RESEARCH POSTER PRESENTATION DESIGN © 2015

www.PosterPresentations.com

ABSTRACT

The essential oil of juniper berries (Juniperus communis L.) is traditionally used for medicinal and flavoring purposes. The Republic of Srpska has a great potential of woody

greenery and juniper berries as a possible raw material for the production of essential oils. Therefore, the subject of this study is to determine the chemical composition of essential oil

obtained from juniper berries (Juniperus communis L.). The essential oil of the Juniperus berry was produced in industrial conditions by the hydrodestilation process, in cooperation

with the PEF "Forest of Republic of Srpska", a.d. Sokolac. A light yellow essential oil, with a pleasant smell, yield of 1.3% was obtained.

Qualitative and quantitative composition of the oil was determined by GC-MS and GC-FID analysis. The obtained results have proved the presence of 58 components, wherein

the highest content of α-pinene (29.3%), myrcene (12.7%), sabinene (9.5%), germacrene D (8.4%), limonene (3.6%) and (E)-Caryophyllene (3.4%) has been determined.

Keywords: Essential oil, Juniperus communis L, GC-MS analysis.

MATERIALS AND METHODS

RESULTS AND DISCUSSION

REFERENCES

[1] E.J. Bowles, The Chemistry of Aromatherapeutic oils (Third Ed.). Crows Nest, NSW, Allen & Unwin Academic, New South Wales, Australia, 2004, 51-58.

[2] M. Lahlou, Essential oils and fragrance commpounds: bioactivity and mechanisms of action, Flavour & Fragrance J., Vol. 19 (2004) 159-165.

[3] R. P. Adams, Junipers of the World: The genus Juniperus. Trafford Publishing Co., Vancouver, British Columbia, 2004, 83-88.

[4] R. Gautam, A. Saklani, S. M. Jachak, Indian medicinal plants as a source of antimycobacterial agents, Journal of Ethnopharmacology, Vol. 110 (2007) 200–234.

[5] M. R. Gonzalez-Tejero, M. Casaresporcel, C. P. Sanchez-Rojas, J. M. Ramiro-Gutierrez, J. Molero-Mesa, A. Pieroni, et al. Medicinal plants in the Mediterranean area:

Synthesis of the results of the project Rubia, Journal of Ethnopharmacology, Vol. 116 (2008) 341–357.

[6] Ž., Marjanović-Balaban, Ekstrakcija eteričnih ulja četinara (jela, smrča, bor, kleka i duglazija) i analiza njihove antimikrobne aktivnosti, Doktorska disetracija,

Univerzitet u Banja Luci, 2007.

[7] A.Y. Gordien, A. I. Gray, S. G Franzblau, V. Seidel, Antimycobacterial terpenoids from Juniperus communis L. and Sanguinaria Canadensis, Journal of

Ethnopharmacology, Vol. 79 (2009) 57–67.

[8] A. Buci, E. Hodaj-Çeliku, H. Manaj, S. Abazi, S., Drushku, D. Lazari, Essential Oil Composition from Juniperus communis Originated from Albania, J. Int.

Environmental Application & Science, Vol. 13 (2018) 15-19.

[9] I. S. StoIlova, J. Wanner, I. Jirovetz, D. TrIfonova, L. Krastev, S. Stoyanova. and I. Krastanov, Chemical composition and antioxidant properties of juniper berry

(Juniperus communis L.) essential oil, Bulgarian Journal of Agricultural Science, Vol. 20-2 (2014) 227-237.

[10] P. Chatzopoulou, and S. Katsiotis, Headspace analysis of the volatile constituents from Juniperus communis L. ‘berries’ (cones) grown wild in Greece, Flavour &

Fragrance J., Vol. 21 (2006) 492-496.

[11] M. Matović, B. Bojović, M. Jusković, Composition of essential oils from three classes of juniper fruit from Serbia. Journal of Medicinal Plants Research, Vol. 5 (2011)

6160-6163.

