Industrial Crops and Products 96 (2017) 120–125

Contents lists available at ScienceDirect

Industrial Crops and Products

journal homepage: www.elsevier.com/locate/indcrop

Phytochemical profiling of volatile components of Lavandula

angustifolia Miller propagated under in vitro conditions
Nese Kirimer a,∗ , Sam Mokhtarzadeh a , Betul Demirci a , Fatih Goger a ,
Khalid Mahmood Khawar b , Fatih Demirci a,c
a
Department of Pharmacognosy, Faculty of Pharmacy, Anadolu University, 26140 Eskisehir, Turkey
b
Tarbiyotek-Agricultural Biotechnology Section, Department of Field Crops, Faculty of Agriculture, Ankara University, 06110 Diskapi, Altindag, Ankara,
Turkey
c
Faculty of Health Sciences, Anadolu University, 26140 Eskisehir, Turkey

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

Article history: The aim of this study was to compare the chemical composition of L. angustifolia volatiles from the field
Received 25 July 2016 crop plant versus in vitro micropropagated plantlets. The rationale of this work was to develop in vitro
Received in revised form culture methods for the fast and independent propagation of medicinal and aromatic plants in particular
17 November 2016
Lavender.
Accepted 28 November 2016
Lavandula angustifolia Miller (Lamiaceae), also known as Lavender, is widely used in herbal medicine,
cosmetic industry, and for culinary purposes. This species is highly valuated for its industrial purposes.
Keywords:
It is a fact that there is a gap in the biotechnological production of industrially valuable crops such as
Lavandula angustifolia
Plant tissue culture
Lavender. As a contemporary approach the plant tissue culture strategy of medicinal and aromatic plants
Volatiles by micropropagation serves as a useful technology to produce sufficient amounts of bioproducts e.g. plant
GC–FID material for multiple purposes.
GC–MS The plant samples were subjected both to hydro- and microdistillation for the isolation of volatiles.
The resulting subsequent volatiles were simultaneously analysed both by GC–FID and GC–MS systems.
The main compounds characterized from the hydrodistillation of micropropagated plantlets were
linalool (22.1%), lavandulyl acetate (15.3%) and linalyl acetate (14.7%). The main compounds identified
through microdistillation of this sample were linalool (14.6%), lavandulyl acetate (12.8%) and linalyl
acetate (4.7%). On the other hand the main compounds of field crop plant from hydro- and microdistilla-
tion were T-cadinol (16.9–9.3%), borneol (8.7–14.0%) and
3-carene (8.7–8.6%), respectively. As a conclusion, the obtained results suggested that volatile compo-
sition of the micropropagated samples resembles to the composition of the officinal Lavender essential
oils, which can be industrially exploited.
© 2016 Elsevier B.V. All rights reserved.

1. Introduction
Lavender or “English Lavender” (Lavandula angustifolia Miller,
Syn L. vera,) of Lamiaceae is an important crop utilized to obtain
both the medicinal and aromatic valuable commercial essential oil.
The plant is found natively in South Europe, Mediterranean coun-
tries of Northern Africa, France, Bulgaria, England, America and
Austria. (Gonçalves and Romano, 2013 and Mokhtarzadeh, 2011).
Fresh or dried tips of L. angustifolia flowering branches are used
for medicinal purposes such as the preparation of herbal drugs,
∗ Corresponding author.
E-mail addresses: nkirimer@gmail.com, nkirimer@anadolu.edu.tr (N. Kirimer).
whereas dry leaves are used in perfumery and cosmetic industry.
As the plant contains linalool and linalyl acetate; it is also used for
the production of skin care/skin cleaning lotions, scented bathing
soap, and foams (Baytop, 1984; Naik, 1998; Lis-Balchin, 2002 and
Cavanagh and Wilkinson, 2002).
Lavender is a perennial evergreen shrub with a lifetime of
approximately 10–20 years. L. angustifolia grows up to 50 cm with
linear-lanceolate leaves grey tomentose, becoming greener with
its age. Inflorence stalk is usually unbranched 10–25 cm long with
a compact spike, with a lower flower cluster distant from the main
spike, blue/mauve, rarely violet pink in colour. Flowering time is
June-July, with a natural variation and a characteristic fragrance
due to its essential oil (Mill, 1982; Lis-Balchin, 2002)

http://dx.doi.org/10.1016/j.indcrop.2016.11.061
0926-6690/© 2016 Elsevier B.V. All rights reserved.
