Document Type : Research Article


Gorgan University of Agricultural Sciences and Natural Resources


The aim of this study is extraction of phenolic compounds from walnut leaves of touyserkani variety with two extraction methods (traditional method and with microwave assisted extraction (MAE)) and evaluation of their antioxidant activity. Total Phenolic contents and antioxidant properties of aqueous, methanol (80%) and ethanol (50%) extracts in different times was determined. At the next stage, effect of methanol extract on retarding of oxidation of soy oil was assessed. All of extracts showed antioxidant activity in a concentration-dependent way. In MAE, methanol extract had highest total phenol whose content is 89.15 ± 0.25 mg/g. the ethanol extract presented highest activity in radical-scavenging activity (EC50 = 27.90 µg/ml), reducing power assay (EC50 = 93.26 µg/ml) and antioxidant activity assay (EC50=68.23 µg/ml). Also extract at 1000 ppm could control oxidation of soy oil and this extract was effective than all of synthesis antioxidant in retarding oil oxidation Hence we can present walnut leaves as a potential source of phenolic compounds and natural antioxidant.


قره خانی، م.، رفیعی، ز.، قربانی، م. و جعفری، س. م.، 1388 . سیستم مایکروویو محفظه باز برای استخراج ترکیبات مؤثره از گیاهان دارویی، 59321.
کامکار، ا. 1388. مطالعه فعالیت آنتی اکسیدانی اسانس و عصاره شوید ایرانی. فصلنامه دانشگاه علوم پزشکی و خدمات بهداشتی درمانی گناباد، 15: 2، ص 11-17.
AOAC, 1990, Official Methods of Analysis. Association of Official Analytical Chemists, 15th ed. Washington, DC., USA.
Arabshahi-Delouee, S. and Urooj, A., 2007, Antioxidant properties of various solvent extracts of mulberry (Morus indica L.) leaves. Food Chemistry, 102(4), 1233-1240.
Barreira, J. C. M., Ferreira, I. C. F. R., Oliveira, M. B. P. P., and Pereira, J. A., 2008, Antioxidant activities of the extracts from chestnut flower, leaf, skins and fruit. Food Chemistry, 107(3): 1106-1113.
Besbes, S., Blecker, C., Deroanne, C., Bahloul, N., Lognay, G., Drira, N.-E., 2004, Date seed oil: Phenolic, tocopherol and sterol profiles. Journal of Food Lipids, 11, 251–265.
Bouaziz, M. and Sayadi, S., 2005, Isolation and evaluation of antioxidants from leaves of a Tunisian cultivar olive tree. European Journal of Lipid Science and Technology, 107: 497–504.
Cacace, J.E., Mazza, G., 2006, Pressurized low polarity water extraction of lignans from
whole flaxseed. J Food Eng, 77:1087-95.
Casazza, A.A., Aliakbarian, B., Mantegna, S., Cravotto, G. and Perego, P., 2010, Extraction of phenolics from Vitis vinifera wastes using non-conventional techniques. Journal of Food Engineering, 100(1), 50-55.
Chemat, S., Aït-Amar, H., Lagha, A. and Esveld, D.C., 2005, Microwave-assisted extraction kinetics of terpenes from caraway seeds. Chemical Engineering and Processing, 44(12), 1320-26.
Chirinos, R., Rogez, H., Campos, D., Pedreschi, R. and Larondelle, Y., 2007, Optimization of extraction conditions of antioxidant phenolic compounds from mashua (Tropaeolum tuberosum Ruiz and Pavon) tubers. Separation and Purification Technology, 55, 217-225.
Chun, S.S., Vattem, D.A., Lin, Y.T. and Shetty, k., 2005, Phenolic antioxidants from clonal oregano (Origanum vulgare) with antimicrobial activity against Helicobacter pylori. Process Biochemistry, 40, 809-816.
Contini, M., Baccelloni, S., Massantini, R. and Anelli, G., 2008, Extraction of natural antioxidants from hazelnut (Corylus avellana L.) shell and skin wastes by long maceration at room temperature. Food Chemistry, 110, 659–669.
Farag, R.S., Mahmoud, E.A.and Basuny, A.M., 2007, Use crude olive leaf juice as a natural antioxidant for the stability of sunflower oil during heating. International Journal of Food Science andTechnology, 42: 107-115.
Goli, A.H., Barzegar, M. and Sahari, M.A., 2005, Antioxidant activity and total phenolic compounds of pistachio (Pistachia vera) hull extracts. Food Chemistry, 92: 521-5.
Isabel, F., Almeida., Eduarda Fernandes., Jose, L. F. C., 2008, walnut (Juglans regia) leaf extracts are strong scavengers of pro-oxidant reactive species. Food Chemistry, 106: 1014-1020
Karpin, A., ska, M., Borowski, J. and Danowska- Oziewicz, M., 2001, Use of natural antioxidants in readyto-serve food. Food Chemistry, 72: 5-9
Kumaran, A. and Karunakaran, R.J., 2007, In vitro antioxidant ctivities of methanol extracts of five Phyllanthus species from India. LWT, 40: 344-352.
Li, B.B., Smith, B. and Hossain, M.M., 2006, Extraction of phenolics from citrus peels: I. Solvent extraction method. Separation and Purification. Technology, 48(2), 182-188.
Li, J.W., Ding, S.D. and Ding, X.L., 2005, Comparison of antioxidant capacities of extracts from five cultivars of Chinese jujube. Process Biochemistry, 40(11), 3607-3613.
