OPTIMIZATION OF THE EXTRACTION CONDITIONS OF PHENOLIC COMPOUNDS FROM ALCHEMILLA VULGARIS USING RESPONSE SURFACE METHODOLOGY

Öz

The object of this study is to determine the best solvent and optimum extraction conditions for extraction of maximum phenolic compounds from Alchemilla vulgaris leaves. Extractions were carried out using solvents with different polarities. Box-Behnken Design was used to optimize extraction conditions including extraction time, temperature, and liquid/solid ratio. In the study, extract obtained with acetone-water indicated the highest total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activity. Optimum extraction conditions for both responses were found as 168 min, 48.5 °C, and liquid/solid ratio of 41:1. The maximum TPC and TFC values were determined as 7.17 mg GAE/gdw and 3.63 mg QE/gdw under optimum extraction conditions. Concentrations of five phenolic compounds analyzed by HPLC increased significantl after optimization. The results indicated that optimizing extraction conditions is critical for quantification of phenolic compounds. The present model can contribute to food industry where phenolic compounds have potential use as biopreservatives.

Anahtar Kelimeler

• Alchemilla vulgaris
• Antioxidant activity
• Box-Behnken Design
• Extraction optimization
• Phenolic compounds

ALCHEMILLA VULGARIS'TEN FENOLİK BİLEŞİKLERİN EKSTRAKSİYON KOŞULLARININ YANIT YÜZEY YÖNTEMİ KULLANILARAK OPTİMİZASYONU

Öz

Bu çalışmanın amacı, Alchemilla vulgaris bitkisinin yapraklarından maksimum fenolik bileşik ekstraksiyonu için en iyi çözücünün ve optimum ekstraksiyon koşullarının belirlenmesidir. Ekstraksiyonlar, farklı polaritelere sahip çözücüler kullanılarak gerçekleştirildi. Box-Behnken tasarımı (BBD), ekstraksiyon süresi, ekstraksiyon sıcaklığı ve sıvı / katı oranı içeren ekstraksiyon koşullarını optimize etmek için kullanıldı. Çalışmada aseton-su ile elde edilen ekstrakt, en yüksek toplam fenolik içerik (TPC), toplam flavonoid içerik (TFC) ve antioksidan aktivite göstermiştir. Her iki yanıt için optimum ekstraksiyon koşulları, 168 dakika, 48.5 ºC ve 41:1 sıvı / katı oranı olarak bulundu. Optimum ekstraksiyon koşulları altında maksimum TPC ve TFC değerleri 7.17 mg GAE / gdw ve 3.63 mg QE / gdw olarak belirlendi. HPLC ile analiz edilen beş fenolik bileşiğin konsantrasyonlarının, optimizasyondan sonra önemli ölçüde arttığı bulundu. Sonuçlar, ekstraksiyon koşullarının optimize edilmesinin fenolik bileşiklerin miktarının belirlenmesi için önemli olduğunu göstermektedir. Mevcut model, fenolik bileşiklerin biyo-koruyucu olarak potansiyel kullanıma sahip olduğu gıda endüstrisine katkıda bulunabilir.

Anahtar Kelimeler

• Alchemilla vulgaris
• Antioksidan aktivite
• Box-Behnken Tasarımı
• Ekstraksiyon optimizasyonu
• Fenolik bileşikler