 N. Kirimer et al. / Industrial Crops and Products 96 (2017) 120–125
Lavender grows naturally in the Mediterranean area mostly
on rocky soils and mild coastal climates. The agricultural prac-
tices directly influence the secondary metabolites produced in
the plants, and thus the quality. Lavender is cultivated mostly in
Europe. The change of volatiles over the year is essential in deter-
mining the appropriate organ type and the period of harvest, which
is another topic for research. Lavadula sp. synthesize and accu-
mulate its volatiles in specialized secretory capitate and peltate
oil glands located abundantly on the surface of the calyx and on
the leaves. Weather parameters such as temperature and rainfall
are known to influence essential oil content and composition of
lavender, along with fertilization (Lis-Balchin, 2002; Hassiotis et al.,
2014).
It is a well-known fact that the commercial essential oil obtained
from Lavandula sp. is produced mainly by steam distillation and to a
lesser extent by hydro-diffusion. Hydro- and steam distillation pro-
cesses can be relatively time consuming, which are performed at
high temperatures also resulting in degradation of thermally unsta-
ble compounds. Consequently, Lavandula volatiles can be extracted
by solvent extraction and by modern techniques such as microwave
and supercritical extraction methods. Nevertheless, Lavandula sol-
vent extracts may contain fats, waxes, pigments, artefacts and
solvent residues along with volatile compounds. Considering the
pros and cons of the extraction methods, new supercritical CO2
extracts are penetrating more and more the markets due to special
natural product requests, which cannot be classified as “essential
oils” by definition (Danh et al., 2013; Lis-Balchin, 2002; Baser and
Buchbauer, 2009).
Among others, the quality of Lavandula essential oils are
regulated by ISO standards (The International Organization for
Standardization). Also various international standards such as
European Pharmacopoeia (PhEur) contain monographs on various
Lavandula sp. preparations securing pharmaceutical grade quality
(Council of Europe, 2014).
According to previous information and applications it is pos-
sible to increase the generation of bioproducts, active ingredients
and secondary metabolites such as volatile constituents of plants
using plant tissue culture techniques (Alfermann and Petersen,
1995; Dornenburg and Knorr, 1996; Ravishankar et al., 1999;
Ramachandra and Ravishankar, 2002 and Zuzarte et al., 2010). Also
it is known that tissue culture techniques have been applied to
several Lavandula species (Zuzarte et al., 2010 and references cited
herein). Micropropagation of rapid multiplying selected Lavandula
sp. genotypes is a modern technique for the modern agricultural
practices (Echeverrigaray et al., 2005; Machado et al., 2013). Out of
approximately billion US-Dollars annual world essential oil trade,
50 million US-Dollars trade consists of lavender essential oil. Turk-
ish lavender oil needs are increasing with the passage of time
and increasing population (Lis-Balchin, 2002; Baser and Buchbauer,
2009).
Fig. 1. L. angustifolia from field conditions in different growth stages.
121
As an initial study, the rationale of this present work was to
develop biotechnological simple and sustainable in vitro culture
methods for the fast and independent propagation of potentially
valuable medicinal and aromatic plants.
The aim of the present work was to appraise the composition
of Lavender essential oil obtained from crops grown in Turkey, and
compare its profile with the volatiles obtained from micropropa-
gation plant tissues as a preliminary study.
2. Materials and methods
2.1. Plant material
The flowering herba and seeds of Lavandula angustifolia Miller
were provided from Prof. Dr. Neset Arslan, from the Section
Medicinal and Aromatic Plants, Department of Field Crops, Fac-
ulty of Agriculture, Ankara University, Ankara, Turkey (for different
growth stages, see Fig. 1.). The flowering herba was collected in the
morning hours at 20 ◦ C in June 2013. The seeds were collected after
the flowering period.
2.2. Lavandula angustifolia micropropagation
The culture conditions for the surface sterilization for 50 seeds
of lavender of field cultivated plant material were determined.
