Document Type : Original Article


Laboratory of Separation Process in Foods, Dep Chemical Engineering, Federal University of Rio Grande do Norte- UFRN, CEP 59072-970 Natal, Brazil


The aim of this study was to quantify the amount of phenolic compounds and flavonoids in extracts from Thymus vulgaris and Origanum vulgare L. and to evaluate the antioxidant activity of these extracts. Extracts were obtained by employing three different techniques: low pressure solvent extraction, Soxhlet, and ultrasound methods. The effects of phenolic and flavonoid concentrations on the percentage of inhibition of free radical activity were also analyzed. The highest amounts of phenolic compounds were obtained with Soxhlet and Low Pressure Solvent Extraction (LPSE) for thyme and oregano respectively. The highest flavonoid levels were obtained by the Soxhlet extracts for both species. All extracts showed strong antioxidant activities.

Graphical Abstract

Extracts from Thymus vulgaris and Origanum vulgare L. obtained by different separation processes: global yield and functional profile


Main Subjects

Aligians, N., Kalpoutzakis, E., Mitaku, S., Chinou, I.B., 2001. Composition and antimicrobial activity of the essential oil of two Origanum species. J. Agric. Food Chem. 49, 4168-4170.
Altunkaya, A., Becker, E.M., Gökmen, V., Skibsted, L.H., 2009. Antioxidant activity of lettuce extract (Lactuca sativa) and synergism with added phenolic antioxidants. Food Chem. 115, 163-168.
Bedin, C., Gutkoski, S.B., Wiest, J.M., 1999. Atividade antimicrobiana das especiarias. Higiene Alimentar. 13, 26-29.
Bonjar, G.H., 2004. Inhibition of clotrimazole-resistant Candida albicans by plants used in Iranian folkloric medicine. Fitoterapia 75, 74-76.
Brasil. Ministery of Agriculture, 2001. Instruction Normative nº 3-ANEXE VI-Technical regulation for fixing identity and quality of própolis. D.O. January 19, 2001.
Buczek, B., Geldart, D., 2000. Determination of density of porous particles using very fine dense powders. Powder Technol. 45, 173-176.
Burt, S.A., Reinders, R.D., 2003. Antibacterial activity of selected plant essential oils against Escherichia coli O157:H7. Lett. Appl. Microbiol. 36, 162-167.
Çabuk, M., Alçiçek, A., Bozkurt, M., Imre, N., 2003. Antimicrobial properties of the essential oils isolated from aromatic plants and using possibility as alternative feed additives. II. Proceedings of National Animal Nutrition Congress, p. 184-187. Konya, Turkey: Organized by Animal Nutrition Science Association.
Chipault, J.R., Mizun, G.K., Hawkins, J.M., Lundberg, W.O., 1952. The oxidant properties of natural species. Food Res. 17, 44-55.
Faiku, F., Haziri, A., Mehmeti, A., Recica, B., 2017. Antioxidant activity of the organic extracts of Origanum vulgare (L.) growing wild in Kosovo. Fresenius Environ. Bull. 26(2A), 1440-1446.
Fernandes, R.P.P., Trindade, M.A., Tonin, F.G., Pugine, S.M.P., Lima, C.G., Lorenzo, J.M., de Melo, M.P., 2017. Evaluation of oxidative stability of lamb burger with Origanum vulgare extract. Food Chem. 233, 101-109.
Fernandez, P.B., Capó, J.T., Cofino, G.M., Dominguez, C.C., Menéndez, A.B., Alarcón, C.M., 2009. Expectorant activity of formulations from Plectrantus amboinicus (Lour) Spreng (French oregano). Rev. Cubana Plant. Med. 14, 1028-4796.
Friedman, M., Henika, P.R., Mandrell, R.E., 2002. Bactericidal activities of plant essential oils and some of their isolated constituents against Campylobacter jejuni, Escherichia coli, Listeria monocytogenes, and Salmonella enterica. J. Food Prot. 65, 1545-1560.
Gebara, E.C.E., Zardetto, C.G.D.C., Mayer, M.P.A., 1996. Study of antimicrobian action of natural substances on S. mutans and S. sobrinus (original title: Estudo in vitro da ação antimicrobiana de substâncias naturais sobre S. mutans e S. sobrinus). Mag. Dent. Univ. São Paulo (original title: Revista Odontologica Universidade de São Paulo). 10, 251-256.
