The Effects of Geographic Region, Cultivar and Harvest Year on Fatty Acid Composition of Olive

The Effects of Geographic Region, Cultivar and Harvest Year on Fatty Acid Composition of Olive Oil Şeyma Şişik Oğraş＊ , Güzin Kaban and Mükerrem Kaya Atatürk University, Faculty of Agriculture, Department of Food Engineering, Erzurum, TURKEY 25240

Abstract: In this study, the effect of cultivar, harvest year (2010 and 2011) and the geographic regions (Mediterranean, Aegean, Southeastern Anatolia, Marmara and Black Sea) on fatty acid compositions of olive oil was investigated. The proportions of palmitic acid and linoleic acid, which comprised most of the amount of fatty acids after oleic acid, showed variations according to the region and cultivar. The amount of linoleic acid was found higher in some cultivars than that of palmitic acid. In the Aegean, Marmara and Mediterranean Regions, no significant difference was observed between olive cultivars for oleic and linoleic acids. The highest linoleic and lowest oleic acid content were determined in the Black Sea Region. One cultivar (Butko) showed an oleic acid proportion below 55% in this region. The effect of the harvest year on the fatty acid composition of the samples was also found significant. In oils harvested in the second year, a decrease in the amount of linoleic acid was observed, while the amount of oleic acid increased. The highest oleic acid proportion was found in olive oils produced from harvests in the Aegean and Mediterranean regions.

Key words: cultivar, fatty acid composition, geographic region, harvest year, olive oil

1 INTRODUCTION the olive cultivar grows6, 7）. The fatty acid composition of The chemical composition of olive oil is evaluated in two olive oil has a key role in olive characterizations. The domi- groups: major and minor components. Triacylglycerols, nant fatty acid is oleic acid which ranges from 55％ to 83％8）. which constitute more than 98％ of the total fat weight, are Furthermore, the olive oil contains some saturated fatty the major components of olive oil. Existing at 0.5 - 1.5％ in acids such as palmitic acid which range 7.5℃ - 20％, the olive oil, many chemical components such as aliphatic and myristic acid which range 0.05％ or less in olive oil9, 10）. triterpenic alcohols, sterols, tocopherols, hydrocarbons and Olive is grown in a large geographic area in Turkey, antioxidants are the minor components of olive oil1, 2）. which is a significant olive producer, and olive oil is eventu- The highest amount of olive is produced in the Aegean ally produced. However, studies on the characterization of region in Turkey, followed by Marmara, Mediterranean, olive oil have only concentrated on specific regions and va- Southeastern Anatolia and Black Sea regions, respective- rieties11－19）. Therefore, this study included exemplification ly3）. More than 80 olive species are grown in Turkey. Al- that considers 5 different regions（Mediterranean, Aegean, though some species like Gemlik, Memecik and Ayvalık can Southeastern Anatolia, Marmara and Black Sea）, exten- be grown in more than one region, species are often specif- sively produced cultivars per region and two harvest years, ic to the region. For example, Domat is a species which is and the olive cultivars were analyzed in terms of fatty acid endemic to the Aegean region, as Saurani to the Mediterra- composition after being processed to the olive oil in agri- nean region, Nizip Yağlık to the Southeastern Anatolia, cultural facilities. Butko to the Black Sea region and Çelebi to Marmara4）. The quality of olive oil depends on a lot of factors such as olive cultivar, climatic and geographic regions where the olives grow, the maturity degree of the olive fruit which will 2 EXPERIMENTAL be processed to the oil, agricultural implementations and In this study, the fruits of five olive cultivars were har- olive oil production process5）. The mostly, olive oil quality vested from five different geographic regions（Black Sea, is related to the olive cultivar and geographic region, where Southeastern Anatolia, Mediterranean, Aegean and

