Multitarget biological activities and RP-HPLC-Based phytochemical profiling of Glaucium leiocarpum Boiss.

Sevgi Altın

Multitarget biological activities and RP-HPLC-Based phytochemical profiling of Glaucium leiocarpum Boiss.

Abstract

This study presents a comprehensive evaluation of the phenolic composition and biological activities of methanolic extracts obtained from the aerial (GLM) and root (GLRM) parts of Glaucium leiocarpum. Phenolic profiling was conducted using reversed-phase high-performance liquid chromatography (RP-HPLC), revealing catechin hydrate, 3-hydroxybenzoic acid, syringic acid, hesperidin, chlorogenic acid, caffeic acid, and trans-cinnamic acid as major constituents in GLM. In contrast, GLRM exhibited a more diverse phenolic profile, including epicatechin, benzoic acid, quercetin, gallic acid, and rosmarinic acid. Antioxidant activities were evaluated using DPPH and FRAP assays, with GLRM demonstrating stronger radical scavenging capacity (IC₅₀: 36.96 µg/mL) and higher ferric reducing power (173.20±0.68 mg TE/g extract). GLRM also contained significantly higher levels of total phenolics (142.81±2.87 mg GAE/g) and flavonoids (76.67±0.72 mg QE/g). Antimicrobial assays demonstrated that only GLRM exerted inhibitory effects against Staphylococcus aureus (8.7±0.57 mm), Enterococcus faecalis (7.7±0.57 mm), and Streptococcus pneumoniae (15.0±0.0 mm). Cytotoxicity studies, assessed by the XTT assay, revealed potent antiproliferative effects against HT-29 colorectal cancer cells (IC₅₀: 3.59 µg/mL for GLM, 2.29 µg/mL for GLRM) and moderate activity against A549 lung carcinoma cells (IC₅₀: 36.01 µg/mL for GLM, 21.99 µg/mL for GLRM). Furthermore, GLRM exhibited strong anticholinesterase activity, with IC₅₀ values of 2.85±0.27 µg/mL for acetylcholinesterase (AChE) and 2.47±0.07 µg/mL for butyrylcholinesterase (BChE). Overall, these findings underscore G. leiocarpum, particularly its root extract, as a valuable source of bioactive secondary metabolites, owing to its rich polyphenolic profile and notable antioxidant, antimicrobial, cytotoxic, and cholinesterase inhibitory activities.

Keywords

Thanks

The authors thank the Erzincan Binali Yıldırım University Scientific Research Center for providing the necessary equipment for cell culture studies and the Aksaray University Science and Technology Application and Research Center for providing the necessary equipment for RP-HPLC analysis.

References

Ahmadinejad, F., Møller, S.G., Hashemzadeh-Chaleshtori, M., Bidkhori, G., & Jami, M.S. (2017). Molecular mechanisms behind free radical scavengers function against oxidative stress. Antioxidants, 6(3), 1–15. https://doi.org/10.3390/antiox6030051
Adıgüzelli, G., Osma, E., Varol, T., Şimşek, S., & Kandemir, A. (2024). Erzincan ili-Ergan dağı’nın farklı rakımlarında yayılış gösteren bazı taksonların antioksidan kapasiteleri [Antioxidant capacities of some taxa distributed at different altitudes of Erzincan province Ergan mountain]. Osmaniye Korkut Ata Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 7(5), 2315-2332.
Akaberi, T., Shourgashti, K., Emami, S.A., & Akaberi, M. (2021). Phytochemistry and pharmacology of alkaloids from Glaucium spp. Phytochemistry, 191(April), 112923. https://doi.org/10.1016/j.phytochem.2021.112923
Al-Qura'n, S. (2015). Glaucium pollen interactions with Chondropyga dorsalis (Coleoptera) in Jordan. Arnaldoa, 22(1), 59-72.
Altın, S., Alp, C., Köksal, E., & Akyüz, S. (2023). Determination of antioxidant, antimicrobial, and antiproliferative activities of Onobrychis argyrea subsp. argyrea extracts. Journal of the Institute of Science and Technology, 13(2), 1134–1141. https://doi.org/10.21597/JIST.1240818
Altın, S., & Köksal, E. (2024). Glaucium cappadocicum : in vitro. Glaucium cappadocicum: In Vitro Antioxidant, Antimicrobial, Anticholinesterase Inhibition Properties, 52(1), 1–9.
Arafa, A.M., Mohamed, M.E.S., & Eldahmy, S.I. (2016). The aerial parts of yellow horn poppy (Glaucium flavum Cr.) growing in Egypt: Isoquinoline alkaloids and biological activities. Journal of Pharmaceutical Sciences and Research, 8(5), 323.
Azwanida, N.N. (2015). A review on the extraction methods use in medicinal plants, principle, strength and limitation. Med Aromat Plants, 4(196), 2167-0412.

