1127 0009-3130/19/5506-1127 ©2019 Springer Science+Business Media, LLC
1) Cumhuriyet University, Faculty of Science, Department of Biology, 58140, Sivas, Turkey, fax: +90 346 2191186, e-mail: makpinar@cumhuriyet.edu.tr; 2) Selcuk University, Faculty of Science, Department of Biology, Konya, Turkey; 3) Cumhuriyet University, Faculty of Science, Department of Molecular Biology and Genetic, 58140, Sivas, Turkey. Published in Khimiya Prirodnykh Soedinenii, No. 6, November–December, 2019, pp. 969–971. Original article submitted November 7, 2017.
Chemistry of Natural Compounds, Vol. 55, No. 6, November, 2019
B R I E F C O M M U N I C A T I O N S
TOTAL LIPID CONTENT AND FATTY ACID COMPOSITION OF SEEDS OF SOME WILD Achillea SPECIES
Nuray Zonuz,1 Nukhet Akpinar,1 Erol Donmez,1 Gokhan Zengin,2 Ali Emre Akpinar,3
and Mehmet Ali Akpinar1*
The Achillea species are known as civanpercemi, barsamotu, marsamotu, binbiryaprakotu, and kandilcicegi in Turkish [1]. The important features of Achillea species are that they have protective activity, antiulcer activity, antispasmodic activity, and biological effects [2, 3]. Most members of the Achillea species are antiseptic and have been used in medicine and cosmetic preparations[4, 5]. Although many studies have been conducted on the chemical and pharmacological properties of this genus [6–8], there is not enough information on the fatty acid composition of the seed oils of Achillea species [9–13]. Therefore, we aim to study the total lipid content and fatty acid composition of the seeds of some wild endemic Achillea species (A. cucullata, A. phrygia, and A. armenorum).
This study is the first report on the total lipid and fatty acid (Table 1) composition of A. cucullata, A. phrygia, and A. armenorum seeds from Turkey′s flora. The total lipid percentages of the seeds were found to be 3.60% (A. phrygia), 4.35 (A. cucullata), and 4.87% (A. armenorum).
The fatty acid compositions of three Achillea species are presented Table 1. Twenty-one fatty acids were identified from the Achillea seed oils in the present study. A. phrygia had the high levels of palmitic (16:0, 28.26%), heptadecanoic (17:0, 5.09%), stearic (18:0, 4.66%), pentadecanoic (15:0, 3.81%), and myristic (14:0, 3.34%) acid. High levels of stearic (18:0), arachidic (20:0), and behenic (22:0) acids were found in A. armenorum. Heneicosanoic acid (21:0) was found at very low levels in all seed samples. It is evident from our results that seed oils of Achillea studied contain 16:0 and 18:0 as the major saturated fatty acids (SFAs). The present results, with respect to SFAs, confirm the earlier reports of other Achillea species [11, 14]. According to Keser et al. [10], the dominant fatty acids in A. millefolium were 16:0 (40.11%) and linoleic acid (18:2, 16.71%) in leaves; linoleic acid (48.70%, 60.85%) and oleic acid (18:1, 19.83%, 18.33%) in flowers and seed extracts, respectively. Also, it has been found that the percentages of ΣSFA and ΣUFA (unsaturated fatty acid) were 15.66% and 84.34% in the seed oil of this species. The differences between fatty acid percentages may be due to environmental factors and geographical zone where the plants were collected [15, 16]. In our study, significant differencse were observed between ΣSFA amounts. ΣSFA in the A. phrygia seed had the highest level (50.32%) while A. cucullata had the lowest level (19.79%).
The major monounsaturated acid (MUFA) in the seed oils of three Achillea species was 18:1 ω-9. The 18:1 ω-9 content was found to be at the highest level in A. cucullata (11.30%) and A. armenorum (10.05%) seeds. But high 18:2 contents were generally associated with lower levels of 18:1. This finding is compatible with the biochemical pathway of desaturation of oleic acid to form 18:2, and further desaturation for the synthesis of linolenic acid (18:3) [17].