＊Correspondence to: Şeyma Şişik Oğraş, Atatürk University, Faculty of Agriculture, Department of Food Engineering, Erzurum, TURKEY 25240 E-mail: [email protected] or [email protected] Accepted June 13, 2016 (received for review November 23, 2015) Journal of Oleo Science ISSN 1345-8957 print / ISSN 1347-3352 online http://www.jstage.jst.go.jp/browse/jos/ http://mc.manusriptcentral.com/jjocs

889 Ş. Ş. Oğraş, G. Kaban and M. Kaya

Table 1 Olive cultivars from different geographic regions. Geographic region Southeastern Mediterranean Aegean Marmara Black sea Anatolia Gemlik Memecik Nizip Yağlık Gemlik Butko Ayvalık Domat Ayvalık Ayvalık Otur Cultivar Saurani Uslu Kilis Yağlık Çelebi Görvele Haşebi Gemlik Halhalı Domat Satı Sarı Ulak Erkence Karamani Memecik Kızıl Satı

Marmara Regions）（ Table 1）. This experiment was repeat- chromatography（GC, Agilent Technologies 6890N）with ed during two years, 2010 and 2011 in November. The olive FID detector. GC system was equipped with a capillary fruits were processed to oil in agricultural facilities of the column（DB23, 60 m×250 µm×0.15 µm）. The oven tem- regions, where the samples were taken. Five hundred mL perature was increased from 100 to 200℃ with rate of 5℃/ oil was taken from each sample, put into brown glass min and from 200 to 250℃ with a rate of 4℃/ min. The in- bottles and kept in dark until the analysis. jection block and detector temperatures were 250℃ and 280℃, respectively. Helium was used as a carrier gas with 2.1 Fatty acid compositions analysis a 1.2 mL/ min flow rate. A fatty acid methyl ester mix Fatty acid methyl esters were prepared from olive oil ac- （Supelco, FAME-mix, 4 - 7801, Bellefonte, PA, USA）was cording to the method described by Metcalfe and used as standard in the system Schmitz20）. 0.05 g of sample was weighed and 1.5 mL of 2 M NaOH（methanolic）was added. The vials were capped 2.2 Statistical analysis under nitrogen gas, and a saponification reaction was The research was established and conducted on the carried out at 80℃ for 1 hour. Samples were cooled and 2 basis of the nested classification model considering 5 dif- ml BF3 - methanol was added. Subsequently, the tubes ferent regions, 5 varieties from each region and 2 harvest were again treated with nitrogen gas and allowed to stand seasons. The research data were subjected to variance for 30 min at 80℃. After cooling, 1 mL of hexane and 1 mL analysis through a packaged software（IBM SPSS Statistics of deionized water was added to the tubes and mixed well. 20）and the variation sources that were found important The tubes were centrifujed for 10 minutes at 6000 rpm. were compared with the Duncan’s multiple comparison Hexane layer was transferred to new tubes containing test. The relationship between harvest year and fatty acid sodium sulfate. After addition of 1 mL of hexane, 2 mL of composition as regions was also evaluated by principal the upper layer were transferred to vials. Vials were sealed component analysis（PCA）using Unscrambler v10. 01. under nitrogen and was maintained until analysis at - 18℃. （Como Process, A. S., Norway）. Fatty acid composition of samples were determined by gas