The lavender seeds were surface sterilized with 5% commercial
bleach (5% NaOCl) for 5 min followed by 3 × 5 min rinsing with
bi-distilled sterilized water. The seeds were germinated on soil
and MS (Murashige and Skoog, 1962) medium supplemented with
30 g/L sucrose 0.65% agar (Sigma type A). The pH of all culture
media was adjusted to 5.7 ± 0.1 before autoclaving at 120 ◦ C for
20 min. About 35 mL of autoclaved medium was poured into each
of the petri dish or culture vessels. All cultures were incubated
at 24 ± 1 ◦ C under 16 h light photoperiod provided by cool white,
fluorescent light (4000 Lux). The cotyledon node explants were
taken from 30 to 45 days old seedlings. Cotyledon nodes of ger-
minating seeds were used as explants that were cultured on MS
medium containing 2 mg/L BAP (6-benzylaminopurine) for eight
weeks. Well-developed micropropagated shoots were rooted on
MS medium containing the 1.25 mg/L IBA (indole-3-butyric acid).
Thereafter, the rooted plants were transferred to peat moss con-
tained in black colored polythene tubes at room temperature. The
peat moss was previously sterilized at 160 ◦ C for 2 h. Microprop-
agated plants were acclimatized under field conditions as seen in
Figs. 2 and 3 (Mokhtarzadeh, 2011).
122 N. Kirimer et al. / Industrial Crops and Products 96 (2017) 120–125
Fig. 2. Micropropagation in cotyledon node explant.
a,b. MS medium with 2.00 mg/L BAP for shoot regeneration on cotyledon node explants of L. angustifolia.
(bar a, b ≈ 1.66 cm).
Fig. 3. The effect of IBA concentrations in rooting for shoots.
2.3. Essential oil extraction
2.3.1. Hydrodistillation
Air-dried flowering herba of field cultivated and micropropa-
gated plants were subjected to hydrodistillation for 3 h using a
Clevenger-type apparatus, according the PhEur 8.0 with modifica-
tions (Council of Europe, 2014).
2.3.2. Microdistillation
The volatiles were obtained after microdistillation of the dried
lavender flowers (400–750 mg) using an Eppendorf MicroDistiller
(Germany) system. Using the distillation program, the sample vial
was heated to 108 ◦ C at a rate of 20 ◦ C/min for 90 min followed
by heating at 112 ◦ C at the rate of 20 ◦ C/min for 30 min (for more
detail, see Tabanca et al., 2010). Thereafter, the organic layer in
the collection vial was separated and analysed by gas chromatog-
raphy (GC) and gas chromatography-mass spectrometry (GC–MS)
simultaneously.
2.4. Analysis conditions
2.4.1. Gas chromatography analysis
The GC-FID analyses were carried out using an Agilent 6890N
GC system using FID detector temperature of 300 ◦ C. To obtain
the same elution order with GC–MS, simultaneous auto-injection
was done on a duplicate of the same column at the same oper-
a,b. MS medium with 1.25 mg/L IBA (bar a, b ≈ 1.5 cm).
ational conditions. Relative percentage amounts of the separated
compounds were calculated from FID chromatograms.
2.4.2. Gas chromatography −mass spectrometry analysis
The GC–MS analyses were carried out using an Agilent 5975
GC–MSD system. Innowax FSC column (60 m x 0.25 mm, 0.25 mm
film thickness) was used with helium as carrier gas (0.8 mL/min).
GC oven temperature was kept at 60 ◦ C for 10 min and programmed
to 220 ◦ C at a rate of 4 ◦ C/min that was kept constant at 220 ◦ C for
10 min and followed by elevating the temperature to 240 ◦ C at a
rate of 1 ◦ C/min. Split ratio was adjusted at 40:1. The injector tem-
perature was set at 250 ◦ C. Mass spectra were recorded at 70 eV,
where the mass range was m/z 35–450.