[12] M. Höferl, I. Stoilova, E. Schmidt, J. Wanner, L. Jirovetz, D. Trifonova, L. Krastev, and A. Krastanov, Chemical composition and antioxidant properties of juniper

berry (Juniperus communis L.) essential oil. Action of the esssential oil on the antioxidant protection of Saccharomyces cerevisiae model organism, Antioxidants, Vol. 3

(2014) 81-98.

[13] Lj. Stanojević, Ž. Marjanović-Balaban, J. Stanojević, D. Cvetković, Lj. Nikolić, A. Milenković, V. Gojković Cvjetković, Chemical composition and antioxidant

activity of Juniper (Juniperus communis L.) berries essential oil, VI International Congress “Engineering, Environment and Materials in Processing Industry“, Zvornik

2019, 718-719.

[14] European Pharmacopoeia, 7th edition, Council of Europe, Strasbourg 2011, 1160-1161.

ACKNOWLEDGMENTS

This study was carried out within a project No. 19.032/961-176/19 of 31/12/2019., financed by the Ministry for Scientific-Technological Development, Higher Education

and Information Society of the Republic of Srpska.

Ž. R. Marjanović-Balaban1, Lj. P. Stanojević2, J. S. Stanojević2, V. S. Gojković Cvjetković3

1University of Banja Luka, Faculty of Forestry, e-mail: zeljka.marjanovic-balaban@sf.unibl.org; 2University of Niš, Faculty of Technology, e-mail: ljilja76@yahoo.com;

jelena_stanojevic@yahoo.com; 3University of East Sarajevo, Faculty of Technology, e-mail: vesna.gokovic@yahoo.com

CHEMICAL COMPOSITION OF INDUSTRIALLY PRODUCED OF

JUNIPER BERRY ESSENTIAL OIL (Juniperus communis L.) FROM THE

REPUBLIC OF SRPSKA

INTRODUCTION

Table 1. Chemical composition of Juniperus communis L. essential oil

CONCLUSION

Medicinal plants present a rich source of new biologically active compounds. The concept of food that combines nutritional and medicinal benefits is particularly popular

today. Essential oils from aromatic and medicinal plants receive particular attention as

potential natural agents with a wide spectrum of biological activities. Essential oils are

proven to have various pharmacological effects, such as: spasmolytic, carminative,

hepatoprotective, antiviral, and anticarcinogenic effects (Bowles, 2004; Lahlou, 2004).

The essential oil of juniper berry is light yellow in color, and the oil yield ranges

from 0.2 to 2%, depending primarily on the geographical origin. The yield of essential

oil of juniper berry originating from Hungary is 1.2%, from Germany only 0.7%, and

from Iran 0.9% (Razvani et al., 2007). It mainly consists of monoterpenes, namely 80%

α- and β-pinene, thujene, sabinen, 5% terpinen-4-ol, α-terpineol, borneol and geraniol.

Sesquiterpenes (α- and β-cadinen, caryophyllene) are present in traces (Rezvani et al.,

2009).

The antioxidant activity of the essential oil of the fruit of different species of Juniperus L. was investigated in vitro (Emami et al., 2007). The anti-radical activity of

essential oil depends on its components (their chemical nature and concentration) (Misharina and Samusenko, 2008; Misharina et al., 2009; Wei and Shibamoto, 2007; Ruberto

and Baratta, 2009). However, regardless of the differences in chemical composition, terpene hydrocarbons are always dominant. In many cases, the antioxidant activity of the

essential oil cannot be attributed to the dominant components of α- and β-pinene. These monoterpene hydrocarbons in juniper berry essential oil do not significantly contribute

to the process of dialdehyde inhibition (Wei and Shibamoto, 2007). Carriers of antioxidant properties are α- and γ-terpinenes, and to a much lesser extent their sesquiterpene

analogues. Mircene, α- and β-pinene inhibit lipid peroxidations in the second phase, and sabinene, limonene, α-pinene and myrcene show anti-radical activity against the DPPH

radical (Bua-in and Paisooksantivatana, 2009; Roberto et al., 2010). The effect of hydroxyl radicals and protection of ribose from degradation is mainly attributed to the content

of β-pinene and limonene (Emami et al., 2007), while the neutralization of superoxide radicals is determined by the presence of germacrene-D (Karioti et al., 2004). The anti-

radical activity of germacrene-D is due to a ring with ten carbon atoms and three double bonds, which is why it acts as a center rich in electrons (Stoilova et al., 2014).

