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Journal of Essential Oil Research

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Volatile fraction composition and biological activity

of lemon oil (Citrus limon L. Burm.): Comparative
study of oils extracted from conventionally grown and
biological fruits
a a b c a
Federica Spadaro , Clara Circosta , Rosaria Costa , Francesco Pizzimenti , Dora Rita
a a
Palumbo & Francesco Occhiuto
a
Pharmaco-Biological Department, School of Pharmacy , University of Messina , Messina ,
Italy
b
Chromaleont s.r.l., a spin-off of the University of Messina , Messina , Italy
c
Dipartimento Farmaco-Chimico, School of Pharmacy , University of Messina , Messina ,
Italy
Published online: 20 Mar 2012.

To cite this article: Federica Spadaro , Clara Circosta , Rosaria Costa , Francesco Pizzimenti , Dora Rita Palumbo & Francesco
Occhiuto (2012) Volatile fraction composition and biological activity of lemon oil (Citrus limon L. Burm.): Comparative study
of oils extracted from conventionally grown and biological fruits, Journal of Essential Oil Research, 24:2, 187-193, DOI:
10.1080/10412905.2012.659518

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 The Journal of Essential Oil Research
Vol. 24, No. 2, April 2012, 187–193

Volatile fraction composition and biological activity of lemon oil (Citrus limon L. Burm.):
Comparative study of oils extracted from conventionally grown and biological fruits
Federica Spadaroa, Clara Circostaa*, Rosaria Costab,c, Francesco Pizzimentia, Dora Rita Palumboa and Francesco
Occhiutoa
a
Pharmaco-Biological Department, School of Pharmacy, University of Messina, Messina, Italy; bChromaleont s.r.l., a spin-off of
the University of Messina, Messina, Italy; cDipartimento Farmaco-Chimico, School of Pharmacy, University of Messina, Messina,
Italy
(Received 21 July 2011; final form 18 October 2011)

In this study, conventional and biological lemon cultivation techniques have been evaluated for their influence on the
composition and biological activity of essential oils extracted from fresh lemon fruits. The essential oils were
Downloaded by [McMaster University] at 20:54 24 December 2014

extracted by hydrodistillation (HD) and analyzed by gas chromatography–flame ionization detector (GC-FID) and
GC–mass spectrometry (GC-MS). The human Ishikawa cell line maintained in culture was used to test the antiprolif-
erative efficacy of essential oils and the paper disc diffusion method was employed to determine the antimicrobial
activity. The results obtained indicated differences in both oil composition, especially in the content of oxygenated
compounds, and biological activity, leading to the conclusion that the biological oils were of higher quality.
Keywords: Citrus limon L. Burm (Rutaceae); biological essential oil; conventional essential oil; hydrodistillation;
GC-FID; GC-MS; antiproliferative activity; antimicrobial activity

