Effect of genotype and extraction method on polyphenols content, phenolic acids, and flavonoids of olive leaves (Olea europaea L. subsp. europaea)

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  • Salma Guebebia University of Gabes, National Engineering School of Gabes, Laboratory of Energy, Water, Environment and Process, LR18ES35, Gabes 6072, Tunisia https://orcid.org/0000-0000-0000-0001
  • Khadija Ben Othman University of Gabes, National Engineering School of Gabes, Laboratory of Energy, Water, Environment and Process, LR18ES35, Gabes 6072, Tunisia https://orcid.org/0000-0000-0000-0001
  • Yassine Yahia University of Gabes, Laboratory of Dry Land Farming and Oasis Cropping, IRA, Medenine 4119, Tunisia https://orcid.org/0000-0002-1088-3287
  • Mehrez Romdhane University of Gabes, National Engineering School of Gabes, Laboratory of Energy, Water, Environment and Process, LR18ES35, Gabes 6072, Tunisia https://orcid.org/0000-0003-0640-1393
  • Walid Elfalleh University of Gabes, Higher Institute of Applied Sciences and Technology of Gabes, Department of Industrial Chemistry and Processes, Gabes 6072, Tunisia https://orcid.org/0000-0002-3493-6451
  • Hédia Hannachi University Tunis El Manar, Faculty of Sciences, Biology Department, Laboratory of Plant Productivity and Environmental Constraint, LR18ES04, Tunis 2092, Tunisia https://orcid.org/0000-0002-7550-2182




Polyphenols, Cultivated olive leaves, Wild olive leaves, Genotype, Method of extraction


Polyphenol’s extraction varied according to various factors. In this study, the effect of genotype and method of polyphenols extraction were investigated using leaves of two cultivated and two wild olive varieties and four hydromethanolic extraction methods. Quantitatively, significant differences were observed according to the extraction method, the genotype, and the interaction genotype-method of extraction. The heat reflux extraction showed the highest polyphenols content in wild olive leaves having an amount of 841.17 mg GAE/100 g DM. The qualitative phytochemical examination using high performance liquid chromatography (HPLC) of olive leaves showed some significant differences of phenolic compounds between genotypes. For the same oleaster genotype, the extraction method seemed to influence qualitatively the polyphenols profiles. The quinic acid was the dominant phenolic acid and the luteolin-7-O-glucoside was the major flavonoid observed in wild olive leaves having, respectively, 618.24 and 3211.44 mg/kg DM. The quinic acid has an amount of 400.15 and 275.39 mg/kg and the luteolin-7-O-glucoside has an amount of 2059.62 and 1214.49 mg/kg in cultivars leaves. The extraction by Soxhlet of wild olive leaves showed the highest quinic acid (1085.80 mg/kg DM) and luteolin-7-O-glucoside (3720.15 mg/kg DM) amounts. The hydromethanolic extraction assisted by Soxhlet of wild olive leaves constituted the optimal method to obtain high polyphenols contents enriched with phenolic acids and flavonoids.


Abdellatif, B., Bouabdellah, B., Samia, M., Mohamed, H., 2019. Comparison of ethanolic extract yield, pH, Polyphenols and flavonoids in algarian Propulus collected from various geographic region. Bionature, 39, 1-6.

Al-Bandak, G., Reopoulou, V., 2007. Antioxidant properties and composition of Majorana syriaca extracts. European Journal of Lipid science and Technology, 109, 247-255. DOI: https://doi.org/10.1002/ejlt.200600234

Andary, C., Longepierre, D., Le Cong, K., Hul, S., Zaremski, A., Michaloud, G., 2019. Study of a chemotaxonomic marker able to identify the genus Aquilaria (Thymelaeaceae). Bois et Forêts des Tropiques, Montpellier, 341, 29-38. DOI: https://doi.org/10.19182/bft2019.341.a31744

Ben Mohamed, M., Guasmi, F., Ben Ali, S., Radhouani, F., Faghim, J., Triki, T., Grati Kammoun, N., Baff, C., Lucini, L., Benincasa, C., 2018. The LC-MS/MS characterization of phenolic compounds in leaves allows classifying olive cultivars grown in South Tunisia. Biochemical Systematics and Ecology, 78, 84-90. DOI: https://doi.org/10.1016/j.bse.2018.04.005

Ben Salah, M., Abdelmelek H., Abderraba, M., 2012. Study of phenolic composition and biological activities assessment of olive leaves from different varieties grown in Tunisia. Medicinal Chemistry, 2, 107-111. DOI: https://doi.org/10.4172/2161-0444.1000124

Besnard, G., Baradat, P., Chevalier, D., Tagmount, A., Bervillé, A., 2001. Genetic differenciation in the olive complex (Olea europaea L.) revealed by RAPDs and RFLPs in the rRNA genes. Genetic Ressources and Crop Evolution, 48, 165-182. DOI: https://doi.org/10.1023/A:1011239308132