Kaynakça

1. Acet, T., Özcan, K. (2018). Aslanpençesi (Alchemilla ellenbergiana) Ekstrelerinin Antioksidan ve Antimikrobiyal Özelliklerinin Belirlenmesi. GUFBED, 8(1): 113-121.
2. Ahmadian-Kouchaksaraie, Z., Niazmand, R., Najafi, M.N. (2016). Optimization of the subcritical water extraction of phenolic antioxidants from Crocus sativus petals of saffron industry residues: Box-Behnken design and principal component analysis. Innov. Food Sci. Emerg. Technol., 36: 234-244, doi: 10.1016/j.ifset.2016.07.005.
3. Al-Farsi, M.A., Lee, C.Y. (2008). Optimization of phenolics and dietary fibre extraction from date seeds. Food Chemistry, 108 (3): 977-985, doi: 10.1016/j.foodchem.2007.12.009.
4. Belwal, T., Dhyani, P., Bhatt, I.D., Rawal, R.S., Pande, V. (2016). Optimization extraction conditions for improving phenolic content and antioxidant activity in Berberis asiatica fruits using response surface methodology (RSM). Food Chemistry, 207:115-124, doi: 10.1016/j.foodchem.2016.03.081.
5. Boroja, T., Mihailović, V., Katanić, J., Pan, S.P., Nikles, S., Imbimbo, P., Bauer, R. (2018).The biological activities of roots and aerial parts of Alchemilla vulgaris L. South African Journal of Botany, 116: 175-184, doi: 10.1016/j.sajb.2018.03.007.
6. Borra, S.K., Gurumurthy, P., Mahendra, J. (2013). Antioxidant and free radical scavenging activity of curcumin determined by using different in vitro and ex vivo models. J Med Plant Res, 7(36): 2680-2690, doi: 10.5897/JMPR2013.5094.
7. Cheok, C.Y., Chin, N.L., Yusof, Y.A., Talib, R.A., Law, C.L. (2012). Optimization of total phenolic content extracted from Garcinia mangostana Linn. hull using response surface methodology versus artificial neural network. Industrial crops and products, 40: 247-253, doi: 10.1016/j.indcrop.2012.03.019.
8. Denev, P., Kratchanova, M., Ciz, M., Lojek, A., Vasicek, O., Blazheva, D., Nedelcheva, P., Vojtek, L., Hyrsl, P. (2014). Antioxidant, antimicrobial and neutrophil-modulating activities of herb extracts. Acta Biochimica Plonica, 61: 359–367.

