Published on May 5, 2017
slide 1: International Journal of Engineering Research Science IJOER ISSN: 2395-6992 Vol-3 Issue-4 April- 2017 Page | 38 The Production Potential of the Olive Oil from Native Cultivars in Albania S. Velo 1 D. Topi 2 Faculty of Natural Sciences University of Tirana Blvd Zogu 1 1010 Tirana Albania Author for correspondence dritan.topiunitir.edu.al Abstract — Albania has an annual production capacity of ca. 50 000 tons of olive fruits and 6000 tons of olive oil. The corresponding cultivation area covers 41000 ha distributed over 90 000 small farms. In total olive trees area covers 6.3 of the total arable land. This paper presents the characterizes the fatty acid profile and total phenolic content of 6 olive varieties namely Kalinjot Ulli i bardhe Tirana Bianco di Tirana Karren Nisiot Kotruvs and Kokerrmadh Berati all from the same harvesting period. Results on fatty acid FA profiles exhibit a variation in concentration of oleic acid 71.53 ± 0.02 Kotruvs to 80.07 ± 0.04 Nisiot. The content of linoleic acid varies from 4.10 ± 0.00 Nisiot to 9.31 ± 0.01 Kotruvs whereas the content of linolenic acid varies from 0.45 ± 0.01 Karren to 0.72 ± 0.02 Kalinjot. Analysed OO from six olive varieties revealed moderate levels of palmitic acid between 9.94 ± 0.01 Nisiot to 12.21 ± 0.01 Kotruvs. From a nutritional point of view it is worth noticing that the Nisiot variety has an n-6/n-3 ratio of 8.11 while the Karren 18.72. The total phenolic content for the studied olive cultivars varied from 89.74 ± 5.47 Karren to 445.03 ± 16.83 mg/kg olive oil Ulli i bardhe Tirana such variation may reflects different antioxidant capacity among olive cultivars. Keywords — Native Olive Cultivars Kalinjot Ulli i Bardhe Tirana Kokerrmadh Berati Nisiot Fatty acid Polyphenol content. I. INTRODUCTION The origin of olive trees in Albania is not different from the road of the distribution of this tree in the Mediterranean area. Archaeological evidences on the agriculture activities such as wheat cobs truss grapes olive lop are stamped in stones and coins of the Illyrian tribes. Olive tree cultivation is mainly concentrated in the Western Plain of the country by penetrating the mainland through the river valleys 1 2. Its distribution is diversified according to climate and geography. It is mainly cultivated in the Mediterranean Climatic Zone which covers 36.2 of the Albanian territory. Albania is a country with an actual annual producing capacity of 50 000 tons of olive fruits and 6000 tons of Olive oil. The olive plantations cover an area of 41000 ha corresponding to 6.3 of the total arable land where the characteristic is its distribution over 90 000 small farms. The incomes from the olive sector are 16 million Euros 3. Recent genetic studies concluded that Albania owns 22 native olive cultivars. All 22 native cultivars are clustered in 7 different groups and the main factor influencing such clustering is the dimension of the olive fruit 4. In what concerns the region from which the native cultivars in this study have been selected and based on RAPD methods cultivars are grouped in three main groups where the main group belongs to the Ulli i bardhe Tirana UbT with a coefficient of similarity that varies from 45 to 84. Hence scientific studies on the native cultivars are very important due to their impact in the future of the olive oil industry. Based on their distribution native olive cultivars are grouped in principal cultivars and secondary cultivars 4. Olive oil is the only vegetable oil that can be consumed without prior refining treatment 5 6. Its major components are triglycerides which represent more than 98 of the total weight. The remaining part belongs to non-saponifiable chemical compounds such as sterols polyphenols alcohols waxes hydrocarbons etc 7 Servilli Montedoro 2003 9. Virgin olive oil composition depends on numerous factors such as the interaction between the cultivar and the environment cultivation techniques fruit ripeness and the oil extraction system 6. The characterisation of fatty acid profiles of olive oils from different olive cultivars is