Journal of Applied Microbiology ISSN 1364-5072

ORIGINAL ARTICLE

Antimicrobial activity of lavender, tea tree and lemon oils

in cosmetic preservative systems
A. Kunicka-Styczynska1, M. Sikora2 and D. Kalemba2
1 Institute of Fermentation Technology and Microbiology, Technical University of Lodz, Lodz, Poland
2 Institute of General Food Chemistry, Technical University of Lodz, Lodz Poland

Keywords
antimicrobial activity, challenge test, cosmetic
formulations, essential oils, preservative
system.
Correspondence
Alina Kunicka-Styczynska, Institute of
Fermentation Technology and Microbiology,
Technical University of Lodz, 90-924 Lodz,
Wolczanska 171 173, Poland.
E-mail: akunicka@p.lodz.pl
2009 0113: received 19 January 2009,
revised 30 March 2009 and accepted 21 April
2009
doi:10.1111/j.1365-2672.2009.04372.x
Abstract
Aims: The aim of the study was to verify the antimicrobial activity of commer-
cial essential oils: lavender, tea tree and lemon as the components of a preser-
vative system in oil in water body milks.
Methods and Results: The inhibition efficacy of essential oils alone (05%), in
mixtures (1%) as well as combined with the synthetic preservative 1,3-dimethy-
lol-5,5-dimethylhydantoin and a 3-iodo-2-propynyl butyl carbamate mixture
(01% and 02%) was tested against Staphylococcus aureus ATCC 6538, Pseudo-
monas aeruginosa ATCC 9027, Candida sp. OCK 0008 and Aspergillus niger
ATCC 16404 in compliance with the standards of the European Pharmacopoeia
Commission. The in vitro activity of oils determined by an impedimetric
method was also compared with their activity in cosmetic preparations. Crite-
rion A for bacteria (reduction in the inoculum by 3 logarithmic units within
7 days with no increase up to the 28th day) and fungi (reduction in the inocu-
lum by 2 logarithmic units within 14 days with no increase up to the 28th
day) was fulfilled for cosmetic formulations containing the tested essential oils
with 02% of the synthetic preservative. The preservative concentration could
be decreased to 01% (with preserving the same efficacy) in combination with
lavender and tea tree oils at a concentration of 05% each.
Conclusions: In all combinations of essential oils with the synthetic preserva-
tive, a synergistic effect of the preservative system components was observed,
which made it possible to reduce the usable level of the synthetic preservative
up to 85 times.
Significance and Impact of the Study: To develop an effective preservative
system in cosmetics in which a synthetic chemical preservative is replaced by
natural essential oils.

good condition. The activity of preservatives in cosmetics

Introduction
Microbiological purity is one of the most important prob-
lems in cosmetic industry, and microbiological impurities
lead to loss of cosmetic properties and also pose a risk of
infection to the users (Wong et al. 2000; Nostro et al.
2004). Moreover, the development of micro-organisms
may cause changes in the consistency, flavour, colour,
phase separation or sediment creation in solutions.
Chemical compounds inhibiting the growth of micro-
organisms are added to a product to ensure its stability.
Referred to as preservatives, they maintain cosmetics in
is usually connected with ensuring that cosmetic prepara-
tions are devoid of impurities during their production
and packaging as well as throughout the entire time of
their use.
For cosmetic goods, sanitary requirements set limits on
maximum permissible concentration of synthetic preser-
vatives (EU Cosmetics Directive 1976) and force cosmet-
ics manufacturers to employ alternative methods of
preservation.
Taking into account the fact that allergies after using
cosmetics occur more and more frequently, which is