Introduction chromatography (GC) composition, the antiproliferative

Citrus limon (L.) Burm. (Rutaceae) is the most impor-
tant fruit tree crop in the world, with an annual produc-
tion of approximately 102 million tonnes. Chemical
industry extracts from lemon, such as essential oils,
flavonoids, vitamins, minerals, dietary fiber, and so on,
are used, as functional ingredients, in the development
of healthy foods (functional foods) (1); in cosmetic and
pharmaceutical industry, for their antimicrobial, antioxi-
dant, anticarcinogenic properties; and in aromatherapy,
for their influence on human mood (2, 3).
Conventional and biological (organic) agriculture
are two methods exploited by farmers: the first is based
on the use of various pesticides, herbicides, antibiotics,
growth hormones, and other chemicals to accelerate
plant growth and increase products size (4). Biological
agriculture does not use any chemicals for these scopes.
Organic farmers use natural feeds and fertilizers and
even though this type of agriculture is more expensive,
in the last few years there has been an increasing inter-
est among consumers towards organic products (5).
This type of cultivation respects the environment,
defends biodiversity, produces healthy food, and pro-
motes quality, tradition and country specificity.
The present study compares the composition
and biological activity of the volatile fractions of lemon
oils obtained from the two cultivation techniques
described above, in particular investigating the gas
and antimicrobial activity.
Experimental
Plant material
In this study, Citrus limon L. Burm. (Rutaceae) from
Femminello white zagara lemon cultivar was used. The
fruits were gathered in November 2010 and came from
the same geographic area, situated in the neighborhood
of Reggio Calabria (farm G. Corigliano, Bovalino,
Italy). Lemon fruits were peeled by hand, separating
the external part of the lemon fruits (flavedo), with a
yield of 29% w/w of lemon peel over the whole fruit.
Fresh plant material was employed for all extractions.
Samples investigated were ten biological and ten
conventional essential oils.
Cell lines, microorganisms, chemicals and biochem-
icals
The Ishikawa line of human endometrial adenocarci-
noma cells was purchased from the ECACC (Porton
Down, Salisbury, UK). Tissue culture medium, fetal
calf serum, penicillin, streptomycin and all other chemi-
cals were purchased from Cambrex Bio Science Milano
S.r.L., Italy. All microorganisms were supplied from
ATCC (American Type Culture Collection) and MTT
Kit from Sigma Aldrich (Milano, Italy).