Myristoleic acid (14:1), pentadecenoic acid (15:1), palmitoleic acid (16:1), c-vaccenic acid (18:1 ω-7), and eicosenoic acid (20:1 ω-9) have been found in low percentages in the seed oils of three Achillea species. Heptadecenoic acid (17:1) was present in only two species (A. phrygia and A. armenorum). With the exception of 18:1 ω-9, the percentages of MUFAs were found to be below 1%.
1128
As far as MUFAs contents are concerned, the present study is supported by previous studies with other Achillea species [9, 10, 12, 13]. Erucic acid (22:1) was not detected in the seed oil of Achillea seed investigated. Erucic acid diet increases doxorubicin toxicity. Bozcali et al. [18] reported that both erucic acid diet and doxorubicin administration can lead to profound renal toxicity or hematological toxicity and cardiotoxicity in rats. The ΣMUFA percentages did not show very significant differences between seed oils of Achillea species. But, ΣMUFA content (13.92 %) in A. armenorum was higher than other species (12.60% in A. cucullata and 11.74% in A. phrygia).
Linoleic acid (18:2 ω-6) was found at the highest percentage in the seed oils of the Achillea species investigated. This acid content was at the highest level in A. cucullata (64.23%) but found to be at lower levels in A. phrygia (33.00%) and A. armenorum (31.95%). The 18:2 ω-6 contents of three Achillea species were richer than 18:3 ω-6 and 18:3 ω-3 contents. Also, the 18:3 ω-3 content in A. armenorum (12.07%) was higher than the in two other species (3.44% in A. cucullata and 4.64% in A. phrygia). Palic et al. [14] reported that the contents of 18:2 and 18:3 were 24.3 and 25.9% in leaves and flowers of A. linguata, 21.6 and 16.8% in A. nobilis, and 41.3 and 7.4% in A. crithmifolia. Also, Ayaz et al. [11] found that the concentrations of 18:2 and 18:3 in the mature achene of A. biebersteinii, A. bisserata, A. multifida, and A. wilhelmsii varied between 32.40–64.43% and between 2.72–16.45%, respectively. The 18:2 and 18:3 affect the fluidity, flexibility, and permeability of membranes. At the same time, they are the precursors of eicosanoids.
The percentages of ΣPUFA ω-6 in A. phrygia (33.22%) and in A. armenorum (32.53%) were significantly lower than that of A. cucullata (64.33%). Also, ΣPUFA ω-6 makes up a larger fraction than ΣPUFA ω-3. It is well known that PUFAs, especially ω-3 and ω-6 fatty acids, are the most important fatty acid compounds in plant and animal. The features of plant and animal oils depend on the rate and composition of the fatty acids which they contain [19, 20]. In our study, any comparison TABLE 1. Contents of Fatty Acid in Seed Oils of Some Achillea Species (mean ± S.E.)*
Fatty acid A. cucullata A. phrygia A. armenorum
10:0A 0.27 ± 0.02 a 0.92 ± 0.01 b 1.64 ± 0.03 c 12:0 0.44 ± 0.00 b 1.30 ± 0.03 a 1.13 ± 0.01 a 14:0 1.78 ± 0.03 a 3.34 ± 0.03 b 1.08 ± 0.01 a 15:0 0.54 ± 0.00 a 3.81 ± 0.03 b 0.53 ± 0.01 a 16:0 11.47 ± 0.09 b 28.26 ± 0.04 a 25.36 ± 0.13 a 17:0 0.89 ± 0.10 a 5.09 ± 0.27 b 1.99 ± 0.02 c 18:0 2.25 ± 0.03 a 4.66 ± 0.09 b 5.01 ± 0.03 b 20:0 1.07 ± 0.03 a 1.26 ± 0.03 a 2.23 ± 0.05 b 21:0 0.06 ± 0.01 b 0.14 ± 0.02 a 0.20 ± 0.00 a 22:0 0.84 ± 0.00 a 1.55 ± 0.04 b 2.29 ± 0.03 c ΣSFA 19.61 ± 0.03 a 50.32 ± 0.06 b 41.46 ± 0.03 c 14:1 0.02 ± 0.00 a 0.18 ± 0.01 a 1.80 ± 0.05 b 15:1 0.21 ± 0.00 a 0.25 ± 0.01 a 0.36 ± 0.01 a 