Hashemi-Moghaddam, H., Mohammadhosseini, M., Basiri, M., 2015. Optimization of microwave assisted hydrodistillation on chemical compositions of the essential oils from the aerial parts of Thymus pubescens and comparison with conventional hydrodistllation. J. Essent. Oil-Bear. Plants 18(4), 884-893.
Herrero, M., Ibañez, E., Señorans, F. J., A. Cifuentes. 2004. Pressurised liquid extracts from Spirulina platensis microalga. Determination of their antioxidant activity and preliminary analysis by micellar electrokinetic chromatography. J. Chromatogr. A 1047, 195-203.
Jacobs, M.B., 1973. The Chemical Analysis of Food and Products, New York: Publishing Co.
Jaine. N.K, Bhagia N., A comparative study of total phenoloc, total flavonoid and antioxidant assays of aqueous and DMSO Extract from leaves of Origanum vulgare. World J. Pharm. Pharm. Sci. 5, 726-736.
Kasrati, A., Alaoui Jamali, C., Abbad, A., 2017. Antioxidant properties of various extract from selected wild Moroccan aromatic and medicinal species. Trends Phytochem. Res. 1(4), 175-182.
Leitão, N.C.M.C.S., Prado, G.H.C., Veggi, P.C., Meireles, M.A.A., Pereira, C.G., 2013. Anacardium occidentale L leaves extraction via SFE: global yields, extraction kinetics, mathematical modeling and economic evaluation. J. Supercrit. Fluids 78, 114-123.
Mazandarani, M., Ghafourian, M., 2017. Influence of extraction methods on total phenolic, flavonoids and antioxidant activity of Thymus kotchyanus L. extract in Semnan Province, Iran. Trends Phytochem. Res. 1(1), 33-38.
Medina, I., Tombo, I., Satué-Gracia, M.T., German, J.B., Frankel, E.N., 2002. Effects of natural phenolic compounds on the antioxidant activity of lactoferrin in liposomes and oil-in-water emulsions. J. Agric. Food Chem. 10, 2392-2399.
Melo, E.A., Mancini-Filho, J., Guerra, N.B., Maciel, G.R., 2003. Antioxidant activity of coriander extracts (Coriandrum sativum L.). Food Sci. Technol. 23, 195-199.
Mohammadhosseini, M., 2016. A Comprehensive Review on New Methods for Processing, Separation and Identification of the Essential Oils. Islamic Azad University of Shahrood Press, Shahrood, Iran.
Mohammadhosseini, M., 2017. The ethnobotanical, phytochemical and pharmacological properties and medicinal applications of essential oils and extracts of different Ziziphora species. Ind. Crops Prod. 105, 164-192.
Mohammadhosseini, M., Sarker, S.D., Akbarzadeh, A., 2017. Chemical composition of the essential oils and extracts of Achillea species and their biological activities: A review. J. Ethnopharmacol. 199, 257-315.
Moura, P.M., Prado, G.H.C., Meireles, M.A.A., Pereira, C.G., 2012. Supercritical fluid extraction from guava (Psidium guajava) leaves: global yield, composition and kinetic data. J. Supercrit Fluid. 62, 116- 122.
Nascimento, G.G.F., Locatelli, J., Freitas, P.C., Silva, G.L., 2000. Antibacterial activity of plant extracts and phytochemicals on antibiotic-resistant bacteria. Braz. J. Microbiol. 31, 247-256.
Noguer, M., Cerezo, A.B., Moyá, M.L., Troncoso, A.M., García-Parrilla, M.C., 2014. Synergism effect between phenolic metabolites and endogenous antioxidants in terms of antioxidant activity. Adv. Chem. Eng. Sci. 4, 258-265.
Novack, J., Christina, B., Langbehn, B., Park, F., Skoula, M., Gorsiou, Y., Franz, C.M., 2000. Ratios of cis- and trans-sabinene hydrate in Origanum marjorana L. and Origanum midrophyllum (Bentham). Biochem. Syst. Ecol. 28, 697-704.
Ozcan, M.M., Ozkan, G., 2015. Antibacterial effects of Istanbul thyme (Origanum vulgare L. subsp hirtum (Link) letsw.) and Karabas thyme (Thymbra spicata L. var. spicata) extracts. J. Food Saf. Food Qual. 66(6), 177-181.
Ozcan, M.M., Ozkan, G., 2015. Antibacterial effects of Istanbul thyme (Origanum vulgare L. subsp hirtum (Link) letsw.) and Karabas thyme (Thymbra spicata L. var. spicata) extracts. J. Food Saf. Food Qual. 66(6), 177-181.
Özkalp, B., Sevgi, F., Özcan,M., Özcan, M.M., 2010. The antibacterial activity of essential oil of oregano (Origanum vulgare L.). J, Food Agric, Environ. 8(2), 272-274.
