Effects of extraction techniques on bioactive compounds and biological properties of Silene behen L.

Abstract

The present research evaluated the maceration and ultrasonication-assisted extraction methods on the phytochemical composition, antioxidant activity, and enzyme inhibition effects of Silene behen. The spectrophotometric methods determined the total bioactive compounds. 22 phenolic compounds were analyzed in S. behen extracts. Benzoic acid was the major phenolic compound (S. behen MAC: 3552.7 µg/g; S. behen UAE: 4215.2 µg/g), followed by gallic acid and epicatechin. S. behen UAE (19.47 mg GAE/g) contained higher total phenolic content compared to S. behen MAC (16.57 mg GAE/g) extract. Similarly, S. behen UAE showed higher antioxidant activity than S. behen MAC. S. behen UAE showed the strongest radical scavenging activity, with DPPH (31.66 mg TE/g) and ABTS (49.54 mg TE/g). The highest reducing power activity was found in the S. behen UAE (CUPRAC: 96.56 mgTE/g; FRAP: 36.29 mgTE/g). The antioxidant results indicated that UAE can be used to extract natural antioxidant agents from S. behen. The tested extracts displayed inhibitory effects against tyrosinase, cholinesterase, α-amylase, and α-glucosidase. S. behen UAE (2.38 mg GALAE/g) demonstrated the highest activity against butyrylcholinesterase enzyme. Inhibition of α-amylase and α-glucosidase was similar across the two extracts. The anti-tyrosinase effect of S. behen MAC (47.49 mgKAE/g) was higher than that of S. behen UAE (45.65 mgKAE/g). These results highlight that S. behen extract may serve as a promising source of natural antioxidants and enzyme inhibitors.

Keywords

• S. behen
• Extraction methods
• Phenolic
• Enzyme inhibition

References

1. Adhikari, L., Kotiyal, R., Pandey, M., Bharkatiya, M., Sematy, A., & Semalty, M. (2020). Effect of geographical location and type of extract on total phenol/flavon contents and antioxidant activity of different fruits extracts of Withania somnifera. Current Drug Discovery Technologies, 17(1), 92-99. https://doi.org/10.2174/1570163815666180807100456
2. Akbulut, S., & Zengin, Z. (2023). Ethnobotanical survey of wild plants used in Gümüşhane province (Turkey). Boletín Latinoamericano y del Caribe de Plantas Medicinales y Aromáticas, 22(2), 237-254. https://doi.org/10.37360/blacpma.23.22.2.18
3. An, S.M., Koh, J.S., & Boo, Y.C. (2010). p‐coumaric acid not only inhibits human tyrosinase activity in vitro but also melanogenesis in cells exposed to UVB. Phytotherapy Research, 24(8), 1175-1180. https://doi.org/10.1002/ptr.3095
4. Angeloni, S., Spinozzi, E., Maggi, F., Sagratini, G., Caprioli, G., Borsetta, G., Ak, G., Sinan, K.I., Zengin, G., Arpini, S., Mombelli, G., Ricciutelli, M. (2021). Phytochemical Profile and Biological Activities of Crude and Purified Leonurus cardiaca Extracts. Plants-Basel,10(2), 195. https://doi.org/10.3390/plants10020195
5. Aygun, R.B., Zengin, G., Yildiztugay, E., Jugreet, S.,Yilmaz, M.A., & Mahomoodally FM. (2022). Chemical characterization, anti-oxidant and anti-enzymatic properties of extracts from two Silene species: A focus on different plant parts and extraction methods. Process Biochemistry, 116, 206-213. https://doi.org/10.1016/j.procbio.2022.03.016
6. Bagci, Y., & Biçer, H. (2015). Analysis of anatomical and morphological characters of the Silene Cappadocica Boiss. & Heldr. and Silene Spergulifolia Bieb. (Caryophyllaceae) species. Scientific Papers-Series B-Horticulture, 59, 293-302.
7. Bechkri, S., Magid, A.A., Khalfallah, A., Voutquenne-Nazabadioko, L., Kabouche, A., Sayagh, C., Harakat, D., & Kabouche, Z. (2022). Antioxidant activity-guided isolation of flavonoids from Silene gallica aerial parts. Phytochemistry Letters, 50, 61-66. https://doi.org/10.1016/j.phytol.2022.05.002
8. Bechkri, S., Magid, A.A., Sayagh, C., Berrehal, D., Harakat, D., Voutquenne-Nazabadioko, L., Kabouche, Z., & Kabouche, A. (2020). Triterpene saponins from Silene gallica collected in North-Eastern Algeria. Phytochemistry, 172, 112274. https://doi.org/10.1016/j.phytochem.2020.112274