16:1 0.27 ± 0.01 a 0.36 ± 0.03 a 0.34 ± 0.02 a 17:1 – 0.09 ± 0.01 0.10 ± 0.00 18:1 ω-9 11.30 ± 0.02 a 9.76 ± 0.06 a 10.05 ± 0.04 a 18:1 ω-7 0.50 ± 0.03 a 0.77 ± 0.07 b 0.98 ± 0.06 b 20:1 ω-9 0.30 ± 0.01 a 0.33 ± 0.00 a 0.21 ± 0.00 a ΣMUFA 12.60 ± 0.01 a 11.74 ± 0.03 a 13.84 ± 0.03 a 18:2 ω-6 64.23 ± 0.30 a 33.00 ± 0.20 b 31.95 ± 0.10 b 18:3 ω-6 0.03 ± 0.01 b 0.05 ± 0.01 b 0.50 ± 0.02 a 20:2 ω-6 0.07 ± 0.00 a 0.16 ± 0.01 b 0.08 ± 0.01 a ΣPUFA ω-6 64.33 ± 0.10 a 33.22 ± 0.07 b 32.53 ± 0.04 b 18:3 ω-3 3.44 ± 0.03 b 4.64 ± 0.14 b 12.07 ± 0.02 a ΣPUFA ω-3 3.44 ± 0.03 b 4.64 ± 0.14 b 12.07 ± 0.02 a ΣFA 1.40 ± 0.05 a 1.60 ± 0.03 a 1.50 ± 0.06 a
______
*Each value represents the mean of three experiments. A(a, b, c, d) Means with the same letter in each row do not significantly differ at P ≤ 0.05. ΣSFA: total saturated fatty acid; ΣMUFA: total monounsaturated fatty acid; ΣPUFA ω-6: total ω-6 polyunsaturated fatty acid; ΣPUFA ω-3: total ω-3 polyunsaturated fatty acid.
1129 could not be made because there were no studies on ω-6 and ω-3 series PUFAs of Achillea species. The novelty of this study is related to the presentation of new information on fatty acid compositions from seed oils of A. cucullata, A. phrygia, and A. armenorum.
Seed Material. The Achillea cucullata Hausskan. & Bornm. used in this study was collected from B6 Sivas region (Sivas-Celalli roadside) in July 2012 (temperature 22°C, altitude about 1300–1400 m), A. phyrgia Boiss.& Balansa was collected from B6 Sivas region (between Gurun-Kangal, Kuskayasi crossroads) in June 2012 (temperature 19–21°C, altitude 1600 m), and A. armenorum Boiss.& Hausskn. was collected from B6 Kahramanmaras region (Goksun, Berit mountain, Yedigardaslar hill) in July 2012 (temperature 30°C, altitude 2800 m). These species are endemic to Turkey′s flora and stored at the Herbarium of the Biology Department of Cumhuriyet University (Sivas).
Total Lipid and Esterification of Fatty Acids from Seeds. The air-dried seed material was ground, and 3 g was taken from each of the milled samples and stored for 2 days in chloroform–methanol (2:1) for 48 h at 4°C. Thereafter, samples were extracted in chloroform–methanol in an ice bath. Autooxidation of PUFAs was minimized by adding 50 μL of butylated hydroxytoluene (2%, w/v in chloroform) to the extraction mixture, and the isolation of the total lipid from seeds was carried out [21]. The total lipids obtained were saponified by refluxing with methanol (50%) containing 6% potassium hydroxide for 1 h at 85°C. The saponifiable lipids were converted to fatty acid methyl esters (FAMEs) for 10 min at 85°C using the standard boron triflouride–methanol (BF3) method [22].
The resultant mixture of FAMEs in hexane–chloroform (4:1) was injected into an HP (Hewlett Packard) Agilent 6890N model GC (gas chromatograph) equipped with a flame ionization detector (FID) and fitted with an HP-88 capillary column (100 m × 0.20 mm i.d., 0.25 μm film). The carrier gas was helium (1 mL min–1), and injector port and detector temperatures were 240 and 250°C, respectively. The FAME solutions (1 μL) were loaded onto the column. The column temperature program was 160°C for the beginning, then a temperature increase at 4°C/min up to 200°C. The FAMEs were identified by comparison of their retention times with external standards (Alltech standard, Lexington, USA).