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Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 107 (2009) 19031911 1903
Antimicrobial action of oils in cosmetics
quite often associated with the presence of synthetic
preservatives (De Groot 1998), it seems justified to search
for their counterparts of natural origin such as essential
oils (Maccioni et al. 2002; Seo et al. 2002; Nostro et al.
2004).
The aim of the study was to test the antimicrobial
activity of commercial essential oils: lavender, tea tree
and lemon oils in oil in water (O W) body milks.
Documented antimicrobial effectiveness usually refers to
in vitro conditions (Inouye et al. 2001; Carson et al. 2006;
Erturk et al. 2006). In the study, in vitro activity of oils was
compared with their activity in cosmetic preparations.
Apart from research on the potential use of essential
oils and their mixtures as sole preservatives in formulas,
research on the activity of preservative systems containing
a combination of essential oils and synthetic preservative
is also presented. The synergistic effects of the activity of
essential oils and 1,3-dimethylol-5,5-dimethylhydantoin
(DMDM hydantoin) in combination with 3-iodo-2-pro-
pynyl butyl carbamate (used in many commercial cos-
metic formulas) are examined. DMDM hydantoin belongs
to a class of preservatives that are formaldehyde donors.
The compound creates strong biocidal and biostatic
effects and is compatible with most cosmetic formula-
tions. The addition of 3-iodo-2-propynyl butyl carbamate
increases the antifungal activity of the preservative.
Materials and methods
Microorganisms
Staphylococcus aureus ATCC 6538, Pseudomonas aerugin-
osa ATCC 9027, Candida sp. OCK 0008 and Aspergillus
niger ATCC 16404 were used. The micro-organisms origi-
nated from ATCC and Collection of Pure Cultures of the
Institute of Fermentation Technology and Microbiology,
Technical University of Lodz, OCK 105. The micro-
organisms were activated through double passaging: bac-
teria on trypticase soy agar medium (TSA for 37C, 48 h;
Oxoid, Basingstoke, UK), yeast and moulds on Sabouraud
dextrose agar medium (SDA for 28C, 72 h yeast and
7 days moulds; bioMerieux, Warsaw, Poland).
Essential oils
Commercial essential oils purchased from the manufac-
turer were used for the experiments. The chemical com-
position of the lavender and tea tree oil fulfilled the
requirements of the European Pharmacopoeia Commis-
sion 5.0 (E.P. 5.0) (2005), while the lemon oil had lower
c-terpinene content, i.e. 23% against the required
60120%. The main components determined with the
use of GC-MS method (Baran et al. 2007) were as fol-
1904 Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 107 (2009) 19031911
A. Kunicka-Styczynska et al.
lows. Lavender oil: linalool 341%, linalyl acetate 333%,
1,8-cineole 25%, terpinen-4-ol 25%, a-terpineol 18%,
lavandulol 11%, lavandulyl acetate 32% and camphor
12%. Tea tree oil: terpinen-4-ol 413%, c-terpinene
191%, a-terpinene 83%, p-cymene 55%, 1,8-cineole
34% and a-terpineol 32%. Lemon oil: limonene 798%,
b-pinene 36%, c-terpinene 23%, geranial 27% and neral
10%.
Antimicrobial activity of essential oils in vitro
The antimicrobial activity of essential oils was determined
by an impedimetric method using Bactometer M64 Sys-
tem (bioMerieux). The suspensions of the tested bacteria
and yeast cells were prepared in a physiological salt solu-
tion (NaCl 85 g l)1), the ones of the mould in a physio-
logical salt solution with the addition of 05 g l)1
polysorbate 80 R and later standardized to the density of
about 108 CFU ml)1. Ethanol was used to disperse the
essential oils. The wells of the impedimeters module were
filled with 01 ml of cell suspension, 0145 ll of oil and
complemented with growth medium to 10 ml. A positive
control was the suspension of micro-organisms in the
medium without essential oils. The negative controls were
the bacteria and fungi cultures with the addition of novo-
biocin (05 lg ml)1) and cycloheximide (02 lg ml)1)
respectively. Samples were incubated for 72 h at tempera-
tures optimal for the growth of individual micro-organ-
isms, as described in the strain activation procedure.
After incubation in the bactometer, the viability of micro-
organisms was controlled by a surface culture on TSA
(bacteria) or SDA (fungi) medium. Plates were incubated
for 3 days for bacteria and yeast and for 5 days
for moulds at temperatures optimal for the growth of
particular micro-organisms.
Minimal inhibitory concentration (MIC) was assigned
as the lowest concentration of an essential oil inhibiting
growth of micro-organisms in the bactometer, at parallel
growth on agar medium. Minimal Bactericidal Concentra-
tion (MBC) or Minimal Fungicidal Concentration (MFC)
was determined as the lowest oil concentration at
which no microbial growth was observed either in the
bactometer wells or on agar medium.
Body milk
Formula ingredients of cosmetics were oil phase: poly-
glyceryl-3 methylglucose distearate 200 g l)1, isopropyl
palmitate 600 g l)1, cyclomethicone 350 g l)1, glyceryl
stearate 250 g l)1; water phase: xanthan gum 15 g l)1,
propylene glycol 700 g l)1, allantoin 20 g l)1, water
7865 g l)1. Body milk without synthetic preservatives and
essential oils acted as a reference sample. The commercial
2009 The Authors
A. Kunicka-Styczynska et al. Antimicrobial action of oils in cosmetics