usually proposed as a methodology to differentiate these products according to their cultivar and geographical origin 6 10. slide 2: International Journal of Engineering Research Science IJOER ISSN: 2395-6992 Vol-3 Issue-4 April- 2017 Page | 39 In this study are presented the fatty acid composition and total polyphenol content of six olive cultivars. Studied cultivars were Kalinjot Ulli i bardhe Tirana UbT Karren Nisiot Kotruvs Kokerrmadh Berati which belong to different regions of Albania with different pedoclimatic conditions. The olives from these cultivars are mainly used in the production of oil except to Kokerrmadh Berati used as Table Olive. Nowadays an attempt to modify the national fund of olive tree is an ongoing process. The scope of the study presented herein is linked with chemical evaluation of the Monovarietal Virgin Olive Oils. Assessment of quality parameters and nutritional value of olive oils from studied cultivars is presented in this paper. Such pioneering study will allow for the identification of native cultivars that produce good quality olive oil and that are very well adapted to the pedoclimatic conditions in Albania. II. MATERIALS AND METHODS 2.1 Sample collection and oil extraction A total of 6 native olive cultivars namely Kalinjot Ulli i bardhe Tirana UbT Karren Nisiot Kotruvs and Kokerrmadh Berati were harvested in their main area of Tirana and Kruja Regions between October and November 2014 and prepared as described in Table 1. The climatic characteristics of the production areas in terms of temperature are reported in Figs. 1. FIGURE 1: TPC OF SIX MONOVARIETAL VIRGIN OLIVE OILS OF NATIVE CULTIVAR. UBT-ULLI I BARDHE TIRANA KMB-KOKERMADH BERATI TABLE 1 OLIVE VARIETIES HARVESTING AND EXTRACTION DAY Code sample Cultivar name Harvesting day Extraction day Nr. 1 Kalinjot 8/11/2014 8/11 Nr. 2 Ulli Bardhe Tirana 10/10/2014 10/10 Nr. 3 Karren 12/11/2014 13/11 Nr. 4 Nisiot 12/11/2014 13/11 Nr. 5 Kotruvs 13/11/2014 13/11 Nr. 6 Kokerrmadh Berati 13/11/2014 14/11 Oil extraction was performed with a Laboratory press under cold extraction conditions and mechanical pressing. Once the olives had been properly cleaned and washed they were poured into the receiving hopper where a screw activated by hand through a handle fed the crusher that was equipped with a fixed grate and a hollow knife impeller. The paste produced fell into the lower mixer where a helicoidally shaped stirrer prepared it. A speed change gear sends the paste to the decanter 285.16 445.03 89.74 203.07 226.97 125.60 20.00 70.00 120.00 170.00 220.00 270.00 320.00 370.00 420.00 470.00 Kalinjot UBT Karren Nisiot Kotruvs KmB mg GA/kg OO Olive cultivar slide 3: International Journal of Engineering Research Science IJOER ISSN: 2395-6992 Vol-3 Issue-4 April- 2017 Page | 40 where separation took place: oil from the front and waste mixed with water from the back of the machine. The oil samples were stored in the dark at 4 o C until analysis. 2.2 Chemicals The chemical reagents were analytical grade from Sigma-Aldrich Chemie Steinheim Germany. Internal standard C15:0 was purchased from Sigma-Aldrich. Gallic acid and Folin-Ciocalteau reagent were supplied by Fluka Chemie GmbH Buchi Switzerland. 2.3 Analytical methods 2.3.1 Determination of fatty acid profiles Fatty acid methyl esters FAME were prepared through direct basic transesterification according to IOOC using pentadecanoic acid C 15:0 as Internal Standard. The assay of FAME was carried out with a HP-6890 Gas chromatograph equipped with a Flame Ionization Detector GC-FID. Separation was achieved in a SP-2380 capillary column 60 m x 0.25 mm x 0.20 m from Supelco. Hydrogen was used as carrier gas at a flow rate of 1.0 ml min -1 . Calculations were performed according to Official Method Ce 1b-89 11. Identification of FA was undertaken with pure standards Sigma-Aldrich Supelco based on the comparison of retention times. Fatty acids were named by using the code i:jn-k where i-indicates the total number of carbons j-the number of double bonds and k-the position of the last double bound counted from the terminal methyl group. 