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Vol. 8(16), pp. 1395-1400, 2 May, 2013
DOI: 10.5897/AJAR11.1837
ISSN 1991-637X ©2013 Academic Journals
http://www.academicjournals.org/AJAR
African Journal of Agricultural
Research
Full Length Research Paper
Chemical composition of Lentisk (Pistacia lentiscus L.)
seed oil
Wissal Dhifi1,2, Nahida Jelali3, Emna Chaabani1,4, Maroua Beji1,4, Saloua Fatnassi5,
Semia Omri5 and Wissem Mnif1,
,4
4*
1Institut Supérieur de Biotechnologie de Sidi Thabet, Biotech Pole de Sidi Thabet, 2020, Université de La
Manouba, Tunisie.
2UR Ecophysiologie Environnementale et Procédés Agroalimentaires, BiotechPole de Sidi Thabet Université de La
Manouba, Tunisia.
3Laboratoire d’Adaptation des Plantes aux Stress Abiotiques, Centre de Biotechnologie, Technopole de Borj-Cedria
(CBBC), B.P. 901, 2050 Hammam-Lif, Tunisie.
4LR11-ES31 Biotechnologie et Valorisation des Bio-Géo Ressources, Institut Supérieur de Biotechnologie de Sidi
Thabet, BiotechPole de Sidi Thabet, 2020, Tunisia Université de la Manouba, Tunisie.
5Laboratory of Lipids, Institute of Research and Physico-Chemical Analysis, Technical Pole of Sidi Thabet, 2020 Sidi
Thabet, L’Ariana, Tunisie.
Accepted 29 April, 2013
We were interested in Pistacia lentiscus fixed oil which was extracted from ripe seeds. The
characterization of this oil was performed on fatty acids, triacylglycerols, tocopherols and sterols
composition. We also evaluated its mineral composition. The results showed that the prominent class
of fatty acids was represented by monounsaturated fatty acids accounting for 52.4% of the whole fatty
acids. It was followed by saturated fatty acids and polyunsaturated fatty acids accounting respectively
for 26.42, 21.18 and 11%. The major fatty acid (FA) was oleic acid with an amount of 51.06%. Linoleic
acid (C18: 2) which is an essential FA accounted for 20.71% of total fatty acids. The majority of
triacylglycerols are in mono and polyunsaturated forms. The major constituents were stearoyl-oleyl-
linoleylglycerol and palmitoyl-dioleylglycerol acounting together for 27.58% of total TAGs. Concerning
sterols, their quantity in Lentisk oil was about4.17 mg/ kg of oil. This quantity is comparable to that of
oil seed rapeseed. We noted the prevalence of β-sitosterol with an amount of 55.5%. Furthermore, P.
lentiscus oil contained 8111.137 mg of tocopherols/kg of oil. α-tocopherol which has the highest
antioxidant activity accounted for 97% of whole tocopherols in Lentisk oil. Lentisk oil was rich in
minerals. The most abundant mineral was Na, followed by K, Ca, Mg, Fe and Cu. These minerals are
essential and indispensable for the human body, for their nutritional value.
Key words: Lentisk, Pistacia lentiscus, ripe seed oil, fatty acids, triacylglycerols, sterols, tocopherols.
INTRODUCTION
Lentisk (Pistacia lentiscus L). is an evergreen shrub or
tree and an aromatic member of the Anacardiaceae
family producing bright red globose berries. It grows in
several Mediterranean region (Bonnier and Douin, 1990).
The fruits, galls, resin and leaves of the Lentisk have a
long tradition in folk medicine dating from the times
*Corresponding author. E-mail: w_mnif@yahoo.fr. Tel: + 216 98 94 73 71. Fax: + 216 70 527 600.
1396 Afr. J. Agric. Res.
of the ancient Greeks (Charef et al., 2008). The same
authors reported that in Algeria the oil of the fruit is used
by the population in traditional medicine in many ways, as
an anti-diarrhoeal and also as constituent of cattle feed.