*Corresponding author. Email: ccircosta@unime.it

ISSN 1041-2905 print/ISSN 2163-8152 online

Ó 2012 Taylor & Francis
http://dx.doi.org/10.1080/10412905.2012.659518
http://www.tandfonline.com
 188 F. Spadaro et al.
Hydrodistillation apparatus and procedure
Fresh lemon peels, 200 g for each sample, were sub-
mitted to hydrodistillation using a Clevenger-type appa-
ratus according to the European Pharmacopoeia (6)
and extracted with 500 mL water for 3 hours (until no
more essential oil was obtained). The essential oil was
collected, dried over anhydrous sodium sulfate and
stored at 4°C until used.
Chemical-physical parameters
The usual physical constants defining the essential oil
(refractive index, optical rotation, specific gravity and
solubility in 95% ethanol) have been determined
according to standard methods (7).
Gas chromatography and gas chromatography–mass
Downloaded by [McMaster University] at 20:54 24 December 2014
spectrometry
GC–flame ionization detector (GC–FID) analyses were
carried out by using a GC Fisons 8560 HRGC Mega 2
series, equipped with an HP-5 column (Hewlett-Pack-
ard), 30 m × 0.25 mm, 0.25 μm film thickness. Oven
temperature was programmed as follows: 60°C (8 min-
utes) to 100°C at 3°C/minute, to 130°C at 2.5°C/min-
ute, to 220°C at 3°C/minute. Neat samples were
injected in split mode, with a split ratio of 1:100. Injec-
tor and FID temperature were set at 280°C. Carrier gas
was He, at a linear velocity of 30.0 cm/s and a pressure
of 99.5 kPa. Data were processed by Thermoquest
Chrom-Card. The quantitative composition of each
sample was obtained from normalized peak areas.
Mass spectra were acquired by means of a Shima-
dzu GCMS-QP2010 system operated with an SLB-
5MS (Supelco, USA) stationary phase (30 m × 0.25
mm, 0.25 μm film thickness). He linear velocity was
30.0 cm/s (pressure 28.9 kPa). Injection took place in
split mode, with a split ratio of 1:100. Oven tempera-
ture program was: 50°C at 4°C/minute to 250°C, to
300°C at 10°C/minute, held for 5 minutes. For the
measurement of Linear Retention Indices, a mix of n-
alkanes ranging from heptane to triacontane (Supelco,
USA) was previously injected at the same experimental
conditions.
Mass spectrometric (MS) conditions were set as fol-
lows: ion source temperature was 200°C, interface tem-
perature was 250°C, scan range was 40 400 m/z, with a
scan interval of 0.20 seconds. For mass spectral identi-
fication, Shimadzu FFNSC 1.3 library was basically
used, along with Adams library, 4th edition (8, 9).
Antiproliferative effect
The Ishikawa line of human endometrial adenocarci-
noma cells maintained in culture was used in order to
test the antiproliferative efficacy of essential oils on
cancer cells.
Cultures of cells
Ishikawa cells, after being thawed from their frozen
stocks, were maintained in Eagle’s Minimum Essential
Medium (MEM) containing 10% (vol/vol) fetal bovine
serum (FBS), supplemented with 100 U/mL penicillin,
and 100 μg/mL streptomycin, 2 mM glutamine and 1
mM non-essential amino acids. Cells were plated at 1.5
× 106 cells/75 cm2 surface area and allowed to replicate
to confluence. Ishikawa cells were passed by standard
methods of trypsinization, plated in 96-well plates (2.5 ×
104 cells/well) and allowed to replicate to preconfluence.
Cell viability
MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazo-
lium bromide] (10) was used for cell viability. Cells
were incubated for 24 hours in the presence and absence
of essential oils at different concentrations (0.1–100 μg/
mL). After this period, a solution of MTT (10 μl of a 5
mg/mL solution in phosphate-buffered saline) was
added and incubated for 3 hours. MTT was reductively
converted into a blue formazan derivative and quantified
photometrically at 570 nm by an ELISA microplate
reader (Bio Rad 550, Bio-Rad Laboratories). Absor-
bance of control cells was considered to have 100% cell
viability and for each treatment group, cell survival was
calculated as a percentage inhibition of controls treated
with vehicle dimethyl sulfoxide (DMSO, 0.2%).
Antimicrobial activity
The paper disc diffusion method was employed to
determine the antimicrobial activity of the essential oils.
For these assays, cultures of the following microorgan-
isms were used: Gram-positive (Staphylococcus aureus,
Staphylococcus epidermis and Bacillus subtilis) and
Gram-negative (Pseudomonas aeruginosa, Escherichia
coli, Serratia marcescens and Proteus mirabilis) bacte-
ria, one yeast (Candida albicans) and one mould
(Aspergillus niger). All microorganisms were supplied
from ATCC. Cultures of the bacteria were maintained
on nutrient agar medium, whereas yeast and mould
were maintained on nutrient Sabouraud agar medium.
The bacterial strains were grown on in Triptic Soy
Broth (TSB) at 37°C for 18 hours, except for the strain
of Serratia marcescens, which was thermostated at a
temperature of 28°C. The yeast and mould were grown
on the surface of Petri dishes containing Sabouraud
Agar (SA) maintained in an incubator at a temperature
of 28°C for 24/48 hours. Briefly, suspensions of the