9. Boga, M. (2017). Chemical constituents, cytotoxic, antioxidant and cholinesterases inhibitory activities of Silene Compacta (Fischer) Extracts. Marmara Pharmaceutical Journal, 21(3), 445-454. https://doi.org/10.12991/marupj.306789
10. Boukhira, S., Bousta, D., El Mansouri, L., Nordine, A., Hamsas El Youbi A, & Daoudi A. (2015). Phytochemical studies, antioxidant activity and protective effect on DNA damage and deoxyribose of Silene vulgaris extract from Morocco. International Journal of Pharmacognosy and Phytochemical Research, 7(6), 1172–1178.
11. Chaachouay, N., Benkhnigue, O., & Zidane, L. (2022). Ethnobotanical and Ethnomedicinal study of medicinal and aromatic plants used against dermatological diseases by the people of Rif, Morocco. Journal of Herbal Medicine, 32, 100542. https://doi.org/10.1016/j.hermed.2022.100542
12. Chandra, S., & Rawat, D.S. (2015). Medicinal plants of the family Caryophyllaceae: a review of ethno-medicinal uses and pharmacological properties. Integrative Medicine Research, 4, 123–131. https://doi.org/10.1016/j.imr.2015.06.004
13. Cui, S.N. (1995). Chaoyaozhi. Yanbian People’s Press, Yanji, 77–79.
14. Elbozan, H., & Korkmaz, N. (2025). An examination of the antioxidant and antibacterial capabilities, as well as the phenolic compounds, of Silene Macrodonta and Silene Chaetodonta. Prospects in Pharmaceutical Sciences, 23(2), 35-41. https://doi.org/10.56782/pps.287
15. Fan, L., Fan, W., Mei, Y., Liu, L., Li, L., Wang, Z., &Yang, L. (2022). Mechanochemical assisted extraction as a green approach in preparation of bioactive components extraction from natural products - A review. Trends in Food Science & Technology, 129, 98 110. https://doi.org/10.1016/j.tifs.2022.09.009
16. Gadjalova, A.V., & Mihaylova, D. (2019). Ultrasound-assisted extraction of medicinal plants and evaluation of their biological activity. Food Research, 3(5), 530 536. https://doi.org/10.26656/fr.2017.3(5).128
17. Gligor, O., Mocan, A., Moldovan, C., Locatelli, M., Crisan, G., & Ferreira, I.C.F.R. (2019). Enzyme-assisted extractions of polyphenols - A comprehensive review. Trends in Food Science & Technology, 88, 302-315. https://doi.org/10.1016/j.tifs.2019.03.029
18. Guan, L., Long, H., Ren, F., Li, Y., & Zhang, H. (2022). Structure-Activity Relationship Study of the Inhibition of α-Amylase by Benzoic Acid and Its Derivatives. Nutrients, 14(9), 1931. https://doi.org/10.3390/nu14091931
19. Hartavi, A., Kargin Solmaz, F.O., Sarikurkcu, C., & Tepe, B. (2025). Extraction technique impacts on phenolic profile, antioxidant capacity, and enzyme inhibition by Anthemis pseudocotula. Chemistry Select, 10(37), e02006. https://doi.org/10.1002/slct.202502006
20. Kilinç, H., Masullo, M., Bottone, A., Karayildirim, T., Alankus, Ö., & Piacente, S. (2019). Chemical constituents of Silene montbretiana. Natural Product Research, 33(3), 335 339. https://doi.org/10.1080/14786419.2018.1451998
21. Kurt, B.Z., Gazioglu, I., Sevgi, E., & Sonmez, F. (2018). Anticholinesterase, antioxidant, antiaflatoxigenic activities of ten edible wild plants from Ordu Area, Turkey. Iranian Journal of Pharmaceutical Research, 17(3), 1047–1056.