Several studies focused on the phytochemical
composition of the resin, the leaves and the galls of
Lentisk and also on its essential oils (Castola et al., 2000;
Duru et al., 2003; Romani et al., 2002) but in contrast,
fewer studies are related to the composition of the fruit oil
(Ucciani, 1995).
It is used as an antibacterial (Iauk et al., 1996) and
antiulcer (Al-Said et al., 1986) agent. The essential oil of
Lentisk are extensively used in the perfumery and in food
and pharmaceutical industries as reported by Calabro
and Curro (1974). Lentisk oil may partially help in the
protection against mercury intoxication, and it could also
be considered a safe nutritional source, at least by
maintaining total cholesterol and LDL-cholesterol in their
normal ranges (Maarouf et al., 2008). Many works were
focused on Lentisk essential oil composition and activities
whereas few studies have focused on its fixed oil. The
fixed oil extracted from mature fruits is commonly used in
Tunisian traditional medicine as an anti-ulcer, wound
healing and antiseptic (Rejeb et al., 2006; Mezni et al.,
2012). The aim of this study was to ascertain fatty acids,
triacylglycerols, tocopherols and sterols composition of
Lentisk fixed oil. We also evaluated its mineral
composition.
MATERIALS AND METHODS
Oil extraction
Oil was extracted from Lentisk mature seeds using hexane; the
ground dried Lentisk seeds (40 g) were placed into a cellulose
paper cone and extracted with 400 ml hexane using a soxhlet
extraction apparatus for 8 h. The solvent was removed via a rotary
vacuum distillation at 40 to 50°C flushing with nitrogen to blanket
the oil during storage. The residue was weighed and stored at
−20°C until it was analysed. Oil weight was determined from 40 g of
the seed powder to calculate the lipid content. The result was
expressed as the lipid percentage in the seed powder dry matter.
Analysis of fatty acids composition
The fatty acid methyl esters (FAME) composition was determined
by the conversion of oil to fatty acid methyl esters prepared by
adding 1 ml of n-hexane to 40 mg of oil followed by 200 µl of
sodium methoxide (2 M). The mixture is heated in the bath at 50°C
for few seconds followed by adding 200 µl HCl (2 N). The top layer
(1 µl) was injected onto a GC (Agilent 6890N, California, USA)
equipped with a flame ionisation detector (FID) and a polar capillary
column (HP-Innowax polyethylene glycol, 0.25 mm internal
diameter, 30 m in length and 0.25 µm film in thickness) to obtain
individual peaks of FAME. The FAME peaks were identified
comparing their retention times with individual standards, FAME
being injected in the same analytical conditions and analysed with
the Agilent Technologies Chemstation A09.01 Software. The
relative percentage of each FA was calculated on the basis of the
peak area of a FA species to the total peak area of whole FA in the
oil sample.
Analysis of triacylglycerols composition
The triacylglycerols (TAGs) profile was obtained by a reverse phase
high performance liquid chromatography (HPLC) (Agilent 1100,
California, USA) equipped with a G1354 quaternary pump, a
G1313A standard auto sampler, and a G1362A refractive index
detector. The chromatogram was carried out using Agilent
Technology Chemstation software. The TAGs were separated using
a commercially packed Hypersil ODS column (125 × 4 mm) with a
particle size of 3 µm and were eluted from the column with a
mixture of acetonitrile/acetone (65/35) at a flow rate of 0.5 ml/min;
the TAGs were detected with a refractive index detector. Ten
microliters of 0.05 g oil diluted in 1ml (chloroform/acetone 50/50,
v/v) was injected into the HPLC. The total run time was 45 min.
TAG peaks were identified by comparison with chromatograms of
sunflower and corn oil obtained in the same analytical conditions.