2.5. Identification of components
Identification of the essential oil components were carried out
by comparison of their relative retention times with those of
authentic samples or by comparison of their relative retention
index (RRI) to series of n-alkanes. Computer matching against com-
mercial (Wiley GC–MS Library, MassFinder 3 Library) (McLafferty
and Stauffer, 1989 and Koenig et al., 2004) and in-house “Başer
Library of Essential Oil Constituents” built up by genuine com-
pounds and components of known oils. Additionally, MS literature
data (Joulain and Koenig, 1998) was also used for the identification.
 N. Kirimer et al. / Industrial Crops and Products 96 (2017) 120–125 123

Table 1
Chemical composition of Lavandula angustifolia volatiles.

No. RRI Compound HD1 % MD1 % HD2 % MD2 % Identification method

1 1014 Tricyclene 0.2 0.3 – – MS

2 1032 ˛-Pinene 4 5.5 – – tR , MS
3 1035 ˛-Thujene 0.8 1 – – MS
4 1072 ␣-Fenchene 0.1 0.2 – – tR , MS
5 1076 Camphene 3.3 4.5 – 2.1 tR , MS
6 1118 ˇ-Pinene 5 6.3 – 0.4 tR , MS
7 1132 Sabinene 0.9 1.1 – – tR , MS
8 1159 ı-3-Carene 8.7 8.6 – – MS
9 1174 Myrcene 1.3 1.1 0.8 0.6 tR , MS
10 1176 ˛-Phellandrene 0.8 1.3 – – tR , MS
11 1187 o-Cymene 0.2 0.2 – – MS
12 1188 ˛-Terpinene 0.2 0.2 – – tR , MS
13 1203 Limonene 4 3.7 0.4 2.6 tR , MS
14 1213 1,8-Cineole 5.9 5.8 – 3.1 tR , MS
15 1246 (Z)-ˇ-Ocimene 0.1 0.1 3.5 1.1 MS
16 1255 -Terpinene 0.3 0.3 – – tR , MS
17 1265 3-Octanone – – 1.2 0.6 tR , MS
18 1266 (E)-ˇ-Ocimene 0.5 0.5 10.4 3.9 MS
19 1278 m-Cymene 1 1.1 – – MS
20 1280 p-Cymene 2.3 2.5 0.2 1.6 tR , MS
21 1282 Hexyl acetate – – 0.8 0.4 tR , MS
22 1286 Isoterpinolene 0.6 0.5 – – MS
23 1290 Terpinolene 0.3 0.2 – – tR , MS
24 1345 3-Octyl acetate – – 0.3 – tR , MS
25 1429 Perillene 0.1 tr – – MS
26 1450 trans-Linalool oxide (Furanoid) – – – 0.6 MS
27 1452 1-Octen-3-ol 0.2 0.2 – – tR , MS
28 1458 cis-1,2-Limonene epoxide 0.1 0.1 – – MS
29 1474 trans-Sabinene hydrate 0.2 0.2 – – MS
30 1478 cis-Linalool oxide (Furanoid) – – – 0.4 MS
31 1483 1-Octyl acetate – – 1.6 – tR , MS
32 1532 Camphor 1.3 2.3 – 1.5 tR , MS
33 1553 Linalool 0.3 0.3 22.1 14.6 tR , MS
34 1556 cis-Sabinene hydrate 0.1 tr – – MS
35 1565 Linalyl acetate – – 14.7 4.7 tR , MS
36 1583 ˛-Santalene 1.4 1.3 – – MS
37 1588 Bornyl formate 0.6 0.6 – 0.5 MS
38 1591 Bornyl acetate 0.5 0.9 – 0.7 tR , MS
39 1611 Terpinen-4-ol 0.6 0.5 – – tR , MS
40 1612 ˇ-Caryophyllene 2.9 2.4 5.1 1.2 tR , MS
41 1617 Lavandulyl acetate 1 1.3 15.3 12.8 tR , MS
42 1639 Cadina-3,5-diene 0.6 0.4 – – MS
43 1668 (Z)-ˇ-Farnesene – – 1.4 – MS
44 1670 trans-Pinocarveol 0.4 0.6 – – tR , MS
45 1686 Lavandulol 0.4 0.6 1.6 1.5 tR , MS
46 1690 Cryptone 0.5 1 – 3.9 MS
47 1704 -Muurolene 5.9 4 – – MS
48 1706 ˛-Terpineol 0.1 0.2 2.1 1.9 tR , MS
49 1719 Borneol 8.7 14 0.8 7.4 tR , MS
50 1722 Bicyclosesquiphellandrene 0.3 0.4 – – MS
51 1733 Neryl acetate – – 0.8 1 tR , MS
52 1751 Carvone 0.2 0.2 – 0.9 tR , MS
53 1765 Geranyl acetate – – 1.6 1.4 tR , MS
54 1776 -Cadinene – – 1.7 2.3 MS
55 1776 Cumin aldehyde 0.5 0.3 – 1.9 tR , MS
56 1808 Nerol – – 0.4 – tR , MS
57 1845 trans-Carveol 0.1 0.4 – – tR , MS
58 1849 Calamenene 0.4 0.4 – – MS
59 1856 m-Cymen-8-ol 0.3 1.3 – – MS
60 1857 Geraniol – – 1.4 1.7 tR , MS
61 1864 p-Cymen-8-ol 0.2 0.8 – 0.7 tR , MS
