mixed type (Hu et al., 1985).
Although the chemical constituents of leaf essential
oils of various C. osmophloeum clones have been stud-
ied, the potential antimicrobial activities of their leaf
essential oils and their constituents has not yet been
evaluated. In this study, the essential oils of leaves
collected from C. osmophloeum clones (A and B) were
extracted and their chemical compositions analyzed,
then the antibacterial activity of the essential oils and
their chemical constituents were investigated. In addi-
tion, the antibacterial activity of cinnamaldehyde con-
geners was examined to help understand the effect of
chemical structure on the antibacterial activity.
2. Materials and methods
2.1. Essential oil distillation
Leaves of two indigenous cinnamon (C. os-
mophloeum) clones (A and B) were collected from
Haw – Lin experimental forest. The essential oils of C.
osmophloeum leaves were obtained by extraction for 6 h
using water distillation.
2.2. GC analysis
GC was performed using a Shimadzu model-14B
equipped with a FID. The column used was 50 m long
by 0.22 mm i. d. glass capillary coated with silica. GC
was programmed from 60 to 220 °C at 2 °C/min.
Identification of the major components of indigenous
cinnamon leaf oils was confirmed by comparsion with
standards, as well as by spiking. The quantity of com-
pounds was obtained by integrating the peak area of
spectrograms.
2.3. Essential oil constituents
The following pure compounds of essential oil con-
stituents were obtained from Acros (Belgium): ben-
zaldehyde, linalool, a-terpineol, geraniol,
cinnamaldehyde, eugenol, cinnamyl alcohol, and cou-
marin. The congeners of cinnamaldehyde, including
4-hydroxybenzaldehyde, cinnamic acid, cinnamyl ace-
tate, and 3-phenylpropionaldehyde, were also obtained
from Acros.
2.4. Bacteria strains
Five strains of Gram-negative bacteria (Escherichia
coli, Pseudomonas aeruginosa, Klebsiella pneumoniae,
Salmonella sp., and Vibrio parahemolyticus) and four
strains of Gram-positive bacteria (Enterococcus faecalis,
Staphylococcus aureus, Staphylococcus epidermidis, and
methicillin-resistant Staphylococcus aureus) were used
in the antibacterial assay. All of these bacteria were
obtained from the bacteriology laboratory of the Na-
tional Taiwan University Hospital.
2.5. Antibacterial susceptibility test
The broth dilution method described in the Manual
of Clinical Microbiology (Jones et al., 1985) was used
to assess the antibacterial activities of the essential oils
and their chemical constituents. The concentrations of
each tested compound incorporated in the Muller–Hin-
ton broth were 100, 250, 500, and 1000 mg/ml. Three
replicas were prepared in each case. The antibacterial
activities were examined after incubation at 37 °C for
18 h. Minimum inhibitory concentration (MIC) was
defined as the lowest concentration of test samples that
resulted in a complete inhibition of visible growth in the
broth. Oxacillin and cefotaxime were used as positive
controls.
3. Results and discussion
The concentration of various ingredients in essential
oils from the same plant species may vary due to
ecological and plant growth factors (Deans and Svo-
boda, 1989; Wang and Yin, 1991; Yin, 1991). In this
study, indigenous cinnamon A and B leaves yielded
6.02 and 7.93 ml of essential oils/kg on water distilla-
tion, respectively. The major chemical constituents of
indigenous cinnamon leaf oils and their relative
amounts were determined by GC analysis. The relative
contents (%) are shown in Table 1. According to the
classification of Hu et al. (1985), indigenous cinnamon
 S.-T. Chang et al. / Journal of Ethnopharmacology 77 (2001) 123–127 125

