An Optimization Study for The Production of Origanum onites Tincture by Response Surface Methodology: Effect of Solid/Liquid Ratio, Ethanol Concentration and Storage Period

Öz

In this study, Origanum onites was used for the production of tincture, a hydroalcholic extract, and manufacturing conditions

were optimized to determine the processing factor variable levels to increase the final quality of samples. For this purpose,

response surface methodology study was applied and Box Behnken design was used for the optimization. Three factors were

selected namely liquid/solid ratio, ethanol concentration and storage period for the preparation of samples. Total phenolic

content, antiradical activity and efficient of concentration (EC50) values were determined to evaluate the bioactivity of tincture

samples. It was observed that the total phenolic content of the tincture samples was affected by ethanol concentration and

liquid/solid ratio. Total phenolic contents of tincture samples were in the range of 964.1-7020.8 mg GAE/ L samples while the

efficient concentration (EC50) ranged from 44.88 to 89.56. Increase of solid level increased the bioactivity values of samples,

but this increment decreased the tincture yield significantly. Optimization results showed that the highest total phenolic content

would be at 50% ethanol concentration, 6 ml ethanol level which means the liquid/solid ratio is 1:5 and 15 days storage for the

tincture samples.

Anahtar Kelimeler

• Origanum onites,tincture,bioactivity,optimization,response surface methodology

Origanum onites Tentürü Üretim Şartlarının Tepki Yüzey Yöntemi ile Optimizasyonu: Katı/Sıvı Oranı, Etanol Konsantrasyonu ve Depolama Süresinin Etkisi

Öz

Bu çalışmada, bir hidroalkolik ekstrakt tipi olan tentür üretimi amacıyla Origanum onites bitkisi kullanılmış ve son ürün

kalitesini artırmaya yönelik olarak üretim şartlarını optimize etmek üzere proses faktör seviyelerinin belirlenmesi amaçlanmıştır.

Bu amaçla, optimizasyon işlemi için tepki yüzey yöntemi tabanlı Box-Behnken deneme tasarımından faydalanılmıştır.

Örneklerin hazırlanması amacıyla sıvı/katı oranı, etanol konsantrasyonu ve depolama süresi faktör olarak seçilmiştir. Tentür

örneklerinin biyoaktif özelliklerinin belirlenmesi amacıyla toplam fenolik madde miktarı, antiradikal kapasite ve EC50 değeri

hesaplanmıştır.Sıvı/katı oranı ve etanol konsantrasyonunun örneklerin toplam fenolik madde miktarı üzerinden etkili olduğu

görülmüştür. Örneklerin toplam fenolik madde değerleri 964.1-7020.8 mg GAE/ L, EC50 değerleri ise 44.88-89.56 aralığında

değişim göstermiştir. Örnek konsantrasyonunun artışı toplam fenolik madde miktarını artırırken, bu artış tentür veriminde

düşüşe yol açmıştır. Optimizasyon sonuçları en yüksek toplam fenolik madde miktarının, %50 etanol konsantrasyonunda, 1:5

katı/sıvı oranında, 15 günlük depolama süresi sonunda elde edilebileceğini göstermiştir.

Anahtar Kelimeler

• Origanum onites,tentür,biyoaktivite,optimizasyon,tepki yüzey yöntemi

Kaynakça

1. Barros, L., Heleno, S. A., Carvalho, A. M., Ferreira, I. C. F. R. (2010). Lamiaceae often used in Portuguese folk medicine as a source of powerful antioxidants: Vitamins and phenolics. LWT – Food Science and Technology, 43: 544–550.
2. Baydar, H., Sagdic¸ O., Ozkan, G., Karadogan, T. (2004). Antibacterial activity and composition of essential oils from Origanum, Thymbra and Satureja species with commercial importance in Turkey. Food Control, 15:169–172.
3. Brand-Williams, W., Cuvelier, M.E., Berset, C. (1995). Use of a free radical method to evaluate antioxidant activity. Food Science and Technology, 28 (1): 25-30.
4. Bone, K. (2003). A clinical guide to blending liquid herbs. Herbal Formulations for the individual patients. Churchill Livingstone, Elsevier, Missouri.
5. Box, G.E.P., Behnken, D. W. (1960). Some new three level designs for the study of quantitative variables. Technometrics, 7: 455–475
6. Castilho, P.C., Savluchinske-Feio, S., Weinhold, T.S., Gouveia, S.C. (2012). Evaluation of the antimicrobial and antioxidant activities of essential oils, extracts and their main components from oregano from Madeira Island, Portugal. Food Control, 23(2): 552-558.
7. Economou, K. D., Oreopoulou, V., Thomopoulos, C. D. (1991). Antioxidant activity of some plant extracts of the family labiatae. Journal of American Oil Chemists Society, 68: 109–113.
8. Gruenwald, J., Brendler, T., Jaenicke, C. (2000). PDR for Herbal Medicines. Medical Economic Co. Inc., Montvale, NJ.

