Documenti di Didattica
Documenti di Professioni
Documenti di Cultura
The International
Journal of
Aromatherapy
intl.elsevierhealth.com/journals/ijar
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.
0962-4562/$ - see front matter c 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.ijat.2006.01.006
44 P. Lertsatitthanakorn et al.
Table 1 Susceptibility of P. acnes to seven Thai herbal essential oils by the disc diffusion method
Essential oils Diameter of inhibition zone (mm)
0.25% v/v 0.50% v/v 1.00% v/v 2.00% v/v
Citronella grass (Cymbopogon nardus L.) 9.5 0.8 10.1 0.8
Lemongrass (Cymbopogon citratus DC)a 9.2 0.2 10.5 0.6 13.2 0.6 14.043 1.5
Kaffir lime (Citrus hystrix DC)b 11.5 0.9 19.6 0.6 25.0 0.5 28.250 0.2
Holy basil (Ocimum sanctum L.) 9.2 0.6 9.750 0.8
Sweet basil (Ocimum basilicum L.) 9.5 0.3 9.750 0.6
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.
46 P. Lertsatitthanakorn et al.
the inhibitory zones of plai and ginger oils were sil oil, linalool in sweet basil and terpinen-4-ol in plai
found to be 4.00% and 1.00% v/v, respectively. oil, may account for their efficacy (Abe et al., 2003;
The diameters of the inhibitory zones of kaffir lime Carson and Riley, 1995; Lee et al., 2005; Sacchetti
oil and lemongrass oil were linear functions within a et al., 2005; Tawil and Yousef, 1988).
range of 0.252.00% v/v, with the relative coeffi-
cient (r2) of 0.9731 and 0.9564, respectively. This Antioxidant activity of the essential oils
suggested that the antibacterial activity of the oils
was dose-dependent. Based on the reduction of DPPH radicals in the
presence of a hydrogen donating antioxidant, the
Susceptibility of P: acnes to the essential oils absorption of DPPH radicals were decreased (Kuli-
sic et al., 2004). The DPPH radical scavenging
In contrast to the antibacterial activity test using activities of the essential oils and references are
the disc diffusion method, the MIC and MBC values shown in Table 3.
of citronella grass oil were in the range of 0.005 K KruskalWallis H, a nonparametric statistical
0.3 and 0.61.2 ll/ml, respectively (see Table 2). analysis, was used to compare the difference among
Citronella grass oil was the most effective essen- samples and positive controls and MannWhitney U
tial oil against P. acnes whereas the MBC value of was used to compare the difference between the
kaffir lime oil (5 ll/ml) indicated that this oil was sample and the positive control. The statistical
less effective than that of lemongrass oil (MBC analysis was interpreted at a P value of 0.05. Among
0.6 ll/ml) and citronella grass oil (MBC 0.6 the IC50 values of the essential oils, those of citro-
1.2 ll/ml). The essential oil of citronella grass had nella grass and holy basil oils (2 and 0.03 ll/ml,
the lowest MIC and MBC, whereas ginger oil had respectively) were notable in that they showed very
the highest (75 ll/ml). The MBC values of holy basil good free radical scavenging activity and were sig-
oil (510 ll/ml) and sweet basil oil (50 ll/ml) indi- nificantly lower than the IC50 value of ascorbic acid
cate that P. acnes was more susceptible to holy ba- (7.9 lg/ml). The IC50 value of plai oil was 6.9 ll/ml,
sil oil than to sweet basil oil. Except for citronella which was lower than that of ginger oil (23.9 ll/ml)
oil, the MICs of the essential oils corresponded to and was significantly lower than ascorbic acid
their MBCs, suggesting a bactericidal effect on P. (7.9 ll/ml), alpha-tocopherol (16.7 ll/ml) and
acnes. butylated hydroxyl toluene (BHT) (31.4 ll/ml). Ex-
The different antimicrobial activity of the essen- cept for kaffir lime oil, all tested essential oils
tial oils may be due to their different chemical com- showed better free radical scavenging activity than
positions. Pauli (2001) analyzed antimicrobial data BHT. The main antioxidant activity from the essen-
of substances mainly found in essential oils and tial oils might be due to the ability of the alcohol,
found that most monoterpenes do not possess a wide phenolic and aldehydric compounds in the oils
spectrum of activity at low dose. Sesquiterpene donating hydrogen to free radicals and thus stopping
alcohols and fatty acids exhibit weak antimicrobial the chain reaction of lipid oxidation at the initial
properties towards Gram-negative bacteria. A small step (King, 1984; Roberts and Caserio, 1965; Shahidi
number of substances in essential oils, the aldehyde and Wanasundara, 1992).
