Chemical composition, antioxidant properties, and enzyme inhibitory activities of methanol extract from Sideritis montana subsp. montana using ultrasound-assisted extraction

Abstract

Natural products are valuable sources of bioactive compounds with therapeutic potential. This study investigated the chemical composition, antioxidant properties, and enzyme inhibitory activities of the methanol extract from Sideritis montana L. subsp. montana L. obtained via ultrasound-assisted extraction. The extraction yielded 5.37%, with a total phenolic content of 63.27 mg GAEs/g extract and a total flavonoid content of 58.32 mg REs/g extract. Chlorogenic acid (563 µg/g extract), luteolin 7-glucoside (513 µg/g extract), and hyperoside (511 µg/g extract) were the most abundant phenolics. Moderate levels of luteolin and hydroxybenzoic acids were also identified. Antioxidant activity was most pronounced in the phosphomolybdenum assay (428.52 mg TEs/g extract), followed by the CUPRAC (217.40 mg TEs/g extract) and FRAP (171.33 mg TEs/g extract) assays, demonstrating strong reducing power. Radical scavenging assays (DPPH: 122.76 mg TEs/g, ABTS: 140.41 mg TEs/g) showed moderate efficacy, while ferrous ion chelation was weak (6.62 mg EDTAEs/g extract). Enzyme inhibition assays indicated potent α-glucosidase (753.81 mg ACEs/g extract) and α-amylase (274.95 mg ACEs/g extract) inhibition, suggesting antidiabetic potential. Tyrosinase inhibition (68.56 mg KAEs/g extract) points to possible dermatological applications, though acetylcholinesterase (2.08 mg GALAEs/g extract) and butyrylcholinesterase (0.45 mg GALAEs/g extract) inhibition was minimal. The results emphasize the bioactive potential of S. montana subsp. montana. Future studies should explore its bioactivity in vivo and identify synergistic effects among its phenolic compounds to further validate its therapeutic applications.

Keywords

• Sideritis montana
• Ultrasound-assisted extraction
• Phytochemical analysis
• Antioxidant activity
• Enzyme inhibition

References

1. Ai-Li, J., & Chang-Hai, W. (2006). Antioxidant properties of natural components from Salvia plebeia on oxidative stability of ascidian oil. Process Biochemistry, 41(5), 1111-1116.
2. Apak, R., Güçlü, K., Özyürek, M., Esin Karademir, S., & Erçaǧ, E. (2006). The cupric ion reducing antioxidant capacity and polyphenolic content of some herbal teas. [Article]. International Journal of Food Sciences and Nutrition, 57(5-6), 292-304.
3. Ashraf, Z., Rafiq, M., Seo, S.-Y., & Babar, M.M. (2015). Synthesis, kinetic mechanism and docking studies of vanillin derivatives as inhibitors of mushroom tyrosinase. Bioorganic & Medicinal Chemistry, 23(17), 5870-5880.
4. Barros, L., Heleno, S.A., Carvalho, A.M., & Ferreira, I.C. (2010). Lamiaceae often used in Portuguese folk medicine as a source of powerful antioxidants: Vitamins and phenolics. LWT-Food Science and Technology, 43(3), 544-550.
5. Bouayed, J., Piri, K., Rammal, H., Dicko, A., Desor, F., Younos, C., & Soulimani, R. (2007). Comparative evaluation of the antioxidant potential of some Iranian medicinal plants. Food Chemistry, 104(1), 364-368.
6. Bouzaiene, N.N., Chaabane, F., Sassi, A., Chekir-Ghedira, L., & Ghedira, K. (2016). Effect of apigenin-7-glucoside, genkwanin and naringenin on tyrosinase activity and melanin synthesis in B16F10 melanoma cells. Life Sciences, 144, 80-85.
7. Cittan, M., & Çelik, A. (2018). Development and validation of an analytical methodology based on Liquid Chromatography–Electrospray Tandem Mass Spectrometry for the simultaneous determination of phenolic compounds in olive leaf extract. Journal of Chromatographic Science, 56(4), 336-343.
8. de Castro, O.n., & Rivera Núñez, D. (1994). taxonomic revision of the section Sideritis (genus Sideritis)(Labiatae): J. Cramer.