A belongs to the mixed type and indigenous cinnamon stronger antibacterial activity than indigenous cinna-
B belongs to the cinnamaldehyde type. mon A. The indigenous cinnamon A oil had inhibitory
The MICs of these two indigenous cinnamon leaf oils properties only on S. epidermidis and V. parahemolyti-
against bacteria are presented in Table 2. The indige- cus at the level of 1000 mg/ml. The indigenous cinna-
nous cinnamon leaf essential oils had demonstrable mon B leaf oil possessed antibacterial activities against
antibacterial properties, and the constituents present in all nine strains of bacteria. The MIC was 500 mg/ml
the oil showed variable levels of inhibition. Further- against K. pneumoniae and 250 mg/ml against the other
more, results from these antibacterial tests also revealed eight strains of bacteria. It appears that the difference
that indigenous cinnamon B leaf oil displayed much in the antibacterial activities between these two indige-
nous cinnamon leaf oils may relate to the content of
Table 1 cinnamaldehyde.
The main components and their relative contents (%) of C. os- There are several reports stating that other leaf oils
mophloeum leaf essential oils
(eg C. zeylanicum Bl.) exhibit antimicrobial activity.
Peak Rt (min)a Compounds % of total Hili and his co-workers (Hili et al., 1997) have assayed
the antimicrobial activity of the C. zeylanicum leaf oil
Cinnamon A against three bacteria and four yeast strains. Their
1 13.8 1,8-Cineol 11.32 results demonstrated that the leaf oil completely inhib-
2 32.9 Benzaldehyde 0.93
3 34.9 Linalool 9.83
ited the growth of E. coli, S. aureus, and P. aeruginosa
4 43.6 Borneol 7.46 at the level of 500 mg/ml. Another report found the
5 44.3 a-Terpineol 4.62 MICs of C. zeylanicum against E. coli, and S. aureus
6 47.0 Neral 12.82 were 0.05 and 0.04%, respectively (Smith-Palmer et al.,
7 47.8 Geranyl acetate 6.03 1998). In comparison with those reports, it is evident
8 53.4 Geraniol 4.73
9 64.6 Cinnamaldehyde 8.35
from our results that the inhibitory activity of C.
10 70.1 Cinnamyl acetate 9.04 osmophloeum leaf oil on bacteria was better than that of
Cinnamon B C. zeylanicum leaf oil. The main constituents of C.
1 33.2 Benzaldehyde 3.20 zeylanicum leaf oil are cinnamaldehyde and eugenol,
2 34.5 Linalool 0.24 but its cinnamaldehyde content (approximately 50–
3 49.5 Geranyl acetate 3.88
4 55.8 Geraniol 2.96
70%) is less than that in C. osmophloeum leaf oil.
5 65.3 Cinnamaldehyde 76.00 Therefore, there is a probability that the difference in
6 69.6 Eugenol 1.05 the antibacterial activities between these two species of
7 71.6 Cinnamyl acetate 0.51 cinnamon leaf oil is related to the content of cin-
8 84.1 Coumarin 0.26 namaldehyde. Furthermore, there was no discernible
a
Retention time
trend of inhibition reflected in the strain characteristics
of bacterium: both Gram-negative and Gram-positive
Table 2 organisms were affected. This is different from Taiwa-
Minimum inhibitory concentration (mg/ml) of two indigenous cinna- nia (Taiwania cryptomerioides Hayata) heartwood es-
mon leaf oils against bacteria sential oil, which had an inhibitory effect against
Gram-positive bacteria only (Chang et al., 2000).
Bacteria Leaf oil A Leaf oil B Oxacillin Cefotaxime
To confirm the relationship of the constituents in
E. coli –a 250 64 0.125 indigenous cinnamon leaf oil and antibacterial activity,
P. – 250 64 16 the eight important constituents of indigenous cinna-
aeruginosa mon leaf oil were tested for antibacterial activity
against E. coli, P. aeruginosa, E. faecalis, and S. aureus.
E. faecalis – 250 16 64
S. aureus – 250 0.25 2
The results of this antibacterial assay are shown in
S. 1000 250 64 32 Table 3. Among the eight constituents, cinnamaldehyde
epidermidi possessed the best antibacterial activity, followed by
s cinnamyl alcohol. Because cinnamaldehyde exhibited
MRSA – 250 64 8 the best antibacterial activity, it was further tested
K. – 500 \128 0.5
pneumonia
against five other strains of bacteria. The MICs of
e cinnamaldehyde against E. coli, P. aeruginosa, E. fae-
Salmonella – 500 \128 1 calis, S. aureus, S. epidermidis, MRSA, K. pneumoniae,
sp. Salmonella sp., and V. parahemolyticus are 500, 1000,
V. 1000 250 \128 1 250, 250, 250, 250, 1000, 500, and 250 mg/ml, respec-
parahemol
yticus
tively. Comparing the antibacterial activity of cin-
namaldehyde with the results for indigenous cinnamon
a
No inhibitory effect at the level of 1000 mg/ml. B leaf oil, the inhibitory effect of cinnamaldehyde was
126 S.-T. Chang et al. / Journal of Ethnopharmacology 77 (2001) 123–127

Table 3
Minimum inhibitory concentration (mg/ml) of the constituents of
indigenous cinnamon leaf oil against bacteria

