Effect of season on yield and composition of the essential oil of Eucalyptus citriodora Hook. leaf grown in sub-tropical conditions of North India

Abstract

Seasonal variation of the essential oil of Eucalyptus citriodora leaves were analysed by Gas chromatography mass spectrometry (GC-MS) analysis in the trees grown at subtropical conditions of North Indian plains for commercial cultivation and determination of proper harvesting time. At this condition, oil yield ranged between 1.0 to 2.1% during different months. Oil yield was observed to be high during April to September, when temperature was high and the yield was low during November to March, when the temperature was relatively low. However, during rainy season, when both temperature and humidity was high, oil yield was also observed to be quite high (1.8 to 2.1%). At this condition, the major constituent of oil was citronellal (69.7 to 87.4%) followed by citronellol (5.1 to 9.9%), linalool (2.1 to 6.4%), isopulegol (0.9 to 3.1%) and citronellyl acetate (0.4 to 1.2). Concentration of citronellal decreased during the summer and rainy seasons, while concentration of rest of the major constituents increased during this period. Yield of oil and concentrations of citronellal was observed to be comparable to the plants grown commercially in South India but concentration of rest of the constituents decreased considerably.

Full text

Journal of Medicinal Plants Research Vol. 6(14), pp. 2875-2879, 16 April, 2012
Available online at http://www.academicjournals.org/JMPR
DOI: 10.5897/JMPR011.1611
ISSN 1996-0875 ©2012 Academic Journals
Full Length Research Paper
Effect of season on yield and composition of the
essential oil of Eucalyptus citriodora Hook. leaf grown
in sub-tropical conditions of North India
N. Manika1, Priyanka Mishra1, Narendra Kumar1, C. S. Chanotiya2 and G. D. Bagchi1*
1Department of Taxonomy and Pharmacognosy, Central Institute of Medicinal and Aromatic Plants (CSIR), P.O.-CIMAP,
Lucknow-226015, India.
2Department of Analytical Chemistry, Central Institute of Medicinal and Aromatic Plants (CSIR), P. O. - CIMAP,
Lucknow-226015, India.
Accepted 22 December, 2011
Seasonal variation of the essential oil of Eucalyptus citriodora leaves were analysed by Gas
chromatography mass spectrometry (GC-MS) analysis in the trees grown at subtropical conditions of
North Indian plains for commercial cultivation and determination of proper harvesting time. At this
condition, oil yield ranged between 1.0 to 2.1% during different months. Oil yield was observed to be
high during April to September, when temperature was high and the yield was low during November to
March, when the temperature was relatively low. However, during rainy season, when both temperature
and humidity was high, oil yield was also observed to be quite high (1.8 to 2.1%). At this condition, the
major constituent of oil was citronellal (69.7 to 87.4%) followed by citronellol (5.1 to 9.9%), linalool (2.1
to 6.4%), isopulegol (0.9 to 3.1%) and citronellyl acetate (0.4 to 1.2). Concentration of citronellal
decreased during the summer and rainy seasons, while concentration of rest of the major constituents
increased during this period. Yield of oil and concentrations of citronellal was observed to be
comparable to the plants grown commercially in South India but concentration of rest of the
constituents decreased considerably.
Key words: Eucalyptus citriodora, seasonal variation, essential oil composition, citronellal, citronellyl acetate,
linalool, citronellol.
INTRODUCTION
Eucalyptus citriodora Hook. (Family- Myrtaceae),
commonly known as ‘Lemon-Scented Eucalyptus’ or
‘Lemon-Scented Gum’, is an evergreen tree native to
Queensland, Australia (Chen et al., 2007). This tree is
highly valued for its citronellal rich essential oil extracted
from its leaves. It has been introduced in different
countries including India for commercial cultivation. Its oil
is widely used in a number of perfumery formulations,
toiletries and as disinfectants. Citronellal obtained from
the oil is used mainly for the production of synthetic
menthol and citronellol. The leaves reported to possess
antiseptic properties and are used in the treatment of
various skin diseases. Oil of E. citriodora reported to
*Corresponding author. E-mail: gd.bagchi@cimap.res.in.
possess antibacterial, antifungal, ascaricidal and insect
repellent activities (Low et al., 1974; Husain et al., 1988;
Ramezani et al., 2002; Singh et al., 2002; Verbel et al.,
2009; Luqman et al., 2008). The oil is also observed to be
phytotoxic and has potential to be used as herbicide
(Batish et al., 2004, 2006a, b, 2007, 2008; Singh et al.,
2005, 2006).
In traditional medicine, essential oil is used as
antispasmodic and to relieve joint pains (Buchman et al.,
1979). This species has been introduced in India during
the middle of 19th century (Shiva et al., 1987). It has been
observed that the location, season, nature of soil, age of
plants and planting density greatly influence the yield of
leaves and oil (Singh et al., 1976; Sefidkon et al., 2009).
Harvesting of leaves for economic recovery of essential
oil during February to June, before the onset of monsoon
has been recommended in tropical weather conditions