Table 1 Preservative system composition in body milk formulations in The prepared suspensions were inoculated on TSA plates
% (v v) for bacteria and SDA plates for fungi, prior to being
Essential oils resuspended so as to obtain 30300 colonies on a plate.
The plates were incubated at 37 and 28C for bacteria
Synthetic Tea tree Lemon: tea tree and fungi respectively. The results were expressed as
Lavender L TTO Lemon C mixture C:TTO
preservative
log CFU ml)1.
01 *
02
03
Statistical analysis of the results
05 Results were analysed using a 3-way anova test at a con-
05
fidence level of P < 005. Results of the population viabil-
05 05
01 05 05
ity were presented as an arithmetic mean of three
05 determinations with standard deviation not exceeding 02
02 05 logarithmic units.
02 05
1 (1C:1TTO)
1 (4C:1TTO) Results
02 1 (4C:1TTO)
Antimicrobial activity of essential oils in vitro
*not added.
In in vitro experiments, the oils exhibited similar inhibi-
synthetic preservative Glydant Plus Liquid (GPL), tory activity against individual micro-organisms and
containing 1,3-dimethylol-5,5-dimethylhydantoin and slight differences in cidal activity (Table 2). However, the
3-iodo-2-propynyl butyl carbamate were presented as sensitivity of the tested micro-organisms to selected oils
active compounds (Lonzagroup, Switzerland); and essen- was very varied. MIC values ranged between 002% and
tial oils: lavender, tea tree and lemon purchased from 004% for Asp. niger and 010% for Ps. aeruginosa. The
FSZ Pollena Aroma (Warsaw, Poland), were added in dif- MBC of oils was usually slightly higher, from 004% to
ferent combinations. The concentrations of essential oils, 020%. The inhibitory activity hierarchy of the tested oils
their mixtures and combinations with the synthetic pre- determined on the basis of MIC values was as follows: tea
servative are presented in Table 1. tree oil > lavender oil > lemon oil. At the same time, the
sensitivity of the micro-organisms was as follows:
Asp. niger > Candida sp. > Staph. aureus > Ps. aeruginosa,
Challenge test
and therefore in in vitro experiments, yeast and moulds
Challenge tests of cosmetics were prepared by introducing were more sensitive than bacteria.
1 ml suspension of bacteria or fungi after activation to
100 ml body milk. The suspensions of test micro-organ-
Synthetic preservative
isms were prepared as it was described in experiments in
the antimicrobial qualities of essential oils in vitro. Body In body milks without preservatives, the cell numbers of
milks were mixed thoroughly and incubated in the dark Ps. aeruginosa and Candida sp. increased by 1 logarithmic
at 20C. The number of viable cells in formulations was unit, but the population of Asp. niger was reduced by
determined by the count plate method immediately after about 1 logarithmic unit (Fig. 1). Staph. aureus cells died,
inoculation and at 2, 7, 14 and 28 days. A sample of and their number decreased by about 4 logarithmic
10 ml body milk was transferred to 90 ml suspending units during the test. The highest sensitivity to the
liquid (peptone K 50 g l)1, NaCl 85 g l)1, pH 72 02). synthetic preservative was exhibited by Staph. aureus. The