9. Emre, I., Kursat, M., Yilmaz, Ö., & Erecevit, P. (2011). Some biological compounds, radical scavenging capacities and antimicrobial activities in the seeds of Nepeta italica L. and Sideritis montana L. subsp montana from Turkey. Grasas Y Aceites, 62(1), 68-75.
10. Erdemoglu, N., Turan, N.N., Cakõcõ, I., Sener, B., & Aydõn, A. (2006). Antioxidant activities of some Lamiaceae plant extracts. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives, 20(1), 9-13.
11. Ertaş, A., Gören, A.C., Boğa, M., Yeşil, Y., & Kolak, U. (2014). Essential oil compositions and anticholinesterase activities of two edible plants Tragopogon latifolius var. angustifolius and Lycopsis orientalis. Natural Product Research, 28(17), 1405-1408.
12. Eruygur, N., & Uçar, E. (2018). Cholinesterase, α-glucosidase, α-amylase, and tyrosinase inhibitory effects and antioxidant activity of Veronica officinalis extracts. Türkiye Tarımsal Araştırmalar Dergisi, 5(3), 253-259.
13. Gao, Y., Fang, L., Wang, X., Lan, R., Wang, M., Du, G., Guan, W., Liu, J., Brennan, M., & Guo, H. (2019). Antioxidant activity evaluation of dietary flavonoid hyperoside using Saccharomyces cerevisiae as a model. Molecules, 24(4), 788.
14. Gião, M.S., González‐Sanjosé, M.L., Rivero‐Pérez, M.D., Pereira, C.I., Pintado, M.E., & Malcata, F.X. (2007). Infusions of Portuguese medicinal plants: Dependence of final antioxidant capacity and phenol content on extraction features. Journal of the Science of Food and Agriculture, 87(14), 2638-2647.
15. Giuliani, C., Maleci Bini, L., Papa, F., Cristalli, G., Sagratini, G., Vittori, S., Lucarini, D., & Maggi, F. (2011). Glandular trichomes and essential oil composition of endemic Sideritis italica (Mill.) Greuter et Burdet from Central Italy. Chemistry & Biodiversity, 8(12), 2179-2194.
16. Gökbulut, A., SatilmiŞ, B., Batçioğlu, K., Cetin, B., & Şarer, E. (2012). Antioxidant activity and luteolin content of Marchantia polymorpha L. Turkish Journal of Biology, 36(4), 381-385.
17. Grzegorczyk, I., Matkowski, A., & Wysokińska, H. (2007). Antioxidant activity of extracts from in vitro cultures of Salvia officinalis L. Food Chemistry, 104(2), 536-541.
18. Huang, Y., Condict, L., Richardson, S.J., Brennan, C.S., & Kasapis, S. (2023). Exploring the inhibitory mechanism of p-coumaric acid on α-amylase via multi-spectroscopic analysis, enzymatic inhibition assay and molecular docking. Food Hydrocolloids, 139, 108524.
19. Islam, M.A., Zaman, S., Biswas, K., Al-Amin, M.Y., Hasan, M.K., Alam, A., Tanaka, T., & Sadik, G. (2021). Evaluation of cholinesterase inhibitory and antioxidant activity of Wedelia chinensis and isolation of apigenin as an active compound. BMC Complementary Medicine and Therapies, 21(1).
20. Khan, M.S., Alokail, M.S., Alenad, A.M.H., Altwaijry, N., Alafaleq, N.O., Alamri, A.M., & Zawba, M.A. (2022). Binding studies of caffeic and p-coumaric acid with α-amylase: Multispectroscopic and computational approaches deciphering the effect on advanced glycation end products (AGEs). Molecules, 27(13), 3992.
21. Kilic, O. (2014). Essential Oil Composition of Two Sideritis L. Taxa from Turkey: A Chemotaxonomic Approach. Asian Journal of Chemistry, 26(8), 2466-2470.
22. Kiliç, I., & Yeşiloğlu, Y. (2013). Spectroscopic studies on the antioxidant activity of p-coumaric acid. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 115, 719-724.