Bauer, A.W., Kirby, W.M., Sherris, J.C., & Turck, M. (1966). Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology, 45(4), 493–496. https://doi.org/10.1093/ajcp/45.4_ts.493
Bektaş, E., Kaltalıoğlu, K., Şahin, H., Türkmen, Z., & Kandemir, A. (2018). Analysis of phenolic compounds, antioxidant and antimicrobial properties of some endemic medicinal plants. International Journal of Secondary Metabolite, 5(2), 75–86. https://doi.org/10.21448/ijsm.392354
Blois, M.S. (1958). Antioxidant determinations by the use of a stable free radical. Nature, 181, 1199–1200. https://doi.org/https://doi.org/10.1038/1811199a0
Bournine, L., Bensalem, S., Peixoto, P., Gonzalez, A., Maiza-Benabdesselam, F., Bedjou, F., Wauters, J.N., Tits, M., Frédérich, M., Castronovo, V., & Bellahcène, A. (2013). Revealing the anti-tumoral effect of Algerian Glaucium flavum roots against human cancer cells. Phytomedicine, 20(13), 1211–1218. https://doi.org/10.1016/j.phymed.2013.06.007
Chang, C.C., Yang, M.H., Wen, H.M., & Chern, J.C. (2002). Estimation of total flavonoid content in propolis by two complementary colometric methods. Journal of Food and Drug Analysis, 10(3), 178–182. https://doi.org/10.38212/2224-6614.2748
Cushnie, T.P.T., & Lamb, A.J. (2005). Antimicrobial activity of flavonoids. International Journal of Antimicrobial Agents, 26(5), 343–356. https://doi.org/10.1016/j.ijantimicag.2005.09.002
Delik, E., Eroğlu, B., Gülben, M., Aykurt, C., & Tefon Öztürk, B.E. (2024). Evaluation of biological activities of various extracts of glaucium alakirensis, Marrubium bourgaei, and Peucedanum alpinum from Türkiye. Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi, 27(6), 1249–1261. https://doi.org/10.18016/ksutarimdoga.vi.1388926
Dikici, E., Altın, S., Alp, C., Işık, M., Köksal, E., & Gülçin, İ. (2024). Determination of secondary metabolites of Cydonia oblonga (Quince) by LC-MS/MS method together with evaluation of its antioxidant and cholinergic potentials. Journal of Chemical Metrology, 18(2), 146–164. https://doi.org/10.25135/jcm.120.2411.3380
Ellman, G.L., Courtney, K.D., Andres, V., & Featherstone, R.M. (1961). A new and rapid colorimetric determinatıon of acetylcholinesterase activity. In Biochemical Pharmacology (Vol. 7). Pergamon Press Ltd.
Gümrükçüoğlu, A. (2025). Glaucıum flavum bitkisinin toprak üstü kısımlarında bulunan fenolik bileşiklerin, uçucu bileşenlerin ve antioksidan kapasitenin analizi [Identification of phenolic and volatile compounds in aerial parts of Glaucium flavum crantz. and determination of their antioxidant activity]. Mus Alparslan University Journal of Science, 13(1), 115 127. https://doi.org/10.18586/msufbd.1681215
Haji, A.F., Ostad, S.N., Khanavi, M., Haji, A.A., Farahanikia, B., & Salarytabar, A. (2013). Cytotoxicity of two species of Glaucium from Iran. 11(43), 85–92.
Hassanpour, S.H., & Doroudi, A. (2023). Review of the antioxidant potential of flavonoids as a subgroup of polyphenols and partial substitute for synthetic antioxidants. Avicenna Journal of Phytomedicine, 13(4), 354–376. https://doi.org/10.22038/AJP.2023.21774
Kalantari, M., & Entezari, M. (2020). Antiproliferative and pro-apoptotic effects of Glaucium flavum extract on A549 Lung Cancer Cells. 11(3), 21–29.