Table 2 The comparison of fatty acid compositions of olive oil which growing in different regions（mean±SD）. Geographic region Fatty acid Southeastern Mediterranean Eagean Marmara Black sea Anatolia Palmitic acid (C16:0) 13.59±1.77b 11.20±1.72a 14.56±1.36b 10.74±1.99a 10.87±1.44a Palmitoleic acid (C16:1) 1.15±0.37bc 0.84±0.36a 1.33±0.22c 0.88±0.31a 1.10±0.20b Heptadecanoic acid (C17:0) 0.07±0.09a 0.05±0.07a 0.09±0.09a 0.17±0.10b 0.08±0.11a Heptadesenoic acid (C17:1) 0.22±0.08a 0.23±0.14a 0.29±0.09ab 0.31±0.10b 0.29±0.16ab Stearic acid (C18:0) 2.41±1.89b* 1.79±2.17ab 2.46±1.31b 0.98±0.89a 1.67±0.71ab Oleic acid (C18:1) 67.20±3.53a 70.68±4.55b 65.59±4.27a 73.31±4.23b 64.52±8.05a Linoleic acid (C18:2) 10.90±3.22ab 10.37±2.34ab 10.52±2.26ab 8.10±1.76a 11.99±2.47c Linolenic acid (C18:3) 0.72±0.15b 0.82±0.12c 0.85±0.11c 0.64±0.11a 0.59±0.12a Arachidic acid (C20:0) 0.29±0.11a 0.25±0.11bc 0.20±0.17b 0.29±0.08c 0.10±0.12a *a - c: Any means in the same column having the same letters are not significantly different (p>0.05), SD; standard devision.

890 J. Oleo Sci. 65, (11) 889-895 (2016) Fatty acid composition of olive oil

Fig. 1 Principal component scores and factor loadings from PCA applied to the fatty acid composition.

891 J. Oleo Sci. 65, (11) 889-895 (2016) Ş. Ş. Oğraş, G. Kaban and M. Kaya

Table 3 T he effect of the harvest year on the fatty acid composition of olive oil（mean±SD）. Harvest year* Fatty Acid First Second Palmitic acid (C16:0) 12.04±2.25a 12.35±2.32a Palmitoleic acid (C16:1) 1.15±0.32b 0.97±0.35a Heptadecanoic acid (C17:0) 0.10±0.11a 0.08±0.09a Heptadecenoic acid (C17:1) 0.32±0.15b 0.22±0.06a Stearic acid (C18:0) 1.61±0.95a 2.12±1.99a Oleic acid (C18:1) 66.74±4.99a 69.78±6.65b Linoleic acid (C18:2) 11.84±2.67b 8.91±1.87a Linolenic acid (C18:3) 0.79±0.14b 0.66±0.14b Arachidic acid (C20:0) 0.19±0.16a 0.26±0.10b *1: 2010 harvest year, 2: 2011 harvest year. a - b: Any means in the same line having the same letters are not significantly different (p > 0.05). SD; standard devision.

