The International Journal of Aromatherapy (2006) 16, 4349

The International
Journal of
Aromatherapy

intl.elsevierhealth.com/journals/ijar

In vitro bioactivities of essential oils used

for acne control
P. Lertsatitthanakorn a, S. Taweechaisupapong b,
C. Aromdee a, W. Khunkitti a,*

a
Department of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Khon Kaen University,
Khon Kaen 40002, Thailand
b
Faculty of Dentistry, Khon Kaen University, Khon Kaen 40002, Thailand

KEYWORDS Summary Essential oils are used in skincare products for perfuming and aroma-
Essential oils; therapy purposes. In this study, the bioactivities of seven essential oils commonly
Propionibacterium used and claimed for skincare namely citronella grass (Cymbopogon nardus L.),
acnes; lemongrass (Cymbopogon citratus DC), Kaffir lime (Citrus hystrix DC), holy
Antioxidant; basil (Ocimum sanctum L.), sweet basil (Ocimum basilicum L.), plai (Zingiber cas-
Anti-inflammatory sumunar Roxb) and ginger (Zingiber officinale Roscoe) were investigated. Investi-
gation of the in vitro susceptibility of the oils against Propionibacterium acnes (P.
acnes) using the broth microdilution technique revealed that citronella grass oil
exhibited the lowest minimum inhibitory concentration (MIC) and minimum bacte-
ricidal concentration (MBC) at 0.0050.3 and 0.61.2 ll/ml, respectively. The MIC
and MBC values of lemongrass oil were 0.6 ll/ml and those of kaffir lime oil and
holy basil oil were 5 ll/ml. Antioxidant activity using the DPPH free radical scav-
enging assay showed that the IC50 values of holy basil oil (0.03 ll/ml), plai oil
(6.9 ll/ml) and citronella grass oil (2 ll/ml) were lower than that of ascorbic acid
(7.9 ll/ml). Anti-inflammatory activity of the oils determined using the 5-lipoxyge-
nase inhibition assay found that IC50 values of holy basil oil (0.04 ll/ml), kaffir
lime oil (0.05 ll/ml) and citronella grass oil (0.15 ll/ml) were less than that of
nordihydroquaretic acid (1.7 lg/ml). Since P. acnes has a role in the inflammation
of acne leading to scar formation, citronella grass oil may help to relieve acne
blemishes. However, further investigation in the form of clinical studies would
be necessary.
c 2006 Elsevier Ltd. All rights reserved.

* Corresponding author. Fax: +66 6643 362 092.

E-mail address: watkhu@kku.ac.th (W. Khunkitti).


