Industrial Crops and Products 34 (2011) 1516–1522

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Industrial Crops and Products

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Growing conditions and postharvest management can affect the essential oil of
Origanum vulgare L. ssp. hirtum (Link) Ietswaart
Giorgio Tibaldi, Emanuela Fontana, Silvana Nicola ∗
AgroSelviTer Dept. – Vegetable Crops & Medicinal and Aromatic Plants, Università degli Studi di Torino, Via Leonardo da Vinci, 44, 10095 Grugliasco (TO), Italy

a r t i c l e i n f o a b s t r a c t

Article history: Origanum vulgare subsp. hirtum (Link) Ietswaart (Greek oregano) has been cultivated since ancient times
Received 27 January 2011 thanks to its herbal and therapeutic properties. Currently, it is added in various commercial prepara-
Received in revised form 28 April 2011 tions thanks to its essential oil composition. This group of secondary compounds is affected in quantity
Accepted 10 May 2011
and quality by biotic and abiotic factors during the cultivating phase and by the postharvest man-
Available online 8 June 2011
agement. Thus, the aims of the work were to study: (1) how the growing conditions (soil full-light
treatment; pot full-light treatment; pot 50%-shade treatment) can affect the essential oil content and
Keywords:
the essential oil profile of Greek oregano branches at full-blossoming stage; and (2) how the posthar-
Greek oregano
Light
vest management (distillation of the essential oil from fresh herbs, dehumidified herbs or oven-dried
Soilless cultivation herbs) can affect the essential oil content and the essential oil profile of this species. The growing con-
Dehumidifying process ditions significantly affected the biometrical parameters. The pot full-light treatment contained the
Terpinene highest dry matter percentage (36.5%) and the highest essential oil content, which was directly cor-
Carvacrol related to the former parameter (r = 0.890**). The postharvest management did not affect the essential
oil content. The growing condition significantly affected the characterizing ␥-terpinene, cis-sabinene
hydrate, 4-terpineol, ␣-terpinene, linalyl acetate, and ␤-bisabolene, while it did not affect carvacrol
(mean value 15.8%), p-cymene (mean value 6.0%), or sabinene (mean value 3.9%). The postharvest
management significantly affected carvacrol, cis-sabinene hydrate, 4-terpineol, p-cymene, ␣-terpinene,
linalyl acetate, and ␤-bisabolene, while it did not affect ␥-terpinene (mean value 14.7%). The response
of some compounds to growing conditions led us to hypothesise an essential oil profile of the plants
grown in pots at 50%-shade to be more of a vegetative stage type oil than the essential oil profile
of the plants grown in soil, although all were at the blossoming stage. The analyses on the individ-
ual compounds indicated that, even though the essential oil content in oregano did not change with a
change in postharvest management, biochemical changes occurred on the profile. These could be due
to enzyme activity, dilution effect or temperature effect during the drying processes. The oven-drying
process and the dehumidifying process changed the essential oil profile in a similar way compared to
the essential oil profile distilled from fresh herbs. Thus, when it is not possible to extract the essential
oil directly from fresh herbs, processors can dry the oregano according to the equipments they have
available.
© 2011 Elsevier B.V. All rights reserved.

1. Introduction and terpenoid compounds. The latter phytochemical group of com-

Greek oregano (Origanum vulgare L. ssp. hirtum (Link) Ietswaart)
is a perennial shrub that is native to dry, rocky calcareous soils in
mountainous areas in South Europe and South-West Asia. It has
been cultivated since ancient times, thanks to its herbal and thera-
peutic properties (Dordas, 2009) and is currently added to various
commercial preparations, mainly as an antioxidant and antimi-
crobial agent (Chorianopoulos et al., 2004), thanks to its phenolic
∗ Corresponding author. Tel.: +39 011 6708773; fax: +39 011 2368773.
E-mail address: silvana.nicola@unito.it (S. Nicola).
pounds, constituent of the EO, is considered to be one of the main
compounds of interest in medicinal plants, and it has been reported
to be affected in quantity and quality by biotic and abiotic fac-
tors because of its physiological implications on plant biological
behaviour (Bianco, 2010).
The EO content is generally known to be affected to a great
extent by environmental factors, while the EO profile is affected
by genetic factors (Wang and Lincoln, 2004). Among the various
abiotic factors, light can play an important role in increasing or
reducing the synthesis and in the accumulation of the EO in peltate
trichome glands of Lamiaceae species. Johnson et al. (2004) found
that in Greek oregano carvacrol is much higher in the summer,

