Jeobp 13 (5) 2010 pp 540 - 544 540

ISSN 0976-5026

Citrus Essential Oil of Nigeria Part I:

Volatile Constituents of Citrus Limon Burm. F. Leaf Oil

Adeleke Adebisi Kasali 1, Oladipupo Adejumobi Lawal 2

and Abayomi Akeem Olaniyan 3

1
Chemistry Department, Lagos State University,
P.M.B. 1087 Apapa Lagos State, Nigeria
2
Department of Chemistry, University of Zululand,
KwaDlangezwa 3886, South Africa
3
National horticultural Research Institute, Idi-Ishin,
P.M.B. 5234, Jericho, Reservation Area, Ibadan Oyo State, Nigeria
Received 21 August 2009; accepted in revised form 11 April 2010

Abstract: The essential oil isolated from leaves of Eureka, and Lisbon (citrus limon Burm.F.)
lemons were analysed by a combination of GC and GC-MS. Twenty eight constituents accounting
for 94.8 % of the Eureka oil and twenty-six constituents accounting for 97.4 % of the Lisbon were
identified. Limonene (30.0 - 34.0 %). and geranial (14.4 - 18.7 %) were found to be the major constituents
in the oils, while other constituents varied.

Key words: Citrus limon Burm. F. Lisbon. Eureka, Rutaceae, essential oil compo-
sition, limonene, neral, geranial, lemon leaf oil, distilled lemon oil.

Introduction: Lemon (Citrus limon Burm. F.) is native to the eastern Himalayan
regions of India 1. It is generally accepted that lemon is a hybrid of citron and sour oranges
2, 3
, it reached China during the song dynasty and Europe following the Persian campaigns of
Alexander the Macedonian 3. Lemon trees are more sensitive to low temperatures 4 and are
not well adapted to humid subtropical or tropical regions due to susceptibility to fungal and
algal diseases 1. In addition, fruit quality, especially peel texture, which becomes coarse, is
generally poorer in humid regions than in Mediterranean-type climates 1.
There are three major groups of lemon: the femminello, verna (Berna) and Sicilian
types 4. The femminello group represents the most widely grown types in Italy; verna is the
major cultivars of Spain 5 while the Sicilian type is important in California and Australia. It

*Corresponding author (Adeleke Adebisi Kasali)

E- mail: < alekekasali@yahoo.com.au >
 Adeleke Adebisi Kasali et al. / Jeobp 13 (5) 2010 pp 540 - 544 541

originated as a seedling from Sicily brought to California in 1858. It is grown in South Africa,
Spain and Israel. The most important cultivars in the Sicilian group are the ‘Eureka’ and
‘Lisbon’. However; ‘Lisbon’ has replaced ‘Eureka’ as a major cultivar’s because of superior
yield 4.
Lemon leaf oils have been used as taxonomic marker as well as in cosmetics and
perfumery. The rind oils are used as flavouring agents in beverages and in foods as well as
cleaning agent and in various industrial applications 3.
The concentration of aliphatic and monoterpenoid aldehydes and the alcohols and
esters are important for the organoleptic quality of cold-pressed lemon oil in addition, several
studies have been carried out on the deterioration of lemon flavour, p-menthadiene-8-ols
and p-menthen-1,8-diols were the major products in the early stages of the decomposition
in addition, citral (neral and geranial) one of the significant components of lemon oil, also
cyclices to give p-cymene and , -dimethylstyrene 6.
A survey of the literature reveals that numerous studies have been carried out to
identify lemon leaf oil components and Lawrence reviewed most of the papers 5. it is only
the chemical composition of the ‘Lisbon’ variety of Lemon of Nigeria origin that has been
reported 7 and to the best of our knowledge there has not been any report about Nigeria
‘Eureka’ lemon leaf oils. This paper reports the volatile constituents of Nigerian varieties of
‘Eureka’, and ‘Lisbon’ lemon leaf oils.

Experimental
Plant material: The leaves were obtained from cultivars growing in the orchards at
the National Horticultural Institute, Idi Ishin-Ibadan, Nigeria.

Essential oil collection: Fresh matured leaves (300g) were subjected to

hydrodistillation for 3hr in accordance with the British pharmacopoeia 8.

GC-analysis: The gas chromatographic (GC) analyses of the oils were performed
on an Orion analytical Micromat 412 double column Gas Chromatograph (GC) fitted with
flame ionization detector (FID), Two capillary columns of differing polarities, CPSil-5 (25 m
x 0.25 mm i.d) equivalent to OV 101 and CPSil-19 (25m x 0.25mm i.d) similar to BP10 (film
thickness 0.15 m), were used. Oven temperature was programmed from 50°C to 230°C at
3°C/min. Injector and detector temperatures were maintained at 200°C and 250°C
respectively. The carrier gas was hydrogen.