0962-4562/$ - see front matter c 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.ijat.2006.01.006
44
Introduction
Acne vulgaris, an inflammatory disease of the seba-
ceous gland, is a common skin disease. Propionibac-
terium acnes (P. acnes) plays an important role in
the inflammation by converting sebum oils into free
fatty acids that stimulate the hair follicle, form
comedones and then induce inflammation. A se-
quence of abnormal wound healing after the damage
that occurs in the sebaceous follicle during acne
inflammation may lead to post-acne scars (Bensoui-
lah, 2002; Holland and Jeremy, 2005). Compounds
targeting acne vulgaris therefore, should be able
to inhibit P. acnes, reduce pro-inflammatory lipids
in sebum as well as reduce post-acne scar formation.
In other words, any essential oils claimed for
acne control should possess antibacterial, anti-
inflammatory and antioxidant activities. Having an
antioxidant activity may be useful for relieving
hypertrophic scar and keloid formation by suppress-
ing the collagen synthesis of the fibroblasts from
hypertrophic scars but not suppressing those of
healthy normal tissue (Taniguchi et al., 1995).
Although topical and oral antibacterials such as
clindamycin, erythromycin, azelaic acid, tetracy-
cline and doxycycline can reduce the population of
P. acnes and also exert anti-inflammatory actions,
they lack antioxidant activity. These conventional
agents also possess adverse effects such as skin irri-
tation, erythrema, dryness and peeling (Musial and
Kubis, 2003). Alternative acne treatments may cause
less adverse effects and possess all the necessary key
activities. Although there are many published stud-
ies on the biological activities of essential oils, little
is known about essential oils used for skincare and
that can control acne and prevent scar formation
(Hirasa and Takemasa, 1998; Koleva et al., 2002;
Mantle et al., 1998; Robertto and Baratta, 2000;
Zygadlo et al., 1995). In this study the antimicrobial,
anti-inflammatory and antioxidant properties of se-
ven essential oils commonly suggested a useful inclu-
sions in skincare products, namely citronella grass
(Cymbopogon nardus L.), lemongrass (Cymbopogon
citratus DC), kaffir lime (Citrus hystrix DC), holy ba-
sil (Ocimum sanctum L.), sweet basil (Ocimum basil-
icum L.), plai (Zingiber cassumunar Roxb) and ginger
(Zingiber officinale Roscoe) were investigated.
Materials and methods
Materials
Samples of pure essential oils obtained via
steam distillation were purchased from Thai China
P. Lertsatitthanakorn et al.
Flavours & Fragrances Industry Co. (Thailand).
Tryptic casein soy agar and thioglycollate broth
were purchased from Hispanlab (Spain). Tween80,
ascorbic acid (vitamin C), butylated hydroxyl tolu-
ene (BHT) and alpha-tocopherol (vitamin E) were
purchased from Sigma Chemicals (USA). Nordihy-
droguaretic acid, 5-Lipoxygenase enzyme: Lyophi-
lized powder (6 units/mg) and linoleic acid were
obtained from Fluka (Switzerland). Dimethylsulfox-
ide (DMSO) and absolute alcohol were from Analar
(UK). Potassium dihydrogen phosphate and dipotas-
sium hydrogen phosphate were from Ajax Finechem
(Australia).
Cultures and growth conditions
P. acnes DMST 14916 strain was obtained from the
Department of Medical Sciences, Ministry of Public
Health, Thailand. Cultures were maintained and
grown in thioglycollate broth at 37 C under anaer-
obic conditions.
Antimicrobial test by disc diffusion
technique
Approximately 106 CFU/ml of P. acnes was seeded
onto 20 ml of tryptic casein soy agar. Various con-
centrations of the essential oil samples were pre-
pared in 95% ethanol and 45 ll amounts were
aseptically impregnated onto 8 mm paper discs in
one application and dried under ambient tempera-
ture for 30 min. Discs containing 45 ll of 95% etha-
nol were used as controls. The discs were then
placed on the surface of the seeded agar. The
plates were then incubated at 37 C under anaero-
bic conditions for 48 h. Antibacterial activity was
defined by measuring the diameter of the growth
inhibition zone. Each concentration of essential
oils was performed in triplicate.
Determination of the minimum inhibitory
concentrations (MICs)
A broth microdilution method was used to deter-
mine the minimum inhibitory concentrations
(MICs) and minimum bactericidal concentrations
(MBCs) of the essential oils. To overcome the
insolubility of the oils in the broth, the thioglycol-
late broth was supplemented with 1% Tween80.
Fifty ll of two folded dilutions of each oil sample