0926-6690/$ – see front matter © 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.indcrop.2011.05.008
 G. Tibaldi et al. / Industrial Crops and Products 34 (2011) 1516–1522
while in the autumn p-cymene predominates. Circella et al. (1995)
found that cis-sabinene hydrate, the main component of the closely
related species O. majorana L., was produced in larger quantities
in plants grown under daylight conditions of 16 h than in those
grown under a daylight length of 13 h or 10 h, while the content of
terpinenes decreased with an increase in daylight length. Kokkini
et al. (1995), studying wild Cretan Mentha spicata L. chemotypes,
found that the oil richness of island plants, compared to other
values found in literature, might be due to the climatic condi-
tions of the area. Other studies have been conducted concerning
how light quality can affect the EO content, the EO compounds
and the antioxidant activity of several medicinal and aromatic
plants in controlled environmental conditions (Maffei et al., 1999;
Nishimura et al., 2009; Shiga et al., 2009). However, there is a lack
of studies on how the solar radiation can affect the EO content and
EO profile. Li et al. (1996) found that Salvia officinalis L., grown
at a 55%-shade treatment, reached the highest total EO content
(0.38%) compared to full-light (0.34%) treatment, and the EO con-
tent decreased in the treatments from 55%- to 85%-shade, while
Thymus vulgaris L. reached the maximum total EO content at full-
light (0.49%).
Cultivating herbs can improve the quality and increase the stan-
dardization of the EO content and profile more than gathering
them from the wild. Among the different cultivation systems, the
soil system in the open-air using a well-defined chemotype or
cultivar is the first method to limit the variability of EO synthe-
sis within the species. Growing the same chemotype or cultivar
in a soilless system leads to a further standardization of the
quality. Recent studies on how the cultivation techniques can
affect nutraceuticals have been conducted on Ocimum basilicum L.,
grown with the floating system, compared to O. basilicum grown
in soil (Sgherri et al., 2010). The results have shown a higher
antioxidant activity of the water and lipid extracts in the for-
mer growing condition than in the latter, for the same plant
age.
EO can vary in quality and quantity according to climate, soil
composition, plant organ, age and vegetative stage (Bakkali et al.,
2008). The EO quantity and quality obtained in herbs at har-
vest should be preserved during the postharvest processes until
the EO is extracted. Convective oven-drying is the most popular
method used to reduce the moisture content of fruit, vegetables,
and herbs; it inhibits microbial growth and prevents some bio-
chemical changes. However, it can give rise to other alterations
that affect herb quality (Hossain et al., 2010), since it requires a
relatively long time and high temperatures to be effective. The
contact between dried material and hot air causes a degradation
in the phytochemicals, as well as colour alterations and a loss
of the volatile compounds, as has been reported for Th. vulgaris
and S. officinalis (Venskutonis, 1997), and for S. sclarea L. (Tibaldi
et al., 2010). Innovative drying systems have been studied to reduce
the loss of volatile compounds; these have mainly been based
on the microwave principle (Szumny et al., 2010), or the freeze-
drying method (Grayer et al., 1996). However, microwave and
freeze-drying systems are hard to implement in small Italian farms
because of the high costs and high technology required, according
to producers and producer associations statements. An alternative
to traditional drying at high temperatures and to the innovative
drying systems previously described could be the dehumidification
process which consists of removing water from plant cells at tem-
peratures close to room temperature. Preliminary studies on mints
(M. spicata L. var. rubra; M. spicata L. var. viridis Casablanca; M. ×
piperita L.) and Calamintha nepeta (L.) Savi have demonstrated that
the dehumidifying process gives positive results on EO quantity and