Bettaieb-Rebey, I., Bourgou, S., Ben Slimen Debbez, I., Jabri-Karoui, I., Hamrouni-Sellami, I., Msaada, K., Limam, F., Marzouk, B., 2011. Effects of extraction solvent and provenances on phenolic contents and antioxidant activities of cumin (Cuminum cyminum L.) seeds. Food and Bioprocess Technology, 5, 2827-2836. DOI: https://doi.org/10.1007/s11947-011-0625-4

Biesaga, M., 2011. Influence of extraction methods on stability of flavonoids. Journal of Chromatography A, 1218, 2505-2515. DOI: https://doi.org/10.1016/j.chroma.2011.02.059

Boldi, A.M., 2004. Libraries from natural product-like scaffolds. Current Opinion in Chemical Biology, 8, 281-286. DOI: https://doi.org/10.1016/j.cbpa.2004.04.010

Brahmi, F., Mechri, B., Dhibi, M., Hammami, M., 2013. Variations in phenolic compounds and antiradical scavenging activity of Olea europaea leaves and fruits extracts collected in two different seasons. Industrial Crops and Production, 49, 256-264. DOI: https://doi.org/10.1016/j.indcrop.2013.04.042

Bucić-Kojić, A., Planinić, M., Tomas, S., Bilić, M., Velic D., 2007. Study of solid–liquid extraction kinetics of total polyphenols from grape seeds. Journal of Food Engineering, 81, 236-242. DOI: https://doi.org/10.1016/j.jfoodeng.2006.10.027

Cui, Z.W., Sun, I.J., Chen, W., Sun, D.W., 2008. Preparation of dry honey by microwave-vacuum drying. Journal of Food Engineering, 84, 582-590. DOI: https://doi.org/10.1016/j.jfoodeng.2007.06.027

Da Porto, C., Poretto, E., Decorti, D., 2015. Comparison of ultrasound-assisted extraction with conventional extraction methods of oil and polyphenols from grape (Vitis vinifera L.) seeds. Ultrasonics Sonochemistry, 20, 1076-1080. DOI: https://doi.org/10.1016/j.ultsonch.2012.12.002

Elfalleh, W., Nasri, N., Marzougui, N., Thabti, I., M'rabet, A., Yahya, Y., Lachiheb, B., Guasmi, F., Ferchichi, A., 2009. Physico-chemical properties and DPPH-ABTS scavenging activity of some local pomegranate (Punica granatum) ecotypes. International Journal of Food Science and Nutrition, 60, 197-210. DOI: https://doi.org/10.1080/09637480903067037

Elfalleh, W., Tlili, N., Nasri, N., Yahia, Y., Hannachi, H., Chaira, N., Ying, M., Ferchichi, A., 2011. Antioxidant pomegranate (Punica granatum L.) fruits. Journal of Food Science, 76, 707-713. DOI: https://doi.org/10.1111/j.1750-3841.2011.02179.x

Gourguillon, L., Destandau, E., Lobstein, A., Lesellier, E., 2016. Comparison of different ways to extract dicaffeoylquinic acids from a halophytic plant. Comptes Rendus Chimie, 113, 1-9.

Hannachi, H., Benmoussa, H., Saadaoui, E., Saanoun, I., Negri, N., Elfalleh, W., 2019. Optimization of ultrasound and microwave-assisted extraction of phenolic compounds from olive leaves by response surface methodology. Research Journal of Biotechnology, 14, 28-37.

Hannachi, H., Breton, C., Msallem, M., Ben El Hadj, S., El Gazzah, M., Bervillé, A., 2008. Differences between local and introduced olive cultivars as revealed by morphology of drupes, oil composition and SSR polymorphisms: a case study in Tunisia. Scientia Horticulturae, 116, 280-290. DOI: https://doi.org/10.1016/j.scienta.2008.01.004

Hannachi, H., Elfalleh, W., Yahia, Y., Laajel, M., Ennajeh, I., Mechlouch, R.F., 2020. Chemicals profiling and antioxidants activities of leaf and fruit of cultivated and wild olive trees (Olea europaea L.). International Journal of Fruit Science, 20, 350-370. DOI: https://doi.org/10.1080/15538362.2019.1644574

Hannachi, H., Nasri, N., Elfalleh, W., Tlili, N., Ferchichi, A., Msallem, M., 2013. Fatty acids, sterols, polyphenols and chlorophylls of olive oils obtained from Tunisian wild olive trees (Olea europaea L. var. sylvestris). International Journal of Food Properties, 16, 1271-1283. DOI: https://doi.org/10.1080/10942912.2011.584201

Horžić, D., Komes, D., Belščak, A., Kovačević, Ganić, K., Iveković, D., Karlović, D., 2009. The composition of polyphenols and methylxanthines in teas and herbal infusions. Food Chemistry, 115, 441-448. DOI: https://doi.org/10.1016/j.foodchem.2008.12.022

Lalas, S., Athanasiadis, V., Gortzi, O., Bounitsi, M., Giovanoudis, I., Tsaknis, J., Bogiatzis, F., 2011. Enrichment of table olives with polyphenols extracter from olive leaves. Food Chemistry, 127, 1521-1525. DOI: https://doi.org/10.1016/j.foodchem.2011.02.009