2.3.2 Determination of Total Polyphenol Content Fractionation of olive oils The method used to perform the fractionation of oils was proposed by 12. Briefly samples were dissolved in n-hexane Sigma Germany and extracted with a methanol/water mixture 60:40 vol/vol. The insoluble fraction non-polar in methanol/water fraction was removed whereas the polar fraction was used as it was for further analysis. Colorimetric determination of total polyphenol content The Folin-Ciocalteau method was used to determine the total polyphenol content TPC of samples according to method proposed by 12. The absorbance of mixture was measured after 1 h of reaction with a UV-VIS Mini-1240 Spectrophotometer Shimadzu at 725 nm. Results were expressed as Gallic acid equivalent mg/kg olive oil calculated from the following calibration curve determined by linear regression: A 725 3.015 GA + 0.005 r 2 0.999 1 where GA was concentration of gallic acid expressed as mg/kg oil. 2.4 Statistical analysis The complete data were evaluated by randomized block design with three replicates from fatty acid analysis and duplicates for TPC values. Results were displayed as mean values and standard error n3. Significance of the differences among the values was determined by analysis of variance using One-way ANOVA test. The level of significance was determined at P0.05. The employed statistical program was SPSS 17.0 Statistics 2008 SPSS Inc. Chicago IL USA. III. RESULTS AND DISCUSSION Fatty acid profiles of the olive cultivars studied are described in Table 2. One-way ANOVA analysis showed that fatty acid profiles of the seven olive cultivars were statistically significantly different. Results revealed that in what concerns palmitic acid PA the cultivars can be grouped in two groups: i those with lower PA content such as Nisiot 9.94 ± 0.01 UBT 10.88 ± 0.01 Kokerrmadh Berati 10.41 ± 0.01 and Kalinjoti 10.92 ± 0.17 cultivars and those with higher PA content such as Kotruvs 12.21 ± 0.01 cultivar. The oleic acid OA content was as follows: 71.53 ± 0.02 Kotruvs 74.61 ± 0.06 UbT 73.94 ± 0.02 Karren 75.1 ± 1.21 Kalinjot 80.07 ± 0.04 Nisiot. Linoleic acid LA content showed high variation among the studied cultivars olive cultivars such as Kalinjot 7.56 ± 0.13 UbT 8.00 ± 0.07 Karren 8.35 ± 0.01 Kotruvs 9.31± 0.01 and Nisiot 4.10 ± 0.00 presented low content of LA. slide 4: International Journal of Engineering Research Science IJOER ISSN: 2395-6992 Vol-3 Issue-4 April- 2017 Page | 41 The alpha-Linolenic acid ALA was found below 1 submitting the quality criteria of the Extra Virgin Olive Oils EVOO. The ALA content varied according to the following ascending order: 0.45 ± 0.01 Karren 0.51 ± 0.01 Nisiot and Kotruvs 0.58 ± 0.01 UbT 0.72±0.02 Kalinjot 0.67 ± 0.01 Kokerrmadh Berati. TABLE 2 TPC OF SIX MONOVARIETAL VIRGIN OLIVE OILS OF NATIVE ORIGIN MG GALLIC ACID/KG OLIVE OIL Variety Mean±SD Kalinjot 285.16±3.29 Ulli i Bardhe Tirana 445.03±16.83 Karren 89.74±0.47 Nisiot 203.07±7.51 Kotruvs 226.97±1.40 Kokerrmadh Berati 125.60±6.09 A high content of ALA contributes to the n-6/n-3 ratio a very important value for the nutritional evaluation of lipids of different origin. Regarding such ratio the Nisiot cultivar shows a n-6/n-3 ratio of 8.11 followed by 10.31 Kokrremadh Berati 10.47 Kalinjot and 13.75 UbT while the remaining cultivars show higher values Karren 18.72 and Kotruvs 18.25. Comparison of the FA profiles of the Albanian native cultivars with those in neighbouring countries 7 13 14 and Northern Africa 15 gives indication that they are comparable with Italian olive cultivars but also with other Mediterranean countries 7 13. For example the level of palmitic acid in the studied native cultivars is comparable with Italian cultivars Leccino 14.3 and Moraiolo 10.5 Spanish cultivars Arbequina 14.3 Lechin 10.5 and Redondilla 12.5 and Greek cultivar 13.3. The level of oleic acid in Albanian cultivars is comparable with Frantoio 78.2 Arbequina 75.3 and Koreiniki 71.9 13. Related to the linoleic acid trends is not as even since the Albanian olive cultivars present content differences. The UbT content is comparable with