All analytical determinations and GC analyses were performed in triplicate, and the mean values were reported. The statistical analyses of total lipid contents and percentages of fatty acid were tested by analysis of variance (ANOVA), and comparisons between means were performed with Tukey′s test. Differences between the mean scores were evaluated as significant if P ≤ 0.05.
ACKNOWLEDGMENT
This work was a part of Project No. F-362 supported by the Scientific Research Foundation of Cumhuriyet University (CUBAP) (Sivas, Turkey).
REFERENCES
1. E. W. Konemen, Gordon Cheers Publication, Hong Kong, 51 (1999).
2. T. Kundakovic, D. N. Mimica, and N. Kovacevic, Fitoterapia, 276, 574 (2005).
3. G. Ozek, M. Tekin, Y. Haliloglu, K. H. C. Baser, and T. Ozek, Rec. Nat. Prod., 12 (2), 142 (2018). 4. E. Yesilada, G. Honda, E. Sezik, M. Tabata, K. Goto, and Y. Ikeshiro, J. Ethnopharmacol., 39, 31 (1993). 5. P. Magiatis, A. L. Skaltsounis, I. Chinou, and S. A. Haroutounian, Z. Naturforsch., 57C, 287 (2002).
6. A. Ertas, N, Boga, N. Hasimi, Y. Yesil, A. C. Goren, G. Topcu, and U. Kolak, Turk. J. Chem., 38, 592 (2014). 7. K. H. C. Baser, Volatiles Essent. Oils, 3 (1), 1 (2016).
8. F. Rezaei, R. Jamei, R. Heidari, and R. Maleki, Int. J. Food Prop., 20 (7), 1522 (2017). 9. S. A. H. Goli, M. Rahimmalek, and B. E. S. Tabatabaei, Int. J. Agric. Biol., 10, 355 (2008).
10. S. Keser, S. Celik, S. Turkoglu, O. Yilmaz, and I. Turkoglu, Turk. J. Pharm. Sci., 12 (2), 133 (2015).
11. F. A. Ayaz, H. Inceer, S. Ayaz-Hayirlioglu, and N. Aksu-Kalmuk, Rom. Biotechnol. Lett., 21 (3), 11576 (2016). 12. N. Zonuz, N. Akpinar, G. Zengin, A. Aktumsek, and M. A. Akpinar, Cumhuriyet Sci. J., 38 (4), 6 (2017). 13. N. Zonuz, N. Akpinar, A. E. Akpinar, E. Donmez, A. Aktumsek, and M. A. Akpinar, Int. J. Curr. Res. Life Sci.,
7 (6), 2286 (2018).
14. R. Palic, G. Stojanovic, N. Randelovic, V. Randelovic, and J. Velickovic, Achillea. Facta Universita, 2 (2), 101 (2000).
1130
15. F. M. Pritchard, H. A. Eagles, R. M. Norton, P. A. Salinbury, and M. Nicolas, Aust. J. Exp. Agric., 40, 679 (2000). 16. N. Bezic, M. Skocibucic, V. Dunkic, and A. Radonic, Phytother. Res., 17, 1037 (2003).
17. A. L. Lehninger, Principles of Biochemistry, Worth Publishers, New York, 1982, pp. 583–614.
18. E. Bozcali, O. Suzer, H. N. Gursoy, P. Atukeren, and K. M. Gumustas, Int. J. Clin. Exp. Med., 2, 337 (2009). 19. A. P. Simopoulos, Biomed. Pharm., 56 (8), 365 (2002).
20. J. D. Buckley and P. R. Howe, Obes. Rev., 10, 648 (2009).
21. J. Folch, M. Lees, and G. H. S. Stanley, J. Biol. Chem., 226, 497 (1957). 22. C. W. Moss, M. A. Lambert, and W. H. Mervin, Appl. Microbiol., 28, 80 (1974).