Analysis of tocopherols composition
Prior to the HPLC analysis, the seed oil 0.5 g was diluted with 5 ml
hexane and 5 µl samples were injected. The tocopherol
composition of M. pomifera seed oils was determined using HPLC
according to norm ISO 9936. The sample was analysed by an
HPLC (Agilent 1100, CA, USA) consisting of a G1354 quaternary
pump, a G1313A standard auto sampler, a G1321A fluorescence
detector set at λ excitation = 295 nm, and λ emission = 330 nm and
a chemstation software. A normal phase column (Pinnacle II silica)
(150 × 3.2 mm × 3 µm) was used with hexane/isopropanol
(99.5/0.5, v/v) as a mobile phase. The system was operated
isocratically at a flow rate of 0.5 ml/min. The separations were
carried out at 30°C. The quantification was based on an external
standard method. The mixed tocopherol standards in a hexane
solution (2 mg/ml) were prepared from the standard compounds: α-,
β-, γ- and the δ-tocopherols (Sigma Chemical Co., St. Louis, MO,
USA).
Analysis of phytosterols composition
Separation of sterols (ST) was performed according to the method
ISO 12228. Lipids (250 mg) were refluxed for 15 min with 5 ml
ethanolic KOH solution (3%, w/v) after addition of cholesterol (1 mg;
Fluka) as an internal standard and a few antibumping granules. The
mixture was immediately diluted with 5 ml of ethanol. The
unsaponifiable part was eluted over a glass column packed with
slurry of aluminium oxide (Scharlau) in ethanol (1:2, w/v) with 5 ml
of ethanol and 30 ml of diethyl ether at a flow rate of 2 ml/min. The
extract was evaporated in a rotary evaporator at 40°C under
reduced pressure, and then ether was completely evaporated under
a steam of nitrogen. For the characterization of sterols, a silica gel
F254 plate (Fluka) was developed in the solvent system n-
hexane/diethyl ether (1:1, v/v). For the detection of sterols, the thin-
layer plate was sprayed with methanol; the sterol bands were
scraped from the plate and recovered by extraction with diethyl
ether. The sterols trimethylsilyl ether (TMS) derivatives were
prepared by adding 100 µl of a silylant reagent N-methyl-N-
(trimethylsilyl) trifluoroacetamide/ pyridine (1/10, v/v) in a capped
glass vial and heated at 105°C for 15 min.
Preparation of standard solutions
A mixture of standard solutions of sterols was prepared by
derivatization (cholesterol, sitosterol, stigmasterol, ergosterol and
campesterol). The sterols trimethylsilyl ether derivatives were
analysed using the GC system (Agilent 6890N, California, USA)
equipped with a FID and the GC chemstation software. A HP-5 (5%
Table 1. Lentisk seed oil fatty acids composition.
FA Amount (% of TFA)
C16 : 0 23.52 ± 3.01b
C16 : 1 1.19 ± 0.12d
C17 : 0 0.10 ± 0.01g
C18 : 0 1.41 ± 0.02c
C18 : 1 51.06 ± 4.37a
C18 : 2 20.71 ± 2.25b
C18 : 3 0.47 ± 0.10e
C20 : 0 0.14 ± 0.02f
C20 : 1 0.15 ± 0.01f
C22 : 0 1.25 ± 0.02d
SFA 26.42 ± 5.09b
MUFA 52.4 ± 7.18a
PUFA 21.18 ± 2.23b
pheynyl methyl polysiloxane column) was used (0.32 mm i.d. × 30
m in length; 0.25 µm film in thickness; an Agilent 19091J-413, CA,
USA). A carrier gas (helium) flow was 1.99 ml/min (split–splitless
injection with a split ratio of 1:200). The detector and the injector
were set at 320°C, and the injected volume was 1_l. The total
analyses were set at 71 min to ensure the elution of all ST. The
operational conditions were: injector temperature 320°C, column
temperature: a gradient of 4°/min from 240 to 255°C. Sterols peak
identification was carried out according to the ISO 12228 reference
method and confirmed by GC–MS (NIST 2002 database) operating
in the same conditions as used for the GC–FID.