quality, and reduces the EO loss, compared to the traditional oven-
drying method. Nevertheless, there is a lack of studies in literature
on the latter system applied to herbs.
1517
Given the lack of studies in literature concerning the influ-
ences of different environmental-abiotic factors on the synthesis
of EO in O. vulgare and concerning the influences of different dry-
ing methods on the modification of the EO content and profile
in Lamiaceae species, and particularly in O. vulgare, the present
work was aimed at studying how growing conditions can affect
the EO content and the EO profile of cultivated Origanum vulgare
ssp. hirtum branches at harvest and postharvest and how posthar-
vest management can affect the EO content and the EO profile of
this species.
2. Material and methods
2.1. Raw material production
The study was carried out in the experimental centre of the
Faculty of Agriculture (44◦ 53 11.67 N; 7◦ 41 7.00 E–231 m a.s.l.
Carmagnola, Italy). The Origanum vulgare ssp. hirtum (hereafter:
oregano) plants were purchased from a nursery in the Albenga
growing area (44◦ 04 N; 8◦ 10 E—13 m a.s.l. Liguria Region, Italy) and
transplanted in the experimental field on 23 April 2009. The field
experiment consisted of testing three different growing conditions
(soil full-light treatment; pot full-light treatment; pot 50%-shade
treatment). A light reduction was obtained in shaded plots using a
100 g m−2 HDPE 50% shading black net (CARRETTA tessitura snc,
Carrè, VI, Italy). The photosynthetic active radiation (PAR) was
recorded weekly in the full-light plots and in the 50%-shade plots
(Table 1) at different times during the day using a line quantum
sensor and an LI-1000 DataLogger (LI-COR inc., Lincoln, NE, USA)
and a reduction index of 84.9–90.0% of PAR was identified in the
50%-shade plots compared to the full-light plots. A long-lasting
grid-plastic black mulch pre-treated to prevent algal growth was
used in both the soil and soilless plots (Manifatture Norberto Par-
dini & Figli S.p.a., Camaiore, LU, Italy); a manual weed control was
carried out according to needs. The plant density was 2.86 plant/m2 ,
with a space grid of 0.50 m × 0.70 m for all the treatments. Each plot
consisted of ca. 100 plants. The soil was a sandy-loam, according
to USDA criteria, while the 4-L black PE pots of the soilless plots
were filled with a mixture in volume of 80% specific peat-based
horticultural medium (‘Neuhaus Huminsubstrat N17’; Klasmann-
Deilmann, Geeste-Groß Hesepe, Germany) and 20% perlite (‘Peralit
25’; Perlite italiana s.r.l., Corsico, MI, Italy). The soilless culture plots
were equipped with a drip irrigation system with a water-flow of
2 L h−1 for each pot, set for 60 min/day in the full-light plots and
15 min/day in the 50%-shade plots. The soil culture plots were not
equipped with any irrigation system and the irrigations were only
conducted at transplanting. Mature vaccine manure was applied to
the soil plots using a 2.0 kg m−2 dosage. The pots were fertigated
with 0.5 L of CIFOUMIC (CIFO s.p.a., San Giorgio di Piano, BO, Italy) at
3 g L−1 every 21 days. The total rainfall (mm), maximum and min-
imum temperatures (◦ C), average relative humidity, and average
atmospheric pressure (mbar) were recorded daily and the weekly
data are reported in Table 1.
Harvest took place in all plots on 17 July 2009. Branches
were collected at the full bloom stage. The branches were then
split into three postharvest managements: (a) fresh samples for
direct EO extraction; (b) dehumidified samples for EO extraction
from dehumidified raw material; (c) oven-dried samples for EO
extraction from dried raw material. Dried samples were obtained
through a dehumidifying or an oven-drying process, which were
conducted in commercial drying structures at the Faculty of Agri-
culture experimental centre. The branches (ca. 2 kg) were arranged
in a 0.10-m layer on perforated steel trays and placed in a
temperature-controlled cell at 25 ◦ C ± 2 ◦ C for 48 h with a dehumid-
ifier (KT-38/S - Tecno.Klima s.r.l., Granarolo Emilia, BO, Italy) for the
1518 G. Tibaldi et al. / Industrial Crops and Products 34 (2011) 1516–1522