Pereira, C.G., 2011. Processes for obtaining extracts and bioactive substances. In Souza, G.H.B., Mello, J.C.P., Lopes, N.P. (Eds). Reviews in Process and Advanced Techniques for Isolation and Structural Determination of Actives in Medicinal Plants, Ed. EDUFOP, Brazil, p. 55-78.
Pereira, C.G., Gualtieri, I.P., Meireles, M.A., 2008. Effect of different extraction processes on the recovery of extracts from Achyrocline satureioides D.C.: An evaluation of antioxidant activity. Sep. Sci. Technol. 43, 1549-1563
Pereira, C.G., Meireles, M.A.A., 2007. Evaluation of global yield, composition, antioxidant activity, cost of manufacturing of extracts from lemon Verbena (Aloysia triphylla L'Herit Britton) and Mango (Mangifera indica L.) leaves. J. Food Proc. Eng. 30, 150-173.
Pina-Vaz, C., Rodrigues, A.G., Costa-de-Oliveira, S., Tavares, C., Salgueiro, L., Cavaleiro, C., Gonçalves, M.J., 2004. Antifungal activity of Thymus oils and their major compounds. J. Eur. Ac. Derm. Vener. 18, 73-78.
Rezaei, P., Mohammadhosseini, M., 2014. Quantitative and qualitative determination of chemical compounds in the essential oil of from the aerial parts of Thymus vulgaris using hydro-distillation combined with gas chromatography-mass spectrometry (GC-MS) and an exhaustive phytochemical investigation on its extract. J. Quantum Chem. Spectrosc. 4(10), 45-59.
Rhayour, K., Bouchikhi, T., Tantaoui-Elaraki, A., Sendide, K., Remmal, A., 2003. The Mechanism of bactericidal action of oregano and clove essential oils and of their phenolic major components on Escherichia coli and Bacillus subtilis. J. Essent. Oil Res. 15, 286-292.
Sagdiç, O., 2003. Sensitivity of four pathogenic bacteria to Turkish thyme and oregano hydrosols. LWT-Food Sci. Technol. 36, 467-473.
Sağdiç, O., Kuşçu, A., Özcan, M., Özçelik, S. 2002. Effects of Turkish spice extracts various concentrations on the growth of Escherichia coli O157:H7. Food Microbiol. 19, 473-480.
Seoudi, D.M., Hewedi, I.H., Medhat, A.M., Osman, S.A., Arbid, M.S., Mohamed, M.K., 2009. Evaluation of the anti-inflammatory, analgesic, and anti-pyretic effects of Origanum majorana ethanolic extract in experimental animals. J. Rad. Res. Appl. Sci. 2, 513-534.
Simões, C.M.O., Schenkel, E.P., Gosmann, G., de Mello, J.C.P., Petrovick, P.R., 2004. Pharmacognosy: The Plant to Medicine (original name: Farmacognosia: da planta ao medicamento), Editora UFRGS, Brazil, p.467-495.
Singleton, V.L., Rossi, J.A., 1965. Colorimetry of total phenolics with phosphomolybdic phosphotungstic acid reagents.  Am. J. Enol. Vitic. 16, 144-158.
Sivropoulou, A., Papanikolau, E., Nicolau, C., Kokkini, S., Lanaras, T., Arsenakis, M., 1996. Antimicrobial and citotoxic activities of Origanum vulgare essential oil. J. Agric. Food Chem. 44, 1202-1205.
Soliman, K.M., Badea, R.I., 2002. Effect of oil extracted from some medicinal plants on different mycotoxigenic fungi. Food Chem. Toxicol. 40, 1669-1675.
Takeuchi, T.M., Pereira, C.G., Braga, M.E.M., Maróstica, Jr M.R., Leal, P.F., Meireles, M.A.A., 2008. Low pressure solvent extraction (Solid-liquid extraction, microwave assisted and ultrasound) from condimentary plants” In M.A.A Meireles (Ed). “Extracting Bioactive Compounds: Theory and Applications”, p. 137-218. EUA: CRC Press.
Vekiari, S.A., Oreopoulou, V., Tzia, C., Thomopoulos, C.D., 1993. Oregano flavonoids as lipid antioxidants. JAOCS 70, 483-487.
Vennat, B., Gross, D., Pourrat, A., Pourrat, H., 1992. Hamamelis virginiana: identification and assay of proanthocyanidins, phenolic acids and flavonoids in leaf extracts. Pharm. Acta Helv. 67, 11-14.
Venugopal, S., Devarajan, S., 2011. Estimation of total flavonoids, phenols and antioxidant activity of local and New Zealand manuka honey. J Pharm. Res. 4(2), 464-466.
Vieira de Melo, S.A.B., Costa, G.M.N., Garau, R., Casula, A., Pittau, B., 2000. Supercritical CO2 extraction of essential oils from Thymus vulgaris Linn. Braz. J. Chem. Eng. 3, 17
Woisky, R.G., Salatino, A., 1998. Analysis of propolis: some parameters and procedures for chemical quality control. J. Apicult. Res. 37, 99-105.