TÜRKİYE’DE BAZI LİKEN TÜRLERİNDEKİ USNİK ASİTİN HPLC YÖNTEMİ İLE DEĞERLENDİRİLMESİ VE ANTİMİKROBİYAL AKTİVİTELERİ

Abstract

Amaç: Bu çalışmanın amacı, Parmeliacea familyasına ait Parmelia saxatilis L. Ach., Parmelia sulcata Taylor, Parmelina tiliacea Hoffm. Hale, Xanthoparmelia conspersa Ach. Hale, ve Flavoparmelia caperata L. Hale liken türlerinin aseton ekstraktlarının Escherichia coli ATCC 35218 , Enterococcus faecalis RSKK 508 , Proteus mirabilis Pasteur Ens. 235 , Staphylococcus aureus, Bacillus subtilis, Bacillus megaterium, Pseudomonas aeruginosa türlerini içeren yedi farklı bakteri türüne karşı antimikrobiyal aktivitelerinin değerlendirilmesidir.Yöntem: İnhibisyon zon çapları her bir ekstrakt için agar difüzyon yöntemi ile belirlenmiştir. Daha sonra, bu türlerdeki usnik asit miktarları HPLC yöntemi kullanılarak tespit edilmiştir.Bulgular: Bu çalışmada elde edilen verilere göre, Türkiye’den toplanan beş liken türünün geniş bir aralıkta değişen oranlarda antimikrobiyal aktivite gösterdikleri sonucuna ulaşılmıştır. En yüksek usnik asit miktarı % 2.38’lik bir oran ile Flavoparmelia caperata liken türünde tespit edilmiştir. İncelenen liken türlerinin tümünün, S. aureus ve P. aeruginosa hariç; E. coli, B. subtilis ve B. megaterium bakterilerine karşı antimikrobiyal aktivite gösterdiği bulunmuştur. F. caperata ’nın aseton ekstraktının B. subtilis ve B. megaterium’a karşı en yüksek antimikrobiyal aktivite gösterdiği belirlenmiştir.Sonuç: Araştırmada likenlerde usnik asit miktarı arttıkça antimikrobiyal aktivitenin de arttığı belirlenmiştir. Bu araştırmanın, Türkiye’de bulunan bazı Parmelia liken türlerinin usnik asit kompozisyonu ve antimikrobiyal aktivitesi üzerine yapılan ilk çalışma olması nedeni ile önemli olduğu düşünülmektedir. Ayrıca, F. caperata başta olmak üzere çalışılan liken türleri tedavi amaçlı ilaç içerisinde antimikrobiyal ajan olarak kullanılabilir

Keywords

• Liken, usnik asit, HPLC

Evaluation of Usnic Acid in Some Likens of Turkey by HPLC Analysis and Screening of their Antimicrobial Activity

Abstract

Objective: The aim of this study was to evaluate the antimicrobial activity of acetone extracts obtained from the Parmelia saxatilis L. Ach., P. sulcata Taylor, Parmelina tiliacea Hoffm. Hale, Xanthoparmelia conspersa Ach. Hale, and Flavoparmelia caperata L. Hale belonging to family Parmeliaceae against seven different bacterial species including Escherichia coli ATCC 35218 , Enterococcus faecalis RSKK 508 , Proteus mirabilis Pasteur Ens. 235 , Staphylococcus aureus, Bacillus subtilis, Bacillus megaterium, Pseudomonas aeruginosa.Method: The inhibition zone diameters were determined for each extract by the agar diffusion method. Afterwards, quantitative analyses of usnic acid amounts found in these species were determined by HPLC method.Results: Based on the data obtained in this research, it is possible to conclude that five species of lichen collected from Turkey exhibited a broad range of antimicrobial activity at varying degrees. In the study, highest amount of usnic acid was determined in Flavoparmelia caperata as; 2.38% of the dry lichen weight. All of the examined lichen species in this study, with the exception of the S. aureus and P. aeruginosa, were found to show antimicrobial activities against E. coli, B. subtilis and B. megaterium. F. caperata was determined have the highest inhibition effect on B. subtilis and B. megaterium.Conclusion: In this research it was determined that, as the amount of usnic acid concentrations were higher, the antimicrobial activities of them also increased. This study is thought to be important as it is the first report on the usnic acid composition and antimicrobial activity of some Parmelia lichen species found in Turkey. Lichens species especially F. caperata could be used as antimicrobial agents in new drugs for therapy