2876 J. Med. Plants Res.
(Muralidharan et al., 1974; Nair et al., 1974).
Leaf oil of E. citriodora plants, growing in different parts
of the world, is characterized by their major constituent’s
citronellal and citonellol (Zini et al., 2003). Chemical
compositions of decanted and recovered oils of E.
citriodora leaves were examined. The decanted oil was
observed to be rich in citronellal, citronellol, citronellyl
acetate and β-caryophyllene, respectively. On the other
hand, recovered oil was rich in isopulegol, borneol,
menthol, neral and eugenol. Citronellal, the major
constituent of the decanted oil, was absent in the
recovered oil (Rao et al., 2003).
However, the oil isolated from the leaves of the plants
growing in Egypt, was reported to be rich in 3- hexen-1-ol
and cis-geraniol (Abd El Mageed et al., 2011). So far,
commercial cultivation of E. citriodora has not been
undertaken in the sub-tropical north Indian plains. For the
determination of proper harvesting time for higher yield
and quality of oil, plants of E. citriodora were
domesticated at this condition and monthly variation in its
leaf oil yield and its constituents has been studied in five
year old trees.
MATERIALS AND METHODS
Plant material
The fresh leaves of E. citriodora were collected every month
(January 2009 to December 2009) from the trees domesticated at
the experimental field of CIMAP, Lucknow and hydro-distilled in
Clevenger-type apparatus for 3 h to extract the essential oil. The
moisture from the oil was removed by anhydrous sodium sulphate,
then measured and stored at 4°C prior to analysis.
Gas chromatography (GC) analysis
For GC, a Perkin-Elmer Auto System XL gas chromatograph was
used fitted with an Equity-5 column (60 m x 0.32 mm i.d., film
thickness 0.25 µm; Supelco Bellefonte, PA, USA). The oven column
temperature ranged from 70 to 250°C, programmed at 3°C/min,
with initial and final hold time of 2 min, using H2 as carrier gas at 10
psi constant pressure, a split ratio of 1:30, an injection size of 0.03
µL neat, and injector and detector (FID) temperatures were 250 and
280°C, respectively.
Gas chromatography mass spectrometry (GC/MS) analysis
GC/MS utilized a Perkin-Elmer Auto System XL GC interfaced with
a Turbo mass Quadrupole mass spectrometer fitted with an Equity-
5 fused silica capillary column (60 m x 0.32 mm i.d., film thickness
0.25 µm; Supelco Bellefonte, PA, USA). The oven temperature
program was the same as described in capillary GC; injector,
transfer line and source temperatures were 250°C; injection size
0.03 µL neat; split ratio 1:30; carrier gas He at 10 psi constant
pressure; ionization energy 70 eV; mass scan range 40 to 450 amu.
Duplicate analysis was performed. Quantitative results are mean
data derived from GC analysis.
Identification of compounds
Characterization was achieved on the basis of retention time,
Kovats Index, relative retention index using a homologous series of
n-alkanes (C8-C25 hydrocarbons, Polyscience Corp. Niles IL),
coinjection with standards in GC-FID capillary column (Aldrich and
Fluka), mass spectra library search (NIST/EPA/NIH version 2.1 and
Wiley registry of mass spectral data 7th edition) and by comparing
with the mass spectral literature data (Adams et al., 2001). The
relative amounts of individual components were calculated based
on GC peak areas without using correction factors.
RESULTS AND DISCUSSION
E. citriodora oil is extensively used in the treatment of
various diseases and also has perfumery value (Husain,
1988). Its oil is usually rich in citronellal and citronellol
(Zini et al., 2003). However, Egyptian oil was found to be
rich in 3- hexen-1-ol and cis-geraniol (Abd El Mageed et
al., 2011). For development of its commercial cultivation,
plants of E. citriodora were grown at sub-tropics of North
India and their oil was evaluated.
Total yield of essential oil from the leaves of E.
citriodora during different months of the year 2009 along
with the weather data of the experimental area have been
shown in Table 1. The constituents of essential oils have
been shown in Table 2. A total of twenty nine compounds
have been identified in the oil by GC and GC–MS, which
constituted 91.9 to 98.5% of the oil. Yield of the oil from
the leaves varied between 1.0 to 2.1% in different
months. Oil yield was low (1.0 to 1.1%) during the months
of November to March when the temperature was
relatively low (min. temp 7 to 16°C; max. temp 23 to
30°C). While, oil yield was high (1.5 to 2.1%) during April
to October, when the temperature was relatively high
(min. temp 18 to 27°C; max. temp 31 to 40°C). However,
during rainy season (July to September), when both
temperature and humidity was high, oil yield was
observed to be quite high (1.8 to 2.1%).
At the sub-tropical conditions also, the major
constituents of the oil were observed to be citronellal
(69.7 to 87.4%) followed by citronellol (5.1 to 9.9%),
linalool (2.1 to 6.4), isopulegol (0.9 to 3.1%) and cironellyl
acetate (5.1 to 9.9%). Rest of the constituents of the oil
was very low in their quantity. Concentration of major oil
constituent citronellal was observed to be maximum
(87.4%) during the month of January. Its concentration
gradually decreased with the advent of summer season.
In May and June, its concentration became 72 to 73%,
while during rainy season (July to August), its
concentration decreased further (69 to 71%).
With the advent of autumn, concentration of citronellal
again picked up and became quite high (83 to 87%)
during winter season, when the day length was also short
(Table 2 and Figure 1). On the other hand, it is interesting
to note that concentration of rest of the constituents
including citronellol, linalool, isopulegol and citronellyl
acetate reduced during the winter season and became
high during summer and rainy seasons (Figure 2).
Concentration of most of the minor constituents like α-
thujone, α-pinene, sabinene, myrcene, α-terpenene, p-