Numerous studies have shown that the monoterpene components contained in juniper essential oil improve, through their antioxidant activity, the oxidative resistance of living

organisms caused by stress. Their anti-radical activity is reflected in the influence of the most important enzymes catalase, peroxidase and transferase (Sepici-Dincel et al., 2007;

Roberto et al., 2010). S. cerevisiae L. is of great importance for a better understanding of cellular protection by ROS (receptor tyrosine kinase) (Jakubowski et al., 2000;

Manfredini et al., 2004; Tsuzi et al., 2004; Inoue et al., 1999).

In this paper, the identification of chemical components of juniper essential oil from the area of the Republic of Srpska is performed.

Figure1. Juniperus berry

Figure 2. Essential oil

As a raw material for the production of essential oil, the fruits of Juniperus L., type Juniperus communis L., were harvested in October 2019 at the Sokolac location, Republic of

Srpska, Bosnia and Herzegovina.

The juniper berry essential oil has a rich history of traditional uses and benefits. This study showed the composition of essential oil juniper berries in Republic of Srpska.

The essential oil was produced in industrial conditions by the hydrodestilation process. The tested juniper berry essential oil is a liquid of light yellow color, with a specific

aroma. Its qualitative and quantitative composition was determined by GC-MS and GC-FID analysis. GC-MS analysis resulted in identifying 58 compounds (97.1% of the total

oil composition). The highest content of α-pinene (29.3%) has been proved, which is in accordance with the results of other researchers, but also with the requirements of the

European Pharmacopoeia, which prescribes a minimum of 20% α-pinene in the essential oil of juniper berries.

Based on the above, it can be concluded that the essential oil of juniper berries from the Republic of Srpska is of good quality, and that further research into its

characteristics should be continued, all with the aim of starting the production process and greater application in all branches of industry.

No. tret, min Compound Molecular formula RIexp RIlit Method of identification Content, %

