The genus Citrus (Rutaseae) comprises various species, varieties
and hybrids, most of them are found in the north, south and
south-eastern parts of Iran (1). Among them, Citrus aurantium L.
is the unique species, with pharmacological applications. The
Hydrolate of its flowers has been used in traditional medicine
as a remedy for the treatment of mild depression, sedation and
as a heart tonic (2, 3).
A search through the literature revealed
some reports on the chemical composition and pharmacological
properties of C. aurantium. The physicochemical indices of neroli oils from
different origins were firstly reported in 1949. GC analysis
was then later made by Prager and Miskeiwicz (4). The
quantitative composition of the flower water absolute was also
studied and it was found that the major components were
linalool (44.2%), a-terpineol (18.5%), and graniol (6.4%) (5).
Neroli is known to be an antidepressant,
antibacterial and cytophylactic agent. Results showed that
patients given Neroli experienced less anxiety compared to
those not receiving it (6). Neroli is also reported to help
promote skin renewal and in addition its smell is helpful in
easing stress and also particularly useful for cases of mild
Although several studies have been
performed on the flower water absolute from other countries, no
research has so far been conducted on the traditional used of
its Hydrolate in Iran. In this work, GC/MS analysis was used
for the Citrus aurantium Hydrolate and its comparison it with the
commercial samples obtained traditionally and industrially.
The flowers of C. aurantium were
collected from Darab on the south of Fars province in Iran at
an altitude of 1107 m, during April 2003. Voucher specimens
have been deposited in the herbarium of Faculty of Pharmacy,
Tehran University of Medical Sciences, Tehran, Iran. The
traditional Hydrolate sample was obtained from the local
producer and the industrial Hydrolate sample was purchased from
the market (Camo Trade Mark).
The oil isolation procedure was performed
via a Clevenger-type apparatus using air-dried flowers (100 g)
by the hydrodistillation method for 3.5 h. Liquid-liquid
extraction for obtaining the aromatic fraction was carried out
on 200 ml of Hydrolate with normal pentane (99+%, Merck). The
organic layer was separated, dried over anhydrous sodium
sulphate and the solvent was evaporated at room temperature.
Identification of oil components
GC/MS was performed on a Thermoquest 2000
with a quadropole detector, on a DB-5 capillary column (30 m x
0.25 mm; 0.25 mm film thickness). The carrier gas was helium with a
flow of 1.5 ml/min, split ratio of 1/25 and a flame ionisation
detector. The column temperature was programmed at 50°C for
1 min and then heated to 260°C with a 2.5°C/min rate
and then kept constant at 260°C for 20 min. The MS operated
at 70eV ionisation energy. Mass range was from m/z 50-300 amu.
Retention indices were calculated by using retention times of
n-alkanes, which were injected at the same chromatographic
conditions. The compounds were identified by comparison of the
relative retention indices (RRI, DB-5) with those in the
literature (8) and by computer searching followed by matching
the mass spectra data with those stored in the computer
Results and Discussion
Three kinds of hydrolates (laboratory
obtained, traditional and industrial samples) were extracted
using pentane by a liquid-liquid extractor. The yield of oils
obtained was 0.015, 0.003 and 0.002 percents, respectively. The
chemical and class composition of the oils are presented in
Tables 1 and 2.
The laboratory obtained Hydrolate was
mainly consisted of alcohol monoterpenes (66.1%) and the major
compounds were geraniol (26.6%), a-terpineol (20.7%) and linalool (15.4%). In the
traditional hydrolate, although the main constituent was
alcohol monoterpenes (50.5%), but the chief components were
linalool (44.1%), methyl anthranilate (11.8%) and linalool
oxide (6.1%), which were different from the laboratory obtained
Hydrolate to some extent. Despite of the other results,
hydrocarbon monoterpenes (49.2%) were the predominant
constituents within the industrial sample, followed by alcohol
monoterpenes (22.3%), ether and oxide monoterpens (16.9%).
The major components in the industrial
Hydrolate were limonene (46.6%) and 1,8-cineol (15.9%).
Limonene, however, was not found in the laboratory obtained and
also traditional hydrolate, as already reported that it was
present in lower than twenty four percent in Neroli oil and
five percent in Hydrolate (9). However, 1,8-cineol was observed
only in this hydrolate.
According to the literature (10), the
percentage of limonene observed in this sample could be a case
of adulteration. It seems that it is contaminated with the
flowers and other parts of the plant. Citrus species peels
normally contain more than 70% limonene (9). The differences
observed in the traditional and laboratory obtained Hydrolate
have presumably risen from the applied fresh flowers in the
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Dictionary of Iranian Plant Names. Farhang Mo'aser Publishers, Tehran (1996) 131
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Pharmacognosy and Medicinal Plant of Iran. Tehran University Publication, Tehran
(4) Prager M and Miskiewicz M. Gas
chromatographic-mass spectrometric analysis, identification and
detection of adulteration of perfumery products from bitter
orange trees. Assos. Off. Anal.
Chem. (1981) 64: 131-138
(5) Peyron L and Bonaccorsi I. Extracts
from the bitter orange flowers (Citrus
aurantium L.): composition and
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Inc., London (2002) 413-424
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fragrance use. Cosmetics and
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(7) Mantel F. The role of alternative
medicine in treating postnatal depression.
Complementary Therapies in Nursing and Midwifery (2002) 8: 197-203
(8) Adams R P.
Identification of Essential Oil Components by Gas
Chromatography/Mass Spectroscopy. Allured Publishing Co., New York (1995)
(9) Dugo G, Cotroneo A, Verzera A and
Bonaccorsi I. Composition of the volatile fraction of
cold-pressed citrus peel oils. In: Dugo G. (ed.)
& Francis Inc., London (2002) 202-231
(10) Juchelka D, Steil A, Witt K and Mosandl
A. Chiral compound of essential oil. XX. Chirality evaluation
and authenticity profiles of neroli and Petitgrain oil.
J. Essent. Oil Res. (1996) 8: 487-497