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JOURNAL OF FOOD COMPOSITION AND ANALYSIS

Journal of Food Composition and Analysis 19 (2006) 676680 www.elsevier.com/locate/jfca

Original Article

Determination of fatty acids and total lipid content in oilseed of 25 pomegranates varieties grown in Iran
Abolfazl Fadavi, Mohsen Barzegar, Mohammad Hossein Azizi
Department of Food Science and Technology, Tarbiat Modarres University, P. O. Box 14115-336, Tehran, Iran Received 3 January 2004; received in revised form 4 September 2004; accepted 6 September 2004

Abstract The fatty acids composition of the seed oils of 25 pomegranates varieties obtained from two different regions of Iran (Punica granatum L.) was qualitatively and quantitatively determined by gas chromatography. The seeds contained oil in the range of about 66.3193 g kg1 dry matter, a notably higher content than western pomegranate cultivars. Levels of lipid content probably could be considered insufcient for economic industrial exploitation except for medical usages and specic consumption. The pre-dominant fatty acid was linolenic acid (C18:3) and its content was about 31.886.6%, followed by linoleic acid (0.724.4%), oleic acid (0.417.4%), stearic acid (2.816.7%), and palmitic acid (0.39.9%). To a lesser extent, the saturated myristic acid (0.14.7%) and behenic acid (0.03.9%) were also found in some cultivars. We have not conrmed the presence of lauric, arachidic and lignoceric acids previously reported in edible and non-edible pomegranate cultivars. Intervarietal differences in fatty acid compositions were shown and they could be useful to establish chemotaxonomic differences. r 2004 Elsevier Inc. All rights reserved.
Keywords: Punica granatum L.; Total lipids; Fatty acids; Oilseed; Pomegranate

1. Introduction Punica granatum L., the pomegranate belongs to the Punicacea family (Harde et al., 1970) and is one of the oldest edible fruits. It has been cultivated extensively in Mediterranean countries, Iran, India and to some extent in the U.S. (California), China, Japan and Russia. Iran is a native land of the pomegranate which is grown in every area, both coastal and mountainous areas. The total pomegranate production in Iran was 665,000 tons in 2003 (Anonymous, 2003). The edible parts of pomegranate fruits are consumed fresh. They are also used in the preparation of fresh juice, canned beverages, jelly, jam and paste and for avoring and coloring drinks, etc. (Ewaida, 1987; Hodgson, 1971; Nagy et al., 1990). Pomegranate fruits contain considerable amounts of seeds, ranging between
Corresponding author. Tel.: +98 21 41965223; fax:+98 21 4196524. E-mail address: mbb@modares.ac.ir (M. Barzegar).

40 and 100 g kg1 of fruit weight depending on cultivar. The seeds of diverse varieties are rich source of total lipids. Data on fatty acid composition in the seed oil of pomegranate also help to establish the chemotaxonomic relationships among the studied varieties. Vegetable oils nowadays are a great source of maintaining oil consumption in families and because of consumers concern with the saturated/unsaturated fatty acid ratio in the diet, the lipid composition of fruit and vegetable has lately received particular attention. Consumers are especially interested in essential fatty acids, with emphasis on the health potential of polyunsaturated fatty acids. It is considered that these fatty acids play a natural preventive role in cardiovascular disease and in alleviation of some other health problem, because they promote the reduction of both total and HDL cholesterol (Melgarejo and Artes, 2000). Also, pomegranate seed oil and extracts might be employed in menopausal women as external and internal phytoestrogen medicaments, as a possible alternative or supplement to conventional hormone replacement therapy (HRT) (Lansky, 1999).

0889-1575/$ - see front matter r 2004 Elsevier Inc. All rights reserved. doi:10.1016/j.jfca.2004.09.002

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Little attention has been paid to the study of chemical composition, and particularly the fatty acid composition, of ripe pomegranate seed oils from fruit varieties growing in two Iranian pomegranate research centers, Yazd and Saveh. El-shaarawy and Nahapetian (1983) showed that 8% of fatty acids of pomegranate seed oil were saturated, 10% monosaturated, 10% disaturated and approximately 70% conjugated acid, most probably punicic acid. El-Nemr et al. (1990) showed that 83.6% of the fatty acid was saturated and 16.3% unsaturated. Melgarejo et al. (1995) reported that (among six cultivars of Spain pomegranate) 3033.8% of the fatty acids were saturated and 66.269.0% of the fatty acids were unsaturated. Melgarejo and Artes (2000) studied seven clones of pomegranate from another part of Spain and found that 4.1626.65% of fatty acids of seed oils were saturated and 73.495.8% of fatty acid was unsaturated. The present study reports the total lipid content as well as the nature of the fatty acids of seed oils from 25 pomegranate cultivars which grow in two important regions of Iran.