Table 1
Meteorological data recorded in the experimental centre of the Faculty of Agriculture during the oregano growing experiment: weekly average solar radiation at full-light
(PAR full-light), weekly average solar radiation at 50%-shade (PAR 50%-shade), total weekly rainfall (mm), weekly average maximum temperature (=Tmax ) (◦ C), weekly average
minimum temperature (=Tmin ) (◦ C), weekly average relative humidity (RH) (%), and weekly average atmospheric pressure (Pressure) (mbar).

Weeka PARb full-light PARb 50%-shade Rainfall Tmax Tmin RH Pressure

(␮mol s−1 m−2 ) (␮mol s−1 m−2 ) (mm) (◦ C) (◦ C) (%) (mbar)

1 1191 149 108.4 15.1 8.6 84.3 985.8

2 1522 175 1.2 23.8 9.2 64.1 989.9
3 1709 204 0.2 24.7 11.4 71.7 991.6
4 1649 235 18.2 24.6 13.0 74.4 987.5
5 1236 167 9.0 31.2 15.8 70.9 989.0
6 1097 115 0.8 25.5 12.4 65.0 989.5
7 1461 147 6.8 26.9 13.0 72.1 980.0
8 1717 259 0.0 31.4 14.5 68.2 989.6
9 1500 189 17.4 29.2 13.2 68.4 986.7
10 1507 165 0.2 30.9 16.5 74.1 983.9
11 1740 259 29.2 30.9 17.0 78.5 983.2
12 1645 246 0.0 29.6 14.9 74.2 987.4
13 1651 220 40.0 32.0 16.9 79.1 988.5
a
Since the transplanting day in the experimental field on 23 April 2009.
b
Data recorded at ca. 15:00 h.

dehumidifying process and at 50 ◦ C for 6 days for the oven-drying When ANOVA was significant, growing condition and postharvest
process. The phytomass yield, dry matter (DM), and EO content management were tested using Tukey’s multiple range test.
were recorded at harvest and then the EO profiles extracted from
fresh, dehumidified, and oven-dried herbs were analysed. 3. Results and discussion

3.1. Raw material and EO content from the fresh phytomass

2.2. Essential oil extraction
The growing condition significantly affected the phytomass
An aliquot of ca. 300 g of fresh, dehumidified or oven-dried
yield, the dry matter (DM), and the EO content (Table 2). The soil
phytomass was steam-distilled using glass distillation equipment,
full-light treatment produced the greatest phytomass yield and sta-
which had been assembled by Exacta + Optech (San Prospero, MO,
tistically differed from both the pot full-light and the pot 50%-shade
Italy), composed of an electrical heating mantle (500 W – Thermo
treatments, which yielded ca. 42% less than the former plants. The
Scientific Electro Thermal, Waltham, MA, USA), a 2-L Pyrex glass
pot full-light treatment contained the highest dry matter percent-
balloon filled with 1.5 L of deionised water with an overlying 4-L
age (36.5%) and statistically differed from the other two growing
modified Pyrex glass balloon filled with the herb material, a cooling
conditions, in which the plants contained ca. 26% of dry matter.
column operating in co-current and a graded burette. Each distilla-
The pot full-light treatment produced the highest EO content and
tion cycle lasted for ca. 105 min, with 45 min of steaming produced
statistically differed from both the soil full-light and the pot 50%-
by the boiling water in the glass balloon. The 45 min distillation
shade treatments, in which the EO was ca. 53% less than the EO
time started when the first drop of liquid condensed in the cooling
distilled from the former.
column and dripped into the graded burette. The EO content was
Of the three growing conditions, the pot full-light treatment
indicated as the ratio in weight (w/w) between the cold oil collected
appeared to suffer from some drought stress and this could explain
from the burette and the phytomass, calculated on a fresh weight
the higher DM than the other two growing conditions. Generally, in
(FW) basis.
stress conditions, plants tend to have more DM than plants grown in
The EO was analysed in a laboratory (Alchim di Roberto Mas-
optimal conditions (Fontana et al., 2010). In this work, the EO con-
ante, Chieri, TO, Italy) which used an Agilent 5973 N GC-MS system
tent was directly correlated to the DM (r = 0.890**), and a significant
(Agilent Technologies Inc., Palo Alto, CA, USA) with an Rtx5 capillary
positive linear regression between the DM and EO content of the
column (30 m × 0.25 mm i.d.). The column was set at a 60 ◦ C hold
pooled data was found (P = 0.001), indicating that the EO content
for 3 min, then warmed by 5 ◦ C m−1 to 250 ◦ C. The auto injection
increases with increasing DM, according to the equation:
volume was 0.5 ␮L, neat, with a split injection (80:1). Peak iden-
tification was made using external standard (Supelco, Bellefonte, EO content = 0.008 DM − 0.131; R2 = 0.762
PA, USA) and single compound standards (Sigma–Aldrich Corp., St.
Thus, it can be concluded that oregano plants subjected to moderate
Louis, MO, USA).
stress can produce more EO, as was also reported by Carrubba et al.
(2002) for S. sclarea. These authors suggested that species subjected
2.3. Statistical design and analyses to water deficiency are prone to producing more EO than species
grown in more suitable conditions, as a reaction to the adverse con-
The statistical experimental design was a randomised com- ditions. Similar considerations, related to environmental stress, the
plete design (RCD). Until harvesting the design was a single EO content and DM were reported by Azizi et al. (2009). The authors,
factor experiment (growing condition) with 3 repetitions. The studying the effect of nitrogen fertilization and drought stress on
total number of treatments during the crop cycle was 3. After oregano grown in greenhouse in pots filled with loess soil:sand
harvest, a two-factor factorial experiment design (growing condi- (1:2, w/w), found that a late water deficiency (close to blossoming
tion × postharvest management) with 3 repetitions was adopted. stage) significantly increased the EO content.
The total number of treatments after harvest was 9 (3 × 3). The
sample unit consisted of ca. 20 plants for the phytomass yield and 3.2. EO profiles at harvest
ca. 7 plants for the other measurements.
The data were submitted to ANOVA, using the Statistical Package The analysis on the EO profiles from fresh oregano samples led to
for Social Science (SPSS Version 17.0, SPSS Inc., Chicago, IL, USA). a detection of 87–89% of the EO profile and it showed that growing
 G. Tibaldi et al. / Industrial Crops and Products 34 (2011) 1516–1522 1519