were prepared in a 96-well plate. Fifty ll of P.
acnes culture was added to each well to make a
final concentration of approximately 106 CFU/ml.
In each test, P. acnes in thioglycollate broth and
thioglycollate broth alone were used as a positive
In vitro bioactivities of essential oils used for acne control
and negative growth control, respectively. The
plates were then incubated at 37 C under anaero-
bic conditions for 96 h before the MICs and MBCs
were determined. The MIC value was defined as
the lowest concentration of essential oil inhibiting
visible growth. To determine MBC value, 10 ll of
broth was removed from each well and spotted
onto tryptic casein soy agar. After incubated at
37 C under anaerobic conditions for 72 h, the
number of surviving organism was determined.
The lowest concentration where less than 0.1%
of the initial inoculum survived was defined as
MBC value. Each experiment was performed in
triplicate.
Free radical-scavenging activity: 2,2-
diphenyl-1-picrylhydrazyl (DPPH) test
The free radical-scavenging activity of essential
oils was measured according to the method modi-
fied from Cuendet et al. (1997). An equal volume
(650 ll) of each essential oil dissolved in ethanol
at various concentrations was mixed with a
0.0004 M DPPH ethanolic solution. Ascorbic acid
(vitamin C) in the range of 0.70814.160 lg/ml,
BHT (butylated hydroxyl toluene) in the range of
0.86634.640 lg/ml and alpha-tocopherol in the
range of 3.96531.720 lg/ml (vitamin E) were used
as the positive controls whilst ethanol alone served
as a negative control. The reaction mixture was
incubated in the dark at room temperature for
30 min before measuring absorbance at 517 nm.
Percentage inhibition was determined by compari-
son with an ethanol treated control group. IC50 val-
ues were defined as the concentration of tested
samples or positive controls required to scavenge
50% DPPH free radicals. Each concentration of
essential oils and positive control was performed
in triplicate.
Table 1 Susceptibility of P. acnes to seven Thai herbal essential oils by the disc diffusion method
Essential oils
Citronella grass (Cymbopogon nardus L.)
Lemongrass (Cymbopogon citratus DC)a
Kaffir lime (Citrus hystrix DC)b
Holy basil (Ocimum sanctum L.)
Sweet basil (Ocimum basilicum L.)
Plai (Zingiber cassumunar Roxb)
Ginger (Zingiber officinale Roscoe)
Data are mean SD; , no inhibition zone; diameter of filter disc is 8.0 mm.
a
P value at 95% confidence limit = 0.002 for dose dependent manner.
b
P value at 95% confidence limit = 0.003 for dose dependent manner.
45
Determination of anti-inflammatory activity
by inhibition of 5-lipoxygenase (5-LOX)
The assay mixture (3 ml) consisted of 0.1 mM lino-
leic acid, 30 ll of each essential oil at various con-
centrations dissolved in dimethylsulphoxide (DMSO)
and 500 ll of freshly prepared 5-lipoxygenase (5-
LOX) (9.44 units/ml) in 0.1 M potassium phosphate
buffer, pH 6.30. The assay mixture without samples
(essential oils) was used as the negative control
while one containing nordihydroguaretic acid was
used as the positive control. The enzyme reaction
was started by adding 5-lipoxygenase and the
absorbance measured at 234 nm. The absorbance
arising from the modification of the unsaturation
of linoleic acid (diene1-4 was converted to
diene1-3) was measured for 10 min at 25 C. The
initial reaction rate was determined from the slope
of the straight line portion of the curve and the
percentage inhibition of enzyme activity was calcu-
lated by comparison with the control. The concen-
tration that gave 50% enzyme inhibition (IC50) for
the sample was calculated. Each concentration of
essential oils and positive control was performed
in triplicate (Baylac and Racine, 2003).
Results and discussion
Antibacterial activity of the essential oils
Due to their solubility in agar and ability to diffuse
from paper discs, the disc diffusion method was
used for screening the antimicrobial activity of the
essential oils (Janssen et al., 1987). As shown in
Table 1, the lowest concentration of citronella, holy
basil and sweet basil oils showing an inhibition zone
were 1.00% v/v, whereas that of lemongrass and
kaffir lime oils was found at 0.25% v/v. However,
Diameter of inhibition zone (mm)
0.25% v/v 0.50% v/v 1.00% v/v 2.00% v/v
9.5 0.8 10.1 0.8
9.2 0.2 10.5 0.6 13.2 0.6 14.043 1.5
11.5 0.9 19.6 0.6 25.0 0.5 28.250 0.2
9.2 0.6 9.750 0.8
9.5 0.3 9.750 0.6
46
the inhibitory zones of plai and ginger oils were
found to be 4.00% and 1.00% v/v, respectively.
The diameters of the inhibitory zones of kaffir lime