3 RESULT and DISCUSSION Sea Region. The ratio of linoleic acid in olive oils produced The effect of the geographic region on the fatty acid in other regions varied between 8.10％ and 10.90％. It has composition of olive oil is given in Table 1. The palmitic also been revealed in other researches related to the topic acid content of oil which obtained from the Aegean, that many factors such as high location, maturity level, cul- Marmara and Black Sea regions was lower than that of the tivar and harvest year influence the fatty acid composition Southeastern Anatolia and Mediterranean regions. The of olive oil14, 17, 22－28）. highest mean palmitic acid value was found in the South- A PCA was performed to summarize the differences in eastern Anatolia region. The lowest stearic acid content fatty acid composition of years for each region. Two princi- （0.98％）, which is another saturated fatty acid, was deter- pal components in Mediterranean, Aegean, Marmara, Black mined in the Marmara region. However, no significant dif- Sea and Southeastern Anatolia were able to explain the ference was found between Marmara region（0.98％）with 91％, 90％, 95％, 100％ and 91％ of the total variance ob- Aegean（1.79％）and Black Sea（1.67％）regions（Table 2）. served, respectively. Oleic acid, the most important fatty In the previous study, olive oils were taken from different acid of olive oil, has strongly positive correlation with PC1 geographic regions of Turkey（Northern and Southern in all of regions except Mediterranean region. In Mediterra- Aegean, Bursa - Akhisar and Southeastern Anatolia）. The nean region, linoleic acid was positively associated with results showed that the highest palmitic acid content was PC1, while oleic acid was highly positive correlated with found in the Southeastern Anatolia Region（18.20％）and PC2（Fig. 1）. the lowest stearic acid（1.61％）in the Manisa - Bursa Harvest year is another factor which has an effect on the regions12）. fatty acid composition and some other qualitative charac- The Aegean and Marmara Regions showed the highest teristics of olive oil. As seen in Table 3, the differences oleic acid content（70.68％ and 73.31％ respectively）. The between the average values of the years and the other re- mean oleic acid content ratio in olive oils produced in the searched fatty acid averages other than palmitic, heptadec- Mediterranean, the Southeastern Anatolia and the Black anoic and stearic acid are quite clear. The oleic acid Sea regions were determined as 67.20％, 65.59％ and amount increased, but the linoleic acid amount decreased 64.52％, respectively. Differences between means as to in the second year. Similarly, Diraman et al.6） stated that region of production were not significant（Table 2）. significant variations occurred on the fatty acid composi- However, the results of all the regions researched comply tion of the olive oils depending on the harvest year. with the Turkish Food Codex‘ Olive Oil and Pomace Oil Cultivar is another important factor influencing the fatty Communiqué’. It has also been determined in some other acid composition of olive oil29）. In this study, the extensive studies that the linoleic acid content is high in the Aegean cultivars of the regions were harvested, processed into oil and Marmara Regions, which have a great share in olive oil and analyzed. Gemlik, an important olive cultivars in production12, 21）. The highest value of linolenic acid, which Turkey, presented a high oleic acid content（＞70％）in the is another unsaturated fatty acid, was found in the Black Mediterranean, Aegean and Marmara regions. But Ayvalık,

892 J. Oleo Sci. 65, (11) 889-895 (2016) Fatty acid composition of olive oil (C20:0) Arachidic . (C18:3) Linolenic （ mean ± SD ） (C18:2) Linoleic 9.53 ± 3.67a 0.69 ± 0.06a 0.31 ± 0.03a 7.90 ± 0.05a 0.77 ± 0.03a 8.93 ± 0.39a 0.13 ± 0.15a 0.88 ± 0.23a 8.10 ± 1.94a 0.67 ± 0.21a 9.34 ± 3.00a 0.18 ± 0.21a 0.70 ± 0.05a 7.63 ± 1.83a 0.22 ± 0.15a 0.62 ± 0.03a 8.10 ± 0.47a 0.35 ± 0.06a 0.65 ± 0.08a 7.33 ± 0.29a 0.28 ± 0.02a 0.58 ± 0.07a 0.33 ± 0.01a 0.28 ± 0.06a 7.71 ± 1.00a 0.58 ± 0.02a 0.24 ± 0.16a Oleic (C18:1) Stearic (C18:0) Fatty acid > 0.05). (C17:1) p Heptadecenoic (C17:0) Heptadecanoic (C16:1) Palmitoleic The interaction effect of geographic regions and cultivars on fatty acid compositions (C16:0) Palmitic