9. Do, Q.D., Angkawijaya, A.E., Tran-Nguyen, P.L., Huynh, L.H., Soetaredjo, F.E., Ismadji, S., Ju, Y.H. (2014). Effect of extraction solvent on total phenol content, total flavonoid content, and antioxidant activity of Limnophila aromatica. J Food Drug Anal, 22(3): 296-302.
10. Duckstein, S.M., Lotter, E.M., Meyer, U., Lindequist, U. (2012). Phenolic Constituents from Alchemilla vulgaris L. and Alchemilla mollis (Buser) Rothm. at Different Dates of Harvest. Z. Naturforsch., C: J. Biosci, 67(11–12): 529–540, doi: 10.1515/znc-2012-11-1201.
11. Fidrianny, I., Natalia, S., Insanu, M. (2015). Antioxidant capacities of various fruit extracts from three varieties of tomato and correlation with total phenolic, flavonoid, carotenoid content. Int J Pharm Clin Res, 7(4): 283-289.
12. Haminiuk, C.W.I., Plata-Oviedo, M.S.V., de Mattos, G., Carpes, S.T., Branco, I.G. (2014). Extraction and quantification of phenolic acids and flavonols from Eugenia pyriformis using different solvents. J Food Sci Technol, 51(10): 2862-2866, doi: 10.1007/s13197-012-0759-z.
13. Hwang, E., Ngo, H.T., Seo, S.A., Park, B., Zhang, M., Yi, T.H. (2018). Protective effect of dietary Alchemilla mollis on UVB-irradiated premature skin aging through regulation of transcription factor NFATc1 and Nrf2/ARE pathways. Phytomedicine, 39: 125-136.
14. Ilić-Stojanović, S., Nikolić, V., Kundaković, T., Savić, I., Savić-Gajić, I., Jocić, E., Nikolić, L.J. (2017). Thermosensitive hydrogels for modified release of ellagic acid obtained from Alchemilla vulgaris L. extract. Int J Poly Mater, 67(9): 553-563, Doi: 10.1080/00914037.2017.1354202..
15. Krishnappa, N.P., Basha, S.A., Negi, P.S., Prasada Rao, U.J.S. (2017). Phenolic acid composition, antioxidant and antimicrobial activities of green gram (vigna radiata) exudate, husk, and germinated seed of different stages. J Food Process Pres, 41(6): e13273, doi: 10.1111/jfpp.13273.
16. Møller, C., Hansen, S.H., Cornett, C. (2009). Characterisation of tannin-containing herbal drugs by HPLC. Phytochemical Analysis, 20: 231–239, doi: 10.1002/pca.1119.
17. Mouratoglou, E., Malliou, V., Makris, D.P. (2016). Novel glycerol-based natural eutectic mixtures and their efficiency in the ultrasound-assisted extraction of antioxidant polyphenols from agri-food waste biomass. Waste and biomass valorization, 7(6): 1377-1387.
18. Murathan, Z.T. (2018). Kuzeydoğu Anadolu Bölgesi ekolojik koşullarında yetişen bazı tıbbi bitkilerin biyokimyasal içeriği ve antioksidan özelliklerinin belirlenmesi. Balıkesir Üniv Fen Bilim Enst Derg, 20(2): 51-60.
19. Nasr, A., Zhou, X., Liu, T., Yang, J., Zhu, G.P. (2019). Acetone-water mixture is a competent solvent to extract phenolics and antioxidants from four organs of Eucalyptus camaldulensis. Turkish J Biochem, 44(3): 231-239, doi: 10.1515/tjb-2018-0438.
20. Neagu, E., Paun, G., Albu, C., Radu, G.L. (2015). Assessment of acetylcholinesterase and tyrosinase inhibitory and antioxidant activity of Alchemilla vulgaris and Filipendula ulmaria extracts. J Taiwan Inst Chem Eng, 52: 1–6, doi: 10.1016/j.jtice.2015.01.026.
21. Oyaizu, M. (1986). Studies on Product of Browning Reaction: Antioxidative Activities of Products of Browning Reaction Prepared from Glucoseamine. Japanese J Nutr Diet, 44(6): 307-315.
22. Pham, N.M.Q., Vuong, Q.V., Bowyer, M.C., Scarlett, C.J. (2019). Optimization of ultrasound-assisted extraction conditions for phenolic compounds and antioxidant capacity from Tuckeroo (Cupaniopsis anacardioides) fruit. Sep Sci Tech, 55:17, 3151-3160, doi: 10.1080/01496395.2019.1673413.
23. Rebey, I.B., Bourgou, S., Debez, I.B.S., Karoui, I.J., Sellami, I.H., Msaada, K., Marzouk, B. (2012). Effects of extraction solvents and provenances on phenolic contents and antioxidant activities of cumin (Cuminum cyminum L.) seeds. Food Bioprocess Tech, 5(7): 2827-2836.