22. Lacaille-Dubois, M.A., Hanquet, B., Cui, Z.H., Lou, Z.C., & Wagner, H. (1999). A new biologically active acylated triterpene saponin from Silene fortunei. Journal of Natural Products, 62(1),133-136. https://doi.org/10.1021/np980172y
23. Mamadalieva, N.Z., Khatib, S., Salohiddinova, N., Jalilov, F.S., Jamalova, D., Albaik, M., & Zengin, G. (2025). Metabolite profiling, in vitro biological activities, and in silico molecular docking analysis of Silene kuschakewiczii. Natural Product Research, 23, 1 9. https://doi.org/10.1080/14786419.2025.2534682
24. Mamadalieva, N.Z., Böhmdorfer, S., Zengin, G., Bacher, M., Potthast, A., Akramov, D.K., Janibekov, A., & Rosenau, T. (2019). Phytochemical and biological activities of Silene viridiflora extractives. Development and validation of a HPTLC method for quantification of 20-hydroxyecdysone. Industrial Crops and Products, 129, 542-548. https://doi.org/10.1016/j.indcrop.2018.12.041
25. Mamadalieva, N.Z., Egamberdieva, D., & Tiezzi, A. (2013). In vitro biological activities of the components from Silene wallichiana. Medicinal and Aromatic Plant Science and Biotechnology, 7(1), 1-6.
26. Mamadalieva, N.Z., Lafont, R., & Wink, M. (2014). Diversity of secondary metabolites in the genus Silene L. (Caryophyllaceae)-Structures, distribution, and biological properties. Diversity, 6(3), 415-499. https://doi.org/10.3390/d6030415
27. Mohti, H., Taviano, M.F., Cacciola, F., Dugo, P., Mondello, L., Zaid, A., Cavo, E., & Miceli, N. (2020). Silene vulgaris subsp. macrocarpa leaves and roots from Morocco: Assessment of the efficiency of different extraction techniques and solvents on their antioxidant capacity, brine shrimp toxicity and phenolic characterization. Plant Biosystems-An International Journal Dealing with all Aspects of Plant Biology, 154(5), 692-699. https://doi.org/10.1080/11263504.2019.1674404
28. Mollica, A., Zengin, G., Sinan, K.I., Marletta, M., Pieretti, S., Stefanucci, A., Etienne, O.K., Jekó, J., Cziáky, Z., Bahadori, M.B., Picot-Allain, C., & Mahomoodally, M.F. (2022). A study on chemical characterization and biological abilities of Alstonia boonei extracts obtained by different techniques. Antioxidants, 11(11), 2171. https://doi.org/10.3390/antiox11112171
29. Perry, L.M. & Metzger, J. (1980). Medical plants of east and southeast Asia. Attributed properties and uses. MIT Press, Cambridge, 74
30. Şahin, B., Cakır, C., Sıcak, Y., & Öztürk, M. (2024). Chemical composition and biological activities of essential oils and extract of Eucalyptus citriodora Hook. International Journal of Secondary Metabolite, 11(2), 394-407. https://doi.org/10.21448/ijsm.1384581
31. Sıcak, Y., Oruç‐Emre, E.E., Öztürk, M., Taşkın‐Tok, T., & Karaküçük‐Iyidoğan, A. (2019). Novel fluorine‐containing chiral hydrazide‐hydrazones: Design, synthesis, structural elucidation, antioxidant and anticholinesterase activity, and in silico studies. Chirality, 31(8), 603-615. https://doi.org/10.1002/chir.23102