oil and lemongrass oil were linear functions within a
range of 0.252.00% v/v, with the relative coeffi-
cient (r2) of 0.9731 and 0.9564, respectively. This
suggested that the antibacterial activity of the oils
was dose-dependent.
Susceptibility of P: acnes to the essential oils
In contrast to the antibacterial activity test using
the disc diffusion method, the MIC and MBC values
of citronella grass oil were in the range of 0.005
0.3 and 0.61.2 ll/ml, respectively (see Table 2).
Citronella grass oil was the most effective essen-
tial oil against P. acnes whereas the MBC value of
kaffir lime oil (5 ll/ml) indicated that this oil was
less effective than that of lemongrass oil (MBC
0.6 ll/ml) and citronella grass oil (MBC 0.6
1.2 ll/ml). The essential oil of citronella grass had
the lowest MIC and MBC, whereas ginger oil had
the highest (75 ll/ml). The MBC values of holy basil
oil (510 ll/ml) and sweet basil oil (50 ll/ml) indi-
cate that P. acnes was more susceptible to holy ba-
sil oil than to sweet basil oil. Except for citronella
oil, the MICs of the essential oils corresponded to
their MBCs, suggesting a bactericidal effect on P.
acnes.
The different antimicrobial activity of the essen-
tial oils may be due to their different chemical com-
positions. Pauli (2001) analyzed antimicrobial data
of substances mainly found in essential oils and
found that most monoterpenes do not possess a wide
spectrum of activity at low dose. Sesquiterpene
alcohols and fatty acids exhibit weak antimicrobial
properties towards Gram-negative bacteria. A small
number of substances in essential oils, the aldehyde
and phenolic compounds, are able to inhibit bacte-
ria, moulds, dermatophytes and yeasts at low con-
centrations. In this study, the key constituents of
citronellal and geranial in citronella grass and kaffir
lime oils, citral in lemongrass oil, eugenol in holy ba-
Table 2 Susceptibility of P. acnes to seven essential oils measured by the broth microdilution method
Essential oils
Citronella grass (Cymbopogon nardus L.)
Lemongrass (Cymbopogon citratus DC)
Kaffir lime (Citrus hystrix DC)
Holy basil (Ocimum sanctum L.)
Sweet basil (Ocimum basilicum L.)
Plai (Zingiber cassumunar Roxb)
Ginger (Zingiber officinale Roscoe)
a
The data is cited as a range due to the replications showing different values.
P. Lertsatitthanakorn et al.
sil oil, linalool in sweet basil and terpinen-4-ol in plai
oil, may account for their efficacy (Abe et al., 2003;
Carson and Riley, 1995; Lee et al., 2005; Sacchetti
et al., 2005; Tawil and Yousef, 1988).
Antioxidant activity of the essential oils
Based on the reduction of DPPH radicals in the
presence of a hydrogen donating antioxidant, the
absorption of DPPH radicals were decreased (Kuli-
sic et al., 2004). The DPPH radical scavenging
activities of the essential oils and references are
shown in Table 3.
K KruskalWallis H, a nonparametric statistical
analysis, was used to compare the difference among
samples and positive controls and MannWhitney U
was used to compare the difference between the
sample and the positive control. The statistical
analysis was interpreted at a P value of 0.05. Among
the IC50 values of the essential oils, those of citro-
nella grass and holy basil oils (2 and 0.03 ll/ml,
respectively) were notable in that they showed very
good free radical scavenging activity and were sig-
nificantly lower than the IC50 value of ascorbic acid
(7.9 lg/ml). The IC50 value of plai oil was 6.9 ll/ml,
which was lower than that of ginger oil (23.9 ll/ml)
and was significantly lower than ascorbic acid
(7.9 ll/ml), alpha-tocopherol (16.7 ll/ml) and
butylated hydroxyl toluene (BHT) (31.4 ll/ml). Ex-
cept for kaffir lime oil, all tested essential oils
showed better free radical scavenging activity than
BHT. The main antioxidant activity from the essen-
tial oils might be due to the ability of the alcohol,
phenolic and aldehydric compounds in the oils
donating hydrogen to free radicals and thus stopping
the chain reaction of lipid oxidation at the initial
step (King, 1984; Roberts and Caserio, 1965; Shahidi
and Wanasundara, 1992).
The chemical constituents of essential oils are
very complex. Lee et al. (2005) found that among
the aroma chemicals from basil, carvacrol, 4-allyl-
phenol and eugenol showed potent antioxidant
activity. Hence those compounds in the essential
MIC (ll/ml) MBC (ll/ml)
a
0.0050.3 0.61.2a
0.6 0.6
5.0 5.0
5.0 5.010.0a
50.0 50.0
25.0 25.0
75.0 75.0
In vitro bioactivities of essential oils used for acne control 47