9. Ivanova, D., Gerova, D., Chervenkov, T., Yankova, T. (2005). Polyphenols and antioxidant capacity of Bulgarian medicinal plants. Journal of Ethnopharmacology, 96: 145-50.
10. Josifovi_c, M., Stjepanovi_c, L., Jankovi_c, M. M., Gaji_c, M., Koji_c, M., Dikli_c, N. (1974).Flora of SR Serbia (in serbian). SANU, 6: 474-475.
11. Kaurinovic, B., Popovic, M., Vlaisavljevic, S., Trivic, S. (2011). Antioxidant capacity of Ocimum basilicum L. and Origanum vulgare L. extracts. Molecules, 16: 7401–7414.
12. Kikuzaki, H., Nakatani, N. (1989). Structure of a new antioxidative phenolic acid from oregano (Oreganum vulgare L). Agricultural and Biological Chemistry, 53: 519–524.
13. Kosar, M., Dorman, H.J.D., Hiltunen, R. (2005). Effect of an acid treatment on the phytochemical and antioxidant characteristics of extracts from selected Lamiaceae species. Food Chemistry, 91: 525–533.
14. Kosar, M., Dorman, H.J.D., Bachmayer, O., Baser, K.H.C., Hiltunen, R. (2003). An improved on-line HPLC-DPPH method for the screening of free radical scavenging compounds in water extracts of Lamiaceae plants. Chemistry and Natural Compounds, 39:161–166.
15. Leporatti, M. L., Ivancheva, S. (2003). Preliminary comparative analysis of medicinal plants used in the traditional medicine of Bulgaria and Italy. Journal of Ethnopharmacology, 87: 123-142.
16. Licina, B.Z., Stefanovic, O.D., Vasic, S.M., Radojevic, I.D., Dekic, M.S., Comic, L.R. (2013). Biological activities of the extracts from wild growing Origanum vulgare L. Food Control, 33: 498-504.
17. Moreno, C.S., Larrauri, J.A.; Calixto, F.S. (1998). A procedure to measure the antiradical efficiency of polyphenols. Journal of the Science and Food Agriculture, 76: 270 – 276
18. Ozbek, T., Gulluce, M., S_ah_In, F., Ozkan, H., Sevsay, S., Baris, O. (2008). Investigation of the antimutagenic potentials of the methanol extract of Origanum vulgare L. subsp. vulgare in the eastern Anatolia region of Turkey. Turkish Journal of Biology, 32: 271–276.
19. Ozkan, G., Baydar, H., Erbas, S. (2010). The influence of harvest time on essential oil composition, phenolic constituents and antioxidant properties of Turkish oregano (Origanum onites L.) Journal of the Science of Food and Agriculture, 90, 205–209.
20. Pizzale, L., Bortolomeazzi, R., Vichi, S., Uberegger, E., Lanfranco, S.C. (2002). Antioxidant activity of sage (Salvia officinalis and S. fruticosa) and oregano (Origanum onites and O. indercedens) extracts related to their phenolic compound content (2002). Journal of the Science of Food and Agriculture, 82:1645–1651.
21. Sahin, F., Gulluce, M., Daferera, D., Sokmen, A., Sokmen, M., Polissiou, M., et al. (2004). Biological activities of the essential oils and methanol extract of Origanum vulgare ssp. vulgare in the eastern Anatolia region of Turkey. Food Control, 15: 549–557.
22. Singleton , V.L., Rossi Jr., J.A. (1965). Colorimetry of total phenolics with phosphomolybdic–phosphotungstic acid reagents. American Journal of Enology and Viticulture, 16: 144 – 158 .
23. Spiridon, I., Bodirlau, R., Teaca, C.A. (2011). Total phenolic content and antioxidant activity of plants used in traditional Romanian herbal medicine. Central European Journal of Biology, 6: 388–396.
24. Seifried, H.E., Anderson, D.E., Fisher, E.I., Milner, J.A. (2007). A review of the interaction among dietary antioxidants and reactive oxygen species. The Journal of Nutritional Biochemistry, 18(9): 567-579.
25. Teixeira, B., Marques, A., Ramos, C., Serrano, C., Matos, O., Neng, N.R., Nogueira, J.M.F., Saraiva, J.A., Nunes, M.L. (2013). Chemical composition and bioactivity of different oregano (Origanum vulgare) extracts and essential oil. Journal of the Science of Food and Agriculture, 93: 2707–2714.
26. Tepe, B., Daferera, D., Sokmen, M., Polissiou, M., Sokmen, A.Q. (2004). The in vitro antioxidant and antimicrobial activities of the essential oil and various extracts of Origanum syriacum L. var bevanii. Journal of the Science of Food and Agriculture 84: 1389–1396.
27. Valko, M., Leibfritz, D., Moncol, J., Cronin, M.T.D., Mazur, M., Telser, J. (2007). Free radicals and antioxidants in normal physiological functions and human disease. The International Journal of Biochemistry & Cell Biology, 39 (1), 44-84.