62 2001 Isocaryophyllene oxide 0.4 0.4 – – MS
63 2008 Caryophyllene oxide 4.3 3.8 0.5 3.4 tR , MS
64 2071 Humulene epoxide-II 0.1 0.1 – – MS
65 2080 Cubenol 1.3 0.8 0.6 1 MS
66 2113 Cumin alcohol 0.1 0.2 – 0.7 tR , MS
67 2131 Hexahydrofarnesyl acetate – – – 0.5 MS
68 2187 T-Cadinol 16.9 9.3 9.6 11.1 MS
69 2232 ˛-Bisabolol 0.4 tr – – tR , MS
70 2349 Cadina-4,10(15)-dien-3-one 0.8 0.4 – – MS
Monoterpene Hydrocarbons 40.5 45 15.3 15.4
Oxygenated Monoterpenes 15.7 24.8 60.8 54.9
Sesquiterpene Hydrocarbons 11.5 8.9 8.2 3.5
Oxygenated Sesquiterpenes 24.2 14.8 10.7 15.5
Others 0.8 1.2 3.9 5.4
Total 92.7 94.7 98.9 94.7

HD1: Hydrodistillation of field cultivar, HD2: Hydrodistillation of micropropagated plants, MD1: Microdistillation of field cultivar, MD2: Microdistillation of micropropagated
plants, RRI: Relative retention indices calculated against n-alkanes, %: Calculated from FID data, tr: Trace (<0.1%). Identification method: tR , identification based on the retention
times (tR ) of genuine compounds on the HP Innowax column; MS: Identified on the basis of computer matching of the mass spectra with those of the Wiley and MassFinder
libraries and comparison with current literature data.
124 N. Kirimer et al. / Industrial Crops and Products 96 (2017) 120–125
Fig. 4. Comparative terpene composition of hydrodistilation (HD1, HD2) and microdistilation (MD1, MD2).
The identified compounds were listed in Table 1. See Fig. 4 for the
distribution of the main components.
3. Results
The yield of the flowering herba and micropropagated plants
showed 0.7% and 2.6% essential oil, which was calculated on a
dry weight base, respectively, suggesting further feasibility stud-
ies. The GC-FID analysis showing the relative percentages (%) of
the separated compounds as calculated from FID chromatograms
are listed in Table 1. GC and GC–MS analyses of the hydrodis-
tilled and microdistilled essential oil of micropropagated plants
were identified as 25 and 35 compounds representing 98.9%
and 94.7% of the obtained essential oils, respectively. The main
compounds obtained by hydrodistillation were linalool (22.1%),
lavandulyl acetate (15.3%), linalyl acetate (14.7%), (E)-␤-ocimene
(10.4%), T-cadinol (9.6%), ␤-caryophyllene (5.1%), (Z)-␤-ocimene
(3.5%), ␣-terpineol (2.1%), and ␥-cadinene (1.7%). Microdistil-
lation results showed presence of 35 compounds. The main
compounds obtained characterized from the microdistillation were
linalool (14.6%), lavandulyl acetate (12.8%), T-cadinol (11.1%), bor-
neol (7.4%), linalyl acetate (4.7%), (E)-␤-ocimene (3.9%), cryptone
(3.9%), caryophyllene oxide (3.4%), and 1,8-cineole (3.1%). When
the relative percentage amounts of compounds were compared,
hydrodistillation showed the maximum percentage of linalool
with 22.1%, whereas the linalool percentage using the microdis-
tillation method was 14.6% only. Next higher percentage was
determined for lavandulyl acetate with 15.3%, which was 12.8%
using the microdistillation method. Same trend was evident in
the third major compound, namely linalyl acetate (14.7%), which
was reduced to 4.7% using the microdistillation process. Moreover,
decrease in the percentage of compounds was not parallel except
for linalool and lavandulyl acetate.