23. Kirimer, N., Baser, K., Demirci, B., & Duman, H. (2004). Essential oils of Sideritis species of Turkey belonging to the section Empedoclia. Chemistry of Natural Compounds, 40, 19-23.
24. Kocak, M.S., Sarikurkcu, C., Cengiz, M., Kocak, S., Uren, M.C., & Tepe, B. (2016). Salvia cadmica: Phenolic composition and biological activity. Industrial Crops and Products, 85, 204-212.
25. Maatouk, M., Mustapha, N., Mokdad-Bzeouich, I., Chaaban, H., Abed, B., Iaonnou, I., Ghedira, K., Ghoul, M., & Ghedira, L.C. (2017). Thermal treatment of luteolin-7-O-β-glucoside improves its immunomodulatory and antioxidant potencies. Cell Stress and Chaperones, 22, 775-785.
26. Masek, A., Chrzescijanska, E., & Latos, M. (2016). Determination of antioxidant activity of caffeic acid and p-coumaric acid by using electrochemical and spectrophotometric assays. International Journal of Electrochemical Science, 11(12), 10644-10658.
27. Miliauskas, G., Venskutonis, P., & Van Beek, T. (2004). Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chemistry, 85(2), 231-237.
28. Mohamed, A.A., Khalil, A.A., & El-Beltagi, H.E. (2010). Antioxidant and antimicrobial properties of kaff maryam (Anastatica hierochuntica) and doum palm (Hyphaene thebaica). Grasas y aceites, 61(1), 67-75.
29. Oboh, G., Agunloye, O.M., Adefegha, S.A., Akinyemi, A.J., & Ademiluyi, A.O. (2015). Caffeic and chlorogenic acids inhibit key enzymes linked to type 2 diabetes (in vitro): a comparative study. Journal of Basic and Clinical Physiology and Pharmacology, 26(2), 165-170.
30. Orhan, I.E. (2021). Cholinesterase inhibitory potential of quercetin towards alzheimer's dis- ease- a promising natural molecule or fashion of the day?- A narrowed review. Current Neuropharmacology, 19(12), 2205-2213.
31. Ozer, M.S., Kirkan, B., Sarikurkcu, C., Cengiz, M., Ceylan, O., Atilgan, N., & Tepe, B. (2018). Onosma heterophyllum: Phenolic composition, enzyme inhibitory and antioxidant activities. Industrial Crops and Products, 111, 179-184.
32. Özcan, M., Chalchat, J., & Akgül, A. (2001). Essential oil composition of Turkish mountain tea (Sideritis spp.). Food Chemistry, 75(4), 459-463.
33. Palá‐Paúl, J., Pérez‐Alonso, M.J., Velasco‐Negueruela, A., Ballesteros, M.T., & Sanz, J. (2006). Essential oil composition of Sideritis hirsuta L. from Guadalajara Province, Spain. Flavour and Fragrance Journal, 21(3), 410-415.
34. Palomino, O., Gomez-Serranillos, P., Carretero, E., & Villar, A. (1996). High-performance liquid chromatography of flavonoids from Sideritis species. Journal of Chromatography A, 731(1-2), 103-108.
35. Park, J.Y., Han, X., Piao, M.J., Oh, M.C., Fernando, P.M.D.J., Kang, K.A., … Hyun, J.W. (2016). Hyperoside induces endogenous antioxidant system to alleviate oxidative stress. Journal of Cancer Prevention, 21(1), 41.
36. Rescigno, A., Casañola‐Martin, G.M., Sanjust, E., Zucca, P., & Marrero‐Ponce, Y. (2011). Vanilloid Derivatives as Tyrosinase Inhibitors Driven by Virtual Screening‐Based QSAR Models. Drug Testing and Analysis, 3(3), 176-181.
37. Sarikurkcu, C., Locatelli, M., Mocan, A., Zengin, G., & Kirkan, B. (2020). Phenolic Profile and Bioactivities of Sideritis perfoliata L.: The Plant, Its Most Active Extract, and Its Broad Biological Properties. Frontiers in Pharmacology, 10, 1642.
38. Sarikurkcu, C., Ozer, M.S., Cakir, A., Eskici, M., & Mete, E. (2013). GC/MS evaluation and in vitro antioxidant activity of essential oil and solvent extracts of an endemic plant used as folk remedy in Turkey: Phlomis bourgaei Boiss. [Article]. Evidence-Based Complementary and Alternative Medicine, 2013, Article ID 293080, 293087 pages.
