ANTIOXIDANT ACTIVITY AND CONTENT OF SELECTED MICRONUTRIENTS IN THE FRONDS OF Asplenium ceterach

Authors

  • Biljana Kukavica University of Banja Luka, Faculty of Natural Sciences and Mathematics, Mladena Stojanovića 2, 78000 Banja Luka, Republic of Srpska, Bosnia and Herzegovina Author
  • Siniša Škondrić University of Banja Luka, Faculty of Natural Sciences and Mathematics, Mladena Stojanovića 2, 78000 Banja Luka, Republic of Srpska, Bosnia and Herzegovina Author
  • Toda Ignjatović University of Banja Luka, Faculty of Natural Sciences and Mathematics, Mladena Stojanovića 2, 78000 Banja Luka, Republic of Srpska, Bosnia and Herzegovina Author
  • Tanja Trifković Vitalwelt Apotheke, Daimlerstraße 70, Michelfeld, Germany Author
  • Dijana Mihajlović University of Banja Luka, Faculty of Agriculture, University City, Vojvode Petra Bojovića Blvd. 1A, 78000 Banja Luka, Republic of Srpska, Bosnia and Herzegovina Author
  • Đura Nakarada University of Belgrade, Faculty of Physical Chemistry, Studentski trg 12-16, 11000 Belgrade, Serbia Author
  • Miloš Mojović University of Belgrade, Faculty of Physical Chemistry, Studentski trg 12-16, 11000 Belgrade, Serbia Author
  • Suzana Živković University of Belgrade, Institute for Biological Research "Siniša Stanković", National Institute of the Republic of Serbia, Bulevar despota Stefana 142, 11108 Belgrade, Serbia Author

DOI:

https://doi.org/10.63356/asb.2026.003

Keywords:

DPPH scavenging activity, reducing power, hydroxyl radical scavenging activity, H2O2 scavenging activity

Abstract

This study aimed to evaluate the total phenolic compound content and antioxidant capacity of the ethanolic extract of Asplenium ceterach fronds, as well as the content of selected micronutrients (Fe, Cu, Zn, and Mn) in fronds collected in the Republic of Srpska (Bosnia and Herzegovina). The extract's total phenolic compound (TPC) concentration was 16.08 ± 0.08 mg GAE/gDW. It demonstrated DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity with an IC₅₀ value of 29.28 ± 2.20 mg/mL. The hydrogen peroxide scavenging capacity was significantly lower (IC₅₀ = 1420.56 ± 35.72 μg/mL), indicating moderate effectiveness against this reactive oxygen species. The extract also showed hydroxyl radical scavenging ability, with 56.26 ± 1.05% residual OH radicals under the tested conditions. Reducing power assays revealed that at 500 µg/mL, the extract reduced Fe³⁺ to 1420.20 ± 53.67 mmol/L Fe²⁺ equivalents, while its Cu²⁺ reducing capacity was 423.14 ± 29.20 µg Trolox/mL at a TPC concentration of 300 µg/mL. The micronutrient levels in A. ceterach were as follows: Fe (164.57 ± 4.08 mg/kg) > Zn (15.61 ± 0.43 mg/kg) > Mn (12.39 ± 0.19 mg/kg) > Cu (1.62 ± 0.12 mg/kg). Our in vitro findings suggest that the ethanolic extract of A. ceterach could serve as a potential natural source of antioxidant compounds.

References

Abbaspour, N., Hurrell, R. & Kelishadi, R. (2014). Review on iron and its importance for human health. Journal of Research in Medical Sciences, 19, 164−174. PMID: 24778671; PMCID: PMC3999603.

Akanni, O. O., Owumi, S. E. & Adaramoye, O. A. (2014). In vitro studies to assess the antioxidative, radical scavenging and arginase inhibitory potentials of extracts from Artocarpus altilis, Ficus exasperate and Kigelia africana. Asian Pacific Journal of Tropical Biomedicine, 4, 492−499. https://doi.org/10.12980/APJTB.4.2014C581

Apak, R., Güçlü, K., Özyürek, M. & Karademir, S. E. (2004). Novel total antioxidant capacity index for dietary polyphenols using their cupric ion reducing capability (CUPRAC). Journal of Agricultural and Food Chemistry, 52, 7970–7981. https://doi.org/10.1021/jf048741x

Benzie, I. F. F. & Strain, J. J. (1996). The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: The FRAP assay. Analytical Biochemistry, 239, 70–76. https://doi.org/10.1006/abio.1996.0292