and phenolic compounds, are able to inhibit bacte- The chemical constituents of essential oils are
ria, moulds, dermatophytes and yeasts at low con- very complex. Lee et al. (2005) found that among
centrations. In this study, the key constituents of the aroma chemicals from basil, carvacrol, 4-allyl-
citronellal and geranial in citronella grass and kaffir phenol and eugenol showed potent antioxidant
lime oils, citral in lemongrass oil, eugenol in holy ba- activity. Hence those compounds in the essential
Table 2 Susceptibility of P. acnes to seven essential oils measured by the broth microdilution method
Essential oils MIC (ll/ml) MBC (ll/ml)
a
Citronella grass (Cymbopogon nardus L.) 0.0050.3 0.61.2a
Lemongrass (Cymbopogon citratus DC) 0.6 0.6
Kaffir lime (Citrus hystrix DC) 5.0 5.0
Holy basil (Ocimum sanctum L.) 5.0 5.010.0a
Sweet basil (Ocimum basilicum L.) 50.0 50.0
Plai (Zingiber cassumunar Roxb) 25.0 25.0
Ginger (Zingiber officinale Roscoe) 75.0 75.0
a
The data is cited as a range due to the replications showing different values.
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 anti-inflammatory activity. In this study, nordihy-
in citronella grass (Mahalwal and Ali, 2002), euge- droguaretic acid, a non-specific lipoxygenase inhib-
nol and methyl eugenol in holy basil (unpublished itor, was used as a positive control. Unfortunately,
data), may play an important role in scavenging the highest concentrations which fall within the lin-
capacity, giving potent antioxidant activity which ear range of the initial rate of sweet basil, plai and
is comparable to ascorbic acid and vitamin E. The ginger oils were less than 50% inhibition. Therefore,
major compounds and amounts of citral in lemon- their IC50 values could not be determined (Table 4).
grass (Baylac and Racine, 2003), citronellal and As reported by Baylac and Racine (2003), a high
neral in kaffir lime, linalool, eugenol and 1,8-cine- citral content interfered with the absorption at
ole in sweet basil (Tomaino et al., 2005), terpinen- 234 nm. Therefore the 5-LOX inhibition of lemon-
4-ol in plai (Giwanon et al., 2000; Masuda et al., grass oil could not be performed. The IC50 values
1993) and zingiberene, b-bisabolene and b-sesquip- of citronella grass oil (0.15 ll/ml), kaffir lime oil
hellandrene in ginger (Sacchetti et al., 2005) may (0.05 ll/ml) and holy basil oil (0.04 ll/ml) were
account for antioxidant activity. less than nordihydroguaretic acid (1.7 lg/ml).
The inhibition of 5-LOX, by decreasing order of
Anti-inflammatory activity of the essential activity, is holy basil, kaffir lime, citronella grass
oils and nordihydroguaretic acid, respectively. The ma-
jor components such as eugenol in holy basil oil and
5-Lipoxygenase (5-LOX) converts arachidonic acid d-limonene in karffir lime oil may contribute to this
into leukotrienes, which are mediators of inflam- activity. These findings were similar to the studies
matory and allergic reactions (Werz and Steinhil- of Raghavenra et al. (2006) and Baylac and Racine
ber, 2005). Reduced IC50 values suggest the better (2003). Even though studies on the anti-inflamma-
inhibitory actions on 5-LOX and hence a greater tory action of sweet basil oil, plai oil and ginger
oil have been reported (Flynn et al., 1986; Kiuchi Cuendet M, Hostettmank K, Potterat O. Iridoid glucosides with
et al., 1992; Masuda and Jitoe, 1995; Jana et al., free radical scavenging properties from Fagraea blumei.
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