Arbequina Frantoio Leccino and Koreiniki. On the other hand the Boçi cultivar presents high content of linoleic acid comparable to the Spanish cultivars Redondilla and Lechin 7 14. TABLE 3 FATTY ACID PROFILE TOTAL POLYPHENOL CONTENT TPC N-6/N-3 AND 18:1/18:2 RATIOS OF THE SEVEN NATIVE OLIVE CULTIVARS Formula Kalinjot Ulli i Bardhe Tirane Karre Nisiot Kotruvs Kokerrmadh Berati 14:0 ND ND ND ND ND ND 16:0 10.92±0.17 10.88±0.01 11.17±0.02 9.94±0.01 12.21±0.01 10.41±0.00 16:1n-9 0.09±0.00 0.07±0.00 ND 0.12±0.00 0.09±0.00 0.13±0.00 16:1n-7 0.48±0.01 0.35±0.01 0.36±0.00 0.41±0.00 0.82±0.00 0.61±0.00 17:0 0.04±0.05 0.13±0.00 0.14±0.00 ND ND ND 17:1 n-7 0.14±0.01 0.19±0.00 0.18±0.00 ND ND ND 18:0 2.31±0.03 2.83±0.01 3.22±0.01 2.56±0.01 1.96±0.01 2.10±0.01 18:1n-9trans ND ND ND ND ND ND 18:1n-9cis 75.11±1.21 74.61±0.06 73.94±0.02 80.07±0.04 71.53±0.02 76.26±0.07 18:1n-7 1.88±0.02 1.53±0.00 1.45±0.01 1.55±0.00 2.87±0.00 2.20±0.01 18:2 n-6cis 7.56±0.13 8.00±0.07 8.35±0.01 4.10±0.00 9.31±0.01 6.92±0.00 20:0 0.36±0.01 0.43±0.01 0.50±0.01 0.45±0.02 0.37±0.01 0.40±0.01 18:3 n-3 0.72±0.02 0.58±0.01 0.45±0.01 0.51±0.00 0.51±0.01 0.67±0.01 20:1 n-9 0.31±0.01 0.28±0.01 0.24±0.02 0.31±0.01 0.34±0.01 0.33±0.01 22:0 0.08±0.04 0.07±0.00 ND ND ND ND n-6/n-3 10.47 13.75 18.72 8.11 18.25 10.31 13.36 14.34 15.04 12.95 14.54 12.92 78.01 77.05 75.94 82.46 75.65 79.53 8.29 8.59 9.03 4.60 9.82 7.59 18:1/18:2 9.93 9.33 8.86 19.54 7.68 11.02 MUFAs/SFAs 5.84 5.37 5.05 6.37 5.20 6.16 MUFAs/PUFAs 9.42 8.97 8.41 17.92 7.71 10.48 slide 5: International Journal of Engineering Research Science IJOER ISSN: 2395-6992 Vol-3 Issue-4 April- 2017 Page | 42 The ratio between monounsaturated and saturated fatty acids of the studied cultivars had an average value of 5.05 - 6.37 whereas the ratio between monounsaturated and polyunsaturated fatty acids presented an average value of 7.71-17.92 Table 3 which are relatively low however the high phenol content could indicate that oil quality was maintained without lipid deterioration. Analysis of the ratio 18:1/18:2 is another indication that refers to the oil oxidation stability. The limit value proposed is 18:1/18:2 ≥ 7.00 17. The results show that the Monovarietal olive oils from 7.68 Kotruvs to 9.33 UbT and 19.54 Nisiot have acceptable oxidation stability. In conclusion all the OO from studied cultivars are above the limit value of 7.00. Quantitative determination of phenolic compounds in olive oil was performed according to the colorimetric method Folin- Ciocalteau 12. The amount of phenolic compounds in olive oil varies from 50 to 1000 mg/kg and depends on several factors such as: climate and extraction technology 6 cultivar 19 and degree of maturation 18. The results for the six olive cultivars Table 3 reveal that the highest value belongs to UbT 445.03 ± 11.90 mg GA/kg olive oil and the lowest value belongs to Karren 63.02 ± 3.9 mg GA/kg olive oil. The results show that the polyphenol content of the studied olive oils had significant differences p0.05 among the cultivars. According to the classification for the olive oils proposed by Montedoro et al 19 regarding to the Total Polyphenol Content the Karren and Kokermadh Berati cultivars can be classified in group of “low” content 50-200 mg/kg while the next four cultivars Kalinjoti UBT Nisioti and Kotruvs can be classified in the group of “medium” content 200-500 mg/kg. The obtained results from the studied cultivars can be related mainly to the cultivar differences. The geographical and pedo-climatical conditions are relatively comparable among regions were collected the olive fruits. The TPC results for UBT are comparable with Koreiniki Greece Picual Spain and Frantoio Italy cultivars 7 13. IV. CONCLUSION The results on the chemical composition of the studied cultivars give for the first time a comprehensive analysis of the Albanian olive cultivars. The variations observed in fatty acid composition and phenolic compounds are probably due to both genetic factors and environmental conditions. By comparison with results from literature it can be concluded that the levels of fatty acids in the oils of the studied cultivars are similar to those found in the group of olive cultivars typical from Northern Mediterranean regions. In general referred to the 18:1/18:2 ratio an index of the oxidative state to the monovarietal olive oils give good consistency. The nutritional profile of Nisioti cultivar is highly interesting as well as the Kokerrmadh Berati cultivar. The total phenolic content of the UBT is particularly of interest with potential to be expanded beyond its territory. REFERENCES 1 Kafazi N. Muço Dh. 1984. Olive Culture. Tirana. Albania: ILB p. 0 In Albanian. 