Abstract
Natural carbonic anhydrase (CA) inhibitors are gaining significant attention for their potential applications in diuretic, antiglaucoma, anti-obesity, and anticancer therapies. This study provides the first comparative evaluation of CA inhibitory potential, antioxidant activity, and LC-MS/MS (Liquid Chromatography Tandem Mass Spectrometry) phytochemical profiles of hydroalcoholic extracts from Satureja hortensis, Juniperus communis, Matricaria chamomilla, Hypericum perforatum, and Valeriana officinalis sourced from Romania and Türkiye. Principal Component Analysis (PCA) was employed to correlate secondary metabolites with observed biological activities. While S. hortensis and J. communis displayed consistent metabolic profiles across both countries, the other species exhibited geographic variation. Correlation analysis indicated that while specific phenolic acids (e.g., gallic, ferulic) supported antioxidant capacity, rosmarinic acid and luteolin were strong contributors to CA inhibition and diuretic potential. Notably, S. hortensis emerged as the most promising source; the Turkish sample demonstrated superior efficacy, attributed to high rosmarinic acid content (49.76 µg/mL) and substantial CA inhibition (64.87 µg acetazolamide equivalent/mL). The highest values of polyphenol and flavonoid content were found in H. perforatum from Türkiye, with 16.95 ± 0.221 and 9.77 ± 0.17, respectively. These findings underscore the therapeutic potential of S. hortensis in managing CA-related disorders.
Data availability
The datasets used and analyzed during the current study are available from the corresponding author upon reasonable request.
References
Çarıkçı, S. et al. Chemical profile of the Anatolian sideritis species with bioactivity studies. Pharm. Biol. 61, 1484–1511 (2023).
Habib, M. R. et al. Thais savignyi tissue extract: bioactivity, chemical composition, and molecular docking. Pharm. Biol. 60, 1899–1914 (2022).
Surma, S., Więcek, A. & Adamczak, M. In Renal Disease Transplantation Forum 81–92 (2023).
Roush, G. C., Kaur, R. & Ernst, M. E. Diuretics: a review and update. J. Cardiovasc. Pharmacol. Therap. 19, 5–13 (2014).
Kehrenberg, M. C. & Bachmann, H. S. Diuretics: a contemporary Pharmacological classification? Naunyn. Schmiedebergs Arch. Pharmacol. 395, 619–627 (2022).
Aswini, E., Vivek, D. & Swathilakshmi, S. Role of phytochemicals in diuresis management. World J. Pharm. Res. 9, 551–562 (2020).
Aydın, F. G., Türkoğlu, E. A., Kuzu, M. & Taşkın, T. In vitro carbonic anhydrase inhibitory effects of the extracts of satureja cuneifolia. Türk Tarım Ve Doğa Bilimleri Dergisi. 8, 1146–1150 (2021).
Foote, J. & Cohen, B. Medicinal herb use and the renal patient. J. Ren. Nutr. 8, 40–42 (1998).
Kaya, E. D., Erğun, B., Demir, Y., Alım, Z. & Beydemir, Ş. The in vitro impacts of some plant extracts on carbonic anhydrase i, Ii and paraoxonase-1. Hacettepe J. Biology Chem. 47, 51–59 (2019).
Masood, N. et al. Antioxidant, carbonic anhydrase Inhibition and diuretic activity of leptadenia pyrotechnica Forssk. Decne Heliyon 9, 639 (2023).
Meharie, B. G. & Tunta, T. A. Evaluation of diuretic activity and phytochemical contents of aqueous extract of the shoot apex of Podocarpus falcactus. J. Exp. Pharmacol. 2020, 629–641 (2020).
Shah, S., Anjum, S. & Littler, W. Use of diuretics in cardiovascular disease:(2) hypertension. Postgrad. Med. J. 80, 271–276 (2004).
Sica, D. A. Thiazide-type diuretics: ongoing considerations on mechanism of action. J. Clin. Hypertens. 6, 661–664 (2004).