tested microorganisms 5 μl of 108 colony-forming units
(CFU/mL) were spread onto solid media plates. Filter
paper discs of 6 mm diameter (Whatman No. 1) were
individually impregnated with 20 μl essential oil, then
laid onto the surfaces of the inoculated plates. At the
end of the incubation time (18 hours at 28°C for
 The Journal of Essential Oil Research
Serratia marcescens, 18 hours at 37°C for all other
bacteria, and 24/48 hours at 28°C for yeasts), positive
antibacterial and antifungal activities were established
by the presence of measurable zones of inhibition. The
antimicrobial activity was recorded as the width (mm,
including the diameter of the disc) of the zone of inhi-
bition after incubation. Each test was performed in
three replicates and repeated twice.
Statistical analysis
Statistical comparison was carried out by one-way anal-
ysis of variance (ANOVA) and Dunnet’s test. The data
represent means±SEM, and values of p<0.05 were con-
sidered statistically significant. In the 96-well plates,
each treatment has five-replicate wells. This number of
replicates allowed to generate a mean and SEM for
Downloaded by [McMaster University] at 20:54 24 December 2014
each experiment, which was assessed in triplicate.
Results and discussion
Essential oils from fresh lemon fruits were extracted by
hydrodistillation using a Clevenger type apparatus,
yielding 0.9% (v/w) of essential oil.
Figure 1. Gas chromatogram of a lemon essential oil from conventional fruits. For identification of components, see Table 1.
189
Figures 1 and 2 and Table 1 show the composition
of the volatile fraction of lemon essential oils (both
conventional and biological) in single components. The
results obtained in this study are in accordance with a
previous work on a similar topic by Verzera et al. (11).
The content of single components and classes of sub-
stances was within the values reported for genuine oils.
Each sample was run in triplicate with coefficient of
variation (CV%) lower than 5%. Comparing the com-
position of the two types of oils, major differences can
be attributed to the content of oxygenated compounds,
mainly of aldehydes and alcohols. The total content of
oxygenated compounds is lower in oils obtained from
conventionally grown fruits (6.96%) than in those
extracted from biological fruits (8.05%). The content of
terpene aldehydes, especially neral and geranial (the so-
called ‘citral’), was higher (4.17%) in oils obtained
from biological fruits. Also, the content of monoterpene
alcohols (linalool, geraniol, α-terpineol and others) was
higher in biological oils (2.86%) rather than in conven-
tional oils (1.81%). The content of monoterpene hydro-
carbons (limonene and others) was lower (88.58%) in
 190 F. Spadaro et al.
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Figure 2. Gas chromatogram of a lemon essential oil from biological fruits. For identification of components, see Table 1.
oils extracted from biological fruits than in those
extracted from conventionally grown fruits (90.06%).
However, it is worth mentioning that monoterpene
hydrocarbons are less valuable than oxygenated com-
pounds in terms of their contribution to the fragrance
of the essential oil. Conversely, oxygenated compounds
have a high impact on the olfactive impression of a
Citrus oil, making it a very valuable natural product.
Table 2 shows the usual chemical–physical con-
stants defining an essential oil (refractive index, optical
rotation, specific gravity and solubility in 95% ethanol),
measured for both conventionally and biologically
grown fruits essential oils. There was no significant dif-
ference between conventional and biological oils.
Antiproliferative effect
It is currently accepted that essential oils could be used
to protect body organs against carcinogenesis. This can-
cer suppressive activity of essential oils observed
in vivo is supported by a number of in vitro assays in
which all their components have been tested on several
human cancer cell lines (12).
In the present study, in order to understand the anti-
proliferative efficacy of lemon essential oils on endo-
metrial cancer cells, experiments were conducted using
cultured human endometrial adenocarcinoma cells
(Ishikawa cells). The results of the viability were mea-
sured using MTT spectrophotometric assay. This
method is based on the quantification of purple-colored
formazan, which was formed by the reduction of MTT.
The reduction of MTT is proportional to the number of
active mitochondria in the live cells. Dose-dependent
inhibition of Ishikawa cells was observed at different
concentrations (0.1–100 μg/mL) of lemon essential oils
(both conventional and biological). At 10 and 100 μg/
mL, inhibition of Ishikawa cells was founds to be more
pronounced for biological oil (39.73% and 49.80%,
respectively, at 24 hours) in comparison with conven-
tional oil (28.70 and 34.20% respectively, at 24 hours)
(Table 3). The antiproliferative effect of the lemon oils
 The Journal of Essential Oil Research 191