Manika et al. 2877
Table 1. Weather data of 2009 in experimental area during collection of Eucalyptus citriodora.
Months
Temperature (Min)
(mean ± SD)
Temperature (Max)
(mean ± SD)
Average day length
Average rainfall
Oil yield
(%)
Jan
07.21 ± 1.439
23.20 ± 1.097
10.40
0.00
1.0
Feb
12.83 ± 1.138
24.71 ± 2.743
11.20
0.47
1.0
Mar
15.74 ± 2.556
26.63 ± 4.705
12.00
0.00
1.0
Apr
22.98 ± 2.020
37.48 ± 1.311
12.40
0.00
1.7
May
25.45 ± 1.573
38.31 ± 3.179
13.20
1.25
1.6
Jun
25.90 ± 1.674
39.47 ± 4.249
14.00
0.00
1.6
Jul
27.00 ± 0.831
32.25 ± 2.392
13.20
5.64
1.8
Aug
26.55 ± 0.830
31.76 ± 2.017
12.00
14.24
1.9
Sep
25.57 ± 1.027
32.91 ± 2.457
12.00
5.11
2.1
Oct
18.43 ± 2.054
31.87 ± 1.487
11.20
0.00
1.5
Nov
14.87 ± 2.704
29.49 ± 2.126
10.40
0.24
1.1
Dec
09.49 ± 1.180
24.65 ± 1.871
10.00
0.00
1.0
Table 2. Chemical composition of essential oils [%] of Eucalyptus citriodora during different months.
Compounds
RI
Jan
Feb
Mar
Apr
May
Jun
Jul
Aug
Sep
Oct
Nov
Dec
α -thujene
932
0.1
0.1
01.
0.1
0.2
0.2
0.1
t
t
t
t
t
α-pinene
936
0.1
0.2
0.3
0.3
0.3
0.2
0.2
0.1
0.1
t
t
t
Sabinene
967
0.3
0.3
0.3
0.4
0.4
0.3
0.3
0.1
0.1
0.2
0.2
0.2
Myrcene
980
t
0.1
0.1
0.2
0.2
0.2
0.1
t
t
0.1
t
t
α-hellandrene
997
t
t
t
0.1
0.1
0.1
0.1
0.1
0.1
0.1
0.1
0.1
α –terpenene
1011
-
-
-
0.1
0.37
0.1
0.1
0.1
0.1
0.1
0.1
t
p-cymene
1015
0.2
0.2
0.2
0.3
0.3
0.3
0.2
0.1
0.1
0.1
0.1
0.1
Limonene
1024
0.1
0.1
0.1
0.1
0.2
0.2
0.2
0.1
0.1
0.1
t
t
1,8-cineole
1031
0.1
0.1
0.1
0.2
0.2
0.5
0.8
0.6
0.3
0.3
0.1
0.1
Z-β-ocimene
1034
t
t
t
0.1
0.2
0.1
0.1
0.1
t
t
t
t
E-β-ocimene
1043
0.1
0.2
0.4
0.4
0.8
0.4
0.4
0.3
0.3
0.3
0.3
0.3
Linalool
1085
2.1
2.4
2.7
2.9
3.5
5.1
6.4
5.7
5.0
3.6
3.2
3.1
Citronellal
1135
87.4
85.6
84.2
81.9
73.2
72.6
71.1
69.7
73.7
81.9
83.2
83.8
Isopulegol
1143
1.0
0.9
1.2
1.3
1.9
2.5
3.1
2.9
2.6
1.8
1.6
1.3
Borneol
1152
0.2
0.2
0.3
0.3
0.5
0.5
0.6
0.6
0.6
0.5
0.5
0.4
Menthol
1161
0.1
0.1
0.1
0.1
0.2
0.2
0.1
0.1
0.2
0.2
0.2
0.2
α-terpeneol
1180
-
0.1
0.1
0.1
0.2
0.1
0.1
0.1
0.3
0.1
0.2
0.1
Citronellol
1208
5.5
5.7
6.1
7.8
7.9
8.7
9.9
9.4
9.2
6.1
5.1
5.2
Nerol
1222
0.2
0.2
0.1
0.1
1.01
0.1
0.1
0.1
0.1