Hullatti, K., Gopikrishna, U. & Kuppast, I. Phytochemical investigation and diuretic activity of cyclea peltata leaf extracts. J. Adv. Pharm. Tech. Res. 2, 241–244 (2011).
Ntchapda, F. et al. Diuretic and antihypertensive activity of the aqueous extract of haematostaphis Barteri stem bark in adrenaline-induced hypertensive Wistar rats. J. Phytopharmacol. 11, 895 (2022).
Alissa, S. A. et al. Inhibition survey with phenolic compounds against the δ-and η-class carbonic anhydrases from the marine diatom Thalassiosira weissflogii and protozoan plasmodium falciparum. J. Enzyme Inhib. Med. Chem. 35, 377–382 (2020).
Innocenti, A., Gülçin, I., Scozzafava, A. & Supuran, C. T. Carbonic anhydrase inhibitors. Antioxidant polyphenols effectively inhibit mammalian isoforms I–XV. Bioorg. Med. Chem. Lett. 20, 5050–5053 (2010).
Karioti, A., Carta, F. & Supuran, C. T. Phenols and polyphenols as carbonic anhydrase inhibitors. Molecules 21, 1649 (2016).
Ibn Sinâ, H. Al-Qânun Fiant-Tibbe (Canon of Medicine). Tehran Iran (Almaʿee Publication, 2015).
Butterweck, V. Mechanism of action of St john’s wort in depression: what is known? CNS Drugs. 17, 539–562 (2003).
Bombardelli, E. & Morazzoni, P. Hypericum Perforatum (Springer, 1995).
Razi, M. Al-Hawi fi’l-tibb (Comprehensive book of medicine). Hyderabad: Osmania Orient. Publications Bureau. 20, 548–553 (1968).
Alahmad, A. et al. Identification of major constituents of hypericum perforatum L. extracts in Syria by development of a rapid, simple, and reproducible HPLC-ESI-Q-TOF MS analysis and their antioxidant activities. ACS Omega. 7, 13475–13493 (2022).
Karpitskiy, D. A., Bessonova, E. A., Shishov, A. Y. & Kartsova, L. A. Handshake of deep eutectic solvent and ionic liquid: two liquid-liquid Microextraction procedures for plant analysis. Talanta 282, 126947 (2025).
Kimáková, K. et al. Phenotyping the genus hypericum by secondary metabolite profiling: Emodin vs. skyrin, two possible key intermediates in hypericin biosynthesis. Anal. Bioanal. Chem. 410, 7689–7699 (2018).
Gumral, N., Kumbul, D. D., Aylak, F., Saygin, M. & Savik, E. Juniperus communis Linn oil decreases oxidative stress and increases antioxidant enzymes in the heart of rats administered a diet rich in cholesterol. Toxicol. Ind. Health. 31, 85–91 (2015).
Seca, A. M. & Silva, A. M. The chemical composition of the Juniperus genus (1970–2004). Recent. Progress Med. Plants. 16, 401–522 (2006).
Raina, R., Verma, P. K., Peshin, R. & Kour, H. Potential of Juniperus communis L as a nutraceutical in human and veterinary medicine. Heliyon 5, 639 (2019).
Das, M. Chamomile: medicinal, biochemical, and Agricultural Aspects (CRC, 2014).
Dai, Y. L. et al. Chamomile: a review of its traditional uses, chemical constituents, Pharmacological activities and quality control studies. Molecules 28, 133 (2023).
Stanic, G. & Samaržija, I. Diuretic activity of satureja Montana subsp. Montana extracts and oil in rats. Phytother. Res. 7, 363–366 (1993).
Bimbiraitė-Survilienė, K. et al. Evaluation of chemical composition, radical scavenging and antitumor activities of satureja hortensis L. herb extracts. Antioxidants 10, 53 (2021).
Nandhini, S., Narayanan, K. B. & Ilango, K. Valeriana officinalis: a review of its traditional uses, phytochemistry and Pharmacology. Asian J. Pharm. Clin. Res. 11, 36–41 (2018).