13.60 ± 1.23a 1.30 ± 0.02ab 0.00 ± 0.00a 0.22 ± 0.07a 2.37 ± 1.18ab 67.64 ± 1.04c 10.80 ± 2.17b 0.87 ± 0.09a 0.26 ± 0.17a 14.06 ± 1.11a 1.06 ± 0.10a 0.23 ± 0.02b 0.38 ± 0.16a 2.98 ± 0.22ab 60.83 ± 3.46a± 1.58bc 11.60 0.86 ± 0.02a 0.27 ± 0.18a 14.04 ± 0.98bc 1.40 ± 0.15b 8.75 ± 1.24a 0.17 ± 0.13a 1.15 ± 0.35a 0.29 ± 0.18a 0.07 ± 0.08a 1.52 ± 0.79a 0.25 ± 0.19a 66.13 ± 2.08a 2.91 ± 4.18a 10.85 ± 2.90a 71.41 ± 4.03a 0.65 ± 0.03a 10.23 ± 0.41a 0.22 ± 0.15a 0.80 ± 0.04bc 0.13 ± 0.15a 13.89 ± 0.22a 1.39 ± 0.03bc16.31 ± 1.10b 0.07 ± 0.08a 1.57 ± 0.00c 0.29 ± 0.01a 0.08 ± 0.06a12.63 ± 2.31a 1.55 ± 0.11a 0.28 ± 0.03a 1.09 ± 0.29a 71.44 ± 1.97d 1.57 ± 0.42a 0.17 ± 0.06a 63.23 ± 0.30ab 13.37 ± 0.33c 0.23 ± 0.03a 0.86 ± 0.07a 1.04 ± 0.59a 0.16 ± 0.19a 69.36 ± 2.63a 8.92 ± 1.68a 0.51 ± 0.21a 0.23 ± 0.11a 0.33 ± 0.08a 0.92 ± 0.98a 73.99 ± 4.07a 14.73 ± 0.46c 1.05 ± 0.18b 0.08 ± 0.10a 0.17 ± 0.03a 2.72 ± 1.23a 64.42 ± 3.79a 10.74 ± 0.28a 0.74 ± 0.05a 0.34 ± 0.05a 9.14 ± 2.67a 0.87 ± 0.27a 0.06 ± 0.12a± 0.41a11.42 0.24 ± 0.16a 1.16 ± 0.16a± 0.37a11.36 1.59 ± 0.86a 1.05 ± 0.14a 0.07 ± 0.09a 54.09 ± 10.95a 0.13 ± 0.11a 10.71 ± 4.60a 0.29 ± 0.13a 0.54 ± 0.22a 0.34 ± 0.19a 1.86 ± 0.53a 0.00 ± 0.00a 1.47 ± 0.78a 68.34 ± 4.40b 12.40 ± 2.23a 68.24 ± 5.67b 0.62 ± 0.10a 12.21 ± 1.91a 0.59 ± 0.11a 0.16 ± 0.12b 0.24 ± 0.04b lık lık ğ ğ Table 4 Table ebi 12.53 ± 1.75ab 1.23 ± 0.13b 0.06 ± 0.07a 0.23 ± 0.01a 3.82 ± 3.18a 66.63 ± 2.78a± 2.25a 11.77 0.79 ± 0.12a 0.33 ± 0.05a ş Satı Otur± 0.59a 11.06 1.20 ± 0.11a 0.04 ± 0.09a 0.21 ± 0.06a 1.53 ± 1.10a 67.01 ± 3.40b± 1.82a 11.95 ± 0.10a 0.59 ± 0.13ab 0.11 Uslu± 0.18b 11.26 0.76 ± 0.17a 0.08 ± 0.10a 0.25 ± 0.19a 0.82 ± 0.60a 70.32 ± 8.17a 10.70 ± 3.76a 0.87 ± 0.10c 0.27 ± 0.04a Çelebi 10.12 ± 0.61a 0.89 ± 0.24a 0.10 ± 0.07a 0.28 ± 0.01a 1.37 ± 1.16a 75.52 ± 0.80a Domat± 0.54b 11.24 0.86 ± 0.16a 0.05 ± 0.06a 0.26 ± 0.07a 2.54 ± 2.48a 68.39 ± 3.89a 10.55 ± 2.24a 0.75 ± 0.07ab 0.25 ± 0.17a Domat 10.37 ± 2.25a 1.00 ± 0.18a 0.15 ± 0.10a 0.28 ± 0.02a 0.61 ± 0.87a 74.95 ± 3.04a Ha Halhalı Gemlik± 0.87b 11.23 0.45 ± 0.50a 0.00 ± 0.00a 0.14 ± 0.16a 1.03 ± 0.69a 73.34 ± 0.88a 10.83 ± 0.66a 0.99 ± 0.05d 0.27 ± 0.04a Saurani± 1.01a 11.51 ± 0.14a 0.65 0.06 ± 0.07a 0.21 ± 0.02a 2.65 ± 2.13a 68.52 ± 4.66a 13.45 ± 5.27a 0.83 ± 0.28a 0.31 ± 0.04a Ayvalık Ayvalık Ayvalık Görvele± 0.60a 11.34 1.21 ± 0.08a± 0.16a 0.11 0.34 ± 0.23a 1.93 ± 0.42a 64.95 ± 6.03b 12.70 ± 1.57a 0.62 ± 0.07a 0.00 ± 0.00a Erkence 13.53 ± 0.94c 0.96 ± 0.15a 0.03 ± 0.07a 0.27 ± 0.12a 1.65 ± 1.00a 69.97 ± 3.74a Kızıl satı Memecik Sarı Ulak Karamani 14.93 ± 1.12ab 1.31 ± 0.34ab 0.05 ± 0.06a 0.27 ± 0.08a 3.82 ± 2.05b 64.79 ± 2.93bc Kilis Ya Nizip Ya Region Cultivar Aegean Memecik Anatolia Marmara Gemlik± 0.74a 11.67 0.93 ± 0.38a 0.18 ± 0.12a 0.42 ± 0.17a 0.97 ± 1.11a 72.72 ± 6.95a Black sea Butko Southeastern Mediterranean Gemlik 15.12 ± 1.48c 1.41 ± 0.48b 0.00 ± 0.00a 0.19 ± 0.03a 1.33 ± 0.69a 70.30 ± 1.90a SD; standard devision. a - d): Any means in the same line having the same letters are not significantly different ( Any means in the same line having letters are not significantly different a - d):