Broadley, M. R., White, P. J., Hammond, J. P., Zelko, I. & Lux, A. (2007). Phylogenetic variation in the shoot mineral concentration of angiosperms. Journal of Experimental Botany, 58(14), 404–416. https://doi.org/10.1093/jxb/erh002

Carter, P. (1971). Spectrophotometric determination of serum iron at the submicrogram level with a new reagent (ferrozine). Analitical Biochemistry, 40, 450−458. https://doi.org/10.1016/0003-2697(71)90405-2

Clemens, S. (2006). Toxic metal accumulation, responses to exposure and mechanisms of tolerance in plants. Biochimie, 88, 1707–1719. https://doi.org/10.1016/j.biochi.2006.07.003

Cobbett, C. & Goldsbrough, P. (2002). Phytochelatins and metallothioneins: Roles in heavy metal detoxification and homeostasis. Annual Review of Plant Biology, 53, 159–182. https://doi.org/10.1146/annurev.arplant.53.100301.135154

Di Sanzo, P., De Martino, L., Mancini, E. & Feo, V. D. (2013). Medicinal and useful plants in the tradition of Rotonda, Pollino National Park, Southern Italy. Journal of Ethnobiology and Ethnomedicine, 9(1), 19. https://doi.org/10.1186/1746-4269-9-19

Dolina, K. & Luczaj, L. (2014). Wild food plants used on the Dubrovnik coast (south-eastern Croatia). Acta Societatis Botanicorum Poloniae, 83, 175–181. https://doi.org/10.5586/asbp.2014.029

Ebrahimzadeh, M. A., Nabavi, S. M., Nabavi, S. F., Bahramian, F. & Bekhradnia, A. R. (2010). Antioxidant and free radical scavenging activity of H. officinalis L. var. angustifolius, V. odorata, B. hyrcana and C. speciosum. Pakistan Journal of Pharmaceutical Sciences, 23, 29−34. PMID: 20067863.

Euro+Med (2026). Euro+Med PlantBase – the information resource for Euro-Mediterranean plant diversity. Retrieved from: http://www.europlusmed.org

Fatima, A., Laila, O., Murtaza, I. & Masoodi, K. (2020). Nutraceutical composition and anti-cancerous potential of an unexplored herb Asplenium ceterach from Kashmir Region. Indian Journal of Pure and Applied Biosciences, 8, 289−297. http://dx.doi.org/10.18782/2582-2845.8036

Froissard, D., Rapior, S., Bessière, J. M., Buatois, B., Fruchier, A., Sol, V. & Fons, F. (2015). Asplenioideae species as a reservoir of volatile organic compounds with potential therapeutic properties. Natural Product Communications, 10, 1079–1083. https://doi.org/10.1177/1934578X1501000671

Güneş, S., Savran, A., Paksoy, M. Y., Koşar, M. & Çakılcıoğlu, U. (2017). Ethnopharmacological survey of medicinal plants in Karaisalı and its surrounding (Adana-Turkey). Journal of Herbal Medicine, 8, 68−75. https://doi.org/10.1016/j.hermed.2017.04.002

Halliwell, B. & Gutteridge, J. M. C. (2015). Free radicals in biology and medicine (5th ed.). Oxford: Oxford University Press. https://doi.org/10.1093/acprof:oso/9780198717478.001.0001

Hurrell, R. F. (1997). Bioavailability of iron. European Journal of Clinical Nutrition, 51 Suppl 1, S4–S8. PMID: 9023471.

Huseinović, S., Bektić, S., Selimović-Ćehajić, S. & Halilović, M. (2024). The use of medicinal plants and phytotherapy in the Konjuh mountain of Bosnia and Herzegovina. European Journal of Medicinal Plants, 35, 296−315. https://doi.org/10.9734/ejmp/2024/v35i61228

Institute of Medicine (2001). Dietary reference intakes for vitamin A, vitamin K, arsenic, boron, chromium, copper, iodine, iron, manganese, molybdenum, nickel, silicon, vanadium, and zinc. Washington, DC: The National Academies Press. https://doi.org/10.17226/10026

Jomova, K., Baros, S. & Valko, M. (2012). Redox active metal-induced oxidative stress in biological systems. Transition Metal Chemistry, 37, 127−134. https://doi.org/10.1007/s11243-012-9583-6

Kabata Pendias, A. (2011). Trace elements in soils and plants (4th ed.). Boca Raton: CRC Press. https://doi.org/10.1201/b10158

Karadeniz, A., Çinbilgel, I., Gün, S. Ş. & Çetin, A. (2015). Antioxidant activity of some Turkish medicinal plants. Natural Product Research, 29, 2308−2312. https://doi.org/10.1080/14786419.2015.1005618

Leporatti, M. L. & Guarrera, P. M. (2007). Ethnobotanical remarks in Capitanata and Salento areas (Puglia, southern Italy). Etnobiología, 5, 51−64.