On the other hand, GC and GC–MS analyses of the hydrodistilled
and microdistilled samples of field cultivated plant a total number
of 57 compounds were identified representing 92.7% and 94.7% of
the volatiles, respectively. The main compounds of cultivated field
plant from hydrodistillation and microdistillation samples were
identified as T-cadinol (16.9 and 9.3%), borneol (8.7 and 14%), ␦-3-
carene (8.7 and 8.6%), ␤-pinene (5.0 and 6.3%), 1,8-cineole (5.9 and
5.8%) and ␥-muurolene (5.9 and 4%), respectively. Both distillation
methods on the field plant resulted in comparable compositions.
However, L. angustifolia samples showed qualitative differences in
the composition of their volatiles. These results are highlighted and
illustrated in Fig. 4, comparatively.
This report is the first investigation and comparison of L.
angustifolia field cultivated and biotechnologically micropropa-
gated sample and the phytochemical profiling of their volatiles by
hyrdro- and microdistillation methods, respectively. To the best of
our knowledge this is the first report on the comparative production
and distillation methods of lavender grown in Turkey.
4. Discussion
According to previous work on L. angustifolia from Xinjiang,
China, 17 compounds were identified in the hydrodistilled oil
with linalool (44.5%), geraniol (11%), lavandulyl acetate (10.8%),
3,7-dimethyl-2,6-octadien-1-ol (10.4%), and isoterpineol (6.8%)
as the main components (Cong et al., 2008). Three major con-
stituents of the leaf oil of the cultivated lavender from Iran were
identified as 1,8-cineole (65.4%), borneol (11.5%) and camphor
(9.5%) (Hajhashemi et al., 2003). Whereas, linalool (35.3%), linalyl
acetate (13.4%) and lavandulyl acetate (10.9%) were main com-
ponents of lavender from Iran (Fakhari et al., 2005). The Brazil
lavender oil main components were reported as borneol (22.4%),
epi-␣-muurolol (13.4%), ␣-prococene I (13%) and 1,8-cineole by
Mantovani et al., 2013. Verma et al. (2010) observed linalyl acetate
(47.6%), linalool (28.1%), lavandulyl acetate (4.3%) and ␣-terpineol
(3.8%) of lavender cultivated in India. In the lavender essential
oil from Trieste-Italy, linalyl acetate (43.1%), linalool (32.7%), and
caryophyllene (4.9%) were reported as main compounds (Evandri
et al., 2005). In another lavender example from Italy, linalool
(35.9–36.5%) and linalyl acetate (21.7–14.4%) were reported as
main compounds (Da Porto et al., 2009).
Zuzarte et al. (2010) suggested that in vitro cultures are helpful
in assuring the quality of the essential oils, and such protocols can
be applied to the propagation of selected chemotypes for further
industrial purposes. There were some recent reports on the chem-
ical composition and quality of micropropagated lavenders from
various groups. Main volatile components of the micropropagated
lavender plants from Italy were identified as caryophyllene (19.3%)
and 1,8-cineole (11.9%) as reported recently by Pistelli et al. (2013).
The results of this present study are in accordance with the
recent work of Machado et al. (2013), who reported that microprop-
 N. Kirimer et al. / Industrial Crops and Products 96 (2017) 120–125
agated lavender (L. angustifolia) contained 4% essential oil during
first and around 5% during second harvest from Southern Brazil. The
major compounds of volatiles were determined as linalool (46.9
and 37.3%) and linalyl acetate (10.1 and 12.2%), respectively.