893 J. Oleo Sci. 65, (11) 889-895 (2016) Ş. Ş. Oğraş, G. Kaban and M. Kaya

another important cultivar, differed as per the regions. Its Acknowledgments proportion was 64.42％ in the Mediterranean region while This research was supported by Research Council of it had a proportion of 69.36％ in the Marmara region. Atatürk University（BAP-2010/ 253）.The financial support Memecik cultivar showed values over 70％ in both Marmara of Atatürk University is gratefully acknowledged. and Aegean regions. Another characteristic of this cultivar was its representation of a lower palmitic acid proportion （Table 4）. However, a higher palmitic acid content was re- ported in another research conducted for the characteriza- References tion of olive oils obtained from the Memecik cultivar. The 1） Bayrak, A.; Kıralan, M. Sızma zeytinyağı ve kalite fak- oleic acid content differed between 73.37％ and 75.64％ in törleri. Hasad yayıncılık, İstanbul（2008）. the same research, similarly to our findings14）. Butko culti- 2） Garcia-Gonzalez, D.; Aparicio-Ruiz, R.; Aparicio, R. var was determined as the cultivar which had the lowest Virgin olive oil - chemical implications on quality and oleic acid content in the Black Sea region and this value is health. Eur. J. Lipid Sci. Tech. 110, 602-607（2008）. below 55 - 83％ that are defined as the limit values for oleic 3） Güler, M.; Cesur, R.; Sarı, N. Zeytinde Bakım İşlemleri. acid in the Turkish Food Codex‘ Olive Oil and Pomace Oil P:38, Adana, Turkey（2010）. Communiqué’. Considering the varieties in the regions, no 4） Food, Agriculture and Livestock Ministry（FALM）of statistically significant difference was found in the Mediter- Turkish Republic, Olive Research Station. Native spe- ranean region in relation to heptadecanoic, heptadecenoic, cies catalogue. arastirma.tarim.gov.tr/izmirzae. Ac- stearic, oleic, linoleic, linolenic and arachidic fatty acids. cessed: August 2015（2012）. 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894 J. Oleo Sci. 65, (11) 889-895 (2016) Fatty acid composition of olive oil

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