Table 1. Percentage composition of conventional and biological hydrodistilled lemon oils.

Conventional Biological
Peak no. Compound LRIexpa LRIlitb Average Min Max Average Min Max
1 Tricyclene 923 923 <0.01 0.00 0.01 0.01 0.00 0.01
2 α-Thujene 926 927 0.26 0.18 0.31 0.33 0.17 0.45
3 α-Pinene 933 933 1.17 1.10 1.19 1.49 0.97 1.68
4 Camphene 955 953 0.04 0.04 0.06 0.05 0.02 0.06
5–6 Sabinene-β-pinene 975 972–978 10.03 8.96 15.32 13.41 10.12 14.57
7 6-methyl-5-hepten-2-one 984 986 <0.01 0.00 0.01 <0.01 0.00 0.01
8 Myrcene 992 991 1.39 1.28 1.42 1.34 0.95 2.05
9–10 Octanal-α-phellandrene 1007 1006–1007 0.15 0.05 0.17 0.11 0.06 0.25
11 δ-3-Carene 1011 1009 <0.01 0.00 0.01 <0.01 0.00 0.01
12 α-Terpinene 1018 1007 0.17 0.08 0.19 0.25 0.12 0.45
13–14 p-Cymene-limonene 1030 1025–1030 68.56 61.52 72.48 61.09 55.04 63.98
15 (Z)-β-Ocimene 1036 1026 0.08 0.06 0.12 0.04 0.02 0.05
16 (E)-β-Ocimene 1048 1046 0.15 0.04 0.18 0.11 0.04 0.23
17 γ-Terpinene 1062 1058 7.73 5.96 8.01 9.89 7.59 12.74
Downloaded by [McMaster University] at 20:54 24 December 2014

18 cis-Sabinene hydrate 1073 1069 0.03 0.01 0.04 0.06 0.05 0.07
19 Octanol 1074 1076 0.09 0.05 0.13 0.03 0.02 0.06
20 Terpinolene 1087 1086 0.34 0.11 1.38 0.47 0.39 0.64
21 trans-Sabinene hydrate 1103 1099 0.03 0.01 0.05 0.08 0.07 0.10
22 Linalool 1104 1101 0.22 0.14 0.32 0.47 0.22 0.56
23 Nonanal 1106 1107 0.14 0.09 0.19 0.19 0.10 0.21
24 cis-Limonene oxide 1136 1137 0.01 0.00 0.02 0.02 0.01 0.03
25 trans-Limonene oxide 1139 1140 0.02 0.00 0.03 0.03 0.01 0.04
26 Camphor 1151 1149 0.01 0.00 0.01 0.02 0.01 0.06
27 Citronellal 1155 1152 0.13 0.06 0.18 0.12 0.09 0.18
28 Borneol 1176 1173 0.05 0.00 0.01 0.08 0.04 0.13
29 Terpinen-4-ol 1183 1180 0.21 0.07 0.32 0.38 0.27 0.55
30 α-Terpineol 1198 1195 0.41 0.26 0.60 0.86 0.74 1.12
31 Decanal 1206 1208 0.07 0.05 0.15 0.02 0.01 0.03
32 Octyl acetate 1213 1214 0.01 0.00 0.01 <0.01 0.00 0.01
33–34 Nerol+citronellol 1226 1230 0.01 0.55 0.86 0.01 0.86 1.25
35 Neral 1240 1238 1.39 0.75 1.59 1.77 1.05 2.12
37 Geraniol 1252 1255 0.85 0.50 1.02 0.92 0.75 1.13
38 Geranial 1272 1268 1.90 1.26 2.13 2.40 2.00 3.05
39 Perillaldehyde 1281 1278 <0.01 0.00 0.01 <0.01 0.00 0.01
40 Bornyl acetate 1285 1285 <0.01 0.00 0.01 <0.01 0.00 0.01
41 Undecanal 1300 1309 0.02 0.00 0.05 0.03 0.02 0.06
42 Nonyl acetate 1317 1313 <0.01 0.00 0.01 0.01 0.01 0.02
43 Methylgeranate 1324 1320 0.01 0.00 0.03 0.03 0.01 0.05
44 Citronellyl acetate 1352 1353 0.06 0.02 0.10 0.02 0.01 0.05
45 Neryl acetate 1359 1361 0.57 0.34 1.08 0.35 0.26 0.56
46 Geranyl acetate 1378 1380 0.66 0.32 1.04 0.12 0.09 0.18
47 Dodecanal 1409 1410 <0.01 0.00 0.01 <0.01 0.00 0.01
48 Decyl acetate 1413 1412 <0.01 0.00 0.02 <0.01 0.00 0.01
49 β-Caryophyllene 1427 1424 0.18 0.06 0.22 0.18 0.05 0.20
50 trans-α-Bergamotene 1439 1432 0.38 0.28 0.56 0.34 0.31 0.45
51 α-Humulene 1463 1454 0.01 0.00 0.01 0.01 0.00 0.01
52 β-Santalene 1465 1459 0.05 0.02 0.07 0.05 0.02 0.06
53 γ-Muurolene 1479 1479 <0.01 0.00 0.01 0.01 0.00 0.02
54 Germacrene D 1481 1480 0.01 0.00 0.01 0.01 0.00 0.01
55 trans-β-Bergamotene 1491 1483 0.02 0.00 0.03 0.02 0.00 0.03
56 Valencene 1500 1496 0.05 0.03 0.10 0.03 0.00 0.03
57 Bicyclogermacrene 1501 1497 0.03 0.01 0.06 0.09 0.08 0.13
58 cis-α-Bisabolene 1505 1503 0.04 0.03 0.08 0.04 0.02 0.05
59 β-Bisabolene 1513 1508 0.55 0.18 0.86 0.50 0.42 0.65
60 trans-α-Bisabolene 1546 1540 0.01 0.00 0.01 0.01 0.00 0.01
61 Spathulenol 1584 1576 0.01 0.00 0.01 0.01 0.00 0.01
62 2,3-Dimethyl-3-(4-methyl-3-pentenyl)-2- 1662 – 0.02 0.00 0.02 0.02 0.00 0.02
norbornanol
63 Campherenol 1675 – 0.02 0.00 0.03 0.02 0.00 0.03
(Continued)
 192 F. Spadaro et al.