Keywords

• Lichen, usnic acid, HPLC

References

1. Vartia KO. Antibiotic in lichens. In: Ahmadjian V, Hale ME (eds). The Lichens. Academic Pres, New York. 1973; 547-61.
2. Proska B, Sturdikova M, Pronayova N, Liptaj T. (-)-Usnic acid and its derivatives. Their inhibition of fungal growth and enzyme activity. Pharmazie 1996; 51:195-6.
3. Wu J, Zhang M, Ding D, Tan T, Yan B. Effect of CIadonia alpestris on Trichmonas in vitro. Chinese J. Parasit. Dis 1995; 13: 126-9.
4. Perry NB, Benn MH, Brennan NJ, Burgess EJ, Ellis G, Galloway DJ, Lorimer SD, Tangney RS. Antimicrobial, antiviral and cytotoxic activity of New Zealand lichens. Lichenologist 1999; 31: 627-36.
5. Yamamoto Y, Miura Y, Kinoshita Y, Higuchi M, Yamada Y, Murakami A, Ohigashi H, Koshimizu K. Screening of tissue culture and thalli of lichens and some of their active constituents for inhibition of tumor promoterinduced Epstein-Barr virus activation. Chem. Pharm. Bull 1995; 43(8):1388-90.
6. Takai M, Uehara Y, Beisler JA. Usnic acid derivatives as potential neoplastic agents. J Med Chem 1979; 22: 1380-4.
7. Kristmundsdottir T, Aradottir RAE, Ingolfsdottir K, Ogmundsdottir RM. Solubilization of the lichen metabolite (+ )-usnic acid for testing in tissue culture. J. Pharm. Pharmacol 2002; 54(11): 1447-52.
8. Cocchietto M, Skert N, Nimis PL, Sava G. A review on usnic acid, an interesting natural compound. Naturwissenchaften 2002; 89: 137-46.

9. Vijayakumar CS, Viswanathan S, Kannappa-Reddy M, Parvathavarthini S, Kundu SB, Sukumar E. Antiinflammatory activity of (+) usnic acid, Fitoterapia 2000; 71: 564-6.
10. Okuyama E, Umeyama K, Yamazaki M, Kinoshita Y, Yamamoto Y. Usnic acid and diffractic acid ass analgesic and antipyretic components of Usnea diffracta, Planta Med. 1995; 61: 113-5.
11. Huneck S. The significance of lichens and their metabolites. Naturwissenschaften 1999; 86: 559-70. Ingolfsdottir K. Usnic acid. Phytochemistry 2002; 61: 729-36.
12. Wunderer H. The garlic dilemma. D Apoth Ztg 1986; 126: 1743-5.
13. Seifert P, Bertram C. Usnic acid natural-preservation from lichens. Seifen Ole Fette Wachse 1995; 121: 480-5.
14. Francolini I, Norris P, Piozzi A, Donelli G, Stoodley P. Usnic Acid, a Natural Antimicrobial Agent Able To Inhibit Bacterial Biofilm Formation on Polymer Surfaces. Antimicrob Agents Chemother 2004; 48: 4360-5.
15. Ingolfsdottir K, Chung GAC, Gissurarson SR, Skulason VG, Vilhelmsdottir M. In vitro antimycobacterial activity of lichen metabolites. Eur J Pharm Sci 1998; 6: 141-4.
16. Duman Cansaran D. Farklı liken örneklerindeki usnik asit miktarlarının yüksek basınçlı sıvı kromatografisi (HPLC) yöntemi ile belirlenmesi ve antimikrobiyal aktiviteleri. Türk Hij Den Biyol Derg 2007; 64(3): 17-21.
17. Cansaran D., Çetin D., Halici G.M., Atakol O. Determination of usnic acid in some Rhizoplaca species from the Middle Anatolia and their antimicrobial activities. Z. Naturforsch 2006; 61(1-2): 47-51.
18. Cansaran D, Atakol O, Halıcı MG, Aksoy A. HPLC analysis of usnic acid in some Ramalina species from Anatolia and investigation of their antimicrobial activities. Pharma Biol 2007; 45-1: 77-81.
19. Gulluce M, Aslan A, Sokmen M, Sahin F, Adiguzel A, Agar G, Sokmen A. Screening the antioxidant and antimicrobial properties of the lichens Parmelia saxatilis, Platismatia glauca, Ramalina pollinaria, Ramalina polymorpha and Umbilicaria nylanderiana. Phytomedicine 2006; 13: 515-21.
20. Candan M, Yilmaz M, Tay T, Kivanc M. Antimicrobial activity of extracts of the lichen Xanthoparmelia pokornyi and its gyrophoric and stenosporic acid constituents. Z Naturforsch C 2006; 61: 319–23
21. Candan M, Yılmaz M, Tay T, Erdem M, Turk-Ozdemir A. Antimicrobial activity of extracts of the lichen Parmelia sulcata and its salazinic acid constituent. Z Naturforsch C 2007; 62: 619-21.
22. Cansaran D, Kahya D, Yurdakulol E, Atakol O. Identification and quantitation of usnic acid from the lichen Usnea species of Anatolia and antimicrobial activity. Z. Naturforsch C 2006; 61c: 773-6.
23. Rankovic B, Music M, Sukdolak S. Antimicrobial activity of extracts of the lichens Cladonia furcata, Parmelia caperata, Parmelia pertusa, Hypogymnia physodes and Umbilicaria polyphylla. Br J Biomed Sci 2007a; 64(4): 143-8.
24. Rankovic B, Music M, Sukdolak S. The antimicrobial activity of substances derived from the lichens Physcia aipolia, Umbilicaria polyphylla, Parmelia caperata and Hypogymnia physodes. W J Micro Bio 2007b; 10: 274-7.