39. Sato, Y., Itagaki, S., Kurokawa, T., Ogura, J., Kobayashi, M., Hirano, T., Sugawara, M., & Iseki, K. (2011). In vitro and in vivo antioxidant properties of chlorogenic acid and caffeic acid. International Journal of Pharmaceutics, 403(1-2), 136-138.
40. Sezen Karaoğlan, E., Hancı, H., Koca, M., & Kazaz, C. (2023). Some bioactivities of isolated apigenin-7-O-glucoside and luteolin-7-O-glucoside. Applied Sciences, 13(3), 1503.
41. Shen, H., Wang, J., Ao, J., Hou, Y., Xi, M., Cai, Y., Li, M., & Luo, A. (2023). Structure-activity relationships and the underlying mechanism of α-amylase inhibition by hyperoside and quercetin: Multi-spectroscopy and molecular docking analyses. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 285, 121797.
42. Tepe, B., Sarikurkcu, C., Berk, S., Alim, A., & Akpulat, H.A. (2011). Chemical composition, radical scavenging and antimicrobial activity of the essential oils of Thymus boveii and Thymus hyemalis. Records of Natural Products, 5(3), 208-220.
43. Thilagam, E., Parimaladevi, B., Kumarappan, C., & Mandal, S.C. (2013). α-Glucosidase and α-amylase inhibitory activity of Senna surattensis. Journal of Acupuncture and Meridian Studies, 6(1), 24-30.
44. Tian, C., Liu, X., Chang, Y., Wang, R., Lv, T., Cui, C., & Liu, M. (2021). Investigation of the anti-inflammatory and antioxidant activities of luteolin, kaempferol, apigenin and quercetin. South African Journal of Botany, 137, 257-264.
45. Venditti, A., Bianco, A., Frezza, C., Serafini, M., Giacomello, G., Giuliani, C., … Lucarini, D. (2016). Secondary metabolites, glandular trichomes and biological activity of Sideritis montana L. subsp. montana from Central Italy. Chemistry & Biodiversity, 13(10), 1380-1390.
46. Venditti, A., Bianco, A., Frezza, C., Serafini, M., Giacomello, G., Giuliani, C., … Maggi, F. (2016). Secondary Metabolites, Glandular Trichomes and Biological Activity of Sideritis montana L. subsp montana from Central Italy. Chemistry & biodiversity, 13(10), 1380-1390.
47. Venditti, A., Bianco, A., Maggi, F., & Nicoletti, M. (2013). Polar constituents composition of endemic Sideritis italica (Mill.) Greuter et Burter from Central Italy. Natural Product Research, 27(15), 1408-1412.
48. Wang, S., Li, Y., Huang, D., Chen, S., Xia, Y., & Zhu, S. (2022). The inhibitory mechanism of chlorogenic acid and its acylated derivatives on α-amylase and α-glucosidase. Food Chemistry, 372, 131334.
49. Wu, L. (2007). Effect of chlorogenic acid on antioxidant activity of Flos Lonicerae extracts. Journal of Zhejiang University Science B, 8, 673-679.
50. Zengin, G., Uren, M.C., Kocak, M.S., Gungor, H., Locatelli, M., Aktumsek, A., & Sarikurkcu, C. (2017). Antioxidant and Enzyme Inhibitory Activities of Extracts from Wild Mushroom Species from Turkey. International Journal of Medicinal Mushrooms, 19(4), 327-336.
51. Zheng, Y., Yang, W., Sun, W., Chen, S., Liu, D., Kong, X., Tian, J., & Ye, X. (2020). Inhibition of porcine pancreatic α-amylase activity by chlorogenic acid. Journal of Functional Foods, 64, 103587.
52. Zhou, J., Tang, Q., Wu, T., & Cheng, Z. (2017). Improved TLC bioautographic assay for qualitative and quantitative estimation of tyrosinase inhibitors in natural products. Phytochemical Analysis, 28(2), 115-124.