Patočka, J. & Jakl, J. Biomedically relevant chemical constituents of Valeriana officinalis. J. Appl. Biomed. 8, 11–18 (2010).
Lung, I., Soran, M. L., Tudoran, C. & Măruţoiu, C. Effect of microwave irradiation on polyphenolic compounds from satureja hortensis L. Cent. Eur. J. Chem. 11, 535–541 (2013).
Shanaida, M., Jasicka-Misiak, I. & Wieczorek, P. P. Chromatographic analysis of polyphenols in the satureja hortensis L.(Lamiaceae Martinov) herb. Pharmacol. OnLine. 2, 1337–1345 (2021).
Gopčević, K. et al. Bioactivity and phenolics profile of aqueous and Ethyl acetate Extracts of satureja kitaibelii Wierzb. Ex Heuff. Obtained by ultrasound-assisted Extraction. Sci. Rep. 12, 21221 (2022).
Elmastaş, M., Gülçin, I., Beydemir, Ş., İrfan Küfrevioğlu, Ö. & Aboul-Enein, H. Y. A study on the in vitro antioxidant activity of Juniper (Juniperus communis L.) fruit extracts. Anal. Lett. 39, 47–65 (2006).
Yaglioglu, A. S. & Eser, F. Screening of some Juniperus extracts for the phenolic compounds and their antiproliferative activities. South. Afr. J. Bot. 113, 29–33 (2017).
Miceli, N. et al. Comparative study of the phenolic profile, antioxidant and antimicrobial activities of leaf extracts of five Juniperus L.(Cupressaceae) taxa growing in Turkey. Nat. Prod. Res. 34, 1636–1641 (2020).
Taviano, M. F. et al. Juniperus Oxycedrus L. subsp. Oxycedrus and Juniperus Oxycedrus L. subsp. Macrocarpa (Sibth. & Sm.) Ball.berries from turkey: comparative evaluation of phenolic profile, antioxidant, cytotoxic and antimicrobial activities. Food Chem. Toxicol. 58, 22–29 (2013).
Catani, M. V., Rinaldi, F., Tullio, V., Gasperi, V. & Savini, I. Comparative analysis of phenolic composition of six commercially available chamomile (Matricaria Chamomilla L.) extracts: potential biological implications. Int. J. Mol. Sci. 22, 10601 (2021).
El Mihyaoui, A. et al. Chamomile (Matricaria Chamomilla L.): a review of ethnomedicinal use, phytochemistry and Pharmacological uses. Life 12, 479 (2022).
Elmastas, M., Çinkiliç, S. & Aboul-Enein, H. Y. Antioxidant capacity and determination of total phenolic compounds in Daisy (Matricaria chamomilla, Fam. Asteraceae). World J. Anal. Chem. 3, 9–14 (2015).
Sotiropoulou, N. S., Megremi, S. F. & Tarantilis, P. Evaluation of antioxidant activity, toxicity, and phenolic profile of aqueous extracts of chamomile (Matricaria Chamomilla L.) and Sage (Salvia officinalis L.) prepared at different temperatures. Appl. Sci. 10, 2270 (2020).
Jürgenliemk, G. & Nahrstedt, A. Phenolic compounds from hypericum perforatum. Planta Med. 68, 88–91 (2002).
Öztürk, N., Tunçel, M. & Potoğlu-Erkara, İ. Phenolic compounds and antioxidant activities of some hypericum species: a comparative study with H. perforatum. Pharm. Biol. 47, 120–127 (2009).
Del Monte, D. et al. Phenolic content, antimicrobial and antioxidant activities of hypericum perfoliatum L. Ind. Crops Prod. 74, 342–347 (2015).
Bhatt, I. D. et al. Characterization of essential oil composition, phenolic content, and antioxidant properties in wild and planted individuals of Valeriana Jatamansi Jones. Sci. Hort. 136, 61–68 (2012).
