SUPPLEMENTARY MATERIAL
Identification and determination of the fatty acid composition of
Quercus brantii growing in southwestern of Iran by GC-MS
S. Khodadousta*, A. Mohammadzadehb, J. Mohammadia, C. Irajiec, M. Ramezanib
a
b
Medicinal plants Research Center, Yasuj University of Medical Science, Yasoj, Iran
Department of Chemistry, Faculty of Science, Islamic Azad University, Arak Branch, Arak, Iran
c
Shiraz University of Medical Science, Shiraz, Iran.
* Corresponding author; Email: khodadoustsaeid@yahoo.com , Tel/Fax: +987412235137
Abstract:
This paper reports the fatty acid composition of the oil extracts from the Quercus brantii fruits growing in
Kohgiloye va Boyer Ahmad province in southwestern of Iran. The Quercus brantii fruit’s oil were extracted with
hexane in soxhlet apparatus and subsequently identified and determined by gas chromatography–mass spectroscopy
(GC–MS). The results revealed that major fatty acid are oleic acid (52.99–66.14%), linoleic acid (10.80-11.11%),
palmitic acid (8.08–10.06%), stearic acid (0.74–1.57%), α-linolenic acid (0.19–0.35%), erucic acid (0.12-0.15%)
and arachidic acid (0.12-0.13%). The total unsaturation and saturation for the oil were 64.60-77.27 % and 9.1711.75 %, respectively. Results indicate that the fruits of Quercus brantii contained 0.19-0.35 % omega-3, 10.9214.77 % omega-6 and 53.14- 66.26 % omega-9. Therefore Quercus brantii can be introduce as rich sources of fatty
acid in food dietary and medical health.
Keywords: Quercus brantii; Fatty acid composition; GC-MS; Fruit
1.
Experimental
1.1. Sample preparations
Fruits of Quercus brantiii collected from the protected areas at Yasouj, Sisakht and Kakan reigns by the
Environmental Protection Agency in Kohgiloyeh and Boyer Ahmad province of Iran in September 2011. In these
regions fruits were collected in 10 points with 1 km distance in all direct. In any sampling points, fruits were
gathered from 10 trees. The plants were identified by taxonomic references (Rechinger, 1963–1998) and Mr. A.
Jafari, Senior Experts, Herbarium of Agricultural Faculty of Yasouj University, Yasouj, Iran (Voucher specimen no.
850). In order to preserve their original quality, the fruit parts were stored in a refrigerator at 4 ± 1 ◦C until drying
experiments. All fruits were cleaned and washed several times with double-distilled water (Altundag and Tuzen
2011; Dundar et al., 2012). To obtain a homogeneous particle size of the grinding sample, the procedure of coning
and quartering (Aberoumand and Deokule 2009) was employed with appropriate amounts of each variety of fruit of
Quercus brantii.
1.2. Fat extraction
The 10 g fruits were milled into powder using a manual mill and extracted with 250 mL hexane in Soxhlet
apparatus in order to determine the amount of fatty acids in the fruits, followed by agitation the sealed Erlenmeyer
flasks (200 rpm) at room temperature for 48 h. After the extraction process, the flask contents were filtered under
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vacuum, and the liquid fraction containing lipid extract and solvent was poured into a 500-mL flask of a rotary film
evaporator to remove the solvent. Then 2 mL KOH (methanolic) and 7 mL hexane was added to the obtained extract
and hold in thermoregulated bath for 15 min at 70 oC in order to methylation of composition. Subsequently the
extract was filtered and dehydrated with anhydrous sodium sulphate and injected to gas chromatography–mass
spectroscopy (GC–MS).
1.3. GC-MS analysis of fatty acids
Gas chromatography-mass spectrometry (GC-MS) analyses were performed using an Agilent gas
chromatograph 7890A (Agilent, little Falls, DE, USA) coupled with an electronically controlled split/splitless
injection port and interfaced to a MSD-5975C mass selective detector. The gas chromatograph was equipped with a
DB-5MS fused silica capillary column (30 m × 0.25 mm × 0.25 µm) purchased from J&W scientific (Folsom, CA,
USA). Helium (99.99%) was used as carrier gas, with a flow rate of 1 ml min-1. The injection was performed at 250
ºC in the split mode (ratio 20:1). The column was first set at 50 ºC, temperature was subsequently increased to 150
ºC at the rate of 25 ºC/min and hold constant for 3 min, then to 230 ºC at the rate of 3 ºC/min and held 1 min. This is
the optimum temperature programming for the conditions of both separation effect and run time. Mass spectrometer
conditions were set as follows: ionisation mode: EI; electron energy 70 eV.
1.4. Statistical analysis
All the analyses were performed in triplicate for each sample and expressed as mean ±SD. Correlations between
parameters were calculated using the simple Pearson correlation coefficient. The significance of differences was
tested by two-way analysis of variance (ANOVA) followed by Duncan’s multiple range tests. The P-values of <
0.05 were considered significant.
References
Aberoumand, A., Deokule, S.S., (2009) Determination of Elements Profile of Some Wild Edible Plants. Food Analytical Methods, 2, 116-119.
Altundag, H., Tuzen, M., (2011) Comparison of dry, wet and microwave digestion methods for the multi element determination in some dried
fruit samples by ICP-OES”, 2011, Food and Chemical Toxicology, 49, 2800-2807.
Dundar, M.S., Altundag, H., Eyupoglu, V., Keskin, C. S., Tutunoglu, C., (2012) Determination of heavy metals in lower Sakarya river sediments
using a BCR-sequential extraction procedure, Environmental Monitoring and Assessment, 184, 33-41.
Rechinger, K.H., (1963–1998) Flora Iranica, vol. 1–173. Akademische Druck und Verlagsanstalt, Graz, Austria.
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