Li, J., Zu, Y.G., Fu, Y.J., Yang, Y.C., Li, S.M., Li, Z.N. and Wink, M., 2010, Optimization of microwave-assisted extraction of triterpene saponins from defatted residue of yellow horn (Xanthoceras sorbifolia Bunge.) kernel and evaluation of its antioxidant activity. Innovative Food Science and Emerging Technologies, 11(4), 637-643.
Lujan‚ R. J., R. J., Rodriguez. and Castro, M. D. L., 2006, Multivariate optimisation of the microwave-assisted extraction of oleuropein and related biophenols from olive leaves. Journal of Analytical and Bioanalytical Chemistry, 385(4), 753-759.
Mandal, V., Mohan, Y. and Hemalatha, S., 2007, Microwave Assisted extraction- An innovative and promising extraction tool for medicinal plant research. Food Chemistry, 92, 144-151.
Mir-Ahmadi, F., Fatemi, H., Sahari, M.A., 2006, Effect of green Tea extract on the inhibition ofsunflower oil oxidation. IJFST, 2 (4): 61 -70.
Olivera, I., Sousa, A., Valentao, P., Andrade, P., Ferreira, F., Bento, A., Seabra, R., Estevinho, L. and Pereira, J. A., 2007, Hazel (Corylus avellana) leaves as source of antimicrobial and antioxidant compounds. Food Chemistry, 105, 1018-1025.
Pan, X., Niu, G. and Liu, H., 2003, Microwave-assisted extraction of tea polyphenols and tea caffeine from green tea leaves. Chemical Engineering and Processing, 42(2), 129-133.
Pan, Y., Wang, K., Huang, S., Wang, H., Mu, X., He, C., Ji, X., Zhang, J. and Huang, F., 2008, Antioxidant activity of microwave-assisted extract of longan (Dimocarpus Longan Lour.) peel. Food Chemistry, 106(3), 1264-1270.
Pereira, J. A., Oliveira, I., Sousa, A., Valento, P., Andrade, P. B., Ferreira, I.C.F.R., Ferreres, F., Bento, A., Seabra, R. and Estevinho, L,, 2007, Walnut (Juglans regia L.) leaves: Phenolic compounds, antibacterial activity and antioxidant potential of different cultivars. Food and Chemical Toxicology, 45(11), 2287-2295.
Prieto, P., Pineda, M. and Aguilar, M., 1999, Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Analytical Biochemistry, 269, 337-341.
Proestos, C. and Komaitis, M., 2008, Application of microwave-assisted extraction to the fast extraction of plant phenolic compounds. LWT-Food Science and Technology, 41(4), 652-659.
Ramos, L., Kristenson, E.M. and Brinkman, U.A.T., 2002, Current use of pressurised liquid extraction and subcritical water extraction in environmental analysis. Journal of Chromatography A, 975, 3-29.
Rumbaoa, R.G.O., Cornago, D.F. and Geronimo, I.M., 2009, Phenolic content and antioxidant capacity of Philippine potato (Solanum tuberosum) tubers. Journal of Food Composition and Analysis, 22(6), 546-550.
Sanchez-Moreno, C., Larrauri, J.A.and Saura-Calixto, F., 1999, Free radical scavenging capacity and inhibition of lipid oxidation of wines, grape juices and related polyphenolic constituents. Food Research International, 32:407–412.
Singh, G. and Marimuthu, P.,2006, Antioxidant and biocidal activities of Carum nigrum (seed) essential oil, oleoresin, and their selected components. Journal of Agricultural and Food Chemistry, 54, 174-81.
Singh, G., Maurya, S. and Delampasona, M.P., 2007, A comparison of chemical antioxidant and antimicrobial studies of cinnamon leaf and bark volatile oils, oleoresins and their constituents. Food and Chemical Toxicology, 45, 1650-1661.
Shahsavari, N., Barzegar, M.A., Sahari, M.A. and Naghdibadi, H., 2008, Antioxidant activity and chemical characterization oil of Bunium persicum. Plant Foods Human Nutrition, 63, 183-88.
Shon, M.Y., Lee, J., Choi, J.H., Choi, S.Y., Nam, S.H., Seo, K.I., Lee, S.W., Sung, N.J., and Park, S.K., 2007, Antioxidant and free radical scavenging activity of methanol extract of chungkukjang. Journal of Food Composition and Analysis, 20(2): 113-118.
Spigno, G. and De Faveri, D.M., 2009, Microwave-assisted extraction of tea phenols: A phenomenological study. Journal of Food Engineering, 93(2), 210-217.
Spigno, G., Tramelli, L. and De Faveri, D.M., 2007, Effects of extraction time, temperature and solvent on concentration and antioxidant activity of grape marc phenolics. Journal of Food Engineering, 81(1), 200-208.
Sutivisedsak, N., Cheng, H.N., Willett, J.L., Lesch, W.C., Tangsrud, R.R. and Biswas, A., 2010, Microwave-assisted extraction of phenolics from bean (Phaseolus vulgaris L.). Food Research International, 43(2), 516-519.
Suzuki, M., Watanabe, T., Miura, A., Harashima, E., Nakagawa, Y. and Tsuji, K., 2002, An extraction solvent optimum for analyzing polyphenol contents by Folin-Denis assay. Nippon Shokuhin Kagaku Kaishi, 49: 507-511
Turkmen, N., Sari, F. and Velioglu, Y.S., 2006, Effects of extraction solvents on concentration and antioxidant activity of black and black mate tea polyphenols determined by ferrous tartrate and Folin-Ciocalteu methods. Food Chemistry, 99(4), 835-841.
Wang, L. and Weller, C.L., 2006, Recent advances in extraction of nutraceuticals from plants. Trends in Food Science and Technology, 17(6), 300-312.