2 Thomaj F. Panajoti Dh. 2003. Olive cultivars. Tirana Albania: ILB p 350 In Albanian. 3 MAFCP. Study on Actual Olive cultivation and its developing perspectives. Ministry of Agriculture Food and Consumer Protection. Tirana 2009 p. 484. 4 Thomaj F. Panajoti Dh. 2005. Variability of Olive varieties according to the multivariate methods. Revista Shqiptare e Shkencave Bujqesore. 64 In Albanian. 5 EUC. 1991. Characteristics of olive oil and olive pomace oil and their analytical methods. Reregulation EEC/2568/91. Official Journal of European Communities L248 1-82. 6 Boskou D. 2006. Olive Oil: Chemistry and Technology. Champaign IL USA: AOCS Press. 7 Aparicio R. Luna G. 2002. Characterization of Monovarietal virgin olive oils. European Journal of Lipid Science and Technology 104 614–627. 8 Servilli M. Montedoro G. 2003. Contribution of phenolic compounds to virgin olive oil quality. European Journal of Lipid Science and Technology. 105 403-408. 9 Pirisi F. M. Cabras P. Falqui C. Migliorini M. Muggelli M. 2000. Phenolic Compounds in Virgin Olive Oil. 2. Reappraisal of the Extraction HPLC Separation and Quantification Procedures. Journal of Agriculture and Food Chemistry. 48 1191-1196. 10 Mannina L. Dugo G. Salvo F. Cicero L. Ansanelli G. Calcagni C. 2003. Study of the cultivar –composition relationship in Sicilian Olive oils by GC NMR and statistical methods. Journal of Agriculture and Food Chemistry. 51 120-127. 11 Official Method Ce 1b-89. 1994. In: Official Methods and Recommended Practices of the American Oil Chemists’ Society 4 th edition AOCS Press: Champaign IL USA. 12 Kalantzakis G. Blekas G. Pegklidou K. Boskou D. 2006. Stability and radical scavenging activity of heated olive oil and other vegetable oils. European Journal of Lipid Science and Technology. 108 329-335. slide 6: International Journal of Engineering Research Science IJOER ISSN: 2395-6992 Vol-3 Issue-4 April- 2017 Page | 43 13 Pinelli P. Galardi C. Mulinacci N. Vincieria F. F. Cimato A. Romani A. 2003. Minor polar compound and fatty acid analyses in monocultivar virgin olive oils from Tuscany. Food Chemistry. 80 331-336. 14 Paz Aguilera M. Beltran G. Ortega D. Fernandez A. Jimenez A. Uceda M. 2005. Characterization of virgin olive oil of Italian olive cultivars: ‘Frantoio’ and ‘Leccino’ grown in Andalusia. Food Chemistry. 89 387–391. 15 Haddada F. M. Krichène Dh. Manai H. Oueslati I. Daoud D. Zarrouk M. 2008. Analytical evaluation of six Monovarietal virgin olive oils from Northern Tunisia. European Journal of Lipid Science and Technology. 110 905–913. 16 Maestri D. M. Labuckas D. O. Meriles J. M. Lamarque A. L. Zyagadlo J. A. Guzman C. A. 1998. Seed composition of soybean cultivars evaluated in different environmental region. Journal of Agriculture and Food Chemistry. 77 494-498. 17 Kiritsakis A. K. Nauos G. G. Polymenoupoulos Z. Thomai T. Sfakiotakis E. Y. 1998. Effect of fruit storage conditions on olive oil quality. J. Amer. Oil Chem. Society 75 721-724. 18 Servilli M. Montedoro G. 2002. Contribution of phenolic compounds to virgin olive oil quality. European Journal of Lipid Science and Technology. 104 602–613. 19 Brenes M. Garcia A. Garcia Prios J. J. Garrido A. 1999. Phenolic compounds in Spanish olive oil. Journal of Agriculture and Food Chemistry. 47 3535–3540. 20 Montedoro G. Servilli M. Miniati E. 1992. Simple and hydrolysable Phenolic Compounds in Virgin Olive oil. 1. Their extraction separation and quantitative and semi quantitative evaluation by HPLC. Journal of Agriculture and Food Chemistry. 40 1571-1576.