Table 2
Effect of the growing conditions on the dry matter (DM) and the essential oil (EO) contents referring to the fresh weight of Origanum vulgare ssp. hirtum grown in Northern
Italy at harvest.

Growing condition Significance (P)

In soil full-light In pot full-light In pot 50%-shade

Phytomass yield (kg m−2 ) 1.595aa 1.045b 0.813b <0.001

DM (%) 26.2b 36.5a 25.5b <0.001
EO content (%, w/w) 0.063b 0.165a 0.093b <0.001
a
Values followed by the same letter, within the same row, are not significantly different according to Tukey’s multiple range test P ≤ 0.050.

oregano in soil at full-light, in pots at full-light, or in pots at
50%-shade leads to different terpene profiles being synthesized
(Table 3). The soil full-light treatment was mainly composed of
4-terpineol, ␥-terpinene, carvacrol, and p-cymene. The pot full-
light treatment was mainly composed of 4-terpineol, ␥-terpinene,
carvacrol, p-cymene and ␣-terpinene. The pot 50%-shade treat-
ment was mainly composed of ␥-terpinene, 4-terpineol, carvacrol,
p-cymene and allocymene. Even though no literature data could
be found to compare with our EO profile, the characterizing com-
pounds of the species were also detected in our study. When the
EO profiles of the fresh oregano were analysed, the carvacrol con-
tent was very low, and this could justify the low EO content that
was obtained. As reported by Esen et al. (2007), a correlation exists
between the oil yield and the carvacrol content and the best oil
yields (2.9–6.5% referring to dry weight) were obtained with the
highest carvacrol contents (up to 78.73%).
3.3. Effect of the pre-harvest and postharvest treatments on the
EO content and profile
The growing condition × postharvest management interaction
did not affect the EO content (P = 0.625) and neither did the
postharvest management (P = 0.880) (data not shown), while the
growing condition instead did (P = 0.001). The pot full-light treat-
ment produced the highest EO content (0.162%) and it statistically
differed from both the soil full-light (0.084%) and the pot 50%-shade
(0.087%) treatments, for which the EO was ca. 47% less than the EO
distilled from the former.
Basically, the EO contents, due to the growing conditions at
harvest alone, or as the main factor after harvest were very sim-
ilar, indicating the limited influence of the dehumidifying and the
oven-drying processes. Azizi et al. (2009) obtained an EO content of
approximately 3.7% on dry weight from oregano, corresponding to
approximately 1.52% on a fresh weight basis (our elaboration). The
10-fold higher EO content than our best result could partially be
due to the different cultural treatments and environmental condi-
tions (see Section 3.1) and also to the different distillation material
used by Azizi et al. (2009). The authors, in fact, extracted the EO from
20 g of dried leaves and inflorescences, after the branches and stems
had been removed. Leaves and inflorescences are known to synthe-
sis most of the EO in Lamiaceae plants, while branches and stems
have less glandular trichomes, and thus less EO. Figiel et al. (2010),
comparing the effect of drying methods (convective drying at 60 ◦ C
and vacuum–microwave drying) on oregano EO compared to the
EO from fresh herbs, found that the EO content decreased with
the convective drying with a decrease in the phytomass moisture,

Table 3
Average essential oil (EO) content and average composition of the EO distilled from fresh Origanum vulgare ssp. hirtum grown in Northern Italy and subjected to different
growing conditions.