GC-MS analysis: GC-MS analysis were carried out on a Hewlett-Packard Gas

Chromatograph (GC) HP 5890A interfaced with a VG Analytical 70-250s double- focusing
mass spectrometer operating at 70eV with an ion source temperature of 230°C. The GC
was fitted with a 25 m x 0.25 i.d. fused silica capillary column coated with CPSil-5. Helium
was the carrier gas at 0.6 bar. The GC operating parameters were identical with those of
the GC analysis.
The constituents of the oils were identified by comparison of their mass spectral
pattern and Kovat’s indices (KI) with those data for original samples in our library built up
 Adeleke Adebisi Kasali et al. / Jeobp 13 (5) 2010 pp 540 - 544 542

from reference oils and authentic sesquiterpenoids. Confirmation of the identification of the
constituents was by matching our MS and KI data with those cited in the literature 16-18.

Result and discussion: Colourless volatile oils were extracted from Eureka and
Lisbon lemon leaves in 0.98 g and 0.85 g yield respectively. The leaf oils were then
comprehensively analysed by means of GC and GC-MS. The volatile constituents identified
from the oils are listed in Table 1, they are arranged in the order of elution on Cpsil-5 column
their retention (Kovat’s) indices are also indicated.
Twenty eight constituents were identified in the Eureka oil sample, accounting for
94.8 % of the total oil. Nine (41.2 %) of these constituents represent monoterpene
hydrocarbons with limonene (30.0 %) as the major constituent. Others include -pinene
(4.5 %), (E)- -ocimene (1.9 %) and sabinene (1.8 %). Twelve oxygenated monoterpenes
(53.2 %) were indentified in the oil sample, geranly acetate (2.4 %), citronellal(1.9 %) and
linalool (1.7 %) were the main oxygenated constituents. Sesquiterpenes were only detected
in trace amount.
However, eight monoterpenes hydrocarbons (49.4 %) were detected in Lisbon volatile
oil, limonene (34.0 %), -pinene (6.2 %) sabinene (3.9%), (E)- -ocimene (2.5 %) and
myrcene (1.5 %) were the major component. Twelve compounds were identified as
oxygenated monoterpenes in Lisbon volatile oils, these accounted for 44.4 % of the oil.
Geranial (14.4 %), neral (12.3 %), neryl acetate (5.7 %) nerol (2.8 %), geranyl acetate (2.5
%), linalool (1.7 %) and citronellal (1.7 %) were the prominent oxygenated components.
The sesquiterpene content was found to be 3.4 % with (E.E.)- -farnesene as the only
noticeable constituent in contrast to trace amount in eureka oil.
The monoterpene contents of the two oil samples is similar to the monoterpene content
of the Indian lemon leaf oil 12 although the concentration of limonene is higher in the Indian
oil however, the concentration of limonene detected in the present study is similar to what
was reported by Malenderas et al 13 and Mondello et al 14. -pinene concentrations of these
samples were similar to those found by Crescimanno et al 15 while (E)- -ocimene, sabinene,
and myrcene differs from what was reported for the Indian 7, Italian, Israel and Spanish
oils 6.
The citral concentration is similar to what was reported by Kumar et al 12, in which
the concentration of geranial is higher than the concentration of neral, a situation that was
observed in Lisbon lemon. Furthermore, the concentration of neral identified in Lisbon was
identical with what was reported by Malenderas et al 13 however, neral concentration
identified in Eureka was similar to the report of Scora et al 16, while the geranial concentration
of both lemon sample agrees with what was reported by Adeishvili et al 17. The volatile
constituents of the two lemon samples were substantially different from the Citrus oil sample
of Benin origin 18.
The present study offer a more detailed description of composition of Nigerian lemon
petitgrain oil than those previously published 7. Even though the data is similar to the previous
data on Nigerian lemon but, some differences are evident. For example, the ester content
was higher, in addition, Ekundayo et al 7 reported the presence of citronellol but it was not
detected in the lemon oil sample analysed in this work. Furthermore, most of the mono-
 Adeleke Adebisi Kasali et al. / Jeobp 13 (5) 2010 pp 540 - 544 543

terpenes identified in this study have not been previously reported as constituents of lemon
volatile oil of Nigerian origin.

Acknowledgement: Authors are grateful to Deutscher Akademischer

Austauschdienst (DAAD) for a fellowship award to A.A Kasali and Lagos State university,
Nigeria for financial support. We also appreciate the assistance of Yvonne Kruepper of the
Institute for Organische Chemie Bibilothek Universitat Hamburg, Germany.