Wu, C. R., Lee, S. Y., Chen, C. H. & Lin, S. D. Bioactive compounds of underground Valerian extracts and their effect on inhibiting metabolic syndrome-related enzymes activities. Foods 12, 636 (2023).
Średnicka-Tober, D. et al. Profile of selected secondary metabolites and antioxidant activity of valerian and lovage grown in organic and low-input conventional system. Metabolites 12, 835 (2022).
Khursheed, A. et al. Efficacy of phytochemicals derived from roots of Rondeletia odorata as Antioxidant, Antiulcer, Diuretic, skin brightening and hemolytic Agents—a comprehensive biochemical and in Silico study. Molecules 27, 4204 (2022).
Topal, M. & GÜLÇİN, İ. Rosmarinic acid: a potent carbonic anhydrase isoenzymes inhibitor. Turk. J. Chem. 38, 894–902 (2014).
Türker, İ. & İşleroğlu, H. Extraction of bioactive compounds from Fenugreek leaves by maceration with D-optimal design. Gıda 48, 305–316 (2023).
Kayir, Ö., Doğan, H., Alver, E. & Bilici, İ. Quantification of Phenolic Component by LC‐HESI‐MS/MS and Evaluation of Antioxidant Activities of Crocus Ancyrensis (Ankara Çiğdemi) Extracts Obtained with Different Solvents. Chemistry & Biodiversity 20, e202201186 (2023).
Doğan, H., Kayir, Ö., Yaylaci, Ö. K., Bi̇li̇ci̇, İ. & Alver, E. Determination of phytochemical content, antioxidant and mineral properties of astragalus Chamaephaca Freyn in different solvents. Sci. Rep. 15, 20886 (2025).
Fernandes, A. J. D., Ferreira, M. R. A., Randau, K. P., de Souza, T. P. & Soares, L. A. L. Total flavonoids content in the raw material and aqueous extractives from Bauhinia monandra Kurz (Caesalpiniaceae). Sci. World J. 2012, 923462 (2012).
Vl, S. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Methods Enzymol. 299, 152–178 (1999).
Erel, O. A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clin. Biochem. 37, 277–285 (2004).
Brand-Williams, W., Cuvelier, M. E. & Berset, C. Use of a free radical method to evaluate antioxidant activity. LWT-Food Sci. Technol. 28, 25–30 (1995).
Armstrong, J. M., Myers, D. V., Verpoorte, J. A. & Edsall, J. T. Purification and properties of human erythrocyte carbonic anhydrases. J. Biol. Chem. 241, 5137–5149 (1966).
Yung-Chi, C. & Prusoff, W. H. Relationship between the Inhibition constant (KI) and the concentration of inhibitor which causes 50 per cent Inhibition (I50) of an enzymatic reaction. Biochem. Pharmacol. 22, 3099–3108 (1973).
Murtagh, F. & Heck, A. Multivariate Data Analysis Vol. 131 (Springer Science & Business Media, 2012).
Acknowledgements
The phenolic compound analyses were carried out by the Hitit University Scientific Research and Application Center (HUBTUAM) in Türkiye, while the biological activity experiments were conducted at the Research Centre for Advanced Chemical Analysis in Romania.
Funding
This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.
Author information
Authors and Affiliations
Contributions
E.B. and D.C.; Conceptualization, methodology, software, validation, formal analysis, investigation, resources, data curation, writing original draft preparation, writing review and editing, supervision.S.C.A.C.: Conceptualization, methodology, software, validation, formal analysis, investigation, resources, data curation, writing—original draft preparation, writing—review and editing.İ.B., Ö.K., and H.D.: Conceptualization, methodology, software, validation, formal analysis, investigation, resources, data curation, writing original draft preparation, writing review and editing.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.
About this article
Cite this article
Büker, E., Casoni, D., Cobzac, S.C.A. et al. LC-MS/MS characterization, biological activity, and carbonic anhydrase inhibitory potential of five medicinal plant extracts from Romania and Türkiye. Sci Rep (2026). https://doi.org/10.1038/s41598-026-38419-5
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-026-38419-5