Table 1. (Continued).
Conventional Biological
a b
Peak no. Compound LRIexp LRIlit Average Min Max Average Min Max
64 α-Bisabolol 1688 1685 0.02 0.00 0.02 0.02 0.00 0.03
65 Nootkatone 1832 1834 0.01 0.00 0.01 0.01 0.00 0.01

98.36 97.96
Monoterpene hydrocarbons 90.06 88.58
Sesquiterpene hydrocarbons 1.33 1.28
Oxygenated monoterpenes 6.54 7.69
Oxygenated sesquiterpenes 0.09 0.08
Monoterpene aldehydes 3.43 4.30
Monoterpene alcohols 1.81 2.86
Monoterpene esters 1.31 0.53
Sesquiterpene alcohols 0.07 0.07
Aliphatic aldehydes 0.24 0.24
Aliphatic alcohols 0.09 0.03
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Aliphatic esters <0.01 0.01

Notes: aLRIexp, experimental linear retention indices calculated on HP-5 column. bLRIlit, linear retention indices retrieved from literature (8,9).

Table 2. Physical properties of lemon essential oil obtained by conventional and biological fruits.
Physical properties Conventional fruits Biological fruits
Specific gravity (d20 20) 0.840 0.838
Refractive index (n20 D) +1.470 +1.475
Optical rotation (degrees) ([a]20 D) +44° +43°