(12), Tabestani (13), Pust Sefeede Torsh (14), Zaghe Yazdi (15), Sefeede Robi Aval Brojen (16), Gorche Shahvar Yazdi(17), Malase Yazdi(18), Malase Porbarij Stahban(19), Khoram Dizin Torsh Gorgan(20), Mesri Torshe Kazeron (21), Jangali Pust Germeze Rodbar Torsh (22), Ardestani Torshe Semnan (23), Torshe Mamoli Lasjer (24) and Pust Syahe Yazd (25). 2.2. Extraction of seed oils Three individual 10 g samples of crushed dry seeds of each pomegranate variety were reuxed with 300 mL of petroleum benzene in weighed asks using a Soxhlet apparatus according to the AACC (1987) method. The oils were recovered by distilling the solvent in a rotary evaporator at 45 1C, then dried to constant weight in a vacuum oven at 90 1C for 1 h and weighed. 2.3. Fatty acid analysis Fatty acid methyl esters (FAMEs) were prepared, following the procedure described by AOAC (1990). Three aliquots of 0.2 g lipid extract for each pomegranate variety were esteried with 10 mL methanolic NaOH solution by reuxing for 10 min at 85 1C. After addition of internal standard (0.1 mL of 2 g/L, C17:0) and 4.4 mL of BF3etherate, the samples were boiled for 2 min.The FAMEs were extracted from a salt saturated mixture with hexane (3.0 mL). For drying FAMEs, anhydride Na2SO4 was added and centrifuged (7 min, 2500 rpm, Kubota, 6900 Japan), then the upper part was poured in specic cell. The esters were separated by GC (Unicam Pu 4550, UK) tted with a capillary column (FFAP, 25 m, 0.25 mm i.d., 0.22 mm lm thickness). Helium was used as carrier gas at inlet pressure of 1.2 kg/cm2. The temperatures of injection port and detector (FID) were maintained at 200 and 240 1C and the temperature programming for the column was applied as follows, 170 1C (4 min), then to 180 1C at 3 1C/min, then to 190 1C at 1 1C/min (25 min). The peaks were identied based on their retention times using authentic standard fatty acids methyl esters and all samples were run in duplicate. 2.4. Materials All solvent/chemicals used were of analytical grade and obtained from Sigma (MO, USA) and Merck (Germany). 2.5. Statistical analysis The statistical examination of the data was performed using MSTATC software package. Mean percentages of each fatty acid in different varieties were compared using the t-test.

2. Materials and methods 2.1. Samples The seeds were obtained from mature fruits growing in Markazi province (10 varieties) and Yazd province (15 varieties) in Iran from the agricultural research centers of Saveh and Yazd, both cultivating specic varieties (more than 100). These two provinces (Markazi and Yazd) besides Khorasan and Fars, represent more than 25% of the total production among 28 provinces (Anonymous, 2001). Commercially ripe fresh fruits were harvested during September and November from different mature trees randomly selected to represent the population of the plantation. Fruits were transported by a ventilated car to the laboratory, where pomegranates with defects (sunburns, cracks, cuts and bruises in husk) were discarded. Physical and chemical parameters at harvest (fruit, seed, skin weights, and titratable acidity, total solid soluble in juice) were analyzed (data are not shown) to characterize the pomegranates. Approximately 2 kg n 10 of pomegranates at harvesting maturity was sampled for each variety, from which varietal composites were prepared. The sweet varieties were Alak-e-Shirin (1), Bihasteh (2), Syah (3), Agha Mohammad Ali Shirin (4), Pust Sefeede Shirin (5), Tabo Larze Aban Mahi (6), Malase Shirin (7) and Shirin Shahvare Yazd (8). The sour sweet varieties were: Malase Torsh (9), Togh Gardan (10), and Syahdane Shahvar Kan (11). The sour varieties were Alak-e-Torsh

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Table 1 Lipid1 content (g kg 1 dry matter) and fatty acid composition2 of oil (fatty esters as % (w/w) total fatty acid esters) from seeds of 25 varieties of pomegranate Lipids C14:0 C16:0 C18:0 C18:1 C18:2 C18:3 C22:0 S Sat. S Unsat. Sat./Unsat.

No.