Oregano in soil full-light Oregano in pot full-light Oregano in pot 50%-shade

RTa EO compound (% EO) (% EO) (% EO)

6.41 ␣-Thujene 0.72 1.47 0.84

6.61 ␣-Pinene 0.46 0.67 0.53
7.60 Sabinene 1.32 3.62 2.42
7.73 ␤-Pinene 0.25 0.29 0.25
7.84 Octan 3-one 0.09 0.12 0.14
7.96 Myrcene 2.08 2.25 2.12
8.42 ␣-Phellandrene 0.37 0.57 0.47
8.76 ␣-Terpinene 2.84 5.33 3.99
8.98 p-Cymene 9.96 8.43 10.07
9.12 Limonene + 1.8-Cineole 1.13 1.38 1.27
9.55 ␤-Ocimene 0.15 0.12 0.15
9.95 ␥-Terpinene 14.58 14.93 14.95
10.20 trans-Sabinene hydrate 0.60 0.99 1.18
10.80 ␣-Terpinolene 2.15 2.35 2.01
11.05 Linalool 1.79 0.92 0.53
11.10 cis-Sabinene hydrate 1.56 4.80 2.99
11.37 Octen 1-ol acetate 1.35 1.32 1.30
11.67 3-Octanyl acetate 0.12 0.11 0.07
12.00 Allocymene 2.98 3.26 4.95
13.44 4-Terpineol 17.51 15.13 13.27
13.80 ␣-Terpineol 1.68 1.77 1.57
15.47 Linalyl acetate 3.23 1.87 3.51
16.82 Carvacrol 14.58 12.31 12.94
20.12 ␤-Caryophyllene 1.70 1.33 1.36
21.65 Germacrene D 0.55 0.51 0.65
22.17 ␤-Bisabolene 2.85 1.63 2.69
Total 87.71 88.80 87.47
a
Retention time expressed in min.
1520 G. Tibaldi et al. / Industrial Crops and Products 34 (2011) 1516–1522
and led only to 31% of the EO from fresh herbs. We found similar
responses when we compared the EO contents of fresh and oven-
dried (50 ◦ C) S. sclarea (ca. 50%) (Tibaldi et al., 2010), the EO contents
from fresh and oven-dried (50 ◦ C) S. officinalis (ca. 24%) (data not
published) and the EO contents of different fresh and oven-dried
(60 ◦ C) mints (ca. 48%) (data not published). No statistical difference
was observed between the EO content of the fresh and the dehu-
midified mints (data not published) or the content of oregano EO in
the present experiment, when the fresh and any drying treatments
were compared.
The growing condition × postharvest management interaction
did not affect the EO profile for any compound (data not shown),
while the main factors did, depending upon the compound. Among
the 27 compounds detected in the EO, the growing condition sig-
nificantly affected 19 of them (Fig. 1 – data partially not shown)
and postharvest management significantly affected 15 (Fig. 2 – data
partially not shown). Only those compounds that were present at
more than the average value of 3% of the total EO are discussed.
Among the main characterizing oregano EO compounds, growing
condition significantly affected ␥-terpinene, cis-sabinene hydrate,
4-terpineol, ␣-terpinene, linalyl acetate, and ␤-bisabolene, while it
did not affect carvacrol (mean value 15.8% EO), p-cymene (mean
value 6.0% EO), and sabinene (mean value 3.9% EO) (Fig. 1).
Among the main characterizing oregano EO compounds, posthar-
vest management significantly affected carvacrol, cis-sabinene
hydrate, 4-terpineol, p-cymene, ␣-terpinene, linalyl acetate, and ␤-
bisabolene, while it did not affect ␥-terpinene (mean value 14.7%
EO) (Fig. 2).
The profile analyses indicated that, even though the EO con-
tent in oregano did not change with the postharvest management
(see Section 3.1), biochemical changes occurred. The biochemi-
cal changes during the drying processes could be due to enzyme
activity, low water activity or the temperature during the drying
processes.
Among the compounds characterizing the EOs of the
oregano evaluated in this study, the most relevant com-
pounds were carvacrol, ␥-terpinene, cis-sabinene hydrate,
4-terpineol, p-cymene, ␣-terpinene, linalyl acetate, sabinene,
and ␤-bisabolene.
Carvacrol and p-cymene, not being affected by the growing con-
ditions, could have a pathway under strong genetic control and
could distinguish the chemotypes. Carvacrol tended to increase and
its precursor p-cymene tended to reduce when the dehumidify-
ing and oven-drying processes were examinated compared to the
EO of fresh oregano. Thus, we could hypothesise that the biosyn-
thetic pathway could also continue after harvest and that the drying
processes, regardless of the technique used, allowed p-cymene
to oxidize into the oxygenated carvacrol. Venskutonis (1997),
studying Th. vulgaris EO constituents, found a similar response
for oxygenated terpenes and hypothesised that the oxidation of
some non-oxygenated compounds could have occurred during dry-
ing.
␥-Terpinene, the characterizing monoterpene of EO along with
carvacrol and p-cymene, and its biogenetic precursors, showed the
highest synthesis in oregano grown in soil and at full-light, while
cis-sabinene hydrate had the highest synthesis in the plants grown
in pots at 50%-shade. The opposite behaviour of these two com-
pounds has led us to hypothesise a more vegetative stage EO profile
of the plants grown in pots than the EO profile of the plants grown