Mineral composition
The mineral constituents (Ca, Na, K, Fe, Mg and Cu) present in
Lentisk seed were analysed, using an atomic absorption
spectrophotometer (NOUVA400, ANALYTIKJENA, Germany) and a
flame ionisation spectrophotometer (Flame Photometer 410,
SCHERWOOD, Germany).
Statistical analyses
All analytical determinations were performed in triplicate. The
values of different parameters were expressed as the mean ±
standard deviation.
RESULTS AND DISCUSSION
Oil characterization
Oil yield of Lentisk was of 35.37%. This yield is
appreciable and is similar to that of some oleaginous
seeds. Lentisk oil is visquous with a green colour. Charef
et al. (2008) reported that the crude fat content of P.
lentiscus varied from 32.8% for black fruits to 11.70% for
red ones. According to Karlenskind (1992), the black fruit
of Letisk can be considered as an oleaginous seed as
peanut, olive, sunflower and cotton seeds whose oil yield
range from 30 to 45%.
Dhifi et al. 1397
Fatty acid and triacylglycerol composition
The prominent class of FA was represented by
monounsaturated FA (MUFA) accounting for 52.4%. It
was followed by saturated FA (SFA) and polyunsaturated
FA (PUFA) and accounting respectively for 26.42, 21.18
and 11% of the whole FA (Table 1). The major FA was
oleic acid (C18: 1) with an amount of 51.06%. This FA is
reputed for its role in preservation of cardiovascular
diseases and its nutritional value (Corbett, 2003). Linoleic
acid (C18: 2) which is an essential FA (EFA) accounted
for 20.71% of whole FA. Furthermore, palmitic acid (C16:
0) was detected at a significant percentage of 23.52%.
C18: 2 had favorable nutritional implications and
beneficial physiological effects in the prevention of
coronary heart disease and cancer (Oomah et al., 2000).
Generally, FA and TG are able to reduce trans epidermal
water loss and so increase skin hydration (Dweck, 2002).
C18: 1 and C18: 2 are known for their anti-inflammatory
properties. Linoleic and alpha linoleic acids provide lipids
necessary for cell membrane repair and cellular
respiration (Loden and Andersson, 1996). Djerrou et al.
(2010) reported that in Lentisk oil, the three dominant FA
were 18: 1 (55.3%), C18: 2 (17.6%) and 16:0 (16.3%).
The FA composition of our sample is similar to that of
Algerian Lentisk oil (2) whose major FA was C18: 1 with
an amount varying from 55.3 to 64%. In Algerian Lentisk
oil, C18: 2 was characterized by a significant percentage
(17.6 to 28.4%). In Pistacia terebinthus oil, the
dominating FA of the oil is C18: 1, which accounted for
43.0 to 51.3% of the total FA (Matthäus and Özcan,
2006). In addition, our results for Lentisk oil agree well
with the data recorded by Ucciani (1995) in his dictionary.
It is to note that the UFA/SFA ratio was equal to 2.78.
Furthermore, the profile of FA confirms the similarity
between Pistacia lentiscus oils and other edible
vegetable oils such as sunflower, peanut, cotton, olive
and avocado.
Furthermore, the low saturated/unsaturated FA ratio
(0.35) reveales a high content in UFA which may give it
nutritional and dietetic virtues. The FA profile of Lentisk
seed oil is similar to that of Pistacia vera (Chahed et al.,
2006) and Pistacia atlantica (Ghalem and BenHassaini,
2007). Lentisk seed oil could be used for nutritional
purposes as an interesting source of omega 6 and
omega 9 FA. The TAGs composition of Lentisk showed
that the majority of TAGs are in mono and
polyunsaturated forms (Table 2). Considering the fatty
acid composition; the major constituents were stearoyl-
oleyl-linoleylglycerol (SOL) and palmitoyl-dioleylglycerol +
(POO) accounting together for 27.58% of total TAGs.