32. Sıcak, Y., Öztürk, İ., Şahin, B., Çam, D., Cakır, C., & Öztürk, M. (2024). Chemical content profile and antioxidant activity of Rhododendron ponticum L. (Ericaceae) extracts. International Journal of Secondary Metabolite, 11(3), 507-513. https://doi.org/10.21448/ijsm.1382522
33. Taleghani, A., Moghimi, R., & Eghbali, S. (2025). Phytochemicals of biologically active fractions from Silene conoidea Leaves and Stems. Current Topics in Medicinal Chemistry, 25(10), 1281-1302. https://doi.org/10.2174/0115680266324758241223063534
34. Unver, A. (2019). Msc Thesis.Etnobotany Researc in the Villages Araund Lamas Stream (Erdemli/ Silifke-Mersin) [Msc Thesis, Balıkesir University], Turkey.
35. Uyen, N.H., Widyowati, R., Sulistyowaty, M.I., Sugimoto, S., Yamano, Y., Kawakami, S., Otsuka, H., & Matsunami, K. (2020). Firmosides A and B: two new sucrose ferulates from the aerial parts of Silene firma and evaluation of radical scavenging activities. Journal of Natural Medicines, 74(4), 796-803. https://doi.org/10.1007/s11418-020-01426-5
36. Uysal, S., Zengin, G., Locatelli, M., Bahadori, M.B., Mocan, A., Bellagamba, G., De Luca, E., Mollica, A., & A., Aktumsek, A. (2017). Cytotoxic and enzyme inhibitory potential of Two Potentilla species (P. speciosa L. and P. reptans Willd.) and their chemical composition. Frontiers in Pharmacology, 8, 290. https://doi.org/10.3389/fphar.2017.00290
37. Valencia, D., Aguilar-González, D.I., Ortega-García, J., Godoy-Hernández, G., Leyva-Peralta, M.A., Moo-Huchín, V.M., Aarland, R.C., Quintero-Vargas, J., Mendoza-Espinoza, J.A., & Zarza-García, A.L. (2023). Phytochemical profile, antioxidant and antiproliferative activity from leaves and seeds of Bixa orellana L. from the Yucatán Peninsula, Mexico. Pharmacognosy Magazine, 19(2), 482-490. https://doi.org/10.1177/09731296231158492
38. Wisal, A., Ullah, A., Khan, B., Sıcak, Y., Öztürk, M., Muhammad, A., Anwar, W., Begum, S., Zafar, S., Carvalho, P.V.S.D., Hassan, S.S., Halim, S.A., & Khan, A. (2025). The cytotoxic, antioxidant, and anticholinesterase activity of steviol and isosteviol analogues together with structure-activity relationships and molecular docking studies. Journal of Molecular Structure, 1345, 143049. https://doi.org/10.1016/j.molstruc.2025.143049
39. Zengin, G., Mahomoodally, M.F., Aktumsek, A., Ceylan, R., Uysal, S., Mocan, A., Yilmaz, M.A., Picot-Allain, C.M.N., Ciric, A., Glamoclija, J., & Sokovic, M. (2018). Functional constituents of six wild edible Silene species: A focus on their phytochemical profiles and bioactive properties. Food Bioscience, 23, 75-82. https://doi.org/10.1016/j.fbio.2018.03.010
40. Zengin, G., Rodrigues, M.J., Abdallah, H.H., Custodio, L., Stefanucci, A., Aumeeruddy, M.Z., Mollica, A., Rengasamy KRR, & Mahomoodally MF. (2018). Combination of phenolic profiles, pharmacological properties and in silico studies to provide new insights on Silene salsuginea from Turkey. Computational Biology and Chemistry, 77, 178 186. https://doi.org/10.1016/j.compbiolchem.2018.10.005