References
1. Davies, F.S. and Gene Albrigo. (1994). Citrus CAB International, Wallingford
UK. P. 30-3.
2. Scura, R.W. (1978). Biochemistry Taxonomy and Evolution of modern cultivated
citrus. Proceedings of the International Society of Citriculture. 1: 277-289.
3. Yelenosky, G. (1985). Cold hardiness in citrus. In Horticultural Reviews Vol VII.
AVI Publishing Co., Westpot, Connecticut. Pp. 201-238.
4. Saunt, J. (1990). Citrus Varieties of the World. Sinclar International UK.
5. Lawrence, B.M. (1993). Progress in essential oils. Perfurm. Flavour, 18: 43-68
6. Mans, H.B. (1991). A critical review on the chemical composition of citrus oils.
Perfum & Flavor., 16: 26-30.
7. Ekundayo, O., Bakare, O., Adesomoju, A., Stahl, B.E. (1990). The Composition
of Lemon petit Grain Oil. (Citrus limon (L) N. L. Burm). J. Essen. Oil Res., 2: 269-
270.
8. British Pharmacopoeia, (1980). Vol II, H.M. Stationary Office, London, pA109.
9. Adams, R.P. (1989) Identification of essential oils by ion trap mass spectroscopy.
Academic Press, New York.
10. Jennings, W., Shibamoto, J. (1980). Qualitative analyses of flavor and fragrance
volatiles by capillary gas chromatography. Academic Press, New York.
11. Davies, N.W. (1990). Gas chromatographic retention indices of monoterpenes and
sesquiterpenes on methyl silicone and carbowax 20M phases. J. Chromatography,
503: 1-24.
12. Kumar, U., Ram, B., Pant A.K., and Gupta, K.C. (1992). Volatile constituents of
the distilled leaf and peel oils of Citrus limon Burm cv. “Plant Lemon-1. J. Essent.
Oil Res. 4: 643-644.
13. Malendra, F.A., Laeneina, A.J., Flores, J., and Guzman, G. (1984). Aceites
essenciales en hojas de variedades de limonero (Citrus limon L. Burm F.) An Edafol,
Agrobiol., 43: 1161-1180.
14. Mondello, L., Cotronco, A., and Giovanni, D. (1997). Italian Citrus Petitgrain
oils. Part iv, Composition of Lemon Petitgrain oil, J. Essent. Oil Res. 9: 495-508.
15. Crescimanno, F.G., Depasquale, F., Geraman, M.A., Bazan, E. and Palazzolo,
E. (1989). Annual variation of essential oil in the leaves of four lemon (Citrus limon
L. Burm. F.) cultivars. Proc. Int. Citrus Congress (Middle East, 1988), 583-588,
Margraf Scientific Books, Weikersheim, Germany. .
16. Scora, R.W., England, A.B., and Chang, D. (1969). Taxonomic affinities within
the rough lemon group (Citrus jambhiri Lush) as aided by gas chromatography of
 Adeleke Adebisi Kasali et al. / Jeobp 13 (5) 2010 pp 540 - 544 544

their essential oils. Proc. First Int. Citrus Symp. 1: 441-450.

17. Adeishili, N. and Khrarehava, L.G. (1987). Volatile components of lemon leaves.
Subtropicheskie Kul’tury, 6: 63-66.
18. Ayedoun, A.M., and Sossou, P.V. (1996). Volatile constituents of the peel and leaf
oils of Citrus limon L. Burm. F. from Benin. J. Essent. Oil Res. 8: 441-444.

Table 1. Volatile Constituents of the leaf oils

of two different Nigerian Citrus Lemons

KI Compounds Eureka Lisbon

941 -Pinene 0.5 0.7

973 Sabinene 1.8 3.9
978 -Pinene 4.5 6.2
988 Myrcene 1.4 1.5
1011 -3-Carene 0.8 0.6
1024 Limonene 30.0 34.0
1038 (E)- -Ocimene 1.9 2.5
1052 -Terpinene 0.3 0.3
1083 Terpinolene 0.3 -
1086 Linalool 1.7 1.7
1129 citronellal 1.9 1.7
1140 Isoneral 0.4 0.5
1158 Isogeranial 1.0 1.1
1165 Terpinolene-4-ol tr 0.7
1175 -Terpineol 0.7 tr
1212 Nerol - 2.8
1217 Neral 19.2 12.3
1236 Geraniol 1.8 1.0
1245 Geranial 18.7 14.4
1337 Citronelly acetate 0.2 -
1342 Neryl acetate 5.3 5.7
1360 Geranyl acetate 2.4 2.5
1391 -Elemene - tr
1423 -Caryophyllene - 0.4
1437 trans- -Begamotene - tr
1456 -Humulene tr 0.4
1499 (E,E)- -Farnesene tr 2.2
1505 -Bisabolene tr 0.3
1522 -Cadinene tr -
1553 (E)-Nerolidol tr -
1579 Caryophyllene oxide tr -
1643 -Cadinol tr -

KI: Elution order and retention indices on a Cpsil-5 column (See Experimental)
tr: trace