Liyana-Pathiranana, C. M. & Shahidi, F. (2005). Antioxidant activity of commercial soft and hard wheat (Triticum aestivum L.) as affected by gastric pH conditions. Journal of Agricultural and Food Chemistry, 53, 2433–2440. https://doi.org/10.1021/jf049320i

Malamas, M. & Marselos, M. (1992). The tradition of medicinal plants in Zagori, Epirus (northwestern Greece). Journal of Ethnopharmacology, 37, 197−203. https://doi.org/10.1016/0378-8741(92)90034-O

Matejić, J. S., Stefanović, N., Ivković, M., Živanović, N., Marin, P. D. & Džamić, A. M. (2020). Traditional uses of autochthonous medicinal and ritual plants and other remedies for health in Eastern and South-Eastern Serbia. Journal of Ethnopharmacology, 261, 113186. https://doi.org/10.1016/j.jep.2020.113186

Milutinović, M., Nakarada, Đ., Božunović, J., Todorović, M., Gašić, U., Živković, S., Skorić, M., Ivković, Đ., Savić, J., Devrnja, N., Aničić, N., Banjanac, T., Mojović, M. & Mišić, D. (2023). Solanum dulcamara L. berries: A convenient model system to study redox processes in relation to fruit ripening. Antioxidants, 12, 346. https://doi.org/10.3390/antiox12020346

Mir, S. A., Mishra, A. K., Reshi, Z. A. & Sharma, M. P. (2013). Preliminary phytochemical screening of some pteridophytes from district Shopian (J & K). International Journal of Pharmacy and Pharmaceutical Sciences, 5, 632−637.

Nikolić, T. (2020). Flora Croatica 2 – vaskularna flora Republike Hrvatske. Zagreb: Alfa d.d.

Nwozo, O. S., Effiong, E. M., Aja, P. M. & Awuchi, C. G. (2023). Antioxidant, phytochemical, and therapeutic properties of medicinal plants: A review. International Journal of Food Properties, 26, 359−388. https://doi.org/10.1080/10942912.2022.2157425

Owen, C. A. (1982). Biochemical aspects of copper: Copper proteins, ceruloplasmin, and copper protein binding. Park Ridge, NJ: Noyes Publications.

Pekgöz, A. K. & Çinbilgel, I. (2019). Phytochemical contents and antioxidant activities of fern, Asplenium ceterach L. in different altitudes. Bangladesh Journal of Botany, 48, 315−320. https://doi.org/10.3329/bjb.v48i2.47674

Pequerul, A., Pérez, C., Madero, P., Val, J. & Monge, E. (1993). A rapid wet digestion method for plant analysis. In M.A.C. Fragoso, M.L. Van Beusichem, & A. Houwers (Eds.), Optimization of plant nutrition (Developments in Plant and Soil Sciences, Vol 53, pp. 3−6). Dordrecht, Netherlands: Springer. https://doi.org/10.1007/978-94-017-2496-8_1

Petkov, V., Slavova, I., Teneva, D., Mladenova, T., Stoyanov, P. & Argirova, M. (2022). Phytochemical study and biological activity of three fern species of the Asplenium genus growing in Bulgaria. The Natural Products Journal, 12, 82−90. https://doi.org/10.2174/2210315511666210512024716

Phaniendra, A., Jestadi, D. B. & Periyasamy, L. (2015). Free radicals: Properties, sources, targets, and their implication in various diseases. Indian Journal of Clinical Biochemistry, 30, 11–26. https://doi.org/10.1007/s12291-014-0446-0

POWO (2026). Plants of the World Online. Facilitated by the Royal Botanic Gardens, Kew. Retrieved from: https://powo.science.kew.org/

Redzic, S. (2010). Wild medicinal plants and their usage in traditional human therapy (southern Bosnia and Herzegovina, W. Balkan). Journal of Medicinal Plants Research, 4(11), 1003−1027. https://doi.org/10.5897/JMPR09.254

Reuter, S., Gupta, S. C., Chaturvedi, M. M. & Aggarwal, B. B. (2010). Oxidative stress, inflammation, and cancer: How are they linked? Free Radical Biology and Medicine, 49, 1603–1616. https://doi.org/10.1016/j.freeradbiomed.2010.09.006

Rice-Evans, C. A., Miller, N. J. & Paganga, G. (1997). Antioxidant properties of phenolic compounds. Trends in Plant Science, 2, 152–159. https://doi.org/10.1016/S1360-1385(97)01018-2

Ross, A. C., Caballero, B., Cousins, R. J. & Tucker, K. L. (2020). Modern Nutrition in Health and Disease. Burlington, MA: Jones & Bartlett Learning.