Kavaz Yüksel, A., Dikici, E., Yüksel, M., Işık, M., Tozoğlu, F., & Köksal, E. (2021). Phytochemical, phenolic profile, antioxidant, anticholinergic and antibacterial properties of Epilobium angustifolium (Onagraceae). Journal of Food Measurement and Characterization, 15(6), 4858-4867.
Kocanci, F.G., Hamamcioglu, B., & Aslim, B. (2017). The anti-AChE and anti-proliferative Activities of Glaucium acutidentatum and Glaucium corniculatum Alkaloid Extracts. Journal of Applied Pharmaceutical Science, 7(8), 191–200. https://doi.org/10.7324/JAPS.2017.70826
Köksal, E., Altay, A., Altın, S., Alp, C., Ulutaş, Y., Şimşek, S., Karakuş, S., Akyüz, S., Aygül, İ., Akşit, H., & Yilmaz, M.A. (2025). Screening of chemical profile by LC–MS/MS and assessment of biological properties of Achillea sintenisii Hub. Mor. From Turkey. Chemistry and Biodiversity, 1–12. https://doi.org/10.1002/cbdv.202500026
Kopustinskiene, D.M., Jakstas, V., Savickas, A., & Bernatoniene, J. (2020). Flavonoids as anticancer agents. Nutrients, 12(2), 1–26. https://doi.org/10.3390/nu12020457
Saxena, M., Saxena, J., Nema, R., Singh, D., & Gupta, A. (2013). Phytochemistry of medicinal plants. Journal of pharmacognosy and phytochemistry, 1(6).
Manoharan, S., Guillemin, G.J., Abiramasundari, R.S., Essa, M.M., Akbar, M., & Akbar, M.D. (2016). The role of reactive oxygen species in the pathogenesis of alzheimer’s disease, parkinson’s disease, and huntington’s disease: A mini review. Oxidative Medicine and Cellular Longevity, 2016. https://doi.org/10.1155/2016/8590578
Mariangela, M. (2017). Medicinal Plants. Plants, 11(1), 449 632. https://doi.org/10.3390/plants10071355
Morteza-Semnani, K., Saeedi, M., Mohammad, &, Mahdavi, R., & Mahdavi, M.R. (2005). Antibacterial Studies on Extracts of Three Species of Glaucium. from Iran. Taylor & Francis, 43(3), 234–236. https://doi.org/10.1080/13880200590928816
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(1), 359–388. https://doi.org/10.1080/10942912.2022.2157425
Oyaizu, M. (1986). Studies on Products of Browning Reaction Antioxidative Activities of Products of Browning Reaction Prepared from Glucosamine. The Japanese Journal of Nutrition and Dietetics, 44(6), 307–315. https://doi.org/10.5264/EIYOGAKUZASHI.44.307
Özcandır, A., Mohammed, F.S., Sevındık, M., Aykurt, C., Selamoğlu, Z., & Akgül, H. (2024). Phenolic composition, total antioxidant, antiradical and antimicrobial potential of endemic Glaucium alakirensis. Sigma Journal of Engineering and Natural Sciences, 42(1), 43–49. https://doi.org/10.14744/sigma.2024.00006
Ozsoy, N., Yilmaz-Ozden, T., Aksoy-Sagirli, P., Şahin, H., & Sarı, A. (2018). Antioxidant, Anti-acetylcholinesterase, Anti-inflammatory and DNA Protection Activities of Glaucium grandiflorum var. grandiflorum. Iranian Journal of Pharmaceutical Research, 17(2), 677–684.
Saraç, H., Daştan, T., Durukan, H., Durna Daştan, S., Demirbaş, A., & Karaköy, T. (2018). Kırmızı gelincik (Fam: Papaveraceae, Glaucium grandiflorum Boiss.&Huet var. grandiflorum) bitkisinin farklı özütlerinin besin elementi içeriğinin ve in vitro antiproliferatif etkilerinin değerlendirilmesi [Nutrient composition and in vitro anti-proliferative effects of different extracts of red poppy (Fam: Papaveraceae, Glaucium grandiflorum Boiss.&Huet var. grandiflorum) plants]. Süleyman Demirel Üniversitesi Ziraat Fakültesi Dergisi, 417–428.