in soil. This hypothesis has been confirmed by Sellami et al. (2009),
who found that O. majorana EO reached the maximum ␥-terpinene
content and the lowest cis-sabinene hydrate content at the bud-
ding stage, while opposite contents were found in the vegetative
stage. The other two terpinenes, 4-terpineol and ␣-terpinene, had
similar behaviour to ␥-terpinene in the growing condition treat-
ment, with the lowest content obtained in the EO distilled in the
oregano grown in pots at 50%-shade. Circella et al. (1995), study-
ing O. majorana, found that the content of terpinenes decreased
with an increase in daylight. The close origin of O. majorana and
oregano has led us to hypothesise that also in oregano the syn-
thesis of terpinenes is related to the length of daylight, with the
lowest contents of terpinenes being obtained during the blossom-
ing stage in summer. However, no data on the effect of daylight
length or the intensity of solar radiation is known in the litera-
ture about oregano. ␥-Terpinene and 4-terpineol showed a similar
response to the growing condition treatments, but the former
was not affected by the postharvest management, while the lat-
ter was significantly reduced, compared to fresh oregano, in both
the dehumidifying and oven-drying processes. These postharvest
responses are in agreement with the results found by Díaz-Maroto
et al. (2002), who studied the effect of processing methods on
Laurus nobilis L. volatiles. The authors found that ␥-terpinene
remained stable to the processing methods tested (air-drying,
oven-drying at 45 ◦ C, freezing, and freeze-drying), while the 4-
terpineol decreased significantly. The increase in the ␣-terpinene
content of the EOs in our dried samples is comparable to that
of the Satureja hortensis found by Sefidkon et al. (2006) who
tested different drying methods, 45 ◦ C oven-drying, sun-drying,
and shade-drying. The authors found that shade-dried samples had
the lowest ␣-terpinene content, which increased in the sun-dried
samples and reached the maximum content in the oven-dried sam-
ples at 45 ◦ C.
The effect of the tested growing conditions on the linalyl acetate
content did not appear to be clear. This compound and its related
form, linalool, are highly volatile, precursors of a wide range of
monoterpenes (Lasky, 2009), and are especially present in the
inflorescences of Lamiaceae species (e.g. S. sclarea in Tibaldi et al.,
2010; Lavandula × intermedia Emeric in Baydar and Erbaş, 2009; S.
officinalis inflorescences in Velickovic et al., 2002). We expected
to find the highest linalyl acetate content in the oregano plants
grown at full-light since they had more inflorescences than the
oregano plants grown in pots at 50%-shade (visual evaluation),
which instead had the highest linalyl acetate content. Apparently,
the latter growing condition reduced the volatility of this com-
pound, and this would seem to imply that the oregano plants
grown at full-light (black mulched and at higher temperature) had
a greater transpiration rate than the oregano plants grown under
shade and thus with less linalyl acetate.
The linalyl acetate was also affected by the postharvest manage-
ment, and showed a similar response to our previous experiment
on S. sclarea (Tibaldi et al., 2010). Zaks et al. (2008) found that
the cell-free extract obtained from glandular thricomes, contain-
ing LAT (linalool acetyl transferase) in M. aquatica var. citrata L. can
efficiently catalyze the formation of linalyl acetate from linalool
and acetyl-coenzyme A. Drying temperatures, with a reduction in
the water activity, may favour the release of the extracts from the
glandular thricomes and also catalyze the same reaction in oregano.
Sabinene, which was not affected by the tested growing con-
ditions, could have a biosynthetic pathway under strong genetic
control. Its content increased with the drying processes and showed
a similar response to the EO of Laurus nobilis (Díaz-Maroto et al.,
2002). Díaz-Maroto et al. (2002) found a slight increase in sabinene
in air-dried samples and a significant increase in oven-dried sam-
ples. Jerković et al. (2001), studying oregano, and Raghavan et al.
(1997), studying O. majorana, found an opposite response to the
drying process to ours, with a decrease in the sabinene content
when the herbs were subjected to the drying process. These dif-
ferent sabinene results found in the present work compared to
similar botanical species studied by other authors should be studied
in more detail in order to understand whether the drying process
could have a different response on terpene modification according
to the chemotype.
 G. Tibaldi et al. / Industrial Crops and Products 34 (2011) 1516–1522 1521