Constituents E. coli P. E. faecalis S. aureus

aeruginosa

Cinnamaldeh 500 1000 250 250

yde
Benzaldehyde –a – – –

Linalool – – – –
a-terpineol 1000 – – –
Geraniol 1000 – – –
Eugenol 1000 – – –
Cinnamyl 1000 1000 1000 500
alcohol
Coumarin 1000 – – 1000
Fig. 1. The structures of cinnamaldehyde congeners.
a
No inhibitory effect at the level of 1000 mg/ml.

less than that of indigenous cinnamon leaf oil against
some species of bacteria. For example, the MICs of
indigenous cinnamon B leaf oil against E. coli, P.
aeruginosa, K. pneumoniae, and Salmonella sp. were less
than those for cinnamaldehyde. So the inhibitory prop-
erties of indigenous cinnamon B leaf oil against these
four strains of bacteria were not solely due to cin-
namaldehyde. It might be that a synergistic effect
derived from some constituents in indigenous cinnamon
B leaf oil endows it with such potent antibacterial
activity. As for the other five strains of bacteria, includ-
ing E. faecalis, S. aureus, S. epidermidis, MRSA, and V.
parahemolyticus, the inhibitory effects of cinnamalde-
hyde were similar to those for indigenous cinnamon B
leaf oil. So the antibacterial activity of indigenous
cinnamon B leaf oil against these five strains of bacteria
probably relates to the presence of cinnamaldehyde. On
the other hand, E. coli, P. aeruginosa, K. pneumoniae,
and Salmonella sp. all belong to Gram-negative bacte-
ria. It is possible that the antibacterial activity of in-
digenous cinnamon leaf oil against Gram-negative
bacteria was enhanced by the presence of constituents
other than cinnamaldehyde.
Table 4
Minimum inhibitory concentration (mg/ml) of cinnamaldehyde congeners against bacteria
Cinnamaldehyde was highly effective in inhibiting the
growth of the nine strains of bacteria. To find out if
some chemical group or structure endows cinnamalde-
hyde with such strong antibacterial activity, four com-
pounds whose chemical structure is similar to
cinnamaldehyde were studied for their antibacterial ac-
tivities. These four compounds were 4-hydroxyben-
zaldehyde, cinnamic acid, cinnamyl acetate, and
3-phenylpropionaldehyde, and their chemical structures
are shown in Fig. 1. The results of this antibacterial
assay are presented in Table 4. It revealed that cin-
namaldehyde also possessed the strongest antibacterial
activity. The antibacterial activity order of four cin-
namaldehyde congeners having both a benzyl ring and
a conjugated double bond was cinnamaldehyde\cin-
namic acid\ cinnamyl alcohol\ cinnamyl acetate. It is
clear that aldehyde group has the best antibacterial
activity. Based on the antibacterial assay of cin-
namaldehyde, 3-phenylpropionaldehyde, benzaldehyde,
and 4-hydroxybenzaldehyde, a conjugated double bond
and a long CH chain outside the ring may result in
better antibacterial activity. In addition, the hydroxyl
group can improve the antibacterial activity.

Compounds Bacteriaa

E.c. P.a. E.f. S.e. S.a. MRSA K.p. S.sp. V.p.

Benzaldehyde –b – – – – – – – –
4-Hydroxybenzal dehyde 1000 1000 – 1000 1000 1000 1000 – 1000
Cinnamyl alcohol 1000 1000 1000 500 – – 1000 1000 1000
Cinnamic acid 1000 1000 1000 1000 1000 1000 – 1000 1000
Cinnamyl acetate – – – – – – – – –
3-Phenylpropion-aldehyde 1000 – 1000 – 500 – – 1000 1000
Cinnamaldehyde 500 1000 250 250 250 250 1000 500 250

a
E.c., Escherichia coli; P.a., Pseudomonas aeruginosa; E.f., Enterococcus faecalis; S.e., Staphylococcus epidermidis; S.a., Staphylococcus aureus;
MRSA, methicillin-resistant Staphylococcus aureus; K.p., Klebsiella pneumoniae; S.sp., Salmonella sp.; V.p., Vibrio parahemolyticus.
b
No inhibitory effect at the level of 1000 mg/ml.
 S.-T. Chang et al. / Journal of Ethnopharmacology 77 (2001) 123–127
In conclusion, this study demonstrated that cin-
namaldehyde type C. osmophloeum leaf essential oil and
its constituents have excellent antibacterial activities.
Both C. osmophloeum leaf essential oil and cinnamalde-
hyde are beneficial to human health, they have the
potential to be used for medical purposes and to be
utilized as anti-bacterial additives in making paper
products.
Acknowledgements
The authors wish to thank Professor H.-T. Chen
(Department of Forestry, Chinese Culture University)
for providing C. osmophloeum leaves.
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