Table 3 Free radical scavenging activity of essential oils compared with standard antioxidants
Substance IC50 (ll/ml),
mean SD
Citronella grass (Cymbopogon nardus L.)a 2.0 0.1
Lemongrass (Cymbopogon citratus DC) 27.0 0.7
Kaffir lime (Citrus hystrix DC) 41.7 0.1
Holy basil (Ocimum sanctum L.)a 0.03 0.0
Sweet basil (Ocimum basilicum L.) 31.0 3.9
Plai (Zingiber cassumunar Roxb)a 6.9 0.01
Ginger (Zingiber officinale Roscoe) 23.9 0.8
Vitamin C (ascorbic acid)b 7.9 0.2
Vitamin E (alpha-tocopherol)b 16.7 1.0
BHT (butylated hydroxyl toluene)b 31.4 0.7
a
P-value <0.05. Significantly different in comparison with the standard antioxidants.
b
The standard antioxidant concentrations are expressed as lg/ml.

oils such as citronellal, geranial and terpinen-4-ol
in citronella grass (Mahalwal and Ali, 2002), euge-
nol and methyl eugenol in holy basil (unpublished
data), may play an important role in scavenging
capacity, giving potent antioxidant activity which
is comparable to ascorbic acid and vitamin E. The
major compounds and amounts of citral in lemon-
grass (Baylac and Racine, 2003), citronellal and
neral in kaffir lime, linalool, eugenol and 1,8-cine-
ole in sweet basil (Tomaino et al., 2005), terpinen-
4-ol in plai (Giwanon et al., 2000; Masuda et al.,
1993) and zingiberene, b-bisabolene and b-sesquip-
hellandrene in ginger (Sacchetti et al., 2005) may
account for antioxidant activity.
Anti-inflammatory activity of the essential
oils
5-Lipoxygenase (5-LOX) converts arachidonic acid
into leukotrienes, which are mediators of inflam-
matory and allergic reactions (Werz and Steinhil-
ber, 2005). Reduced IC50 values suggest the better
inhibitory actions on 5-LOX and hence a greater
anti-inflammatory activity. In this study, nordihy-
droguaretic acid, a non-specific lipoxygenase inhib-
itor, was used as a positive control. Unfortunately,
the highest concentrations which fall within the lin-
ear range of the initial rate of sweet basil, plai and
ginger oils were less than 50% inhibition. Therefore,
their IC50 values could not be determined (Table 4).
As reported by Baylac and Racine (2003), a high
citral content interfered with the absorption at
234 nm. Therefore the 5-LOX inhibition of lemon-
grass oil could not be performed. The IC50 values
of citronella grass oil (0.15 ll/ml), kaffir lime oil
(0.05 ll/ml) and holy basil oil (0.04 ll/ml) were
less than nordihydroguaretic acid (1.7 lg/ml).
The inhibition of 5-LOX, by decreasing order of
activity, is holy basil, kaffir lime, citronella grass
and nordihydroguaretic acid, respectively. The ma-
jor components such as eugenol in holy basil oil and
d-limonene in karffir lime oil may contribute to this
activity. These findings were similar to the studies
of Raghavenra et al. (2006) and Baylac and Racine
(2003). Even though studies on the anti-inflamma-
tory action of sweet basil oil, plai oil and ginger