Shahidi, F. & Ambigaipalan, P. (2015). Phenolics and polyphenolics in foods, beverages and spices: Antioxidant activity and health effects – A review. Journal of Functional Foods, 18, 820–897. https://doi.org/10.1016/j.jff.2015.06.018

Sidhoum, R. Halli, L., Yalaoui, O., & Belkadi, A. (2020). Phenolic profile, anti-inflammatory and diuretic properties of Asplenium ceterach tested on albino mice and Wistar albino rats. Asian Journal of Research in Chemistry, 13, 449−454. DOI: 10.5958/0974-4150.2020.00080.2

Simonetti, P., Gardana, C. & Pietta, P. (2001). Plasma levels of caffeic acid and antioxidant status after red wine intake. Journal of Agricultural and Food Chemistry, 49, 5964−5968. https://doi.org/10.1021/jf010546k

Singleton, V.L. & Rossi, J.A. (1965). Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. American Journal of Enology and Viticulture, 16, 144–158. DOI: 10.5958/0974-4150.2020.00080.2

Škondrić, S. (2019). Ljekovite biljke Nevesinjskog polja. Banja Luka: Univerzitet u Banjoj Luci, Prirodno–matematički fakultet.

Szabo, R., Bodolea, C. & Mocan, T. (2021). Iron, copper, and zinc homeostasis: Physiology, physiopathology, and nanomediated applications. Nanomaterials, 11, 2958. https://doi.org/10.3390/nano11112958

Tatipamula, V. B. & Kukavica, B. (2021). Phenolic compounds as antidiabetic, anti‐inflammatory, and anticancer agents and improvement of their bioavailability by liposomes. Cell Biochemistry and Function, 39, 926−944. https://doi.org/10.1002/cbf.3667

Tomou, E. M. & Skaltsa, H. (2018). Phytochemical investigation of the fern Asplenium ceterach (Aspleniaceae). Natural Product Communications, 13, 849−850. https://doi.org/10.1177/1934578X180130071

Valko, M., Morris, H. & Cronin, M. T. D. (2005). Metals, toxicity and oxidative stress. Current Medicinal Chemistry, 12, 1161–1208. DOI: 10.2174/0929867053764635

Veljović Jovanović, S., Kukavica, B., Vidović, M., Morina, F. & Menckhoff, L. (2018). Class III peroxidases: functions, localization and redox regulation of isoenzymes. In D. Gupta, J. Palma, & Corpas, F. (Eds.), Antioxidants and antioxidant enzymes in higher plants (pp. 269-300). Cham: Springer International Publishing. https://doi.org/10.1007/978-3-319-75088-0_13

Wróblewski, M., Wróblewska, W. & Sobiesiak, M. (2024). The role of selected elements in oxidative stress protection: Key to healthy fertility and reproduction. International Journal of Molecular Sciences, 25, 9409. https://doi.org/10.3390/ijms25179409

Zhou, Y., Chang, S., Huang, X., Wang, W., Hou, F., Wang, Y. & Nan, Z. (2025). Climate and human activities alter coupling of soil macro-and micronutrients: Evidence from a long-term experiment in typical steppes. Geoderma, 456, 117250. https://doi.org/10.1016/j.geoderma.2025.117250

Živković, S., Milutinović, M. & Skorić, M. (2022). Phytochemicals and biological activities of Asplenium ceterach. In H. N. Murthy (Ed.), Bioactive Compounds in Bryophytes and Pteridophytes (pp. 1−29). Cham: Springer. https://doi.org/10.1007/978-3-031-23243-5_19

Živković, S., Skorić, M., Šiler, B., Dmitrović, S., Filipović, B., Nikolić, T. & Mišić, D. (2017). Phytochemical characterization and antioxidant potential of rustyback fern (Asplenium ceterach L.). Lekovite sirovine, 37, 15−20. https://doi.org/10.5937/leksir1737015z

Downloads

Published

2026-03-25

Issue

Section

Articles