Table 2 Antimicrobial activity of tested

essential oils at Minimal Inhibitory Concen- Essential oils
tration (MIC) and Minimal Bactericidal Lavender L Tea tree TTO Lemon C
Concentration (MBC) or Minimal Fungicidal
Concentration (MFC) in % (v v) Micro-organisms MIC MBC MFC MIC MBC MFC MIC MBC MFC

Staphylococcus aureus 005 010 004 015 005 020

Pseudomonas aeruginosa 010 010 010 020 010 020
Candida sp. 004 007 003 005 004 010
Aspergillus niger 003 004 002 005 004 004

2009 The Authors

Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 107 (2009) 19031911 1905
Antimicrobial action of oils in cosmetics A. Kunicka-Styczynska et al.

7 7

Viable cells in log (CFU ml1)

Viable cells in log (CFU ml1)

Staph. aureus Ps. aeruginosa
6 6
5 5
4 4
3 3
2 2
1 1
0 0
0 5 10 15 20 25 30 0 5 10 15 20 25 30
Time (days) Time (days)

7 7
Viable cells in log (CFU ml1)

Viable cells in log (CFU ml1)

Asp. niger
6 6
Candida sp.
5 5
4 4
3 3
2 2
1 1
0 0
0 5 10 15 20 25 30 0 5 10 15 20 25 30
Time (days) Time (days)

Figure 1 Growth inhibition of Staphylococcus aureus, Pseudomonas aeruginosa, Candida sp. and Aspergillus niger in body milk formulations
without and with the synthetic preservative [Glydant Plus Liquid (GPL)]; body milk ( ), body milk with 01% GPL ( d ), body milk with 02% GPL
( ), body milk with 03% GPL ( . ).

preservative with the lowest concentration tested, i.e.

Lemon and tea tree oil
01%, eliminated Staph. aureus and Asp. niger from the
formulation after 7 and 14 days of incubation respec- As predicted, lemon oil added to body milk at a concen-
tively. However, it was not effective against Ps. aeruginosa tration of 05% exhibited weaker inhibitory activity
and Candida sp. An increase in the preservative concen- against Staph. aureus in comparison with tea tree oil
tration to 03% resulted in body milk sanitization within (Fig. 3). However, no statistically significant differences in
14 days. the activity of lemon and tea tree oils against the remain-
ing micro-organisms were observed. A combination of a
synthetic preservative at a concentration of 02% and
Lavender and tea tree oil
both of the oils at a concentration of 05% inhibited all
Lavender and tea tree oils added at a concentration of the micro-organisms present in the milk as early as after
05% to cosmetic formulations killed Staph. aureus as 2 days of incubation. The addition of lemon or tea tree
early as after 2 days of incubation (Fig. 2). At the same oils enhanced the fungistatic activity of the synthetic
time, lavender oil did not affect the remaining micro- preservative (Figs 1 and 3).
organisms, and tea tree oil only slightly inhibited the Consumer fragrance assessment of cosmetics (M. Sikora
growth of Asp. niger (by about 1 logarithmic unit). After and A. Kunicka-Styczynska, unpublished data) showed
7 days of a challenge test, an inhibitory effect against acceptance for the concentrations of individual oils that
Ps. aeruginosa was observed in the body milk with a mix- did not exceed 05%. At the same time, it revealed accep-
ture of both oils, at a concentration of 05% each. The tance for the fragrance of the mixture of lemon and tea tree
mixture, however, did not exhibit fungistatic activity. The oils at a summary concentration of 1%, with preference
introduction of a synthetic preservative at a concentration shown to lemon oil note. On that basis, a formulation con-
of 01% to the formulation containing 05% of lavender taining lemon and tea tree oil, introduced to the cosmetic
and tea tree oil triggered an unexpected inhibitory effect as mixtures at 1 : 1 or 4 : 1, was created. As expected, in
against all the tested strains as early as after 2 days. The body milks containing 1% of those oils mixtures (irrespec-
mixture of oils significantly enhanced the activity of the tive of their volume ratio), Staph. aureus was not detected
preservatives. from the second day, whereas the growth inhibition of