Varieties

Sweet 1 2 3 4 5 6 7 8 82.072.5 12371.5 66.371.8 84.373.0 13472.8 72.371.1 95.070.9 12272.4 97.4725.6a ND ND ND 3.9 ND ND ND ND 0.5a 0.03 ND ND 0.01a 0.7 ND ND ND 0.5 ND 0.2 ND 0.1 ND 0.1 ND ND ND 0.1a 12.170.6 6.470.2 9.270.4 9.272.9a 9.870.9 6.170.6 8.470.5 7.670.8 9.570.3 26.871.3 10.571.0 5.770.8 4.870.2 6.770.4 6.870.3 5.670.6 8.770.9 4.970.4 8.776.6a 12773.1 12472.2 19371.5 148729.0b 5.970.4 5.370.2 7.570.4 6.271.1a 5.870.7 4.870.6 6.070.5 4.970.8 4.070.3 16.771.2 6.171.0 4.670.8 4.070.2 4.570.3 6.070.3 3.870.5 5.270.6 3.770.4 5.774.1a 3.270.5 0.570.2 2.170.1 2.370.2 0.370.07 9.970.6 3.970.3 0.670.1 0.370.04 1.670.2 0.570.05 1.770.3 2.770.6 0.470.03 2.173.1a 10.171.2 4.870.4 10.570.5 8.170.6 11.770.3 17.472.5 16.171.7 0.470.1 4.670.2 8.771.1 11.371.4 7.670.8 8.870.6 6.470.5 9.075.6a 9.471.3 4.770.2 10.470.5 8.871.0 13.271.2 24.471.5 21.071.5 7.370.9 8.370.5 7.770.2 11.871.0 8.070.5 9.570.3 6.170.7 10.876.9a 70.870.5 84.571.6 70.771.5 75.572.1 65.672.3 31.872.4 52.473.5 86.671.0 82.374.1 76.873.2 70.071.5 78.871.1 73.171.0 82.672.0 71.5717.9a 5.770.5 0.370.1 0.770.1 2.273.0a 6.970.6 1.570.5 10.771.0 6.474.6a 9.070.5 8.270.5 13.872.0 10.373.0a 72.071.0 84.071.5 66.370.8 74.179.0a 17674.2 12271.2 10771.3 15172.7 12772.0 11271.3 12671.5 13572.6 15373.2 15272.4 13472.1 14271.8 11171.5 12573.0 134719.4b 0.170.02 0.870.02 0.370.01 0.470.04 4.770.08 0.270.04 0.370.06 0.570.02 0.470.01 0.670.05 0.270.03 0.170.00 0.870.07 0.870.05 0.771.5a 0.570.06 0.870.01 1.070.05 0.7670.3a 0.170.01 0.370.02 1.270.01 3.470.04 0.270.00 0.470.05 0.370.02 0.870.03 0.871.0a 6.171.0 5.370.8 6.070.5 2.870.2 12.370.3 5.270.5 5.170.4 5.570.3 6.072.7a 0.670.1 2.970.0 2.770.2 1.170.1 0.870.1 2.470.3 2.070.4 2.270.1 1.870.9a 12.771.2 10.470.9 6.370.5 9.072.0 16.272.1 10.371.5 9.071.0 10.170.8 10.572.9a 12.070.5 5.470.4 0.770.2 10.470.8 14.671.2 10.971.5 8.970.9 9.871.0 9.174.3a 68.572.5 75.773.5 83.173.7 69.472.0 55.872.1 70.871.9 74.671.3 71.670.9 71.277.7a 6.871.0 8.570.8 9.970.5 11.270.2 13.370.3 8.070.6 7.470.6 8.570.3 9.272.2a

Alak-e-Shirin Bihasteh Syah Agha Mohammad Ali Shirin Shirin Pust Sefeed Larze Aban Mahi Tabo Shirin Malase Shirin Shahvare Yazd Average

93.272.8 91.573.6 90.173.7 88.872.9 86.773.2 92.072.8 92.671.9 91.571.6 90.872.1a 87.971.3 93.671.7 90.872.4 90.872.8a 90.271.8 93.971.7 91.671.7 92.472.4 90.572.6 73.273.8 89.574.2 94.371.3 95.274.1 93.373.4 93.272.3 94.471.5 91.371.2 95.172.2 91.376.8a

0.0770.01 0.0970.01 0.1170.01 0.1370.00 0.1570.01 0.0970.01 0.0870.01 0.1070.00 0.1070.03a 0.1070.01 0.0870.00 0.1070.01 0.1070.01a 0.1170.01 0.0670.01 0.0970.01 0.0870.01 0.1070.00 0.3770.03 0.1270.01 0.0670.01 0.0570.00 0.0770.00 0.0770.00 0.0670.01 0.0970.01 0.0570.00 0.0970.01a

A. Fadavi et al. / Journal of Food Composition and Analysis 19 (2006) 676680

Sour sweet 9 10 11

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Torsh Malase Gardan Togh Syahdane Shahvar Kan Average

Sour 12 13 14 15 16 17 18 19 20 21 22 23 24 25

Alak-e-Torsh Tabestani Pust Sefeede Torsh Zaghe Yazdi Sefeede Robi Aval Brojen Gorche Shahvar Yazdi Malase Yazdi Malase Porbarij Stahban Khoram Dizin Torsh Gorgan Torshe Kazeron Mesri Jangali Pust Germeze Rodbar Ardestani Torshe Semnan Torshe Mamoli Lasjer Pust Syahe Yazd Average

a,b means in a column with the same letter are not signicantly different, Po0.05. 1,2 Values are average of three individual samples each analyzed in duplicate, 7 standard deviation.