Stearoyl-dilinoleoylglycerol (SLL) and palmitoyl-oleyl-
linoleoylglycerol (POL) represented 21.5% of total TAGs
whereas Trioleylglycerol (OOO), dioleyl-linoleylglcerol
(OOL) and dipalmitoyl-oleylglycerol (PPO) were signify-
cantly represented with respective amounts of 12.04,
9.83 and 8.51%. Furthermore, the trilinoleyl-glycerol (LLL)
1398 Afr. J. Agric. Res.
Table 2. Lentisk seed oil triacylglycerols composition
Tag Amount (% of total tags)
LLLn -
LLL 1.32 ± 0.28f
OLLn -
OLL 5.67 ± 1.62e
PLL 7.97 ± 1.86de
OOL 9.83 ± 2.03cd
SLL + POL 21.50 ± 2.06b
PPL 5.58 ± 1.12e
OOO 12.04 ± 1.43c
SOL + POO 27.58 ± 2.36a
PPO 8.51 ± 1.09d
LLL: Trilinoleoyl-glycerol, OLLn: Oleyl-linoleoyl-
linolenoylglycerol, OLL: Oleyl-dilinoleoyl-glycerol, PLL:
Palmitoyl-dilinoleoyl-glycerol, OOL: Dioleyl-
linoleoylglcerol, SLL: Stearoyl-dilinoleoylglycerol, POL:
Palmitoyl-oleyl-linoleoylglycerol, PPL: Dipalmitoyl-
linoleoylglycerol, OOO: Trioleylglycerol, SOL: Stearoyl-
oleyl-linoleoylglycerol, POO: Palmitoyl-dioleylglycerol,
PPO: Dipalmitoyl-oleylglycerol
Table 3. Lentisk seed oil tocopherols composition.
Tocopherol Quantity
(mg/ g of oil)
Amount (% of
total tocopherols)
α-tocophérol 7.59 ± 0.61a 93.62a
β-tocophérol 0.47 ± 0.02b 5.79b
γ-tocophérol 0.48 ± 0.04b 0.59c
δ-tocophérol - -
was characterized by a low percentage (1.32%)
Unsaponifiable compostion of P. lentiscus seed oil
The unsaponifiable fraction of oils contains tocopherols,
sterols and phenolic components. However, no study has
been published which concern this fraction. It is important
to mention that Mattähaus and Özcan (2006) quantified
FA, tocopherols and sterols in Pistacia terebinthus Chia.
The stability of vegetable oils under the conditions of
oxidation is due to the presence of high levels of natural
antioxidants, the most important are the tocopherols,
which come in four isomeric forms: α, β, γ and δ. Lentisk
oil contained 8111.137 mg of tocopherols/ kg of oil. It
should be noted that this oil is rich in tocopherols some
vegetable oils such as corn oil (1111.4 mg/ kg of oil) and
rapeseed oil (820 mg/ kg of oil) and sunflower seed oil
(734 mg/ kg of oil) according to Ayerdi (2008). α-
tocopherol which had the highest antioxidant activity
accounted for 93.62% of whole tocopherols in Lentisk oil.
The isomers β and γ were detected with respective
amounts of 5.79 and 0.59% (Table 3). δ-tocopherol was
not detected. Matthäus and Özcan (2006) reported that
seed oil of P. terebinthus was characterized by the
predominance of α- and γ isomers of tocopherols. It also
contained different tocotrienols, with γ-tocotrienol as the
dominate compound of this group. The effect of the
environment on the temperature seems to be a key factor
in the accumulation of tocopherols (Ayerdi, 2008).
According to this author, the amount of α–tocopherol
increases in sunflower oil during the maturation parallel
with a decrease in that γ-tocopherol which is the
precursor of α-tocopherol. The results of quantitative
determinations of α-tocopherol in P. lentiscus var. chia, in
and P. terebinthus leaves by TLC-densitometry and
colorimetry. The maximum content of α-tocopherol was
found in the leaves of P. lentiscus var. chia (Kivçak and
Akay, 2010). Consumption of food rich in natural
antioxidants is protective against some types of cancer
and may also reduce the risk of cardiovascular and
cerebrovascular events Aruoma (1998). These actions of
antioxidants have been attributed to their ability to
scavenge free radicals, thereby reducing oxidative
damage of cellular biomolecules such as lipids, proteins,
and nucleic acids (Ferguson, 1995).