The micropropagated Lavandula plantlets from this present
study were relatively low in linalool content, as it would be
expected in higher percentages in regard to the pharmaceutical
quality (Council of Europe, 2014). Also it is known that rainfalls may
decrease linalool production and the quality of lavender oil, which
can be recovered after several days of growth, suggesting that
linalool biosynthesis is regulated during specific growth periods by
both environmental and developmental conditions (Hassiotis et al.,
2014).
5. Conclusion
According to the micropropagation results of the phytochemi-
cal profile of L. angustifolia, which were in promising compliance
of the standard essential oil composition of the Pharmacopoeia,
more detailed in vitro experimental studies can be performed.
The cultivated field plant samples did not meet the level of the
expected standards contrary to the micropropagated samples. The
phytochemical variation of the cultivated field plant versus the
micropropagated material may be influenced by environmental,
climatic stress factors as well as collection period and techniques.
Further biotechnological studies as well as agronomical field com-
parisons would give more insight to the production valuable of
secondary metabolites such as essential oils.
Acknowledgement
The authors greatly acknowledge the Scientific and Techno-
logical Research Council of Turkey (TÜBİTAK) for support part of
this work as a post-doctorate research project (2216-BIDEB) of S.
Mokhtarzadeh.
References
Alfermann, A.W., Petersen, M., 1995. Natural products formation by plant cell
biotechnology. Plant Cell Tissues Organs 43, 199–205.
Baser, K.H.C., Buchbauer, G. (Eds.), 2009. and Applications, CRC Press.
Baytop, T., 1984. Therapy with Medicinal Plants in Turkey (Past and Present).
Istanbul University, pp. 316.
Cavanagh, H.M., Wilkinson, J.M., 2002. Biological activities of lavender essential oil.
Phytother. Res. 16, 301–308.
Cong, Y., Abulizi, P., Zhi, L., Wang, X., Mirensha, M., 2008. Chemical composition of
the essential oil of Lavandula angustifolia from Xinjiang, China. Chem. Nat.
Compd. 44, 810.
Council of Europe, 2014. European Pharmacopoeia (PhEur), vol. 8.0. Council of
Europe, Strasbourg.
Da Porto, C., Decorti, D., Kikic, I., 2009. Flavour compounds of Lavandula angustifolia
L. to use in food manufacturing: comparison of three different extraction
methods. Food Chem. 112, 1072–1078.
Danh, L.T., Han, L.N., Triet, N.D.A., Zhao, J., Mammucari, R., Foster, N., 2013.
Comparison of chemical composition, antioxidant and antimicrobial activity of
Lavender (Lavandula angustifolia L.) essential oils extracted by supercritical
CO2 , hexane, hydrodistillation. Food Bioprocess Technol. 6, 3481–3489.
125
Dornenburg, H., Knorr, D., 1996. Production of the phenolic flavour compounds
with cultured cells and tissue of Vanilla planifolia species. Food Biotechnol. 10,
75–92.
Echeverrigaray, S., Basso, R., Andrade, L.B., 2005. Micropropagation of Lavandula
dentata from axillary buds of field-grown adult plants. Biol. Plant. 49, 439–442.
Evandri, M.G., Battinelli, L., Daniele, C., Mastrangelo, S., Bolle, P., Mazzanti, G., 2005.
The antimutagenic activity of Lavandula angustifolia (lavender) essential oil in
the bacterial reverse mutation assay. Food Chem. Toxicol. 43, 1381–1387.
Fakhari, A.R., Salehi, P., Heydari, R., Ebrahimi, S.N., Haddad, P.R., 2005.
Hydrodistillation-headspace solvent microextraction, a new method for
analysis of the essential oil components of Lavandula angustifolia Mill. J.
Chromatogr. A 1098, 14–18.
Gonçalves, S., Romano, A., 2013. Protocols for micropropagation of selected
economically-important horticultural plants (Micropropagation of Lavandula
spp.). Methods Mol. Biol. 994, 189–198.
Hajhashemi, V., Ghannadi, A., Sharif, B., 2003. Anti-inflammatory and analgesic
properties of the leaf extracts and essential oil of Lavandula angustifolia Mill. J.