Liazid, A., Palma, M., Brigui, J., Barroso, C.G., 2007. Investigation on phenolic compounds stability during microwave-assisted extraction. Journal of Chromatograohy A, 1140, 29-43. DOI: https://doi.org/10.1016/j.chroma.2006.11.040

Miguel, M.G., Nunes, S., Dandlen, S.A., Cavaco, A.M., Antunes, M.D., 2010. Phenols and antioxidant activity of hydro-alcoholic extracts of propolis from Algarve saouth of Portugal. Food and Chemical Toxicology, 48, 3418-3423. DOI: https://doi.org/10.1016/j.fct.2010.09.014

Nayak, B., Dahmoun, F., Moussi, K., Dain, S., Aoun, O., Khodir, M., 2015. Comparison of microwave, ultrasound and accelerated-assisted solvent extraction for recovery polyphenols from Citrus sinensis peels. Food Chemistry, 187, 507-516. DOI: https://doi.org/10.1016/j.foodchem.2015.04.081

Papoti, V.T., Tsimidou, M.Z., 2009. Impact of sampling parameters on the radical scavenging potential of olive (Olea europaea L.) leaves. Journal of Agricultural and Food Chemistry, 57, 3470-3477. DOI: https://doi.org/10.1021/jf900171d

Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., Rice-Evans, C., 1999. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine, 26, 1231-1237. DOI: https://doi.org/10.1016/S0891-5849(98)00315-3

Ryan, D., Antolovich, M., Prenzler, P., Robards, K., Lavee, S., 2002. Biotransformations of phenolic compounds in Olea europaea L. Scientia Horticulturae, 92, 147-176. DOI: https://doi.org/10.1016/S0304-4238(01)00287-4

Singh, R., 2016. Chemotaxonomy: A Tool for Plant Classification. Journal of Medicinal Plants Studies, 4, 90-93.

Siracusa, L., Patanè, C., Avola, G., Ruberto, G., 2012. Polyphenols as Chemotaxonomic Markers in Italian “Long-Storage” Tomato Genotypes. Journal of Agricultural and Food Chemistry, 60, 309-314. DOI: https://doi.org/10.1021/jf203858y

Trusheva, B., Trunkov, D., Bankova, V., 2007. Different extraction methods of biologically active components from propolis: a preliminary study. Chemistry Central Journal, 1, 1-4. DOI: https://doi.org/10.1186/1752-153X-1-13

Tsakona, S., Galanakis, C.M., Gekas, V., 2012. Hydro-ethanolic mixtures for the recovery of phenols from Mediterranean plant materials. Food and Bioprocess Technology, 5, 1384-1393. DOI: https://doi.org/10.1007/s11947-010-0419-0

Vermerris, W., Nicholson, R.L., 2006. Phenolic compound biochemistry, (Dordrecht, Netherlands: Springer. Vinha, A.F., Ferreres, F., Silva, B.M., Valentaõ, P., Gonçalves, A., Pereira, J.A., Oliveira, M.B., Seabra, R.M., Andrade, P.B., 2005. Phenolic profiles of Portuguese olive fruits (Olea europaea L.): influences of cultivar and geographical origin. Food Chemistry, 89, 561-160. DOI: https://doi.org/10.1016/j.foodchem.2004.03.012

Vinha, A., Silva, B., Andrade, P., Eabra, R., Pereira, J., Oliveira, B., 2002. Development and evaluation of an HPLC/DAD method for analysis of phenolic compounds from olive fruits. Journal of Liquid Chromatogray and Related Technologies, 25, 151-160. DOI: https://doi.org/10.1081/JLC-100108546

Xiang, G., Yang, H., Yang, L., Zhanga, X., Cao, Q., Miao M., 2010. Multivariate statistical analysis of tobacco of different origin, grade and variety according to polyphenols and organic acids. Microchemical Journal, 95, 198-206. DOI: https://doi.org/10.1016/j.microc.2009.12.001

Yahia, Y., Benabderrahim, M.A., Tlili, N., Hannachi, H., Ayadi, L., Elfalleh, W., 2020. Comparison of Three Extraction Protocols for the Characterization of Caper (Capparis spinosa L.) Leaf Extracts: Evaluation of Phenolic Acids and Flavonoids by Liquid Chromatography – Electrospray Ionization – Tandem Mass Spectrometry (LC–ESI–MS) and the Antioxidant Activity. Analytical Letters, 53 1366-1377. DOI: https://doi.org/10.1080/00032719.2019.1706546




How to Cite

Guebebia, S., Othman, K. B., Yahia, Y., Romdhane, M., Elfalleh, W., & Hannachi, H. (2021). Effect of genotype and extraction method on polyphenols content, phenolic acids, and flavonoids of olive leaves (Olea europaea L. subsp. europaea). International Journal of Plant Based Pharmaceuticals, 2(1), 17–24. https://doi.org/10.62313/ijpbp.2022.9



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