2009 The Authors

1906 Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 107 (2009) 19031911
A. Kunicka-Styczynska et al. Antimicrobial action of oils in cosmetics

7 7

Viable cells in log (CFU ml1)

Viable cells in log (CFU ml1)

Staph. aureus
6 6
5 5 Ps. aeruginosa
4 4
3 3
2 2
1 1
0 0
0 5 10 15 20 25 30 0 5 10 15 20 25 30
Time (days) Time (days)

7 7
Viable cells in log (CFU ml1)

Viable cells in log (CFU ml1)

Asp. niger
6 6
5 Candida sp. 5
4 4
3 3
2 2
1 1
0 0
0 5 10 15 20 25 30 0 5 10 15 20 25 30
Time (days) Time (days)

Figure 2 Growth inhibition of Staphylococcus aureus, Pseudomonas aeruginosa, Candida sp. and Aspergillus niger in body milk formulations with
different preservative systems containing lavender oil (L), tea tree oil (TTO) and these oils combined with the synthetic preservative [Glydant Plus
Liquid (GPL)]; body milk with 05% L ( ), body milk with 05% TTO ( d ), body milk with 05% L and 05% TTO ( ), body milk with 01% GPL
and 05% L and 05% TTO ( . ).

Ps. aeruginosa, Candida sp. and Asp. niger was not
observed (Fig. 4). Nevertheless, the enhancing effect on the
activity of the synthetic preservative (02%) in the presence
of the last mixture of oils was observed (Figs 1 and 4).
Discussion
Antimicrobial activity of lavender and tea tree oil in
in vitro conditions is well documented. However, varied
methods of research do not allow making explicit refer-
ences (Kalemba and Kunicka 2003). MIC values for tea
tree oil against Asp. niger determined by different
researchers vary almost 50 times, from 0016% (Beylier
1979) to 075% v v (Peciulyte 2004). Similarly, the inhibi-
tory effect of lavender oil against Staph. aureus was
observed at minimum concentrations of both 001% (Ino-
uye et al. 2001) and 032% (Cavanagh and Wilkinson
2002). Results obtained by us fall into those ranges. Both
of the tested oils were characterized by higher activity
against Ps. aeruginosa (Viljoen and Van Vuuren 2006)
and Candida spp. (Hammer et al. 1998; Schwiertz et al.
2006) than observed in the literature. The lemon oil
tested by us exhibited approximately ten times higher
activity than previously described (Devkatte et al. 2005;
Erturk et al. 2006). Simultaneously, the other findings
(Fisher and Phillips 2006) showed up to three times
higher MIC values. A disagreement of the presented
results and literature data are because of a different test
system used in the study. The wells of the impedimeter
are tightly closed, which prevents the essential oils from
evaporating, and the amount of oil do not change during
the test. Moreover, the vapours probably contribute to
the antimicrobial effect of the essential oil in this system.
Recent studies highlight the role of water-soluble and
-vaporized components in the assessment of antimicrobial
activity of essential oils (Inouye et al. 2006; Fisher and
Phillips 2008). The answer of micro-organisms is also
quicker because of the small volume of the probe, which
makes the system less inert.
According to the criteria of the European Pharmaco-
poeia Commission 5.0 (E.P. 5.0) (2005) set for topical
preparations, Criterion A for bacteria and fungi was
fulfilled for cosmetic formulations containing minimum
02% of the synthetic preservative used as a sole inhibi-
tor or in combination with the tested essential oils.
Criterion A for bacteria signifies a reduction in the
inoculum by 3 logarithmic units within 7 days of a
challenge test with no increase up to the 28th day, and

2009 The Authors

Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 107 (2009) 19031911 1907
Antimicrobial action of oils in cosmetics A. Kunicka-Styczynska et al.