24. Renda, G., Özel, A., Barut, B., Korkmaz, B., Šoral, M., Kandemir, Ü., Liptaj, T. (2017). Bioassay Guided Isolation of Active Compounds from Alchemilla barbatiflora Juz. Rec Nat Prod, 12: 1-7.
25. Sánchez-Moren, C., Plaza, L., de Ancos, B., Cano, M.P. (2003). Quantitative bioactive compounds assessment and their relative contribution to the antioxidant capacity of commercial orange juices. J. Sci. Food Agric, 83: 430–439.
26. Sepahpour, S., Selamat, J., Abdul Manap, M.Y., Khatib, A., Abdull Razis, A.F. (2018). Comparative analysis of chemical composition, antioxidant activity and quantitative characterization of some phenolic compounds in selected herbs and spices in different solvent extraction systems. Molecules, 23(2): 402, doi: 10.3390/molecules23020402.
27. Shelembe, J.S., Cromarty, D., Bester, M.J., Minnaar, A., Duodu, K.G. (2012). Characterisation of phenolic acids, flavonoids, proanthocyanidins and antioxidant activity of water extracts from seed coats of marama bean [Tylosema esculentum]–an underutilised food legume. Int J Food Sci Tech, 47(3): 648-655.
28. Shi, J., Yu, J., Pohorly, J., Young, J.C., Bryan, M., Wu, Y. (2003). Optimization of the extraction of polyphenols from grape seed meal by aqueous ethanol solution. J Food Agric Environ, 1(2): 42-7.
29. Silva, E.M., Rogez, H., Larondelle, Y. (2007). Optimization of extraction of phenolics from Inga edulis leaves using response surface methodology. Sep Pur Tech, 55(3): 381-387.
30. Sousa, M.S.B., Júnior, J.M.L., de Souza Buarque, D. (2019). Optimization of the extraction of polyphenols and antioxidant capacity from Byrsonima crassifolia (L.) kunth fruit by response surface methodology. In Plant Physiological Aspects of Phenolic Compounds. IntechOpen, doi:10.5772/intechopen.83457.
31. Tabaraki, R., Rastgoo, S. (2014). Comparison between conventional and ultrasound-assisted extractions of natural antioxidants from walnut green husk. Korean J Chem Eng, 31(4): 676-683.
32. Tasić-Kostov, M., Arsić, I., Pavlović, D., Stojanović, S., Najman, S., Naumović, S., Tadić, V. (2019). Towards a modern approach to traditional use: in vitro and in vivo evaluation of Alchemilla vulgaris L. gel wound healing potential. Journal of ethnopharmacology, 238: 111789, doi: 10.1016/j.jep.2019.03.016.
33. Usta, C., Yildirim, A.B., Turker, A.U. (2014). Antibacterial and antitumour activities of some plants grown in Turkey. Biotech & Biotech Equip, 28(2): 306-315.
34. Vlaisavljević, S., Jelača, S., Zengin, G., Mimica-Dukić, N., Berežni, S., Miljić, M., Stevanović, Z.D. (2019). Alchemilla vulgaris agg.(Lady's mantle) from central Balkan: antioxidant, anticancer and enzyme inhibition properties. RSC Advances, 9(64): 37474-37483.
35. Yazici, S.O., Ozmen, I., Yildirim, B., Genc, H., Ozeloglu, B., Gülsün, M., Ozcaka, S. (2020). Biochemical Composition of Lathyrus L. Seeds: Antioxidant Activities, Phenolic Profiles, β-ODAP and Protein Contents. Legume Research-An International Journal, 43: 723-727, doi: 10.18805/LR-516.
36. Yılmaz, Y., Toledo, R.T. (2006). Oxygen radical absorbance capacities of grape/wine industry byproducts and effect of solvent type on extraction of grape seed polyphenols. J Food Comp Anal, 19(1): 41-48, doi: 10.1016/j.jfca.2004.10.009.
37. Zhang, L.L., Xu, M., Wang, Y.M., Wu, D.M., Chen, J.H. (2010). Optimizing ultrasonic ellagic acid extraction conditions from infructescence of Platycarya strobilacea using response surface methodology. Molecules, 15(11):7923-7932, doi: 10.3390/molecules15117923.
38. Zhang, Y., Zhang, X., Qi, W., Xu, J., Yuan, Z., Wang, Z. (2018). Ann and Rsm Based Optimization of Cellulase Production By Hypocrea Sp Z28 By Submerged Fermentation. Cell Chem Technol, 52(3-4): 259-264.
39. Zhao, H.X., Zhang, H.S., Yang, S.F. (2014). Phenolic compounds and its antioxidant activities in ethanolic extracts from seven cultivars of Chinese jujube. Food Sci Humn Well, 3(3-4), 183-190.