Ethnopharmacol. 89, 67–71.
Hassiotis, C.N., Ntana, F., Lazari, D.M., Poulis, S., Vlachonasios, K.E., 2014.
Environmental and developmental factors affect essential oil production and
quality of Lavandula angustifolia during flowering period. Ind. Crops Prod. 62,
359–366.
Joulain, D., Koenig, W.A., 1998. The Atlas of Spectral Data of Sesquiterpene
Hydrocarbons. EB-Verlag, Hamburg.
Koenig, W.A., Joulain, D., Hochmuth, D.H. 2004. Terpenoids and Related
Constituents of Essential Oils. MassFinder 3. Hamburg, Germany.
Lis-Balchin, M., 2002. Lavender: The Genus Lavandula (Medicinal and Aromatic
Plants − Industrial Profiles). CRC Press, Taylor & Francis, London.
Machado, M.P., Ciotta, M.N., Deschamps, C., Zanette, F., Côcco, L.C., Biasi, L.A., 2013.
In vitro propagation and chemical characterization of the essential oil of
Lavandula angustifolia cultivated in Southern Brazil. Cienc. Rural 43, 283–289.
Mantovani, A.L.L., Vieira, G.P.G., Cunha, W.R., Groppo, M., Santos, R.A., Rodriques,
V., Magalhaes, L.G., Crotti, A.E.M., 2013. Chemical composition,
antischistosomal and cytotoxic effects of the essential oil of Lavandula
angustifolia grown in Southeastern Brazil. Rev. Bras. Farmacogn. 23, 877–884.
McLafferty, F.W., Stauffer, D.B., 1989. The Wiley/NBS Registry of Mass Spectral
Data. J. Wiley and Sons New York.
Mill, R.R., 1982. Lavandula L. In: Davis, P.H. (Ed.), Flora of Turkey and the East
Aegean Islands, vol. 7. Edinburgh Univesity Press, pp. 76–78.
Mokhtarzadeh, S., 2011. Optimisation of tissue culture and genetic transformation
in Lavandula angustifolia Miller subsp. angustifolia Miller and L. stoechas L.
subsp. stoechas. PhD Thesis. Ankara University, pp. 114.
Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassay
with tobacco tissue culture. Physiol. Plant 15, 473–497.
Naik, G.R., 1998. Micropropagation studies in medicinal and aromatic plants. In:
Khan, I.A., Khanun, A. (Eds.), Role of Biotechnology in Medicinal and Aromatic
Plants. Ukaz publication, Hyderabad, pp. 50–56.
Pistelli, L., Noccioli, C., D’Angiolillo, F., Pistelli, L., 2013. Composition of volatile in
micropropagated and field grown aromatic plants from Tuscany Islands. Acta
Biochim. Pol. 60, 43–50.
Ramachandra, R.S., Ravishankar, G.A., 2002. Plant cell cultures: chemical factories
of secondary metabolites. Biotechnol. Adv. 20, 101–153.
Ravishankar, G.A., Bhyalakshmi, N., Ramachandra, R.S., 1999. Production of food
additives. In: Ramawat, K.G., Merillon, J.M. (Eds.), Biotechnology: Secondary
Metabolites. Oxford IBH, New Delhi, pp. 89–110.
Tabanca, N., Demirci, B., Turner, J.L., Pounders, C., Demirci, F., Baser, K.H.C., Wedge,
D.E., 2010. A rapid microdistillation method for eight Salvia taxa from the
Dallas Arboretum and Botanical Garden. Nat. Prod. Commun. 9, 1421–1426.
Verma, R.S., Rahman, L.U., Chanotiya, C.S., Verma, R.K., Chauhan, A., Yadav, A.,
Singh, A., Yadav, A.K., 2010. Essential oil composition of Lavandula angustifolia
Mill cultivated in the mid hills of Uttarakhand, India. J. Serb. Chem. Soc. 75,
343–348.
Zuzarte, M.R., Dinis, A.M., Cavaleiro, C., Salgueiro, L.R., Canhoto, J.M., 2010.
Trichomes, essential oils and in vitro propagation of Lavandula pedunculata
(Lamiaceae). Ind. Crop. Prod. 32, 580–587.