0.2
0.4
0.2
Geraniol
1240
0.1
0.1
0.2
0.3
0.9
0.1
0.1
0.1
0.1
0.2
0.2
0.3
Geranial
1250
0.1
t
t
t
-
0.1
-
-
0.1
0.1
0.1
0.1
Unidentified
1275
0.2
0.2
0.3
0.3
1.1
0.8
0.6
0.6
0.6
0.2
0.2
0.2
Citronellyl acetate
1331
0.4
0.5
0.8
0.8
0.8
0.8
0.9
1.2
0.6
0.6
0.6
0.4
Eugenol
1342
t
t
-
0.1
0.3
0.1
t
t
-
t
-
t
Geranyl acetate
1365
t
t
-
0.1
0.2
0.1
t
-
-
-
-
t
β-caryophyllene
1425
-
t
0.2
0.2
0.2
0.1
0.1
0.1
0.1
0.1
-
-
Aromadendrene
1446
-
t
t
0.1
0.1
0.2
0.2
0.2
0.3
0.2
0.1
0.1
Cadinene-γ
1509
-
-
-
-
t
t
t
-
-
t
-
-
Cadinene-δ
1516
-
-
-
-
0.1
t
t
-
-
0.1
-
-
Caryophyllene oxide
1571
0.1
0.1
t
t
t
0.1
0.1
0.1
0.1
0.1
0.1
0.1
Total identified
compounds (%)
98.2
97.2
98.5
98.4
94.28
94.0
95.9
91.9
94.2
97.1
96.4
96.1

2878 J. Med. Plants Res.
Figure 1. Seasonal variations in oil yield of Eucalyptus citriodora throughout the year.
Percentage (%)
Figure 2. Variation in the content of different major oil constituents of Eucalyptus citriodora throughout the year.
cymene, limonene, Z-β-ocimene, E-β-ocimene, menthol,
geraniol, eugenol, geranyl acetate and β-caryophyllene
also increased during summer season. While concen-
trations of some minor constituents like 1, 8-cineole and
borneol increased during rainy season.
Growing of E. citriodora at sub-tropical conditions
showed that at this situation where there is wide variation
in temperature and humidity, there is comparable yield of
oil almost equivalent to commercial cultivation at South
India that is, between 1.0 to 2.1% and the main
constituent citronellal between 69.7 to 87.4%. However,
concentration of citronellol and geraniol reduced
considerably. Therefore, for higher yield of oil, the leaves
should be harvested during rainy season (July to
September) and for the higher yield of citronellal, it
should be harvested during winter months (November to
March) particularly in the month of January.
ACKNOWLEDGEMENTS
The authors are thankful to the Director, Central Institute
of Medicinal and Aromatic Plants (CSIR-CIMAP),
Lucknow, India for facilities during the present work.
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