7 7

Viable cells in log (CFU ml1)

Viable cells in log (CFU ml1)

Staph. aureus Ps. aeruginosa
6 6
5 5
4 4
3 3
2 2
1 1
0 0
0 5 10 15 20 25 30 0 5 10 15 20 25 30
Time (days) Time (days)

7 7
Viable cells in log (CFU ml1)

Viable cells in log (CFU ml1)

Asp. niger
6 Candida sp. 6
5 5
4 4
3 3
2 2
1 1
0 0
0 5 10 15 20 25 30 0 5 10 15 20 25 30
Time (days) Time (days)

Figure 3 Growth inhibition of Staphylococcus aureus, Pseudomonas aeruginosa, Candida sp. and Aspergillus niger in body milk formulations with
different preservative systems containing lemon oil (C) and tea tree oil (TTO) and these oils combined with the synthetic preservative [Glydant Plus
Liquid (GPL)]; body milk with 05% C ( ), body milk with 05% TTO ( d ), body milk with 02% GPL and 05% C ( ), body milk with 02%
GPL and 05% TTO ( . ).

for fungi, a reduction in the inoculum by 2 logarithmic The substantially lower activity of essential oils in cos-
units within 14 days with no increase up to the 28th metic formulations in comparison with in vitro condi-
day. Preservative concentration could be decreased to tions may result from the presence of lipid compounds.
01% with fulfilment of criterion A for a combination The high affinity of essential oils for the oily cosmetic
with lavender and tea tree oils at a concentration of ingredients limits their accessibility in the water phase
05% each. At the same time, in all the combinations resulting in antimicrobial activity reduction (Manou et al.
of essential oils with the synthetic preservative, the 1998; Nostro et al. 2002, 2004). It therefore seems justi-
synergistic effect of the components of the preservative fied to increase the concentration of essential oil in a for-
system was observed. Synergism between essential oils mulation. There is a possibility of a substantial increase
and methyl p-hydroxybenzoate (MPB) was observed in in essential oil concentration to 5% in hygienic skin wash
skin cream research conducted by Maccioni et al. meant for short contact with the skin (Messager et al.
(2002), where Laurus nobilis, Eucalyptus globulus and 2005a,b). The concentration of Artemisia afra, Lavandula
Salvia officialis oils added at a concentration of 0025% officinalis, Rosmarinus officinalis and Pteronia incana oils
and 00125% were active only if combined with MPB. in aqueous cream increased to 15% contributed to better
A similar phenomenon was observed for tea tree oil in antimicrobial effectiveness of these preparations (Muyima
formulations with ethanol addition (Messager et al. et al. 2002). However, an increase in the summary con-
2005a). A synergistic effect between different active centration of lemon and tea tree oils to 1% in the body
chemical substances with antiseptic properties has been milks tested by us did not bring about the desired effect.
thoroughly described (Jeng and Severin 1998; Herrera On the other hand, a combination of lavender and tea
et al. 2003; Messager et al. 2005b), which, given the tree oil, both at a concentration of 05%, triggered visible
chemical complexity of essential oils, justifies the results synergistic effect against Ps. aeruginosa and much weaker
of this study. It has also been concluded that both syn- against fungi. The research presented confirmed the possi-
ergism and antagonism can occur between components bility of the antagonistic or synergistic activity of the
of tea tree oil and the components of formulations components depending on the introduced oils (Cox et al.
(Cox et al. 2001). 2001). Adding relatively small quantities of essential oils

2009 The Authors

1908 Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 107 (2009) 19031911
A. Kunicka-Styczynska et al. Antimicrobial action of oils in cosmetics

7 7

Viable cells in log (CFU ml1)

Viable cells in log (CFU ml1)

Staph. aureus
6 6
5 5
Ps. aeruginosa
4 4
3 3
2 2
1 1
0 0
0 5 10 15 20 25 30 0 5 10 15 20 25 30
Time (days) Time (days)

7 7

Viable cells in log (CFU ml1)

Viable cells in log (CFU ml1)

Asp. niger
6 6
Candida sp.
5 5
4 4
3 3
2 2
1 1
0 0
0 5 10 15 20 25 30 0 5 10 15 20 25 30
Time (days) Time (days)