Table 3. Effect of essential oil of biological and citral as a cancer chemopreventive agent targeted
conventional lemon on proliferation Ishikawa cells measured towards inflammation related carcinogenesis, such as
by MTT assay after 24 and 48 hours. skin and colon cancer, has already been reported (14).
% Inhibition±SD
Conc. Antimicrobial activity
Treatment (μg/mL) 24 hours 48 hours
Essential oils can be used as antimicrobial and anti-
Control 0 0 fungal agents (15–18). Several components of lemon
Biological oil are responsible for the different targets in the infectious
0.1 10.20±3.10 12.15±2.70
entities. Linalool and citral are the main components
1 16.15±2.85⁄ 19.60±1.20⁄
10 39.73±2.12⁄ 42.45±3.75⁄ from lemon oils with antimicrobial effects in both
100 49.80±1.48⁄ 43.20±2.67⁄ direct oil and vapor form (2). The antimicrobial effect
Conventional oil of essential oils should be considered with care,
0.1 7.15±1.60 9.40±1.80 because, although essential oils may constitute a good
1 13.35±1.55 14.10±2.77
alternative to combat the increasing resistance patho-
10 28.70±2.67⁄ 29.18±3.15⁄
100 34.20±2.40⁄ 35.75±2.68⁄ gens (19), they could also produce an imbalance in gut
microflora (20).
Note: ⁄p<0.05 compared with control culture.
The results obtained from this study by the disc
diffusion method using lemon essential oils showed
inhibition zones against all microorganisms tested
could be related to monoterpene hydrocarbons, since (Table 4). Also, it was demonstrated that E. coli, B.
they have been reported to prevent lung, liver, mam- subtilis, C. albicans and A. niger were the most
mary and stomach cancers (13). However, a close sensitive microorganisms tested, whereas the bacteria
inspection of the results suggested that other active S. epidermis and S. marcescens were resistant to these
components, e.g. aldehydes (citral), might boost the oils. The antimicrobial activities of oils extracted from
activity of monoterpene hydrocarbons. Exploration of biological and conventionally grown fruits were similar,
 The Journal of Essential Oil Research 193

Table 4. Antimicrobial activity of lemon essential oil obtained by conventional and biological fruits.
Inhibition zone diameter (mm)
Tested microorganism Conventional fruits Biological fruits
Staphylococcus aureus (Staphyllococcaceae), Gram-positive bacterium (ATCC6538) 11.0±0.5 12.5±0.2⁄
Stapylococcus epidermidis (Staphyllococcaceae), Gram-positive bacterium (ATCC 12228) 10.0±1.5 10.0±1.7
Bacillus subtilis (Bacillaceae), Gram-positive bacterium (ATCC 8037) 32.0±2.5 33.0±1.7
Pseudomonas aeruginosa (Pseudomonadaceae), Gram-negative bacterium (ATCC9027) 12.5±2.0 13.0±1.4
Escherichia coli (Enterobacteriaceae), Gram-negative bacterium (ATCC4197) 14.0±0.2 18.5±1.5⁄
Serratia mercescens (Enterobacteriaceae) Gram-negative bacterium (ATCC 10759) 10.5±1.8 10.5±1.3
Proteus mirabilis (Enterobacteriaceae), Gram-negative bacterium (ATCC 4630) 10.5±0.5 14.0±0.7⁄
Candida albicans (Cryptococcaceae), yeast (ATCC 24433) 22.0±3.0 30.0±2.8⁄
Aspergillus niger (Trichocomaceae), mould (ATCC 1004) 20.0±1.4 26.5±1.6⁄
Note: ⁄p<0.05 compared with conventional fruits.

although the data obtained pointed out a major activity 7. Agence Francaise de Normalisation. Huiles Essentielles.
for biological oils, a finding that could be ascribed to Editions Afnor, Paris (2007).
Downloaded by [McMaster University] at 20:54 24 December 2014

8. FFNSC 1.3 – Flavour and Fragrance Natural and

the higher content of oxygenated compounds, such as
citral.
In summary, this study has shown that the different
agriculture techniques cause changes in the oil compo-
sition and, consequently, in the antiproliferative and
antimicrobial activity. The biological oil samples here
investigated reported a higher content of aldehydes and
alcohols compared with the oils obtained from conven-
tionally cultivated fruits. Citral content in lemon oil is
an important parameter for assessing its quality, to the
extent that the market value of the oil is defined
according to its citral content.
In conclusion, the above reported findings indicate
that the biological oils are of higher quality in terms of
their composition and biological activity compared with
oils extracted from conventionally grown fruits.
References
1. F.R. Marìn, M. Martinez, T. Uribesalgo, S. Castillo and
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