1. 9.50 Tricyclene C10H16 917 921 RI, MS 0.1

2. 9.67 α-Thujene C10H16 922 924 RI, MS 1.2

3. 9.99 α-Pinene C10H16 931 932 RI, MS, Co-I 29.3

4. 10.35 α-Fenchene C10H16 941 945 RI, MS tr

5. 10.42 Camphene C10H16 943 946 RI, MS, Co-I 0.2

6. 10.60 Thuja-2,4(10)-diene C10H14 948 953 RI, MS 0.1

7. 10.97 Verbenene C10H16 958 961 RI, MS tr

8. 11.32 Sabinene C10H16 968 969 RI, MS 9.5

9. 11.42 β-Pinene C10H16 971 974 RI, MS, Co-I 2.0

10. 11.93 Myrcene C10H16 986 988 RI, MS 12.7

11. 12.37 α-Phellandrene C10H16 998 1002 RI, MS 0.1

12. 12.58 δ-3-Carene C10H16 1004 1008 RI, MS 0.1

13. 12.81 α-Terpinene C10H16 1010 1014 RI, MS 0.7

14. 13.10 p-Cymene C10H14 1018 1020 RI, MS 0.5

15. 13.27 Limonene C10H16 1023 1024 RI, MS, Co-I 3.6

16. 13.92 (E)-β-Ocimene C10H16 1042 1044 RI, MS tr

17. 14.34 γ-Terpinene C10H16 1053 1054 RI, MS 1.4

18. 14.66 cis-Sabinene hydrate C10H16 1062 1065 RI, MS 0.1

19. 15.41 Terpinolene C10H16 1084 1086 RI, MS 1.1

20. 16.78 α-Campholenal C10H16O 1122 1122 RI, MS 0.1

21. 17.47 trans-Sabinol C10H16O 1142 1137 RI, MS 0.1

22. 18.25 Borneol C10H18O 1164 1165 RI, MS, Co-I 0.1

23. 18.53 cis-Pinocamphone C10H16O 1172 1172 RI, MS tr

24. 18.66 Terpinen-4-ol C10H18O 1175 1174 RI, MS 2.4

25. 19.10 α-Terpineol C10H18O 1187 1186 RI, MS, Co-I 0.2

26. 19.31 Myrtenal C10H16O 1194 1195 RI, MS 0.1

27. 20.58 trans-Chrysanthenyl acetate C12H18O2 1231 1235 RI, MS tr

28. 21.29 Piperitone C10H16O 1252 1249 RI, MS 0.1

29. 22.35 Isobornyl acetate C12H20O2 1283 1283 RI, MS, Co-I 0.2

30. 24.06 δ-Elemene C15H24 1335 1335 RI, MS 0.1

31. 24.42 α-Terpinyl acetate C12H20O2 1346 1346 RI, MS tr

32. 24.47 α-Cubebene C15H24 1347 1345 RI, MS 0.8

33. 25.34 α-Copaene C15H24 1375 1374 RI, MS 0.3

34. 25.44 Geranyl acetate C12H20O2 1378 1379 RI, MS tr

35. 25.86 β-Elemene C15H24 1391 1389 RI, MS 2.3

36. 26.78 (E)-Caryophyllene C15H24 1421 1417 RI, MS 3.4

37. 27.04 β-Copaene C15H24 1429 1430 RI, MS tr

38. 27.13 γ-Elemene C15H24 1432 1434 RI, MS 1.9

39. 27.77 (E)-β-Farnesene C15H24 1453 1454 RI, MS tr

40. 27.83 α-Humulene C15H24 1455 1452 RI, MS 3.2

41. 28.09 cis-Muurola-4(14),5-diene C15H24 1464 1465 RI, MS 0.2

42. 28.50 γ-Muurolene C15H24 1477 1478 RI, MS 0.6

43. 28.71 Germacrene D C15H24 1484 1484 RI, MS 8.4

44. 28.83 β-Selinene C15H24 1488 1489 RI, MS 0.5

45. 29.00 trans-Muurola-4(14),5-diene C15H24 1493 1493 RI, MS tr

46. 29.11 Bicyclogermacrene C15H24 1497 1500 RI, MS 2.0

47. 29.17 α-Muurolene C15H24 1499 1500 RI, MS tr

48. 29.37 δ-Amorphene C15H24 1506 1511 RI, MS 0.2

49. 29.61 γ-Cadinene C15H24 1514 1513 RI, MS 0.9

50. 29.86 δ-Cadinene C15H24 1523 1522 RI, MS 1.7

51. 30.11 trans-Cadina-1,4-diene C15H24 1532 1533 RI, MS 0.1

52. 30.28 α-Cadinene C15H24 1537 1537 RI, MS 0.3

53. 30.43 Selina-3,7(11)-diene C15H24 1543 1545 RI, MS 0.2

54. 30.92 Germacrene B C15H24 1560 1559 RI, MS 2.9

55, 31.48 Spathulenol C15H24O 1579 1577 RI, MS 0.2

56. 31.66 Caryophyllene oxide C15H24O 1585 1582 RI, MS 0.2

57. 33.24 epi-α-Cadinol C15H26O 1642 1638 RI, MS 0.3

58. 33.61 α-Muurolol (Torreyol) C15H26O 1655 1644 RI, MS 0.4

Total identified (%) 97.1

Grouped components (%)

Monoterpene hydrocarbons (1-19) 62.7

Oxygen-containing monoterpenes (20-29, 31, 34) 3.3

Sesquiterpene hydrocarbons (30, 32, 33, 35-54) 30.0

Oxygen-containing sesquiterpenes (55-58) 1.1

tret.: Retention time; RIlit-Retention indices from literature (Adams, 2007); RIexp: Experimentally determined retention indices using a homologous series of n-alkanes (C8-C20) on the HP-5MS column. MS: constituent identified by mass-spectra comparison; RI:

constituent identified by retention index matching; Co-I: constituent identity confirmed by GC co-injection of an authentic sample; tr = trace amount (