Sauceda, A.E.Q., Sáyago-Ayerdi, S.G., Ayala-Zavala, J.F., Wall-Medrano, A., de la Rosa, L.A., González-Aguilar, G.A., & Álvarez-Parrilla, E. (2017). Biological actions of phenolic compounds. Fruit and Vegetable Phytochemicals: Chemistry and Human Health: Second Edition, 1, 125–137. https://doi.org/10.1002/9781119158042.ch6
Slinkard, K., & Singleton, V.L. (1977). Total phenol analysis: Automation and comparison with manual methods. American Journal of Enology and Viticulture, 28(1), 49–55. https://doi.org/10.5344/AJEV.1977.28.1.49
Sun, W., & Shahrajabian, M.H. (2023). Potencial terapêutico de compostos fenólicos em plantas medicinais – produtos naturais de saúde para a saúde humana. Molecules, 28(4), 1845.
Tavsan, Z., & Kayali, H.A. (2019). Flavonoids showed anticancer effects on the ovarian cancer cells: Involvement of reactive oxygen species, apoptosis, cell cycle and invasion. Biomedicine and Pharmacotherapy, 116(May), 109004. https://doi.org/10.1016/j.biopha.2019.109004
Thebti, A., Meddeb, A., Ben Salem, I., Bakary, C., Ayari, S., Rezgui, F., Essafi-Benkhadir, K., Boudabous, A., & Ouzari, H.I. (2023). Antimicrobial Activities and Mode of Flavonoid Actions. Antibiotics, 12(2). https://doi.org/10.3390/antibiotics12020225
Torunoğlu, E.I., & Akdağ, T. (2025). Investigation of biological interactions in Euphorbia rigida extract using molecular docking. International Journal of Secondary Metabolite, 12(2), 271–288. https://doi.org/10.21448/ijsm.1537429
Toul, F., Ballou, M., & Kessou, M. (2025). A comparative analysis of the effects of different drying and storage techniques on the phenolic content of Pistacia atlantica leaves. International Journal of Secondary Metabolite, 12(2), 261–270. https://doi.org/10.21448/ijsm.1497914
Vaou, N., Stavropoulou, E., Voidarou, C., Tsakris, Z., Rozos, G., Tsigalou, C., & Bezirtzoglou, E. (2022). Interactions between medical plant-derived bioactive compounds: Focus on antimicrobial combination effects. Antibiotics, 11(8), 1014.
Yagi, S., Zengin, G., Uba, A.I., Maciejewska-Turska, M., Sieniawska, E., Świątek, Ł., Rajtar, B., Bahşi, M., Guler, O., Dall’Acqua, S., & Polz-Dacewicz, M. (2024). Exploring chemical composition, antioxidant, enzyme inhibitory and cytotoxic properties of Glaucium acutidentatum Hausskn. & Bornm. from Turkey flora: A novel source of bioactive agents to design functional applications. Antioxidants, 13(6). https://doi.org/10.3390/antiox13060643
Yeniçeri, E., Altay, A., Koksal, E., Altın, S., Taslimi, P., Yılmaz, M.A., Cakir, O., Tarhan, A., & Kandemir, A. (2024). Phytochemical profile by LC-MS/MS analysis and evaluation of antioxidant, antidiabetic, anti-Alzheimer, and anticancer activity of Onobrychis argyrea leaf extracts. European Journal of Integrative Medicine, 66(October 2023). https://doi.org/10.1016/j.eujim.2024.102337
Zhang, Y., Cai, P., Cheng, G., & Zhang, Y. (2022). A brief review of phenolic compounds identified from plants: Their extraction, analysis, and biological activity. Natural Product Communications, 17(1). https://doi.org/10.1177/1934578X211069721