Table 4 Inhibition of 5-lipoxygenase activity of essential oils

Substance IC50 (ll/ml), mean SD
Citronella grass (Cymbopogon nardus L.) 0.15 0.0
Lemongrass (Cymbopogon citratus DC) Nda
Kaffir lime (Citrus hystrix DC) 0.05 0.0
Holy basil (Ocimum sanctum L.) 0.04 0.0
Sweet basil (Ocimum basilicum L.) More than 0.06b
Plai (Zingiber cassumunar Roxb) More than 0.15b
Ginger (Zingiber officinale Roscoe) More than 0.10b
Nordihydroguaretic acid (lg/ml) 1.7 0.1
a
For lemongrass, constituents within the oil interfered with the absorbance at 234 nm and the enzymes activity could not
therefore be measured.
b
More than concentration indicated, the initial reaction rate was not in the linear range and the concentration lower than that
indicated showed less than 50% inhibition.
48
oil have been reported (Flynn et al., 1986; Kiuchi
et al., 1992; Masuda and Jitoe, 1995; Jana et al.,
1999; Ozaki et al., 1991; Pongprayoon et al.,
1996; Singh, 1998; Sharma et al., 1994), the anti-
inflammatory action of these oils on the leukotri-
ene synthetic pathway via a 5-LOX enzyme could
not be evaluated in this study. In addition, apart
from inhibition of the 5-LOX enzyme, the anti-
inflammatory action can also be attributed to the
inhibition of cyclooxygenase enzyme (COX) (Fio-
rucci et al., 2001). Therefore, this preliminary
study on the anti-inflammatory action of the essen-
tial oils could suggest an action via the inhibition of
5-LOX enzyme only. The other pathways of inflam-
matory responses are not elucidated and thus
further studies are needed to clarify the issue.
Conclusion
As previously above, compounds targeting acne
vulgaris should be able to inhibit P. acnes, reduce
inflammation as well as reduce the post-acne scar
formation. In the present study, the essential oil
of citronella grass possessed pronounced antibac-
terial activity against P. acnes, had a very good
free radical scavenging activity and inhibited 5-
LOX enzyme at a practical low concentration,
which could be responsible for the anti-inflamma-
tory activity. Thus, the citronella grass oil may
have a potential for acne control. In addition,
having the most effective antioxidant activity
and 5-LOX inhibitory action, holy basil oil may
be a good candidate for the prevention of scar
formation. This study suggests possible options
for blending aromatic oils for aromatherapy and
cosmetic purposes. However, for safety use it is
important to perform skin irritation tests and their
efficacy must be further studied in an in vivo
model.
References
Abe SSY, Inoue S, Ishibashi H, Mayuyama N, Takizawa T, Oshima
H, Yamaguchi H. Anti- Candida albicans activity of essential
oils including Lemongrass (Cymbopogon citratus) oil and its
component, citral. Nippon Ishinkin Gakkai Zasshi
2003;44:28591.
Baylac S, Racine P. Inhibition of 5-lipoxygenase by essential oils
and other natural fragrant extracts. Int. J. Aromather.
2003;13:13843.
Bensouilah J. Aetiology and management of acne vulgaris. Int. J.
Aromather. 2002;12:99104.
Carson CF, Riley TV. Antimicrobial activity of the major
components of the essential oil of Melaleuca alternifolia.
J. Appl. Bacteriol. 1995;78:2649.
P. Lertsatitthanakorn et al.
Cuendet M, Hostettmank K, Potterat O. Iridoid glucosides with
free radical scavenging properties from Fagraea blumei.
Helv. Chim. Acta 1997;80:114452.
Fiorucci S, Meli R, Bucci M, Cirino G. Dual inhibitors of
cyclooxygenase and 5-lipoxygenase. A new avenue in anti-
inflammatory therapy? Biochem. Pharmacol. 2001;62:
14338.
Flynn DL, Rafferty MF, Boctor AM. Inhibition of human neutro-
phil 5-lipoxygenase activity by gingerdione, shogaol, capsa-
icin and related pungent compounds. Prostaglandins Leukot.
Med. 1986;24:1958.
Giwanon R, Thubthimthed S, Suntorntanasart UR. Antimicrobial
activity of terpenen-4-ol and sabinene. Thai. J. Pharm. Sci.
2000;24:27.
Hirasa K, Takemasa M. Spice science and technology. New
York: Dekker Inc; 1998.
Holland DB, Jeremy AHT. The role of inflammation in the
pathogenesis of acne and acne scarring. Semin. Cutan. Med.
Surg. 2005;24:7983.
Jana U, Chattopadhyay RN, Shaw BP. Preliminary studies on
anti-inflammatory activity of Zingiber officinale Rosc., Vitex
negundo Linn and Tinospora cordifolia (Willid) Miers in albino