Fig. 1. Effect of the growing conditions on the characterizing EO compounds of Origanum vulgare ssp. hirtum grown in Northern Italy. * Values followed by the same letter,
within the same histogram group, are not significantly different according to Tukey’s multiple range test P ≤ 0.050. P = statistical significance values.

Fig. 2. Effect of the postharvest management on the characterizing EO compounds of Origanum vulgare ssp. hirtum grown in Northern Italy. * Values followed by the same
letter, within the same histogram group, are not significantly different according to Tukey’s multiple range test P ≤ 0.050. P = statistical significance values.

␤-Bisabolene characterises some Origanum species, but is 4. Conclusions

absent in O. majorana EO, and thus can be considered a distin-
guishing key-mark between the two main commercial species.
The response in oregano to the growing conditions tested in our
work is unclear, although we could hypothesise that the presence
of this compound, like other sesquiterpenes, such as germacrene
D, decreases with an increase in plant stress. During postharvest
management, the ␤-bisabolene content increased for both drying
techniques, compared to the fresh herbs, like the other sesquiter-
penes in the present experiment and in our other experiments
on Lamiaceae species (different mints; S. officinalis: data not pub-
lished). These results are in agreement with those of Jerković et al.
(2001), who found a constant increase in ␤-bisabolene in dried
oregano samples throughout the growing season compared to their
respective fresh samples.
Our study have indicated that the concentration of monoter-
penes in oregano plants is affected by several growing factors
and light, in terms of solar radiation, has been confirmed to
play an important role. Soilless growing systems resulted to
be a suitable growing system for herbs, offering the advan-
tages of raising temperate–climate demanding species in colder
areas. A defined cultivation management, tuning water and
light stresses during the summer, allows to increase the essen-
tial oil content, while covering the plants in small greenhouse
during their winter quiescence, increasing temporary plant
density, can reduce crop protecting costs and free available
space for winter crops. Further studies on the influence of
other abiotic factors are needed to understand how to fur-
1522 G. Tibaldi et al. / Industrial Crops and Products 34 (2011) 1516–1522
ther enhance the EO content and EO compounds of Greek
oregano.
The oven-drying process and the dehumidifying process
changed the EO in a similar way compared to the EO distilled
from fresh herbs. Thus, when extracting the EO directly from fresh
herbs is not possible, processors can dry the oregano according to
the equipments they have available. Further studies are needed
concerning the postharvest management of oregano in order to
understand to what extent the different drying techniques can
affect the phenolic fraction, which along with the EO, represents
the nutraceutical complex of the species.
Acknowledgement
The authors have contributed equally to the research and to the
manuscript.
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