Figure 4 Growth inhibition of Staphylococcus aureus, Pseudomonas aeruginosa, Candida sp. and Aspergillus niger in body milk formulations with
different preservative systems containing mixture of lemon oil (C) and tea tree oil (TTO) and these oils combined with the synthetic preservative
[Glydant Plus Liquid (GPL)]; body milk with 1% 1C:1TTO ( ), body milk with 1% 4C:1TTO ( d ), body milk with 02% GPL and 1% 4C:1TTO ( ).

to cosmetics results from its potential irritation and aller-
gic influence and also depends on the consumers accep-
tance of the fragrance. The accepted level of oils in
formulations did not exceed 05% for ones tested
individually and 1% for mixtures (M. Sikora and
A. Kunicka-Styczynska, unpublished data).
Essential oils without a synthetic preservative acted as
an active preservative system only against Gram-positive
bacteria Staph. aureus, parallel to experiments in L. nobi-
lis, E. globulus and S. officialis oils in skin cream formula-
tions (Maccioni et al. 2002). The effect was not observed
in Gram-negative bacteria Ps. aeruginosa. A higher sensi-
tivity of Gram-positive bacteria to the tested preservative
systems in all of the examined formulations results from
a less complex structure and composition of their cell
wall resulting in better permeability of the components of
the essential oils (Inouye 2003; Kalemba and Kunicka
2003; Bakkali et al. 2008). Growth of Ps. aeruginosa and
Candida sp. in formulations with essential oils addition
after the adaptation phase proves the low sensitivity of
those organisms and their ability to use body milk com-
ponents as a carbon source. Similar phenomenon was
documented for Ps. aeruginosa in W O formulations
containing Thymus vulgaris oil (Manou et al. 1998). The
growth of Candida albicans in the presence of 05%
A. afra, L. officinalis, R. officinalis and P. incana oils in
the cream formulation also confirms our observations
(Muyima et al. 2002). Despite the high fungistatic activity
of the oils tested in our study, Asp. niger was not sensitive
to them in the cosmetic formulations. The same results
were obtained for thyme oil in O W and W O creams
(Manou et al. 1998) as well as for fir, lavender, rosemary
and P. incana oils added to aqueous cream at a concen-
tration of 05% (Muyima et al. 2002).
The application of essential oils for preserving cosmetic
formulations is still doubtful because of their relatively
low antimicrobial activity in comparison with the com-
monly used chemical preservatives (Kabara 1984). A fur-
ther limitation is the acceptance of the organoleptic
qualities of the cosmetic with more than 2% of essential
oil. Preservative systems consisting of an essential oil and
a chemical preservative seem to be a compromise solu-
tion. The combination of the GPL preservative with
lemon and tea tree oils, presented in this study, allows
reducing the usable level of GPL from 085%, recom-
mended by the producer, to 02%. Moreover, substituting
lemon oil with lavender oil allows reducing the GPL dose
in the examined formulations by 85 times. Presumably,
the action of the proposed systems implemented in O W
lotions and washing liquids seems to be quite promising.
However, the possibility of interactions between the oil
components, the synthetic preservative and the cosmetic

2009 The Authors

Journal compilation 2009 The Society for Applied Microbiology, Journal of Applied Microbiology 107 (2009) 19031911 1909
Antimicrobial action of oils in cosmetics A. Kunicka-Styczynska et al.

components, which are difficult to predict, makes it nec- Hammer, K.A., Carson, C.F. and Riley, T.V. (1998) In vitro
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specific formulations. (tea tree) oil and tea tree products, against Candida spp. J
Antimicrob Chemother 42, 591595.
Herrera, D., Roldan, S., Santacruz, I., Santos, S., Masdevall, M.
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The authors express their thanks to the Polish Ministry of of four commercial 0.12% chlorhexidine mouthrinse
Science and Higher Education for supporting this work formulations: an in vitro contact test and salivary bacterial
counts study. J Clin Periodontol 30, 307314.
(Grant MNiSW N405 055 31 3865).
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