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3. Results and discussion The amount of total lipid on dry matter ranged between 66.3 and 193 g kg1 in all varieties, but in sweet varieties lipid content was between 66.3 and 134 g kg1, whereas in sour sweet varieties, it was between 124 and 193 g kg1, and in sour varieties, between 112 and 176 g kg1 (Table 1). The observation on sour and sweet varieties is in agreement with results obtained by Melgarejo et al. (1995). Our results on lipid content of sour sweet varieties are in contrast with those reported by Melgarejo et al. (1995). Our results on lipid content of sweet varieties are in contrast with those reported by El-shaarawy and Nahapetian (1983). They found a higher lipid content (161 g kg1) in a sweet variety grown in Iran. El-Nemr et al. (1990) reported a high level of lipid content (272 g kg1) in an unidentied Egyptian variety which contradicts our results. Our results show that lipid contents of different varieties are as follows: sweetosourosour sweet. Further, an ornamental non-edible sour variety (Nana cv) was shown by Tsuyaki et al. (1981) to contain 78 g kg1 of total lipid. Thus it can be seen that lipid content of different pomegranate varieties seed oil can differ from some previously studied varieties. Finally, these lipid contents of pomegranate seeds could probably be considered insufcient for economic industrial exploitation compared with those of conventional oilseeds (Al-Maiman and Dilshad, 2002). Seven fatty acids were identied in seed oils of 25 pomegranates varieties. Individual percentages of each fatty acid are given in Table 1. Linolenic acid was determined to be the pre-dominant fatty acid in 25 pomegranates varieties. Its amounts ranged between 31.8 and 86.6%. Linoleic acid was determined to be the second most abundant in these samples. Percent of linoleic acid was between 0.7 and 24.4. Oleic, stearic, and palmitic acids were determined in all pomegranate varieties. Their amounts ranged between 0.417.4, 2.816.7, and 0.39.9%, respectively. To a lesser extent, the saturated myristic (0.14.7%) and behenic acid (0.03.9%) were also found in these samples. It has been conrmed that the unsaturated fatty acids were pre-dominant in all varieties. However, linolenic acid was the main fatty acid in the present study. We could not conrm caprylic and stearic acids as the major fatty acids reported in a sweet Egyptian pomegranate cultivar (El-Nemr et al., 1990). Also, our results did not conrm the presence of caproic, capric, myristoleic acids as reported by El-Nemr et al. (1990). Neither could we conrm the presence of C12:1, C15:0, C14:1, C20:1, C20:2 and C22:1 found by Tsuyuki et al. (1981) in the ornamental Nana pomegranate. However, our results conrm the presence of linolenic (C18:3) and linoleic (C18:2) acid in all seed oils of pomegranates as previously reported (Melgarejo and Artes, 2000).

The saturated/unsaturated acid ratio was generally very low (Table 1). The lowest and highest are 0.05 (varieties No. 20 and 25) and 0.37 (variety No. 17), respectively. These ratios were the same as previously reported (Melgarejo and Artes, 2000) and considerably lower than those reported by these researchers earlier (Melgarejo et al., 1995).

4. Conclusion Based on our study, the seeds of all 25 varieties so far had similar (but not identical) fatty acid compositions and contained low amounts of saturated fatty acids (except for variety Gorche Shahvar Yazdi (No. 17). Intervarietal differences in fatty acids composition were found and they can be used to establish chemotaxonomic differences, which have also been shown in some other species (Melgarejo et al., 1995; Sundar and Sino, 1992). The lowest saturated/unsaturated acid ratio was found in varieties No. 20 and 25, whilst variety No. 17 showed the highest (0.37). These ratios were considerably lower than those reported by Melgarejo et al. (1995), because saturated lipids were lower and unsaturated acids were higher in the present work. The saturated/unsaturated acid ratios were in strong contrast with those (5.1) reported earlier (El-Nemr et al., 1990).

Acknowledgements The authors wish to thank Tarbiat Modarres University Research Council for its nancial support.

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