rats. Indian J. Pharmacol. 1999;31:2323.
Janssen AM, Scheffer JJ, Baerheim-Svendsen A. Antimicrobial
activities of essential oils: a 19761986 literature review.
Aspects of the test methods. Planta Med. 1987;53:3958.
King RE. Dispensing of medication. Pensylvania: Mack Publish-
ing Company; 1984.
Kiuchi F, Iwakami S, Shibuya M, Hanaoka F, Sankawa U.
Inhibition of prostaglandin and leukotriene biosynthesis by
gingerols and diarylheptanoids. Chem. Pharm. Bull. (Tokyo)
1992;40:38791.
Koleva II, van Beek TA, Linssen JPH, de Groot A, Evstatieva LN.
Screening plant extracts for antioxidant activity: a compar-
ative study on three testing methods. Phytochem. Anal.
2002;13:817.
Kulisic T, Radonic A, Katalinic V, Milos M. Use of different
methods for testing antioxidative activity of oregano essen-
tial oil. Food Chem. 2004;85:63340.
Lee S, Umano K, Shibamoto T, Lee K. Identification of volatile
components in basil (Ocimum basilicum L.) and thyme leaves
(Thymus vulgaris L.) and their antioxidant properties. Food
Chem. 2005;9:1317.
Mahalwal VS, Ali M. Volatile constituents of Cymbopogon nardus
(Linn.) Rendle. Flav. Frag. J. 2002;18:736.
Mantle D, Anderton JG, Falkous G, Barnes M, Jones P, Perry EK.
Comparison of methods for determination of total antioxi-
dant status: application to analysis of medicinal plant
essential oils. Comp. Biochem. Physiol. B 1998;121:38591.
Masuda T, Jitoe A. Phenylbutenoid monomers from the rhizomes
of Zingiber cassumunar. Phytochem. 1995;39:45961.
Masuda T, Jiote A, Nakatani N. Structures of cassumunin A, B,
and C, new potent antioxidants from Zingiber cassumunar.
Chem. Lett. 1993:18992.
Musial W, Kubis A. Preliminary assessment of alginic acid as a
factor buffering triethanolamine interacting with artificial
skin sebum. Eur. J. Pharm. Biopharm. 2003;55:23740.
Ozaki Y, Kawahara N, Harada M. Anti-inflammatory effect of
Zingiber cassumunar Roxb. and its active principles. Chem.
Pharm. Bull. (Tokyo) 1991;39:23536.
Pauli A. Antimicrobial properties of essential oil constituents.
Int. J. Aromather. 2001;1:12633.
Pongprayoon U, Tuchinda P, Claeson P, Sematong T, Reutrakul
V, Soontornsaratune P. Topical anti-inflammatory activity of
the major lipophilic constituents of the rhizome of Zingiber
cassumunar. 1. The essential oil. Phytomedicine 1996;3:
S319.
In vitro bioactivities of essential oils used for acne control
Raghavenra H, Diwakr BT, Lokesh BR, Naidu KA. Eugenol the
active principle from cloves inhibits 5-lipoxygenase activity
and leukotriene-C4 in human PMNL cells, Prostaglandins.
Prostaglandins Leukot Essent Fatty Acids 2006;74:237.
Roberts JD, Caserio MC. Basic principles of organic chemis-
try. New York: WA Benjamin, Inc.; 1965.
Robertto G, Baratta MT. Antioxidant activity of selected
essential oil components in two lipid model systems. Food
Chem. 2000;69:16774.
Sacchetti G, Maietti S, Muzzoli M, Scaglianti M, Manfredini S,
Radice M, Bruni R. Comparative evaluation of 11 essential
oils of different origin as functional antioxidants, antiradicals
and antimicrobials in foods. Food Chem. 2005;91:62132.
Shahidi F, Wanasundara PKJPD. Phenolic antioxidants. Crit. Rev.
Food Sci. Nutr. 1992;32:67103.
Sharma JN, Srivastava KC, Gan EK. Suppressive effects of
eugenol and ginger oil on arthritic rats. Pharmacology
1994;49:3148.
49
Singh S. Comparative evaluation of anti-inflammatory potential
of fixed oil of different species of Ocimum and its possible
mechanism of action. Indian J. Exp. Biol. 1998;36:102831.
Taniguchi S, Kono T, Ishii M. Difference of collagen biosynthesis
between hypertophic scar and normal tissue by some antiox-
idants. J. Dermatol. Sci. 1995;10:61102 (JSID Abstract).
Tawil GG, Yousef RT. Activity of volatile oil components against
Candida albicans. Alexandria J. Pharm. Sci. 1988;2:236.
Tomaino A, Cimino F, Zimbalatti V, Venuti V, Sulfaro V, De
Pasquale A, Saija A. Influence of heating on antioxidant
activity and the chemical composition of some spice essen-
tial oils. Food Chem. 2005;89:54954.
Werz O, Steinhilber D. Development of 5-lipoxygenase inhibitors
lessons from cellular enzyme regulation. Biochem. Phar-
macol. 2005;70:32733.
Zygadlo JA, Lamarque AL, Maestri DM, Grosso NR. Use of
essential oils as natural antioxidants. Grasas Y Aceites
1995;46:2858.