Details

Primary Language

English

Subjects

Natural Products and Bioactive Compounds

Journal Section

Research Article

Authors

Sevgi Altın ^*
0000-0002-0820-912X
Türkiye

Early Pub Date

August 4, 2026

Publication Date

-

Submission Date

August 20, 2025

Acceptance Date

May 23, 2026

Published in Issue

Year 2026 Number: Advanced Online Publication

IZ

https://izlik.org/JA92TE59CA

Cite

RIS / Bibtex

APA

Altın, S. (2026). Multitarget biological activities and RP-HPLC-Based phytochemical profiling of Glaucium leiocarpum Boiss. International Journal of Secondary Metabolite, Advanced Online Publication. https://izlik.org/JA92TE59CA

AMA

1.Altın S. Multitarget biological activities and RP-HPLC-Based phytochemical profiling of Glaucium leiocarpum Boiss. Int. J. Sec. Metabolite. 2026;(Advanced Online Publication). https://izlik.org/JA92TE59CA

Chicago

Altın, Sevgi. 2026. “Multitarget Biological Activities and RP-HPLC-Based Phytochemical Profiling of Glaucium Leiocarpum Boiss”. International Journal of Secondary Metabolite, no. Advanced Online Publication. https://izlik.org/JA92TE59CA.

EndNote

Altın S (August 1, 2026) Multitarget biological activities and RP-HPLC-Based phytochemical profiling of Glaucium leiocarpum Boiss. International Journal of Secondary Metabolite Advanced Online Publication

IEEE

[1]S. Altın, “Multitarget biological activities and RP-HPLC-Based phytochemical profiling of Glaucium leiocarpum Boiss”., Int. J. Sec. Metabolite, no. Advanced Online Publication, Aug. 2026, [Online]. Available: https://izlik.org/JA92TE59CA

ISNAD

Altın, Sevgi. “Multitarget Biological Activities and RP-HPLC-Based Phytochemical Profiling of Glaucium Leiocarpum Boiss”. International Journal of Secondary Metabolite. Advanced Online Publication (August 1, 2026). https://izlik.org/JA92TE59CA.

JAMA

1.Altın S. Multitarget biological activities and RP-HPLC-Based phytochemical profiling of Glaucium leiocarpum Boiss. Int. J. Sec. Metabolite. 2026. Available at https://izlik.org/JA92TE59CA.

MLA

Altın, Sevgi. “Multitarget Biological Activities and RP-HPLC-Based Phytochemical Profiling of Glaucium Leiocarpum Boiss”. International Journal of Secondary Metabolite, no. Advanced Online Publication, Aug. 2026, https://izlik.org/JA92TE59CA.

Vancouver

1.Sevgi Altın. Multitarget biological activities and RP-HPLC-Based phytochemical profiling of Glaucium leiocarpum Boiss. Int. J. Sec. Metabolite [Internet]. 2026 Aug. 1;(Advanced Online Publication). Available from: https://izlik.org/JA92TE59CA