This richness in tocopherols, including the
predominance of α-tocopherol, which is a very good
antioxidant fatty phases, contributes to the natural
protection and conservation of the oil against oxidation. It
is important to mention that Lentisk oil has a good
vitaminic activity due to its high content of vitamin E.
According to Table 4, the oil of Lentisk contained β-
sitosterol as the major phytosterol (55.55%), followed by
cholesterol (44.45%). However, stigmasterol and other
sterols were not detected. They may disappear during
maturation. It should be noted that the respective
quantities of β-sitosterol and cholesterol in our sample
were of 231.67 mg/ 100 g of oil and 185.35 mg/ 100 g of
oil.
The whole quantity of sterols in Lentisk oil was of 4.17
mg/kg of oil. This quantity is comparable to that of oilseed
rape which is one of the most important oil seed crops in
the world. Phytosterols content in new oilseed rape
varieties ranges between 5.13 and 9.79 g/kg oil. It is
important to note that the phytosterols content ranges
between 1.41 and 15.57 g/ kg oil and depends of plant
species (Gul and Amar, 2006).
In P. terebinthus L. seed oil, the total content of sterols
ranged between 1341.3 and 1802.5 mg/kg, with β-
sitosterol as the predominant sterol accounting for more
than 80% of the total amount of sterols (Matthäus and
Özcan, 2006). The levels of tocopherols and phytosterols
in oil seeds (rapeseed and soybean) are highly
dependent on both genetic factors and plant
environmental conditions of cultivation (Abidi, 2003). In
recent years increased interest in phytosterols lies in their
potential to reduce plasma low-density lipoprotein
cholesterol level, decreasing coronary mortality and
therefore acting as naturally preventive dietary product
(Gul and Amar, 2006). It has been found that plants that
Table 4. Lentisk seed oil sterols composition.
Sterol Quantity (mg/ 100 g of oil) Amount (% of
total ST)
β-sitostérol 231.67 ± 10a 55.55
Cholesterol 185.35 ± 22b 44.45
Table 5. Lentisk seed oil mineral composition.
Mineral Quantity (mg/ 100 g of oil)
Na 25.36 ± 3.25a
K 2.17 ± 0.05b
Ca 0.25 ± 0.04c
Mg 0.19 ± 2.23d
Fe 0.004 ± 0.00tr
Cu 0.0001 ± 0.00tr
have cicatrizing and vulnerary properties often have a
high level of plant sterols (Dweck, 2002). Nuts contain
bioactive constituents that elicit cardio-protective effects
including phytosterols, tocopherols and squalene.
Mineral composition of the seed of P. lentiscus
The mature seeds of Lentisk are rich in minerals. The
most abundant mineral is Na, followed by K, Ca, Mg, Fe
and Cu (Table 5). These minerals are essential and
indispensable to the human body, for their nutritional
value. The mineral composition of P. lentiscus seed
revealed its nutritional value for human and/or animal
consumption. According to Ferguson (1995), pistachio is
a rich source of phosphorus, potassium, magnesium,
calcium and iron. The importance of mineral composition
is due to their nutritional properties and beneficial health
effects, as well as their meeting of dietary guidelines
required for a healthy diet (Welna et al., 2008).
According to this study, the seeds of Lentisk seemed to
be a good source of oil. This oil had an interesting FA
composition. It is rich in C18: 1 which is reputed for its
role in prevention of cardiovascular diseases and also in
C18: 2 which is an essential FA with beneficial
physiological effects in the prevention of coronary heart
disease and cancer. This oil is characterized by an
interesting composition of nutritional point of view. In a
further work, we will try to complete this research by
exploring phenols composition and some physico-
chemical properties of Lentisk oil. Such study could lead
to industrial applications particularly in cosmetics
industry.
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