Total phenolic content, antimicrobial activity, and LC-MS analysis of Limonium otolepis with isolation of apigenin

Dadahon Urmonov; Ikbol Zhalolov; Uchkun Ishimov; Dilmurodjon Turdaliev; Shakhnoz Azimova; Alidjan Ibragimov

Total phenolic content, antimicrobial activity, and LC-MS analysis of Limonium otolepis with isolation of apigenin

Abstract

Limonium otolepis (Schrenk) Kuntze, a member of the Plumbaginaceae family, was investigated for its phenolic composition and antimicrobial activity. The ethyl acetate extract showed the highest total phenolic content (123.48 ± 10.04 mg GAE/g extract), followed by ethanol and water extracts. Chromatographic separation of the ethanolic extract led to the isolation of apigenin, reported for the first time in this species. LC–MS analysis of the ethyl acetate subfractions predicted several phenolic acids (gallic, caffeic, protocatechuic, syringic acids) and flavonoids (myricetin, hesperetin). Antimicrobial assays at 2 mg/disc concentration revealed that the ethyl acetate fraction displayed the strongest inhibition, particularly against Pseudomonas aeruginosa (14±0.13 mm), whereas the 60% ethanol fraction was less active and the chloroform fraction showed minimal effects. These results highlight L. otolepis as a potential source of bioactive phenolics, with apigenin as one of the key constituents warranting further pharmacological evaluation.

Keywords

References

Abdel-Sattar, E., Shams, M.M., Abd-Rabo, M.M., Mahmoud, N., & Mahrous, E.A. (2021). Chemical and biological investigations of Limonium axillare reveal mechanistic evidence for its antidiabetic activity. PLOS ONE, 16(8), e0255904. https://doi.org/10.1371/journal.pone.0255904
Ahmed, K.M., Kandil, F.E., & Mabry, T.J. (1999). An anticancer tannin and other phenolics from Limonium axillare (Fam. Plumbaginaceae). Asian Journal of Chemistry, 11(1), 261–263.
Allemailem, K.S., Almatroudi, A., Alharbi, H.O.A., AlSuhaymi, N., Alsugoor, M.H., Aldakheel, F.M., Khan, A.A., & Rahmani, A.H. (2024). Apigenin: A bioflavonoid with a promising role in disease prevention and treatment. Biomedicines, 12(6), 1353. https://doi.org/10.3390/biomedicines12061353
Al-Madhagi, W.M., Hashim, N.M., Ali, N.A.A., & Othman, R. (2019). Phytochemical screening, cytotoxic and antimicrobial activities of Limonium socotranum and Peperomia blanda extracts. Tropical Biomedicine, 36(1), 11–21.
Apel, K., & Hirt, H. (2004). Reactive oxygen species: Metabolism, oxidative stress, and signal transduction. Annual Review of Plant Biology, 55, 373 399. https://doi.org/10.1146/annurev.arplant.55.031903.141701
Avaz, S., Korcan, S.E., Kargioglu, M., Serteser, A., Konuk, M., & Erol, I. (2013). Antimicrobial activities in root extracts of Limonium spp. growing in Afyonkarahisar, Turkey. Journal of Environmental Biology, 34(3), 561–568.
Bashir, A.K., Abdalla, A.A., Wasfi, I.A., Hassan, E.S., Amiri, M.H., & Crabb, T.A. (1994). Flavonoids of Limonium axillare. Pharmaceutical Biology, 32(4), 366 372. https://doi.org/10.3109/13880209409083017
Baysal, I., Ekizoglu, M., Ertas, A., Temiz, B., Agalar, H.G., Yabanoglu-Ciftci, S., Temel, H., Ucar, G., & Turkmenoglu, F.P. (2021). Identification of phenolic compounds by LC-MS/MS and evaluation of bioactive properties of two edible halophytes: Limonium effusum and L. sinuatum. Molecules, 26(13), 4040. https://doi.org/10.3390/molecules26134040

Blainski, A., Gionco, B., Oliveira, A.G., Andrade, G., Scarminio, I.S., Silva, D.B., Lopes, N.P., & Mello, J.C.P. (2017). Antibacterial activity of Limonium brasiliense (Baicuru) against multidrug-resistant bacteria using a statistical mixture design. Journal of Ethnopharmacology, 198, 313–323. https://doi.org/10.1016/j.jep.2017.01.013
Carius, B., Silva, H., Silva, A.M.S., & Pinto, D.C.G.A. (2022). Chemical Profiling of Limonium vulgare Mill. Using UHPLC-DAD-ESI/MS2 and GC-MS Analysis. Applied Sciences, 12(13), 6384. https://doi.org/10.3390/app12136384
Ciupei, D., Colişar, A., Leopold, L., Stănilă, A., & Diaconeasa, Z.M. (2024). Polyphenols: From classification to therapeutic potential and bioavailability. Foods, 13(24), 4131. https://doi.org/10.3390/foods13244131
Eshboev, F., Karakozova, M., Abdurakhmanov, J., Bobakulov, K., Dolimov, K., Abdurashidov, A., Baymirzaev, A., Makhnyov, A., Terenteva, E., Sasmakov, S., Piyakina, G., Egamberdieva, D., Nazarov, P.A., & Azimova, S. (2023). Antimicrobial and cytotoxic activities of the secondary metabolites of endophytic fungi isolated from the medicinal plant Hyssopus officinalis. Antibiotics (Basel, Switzerland), 12(7), 1201. https://doi.org/10.3390/antibiotics12071201
Gadetskaya, A.V., Tarawneh, A.H., Zhusupova, G.E., Gemejiyeva, N.G., Cantrell, C.L., Cutler, S.J., & Ross, S.A. (2015). Sulfated phenolic compounds from Limonium caspium: Isolation, structural elucidation, and biological evaluation. Fitoterapia, 104, 80 85. https://doi.org/10.1016/j.fitote.2015.05.017
Gancedo, N.C., Isolani, R., de Oliveira, N.C., Nakamura, C.V., de Medeiros Araújo, D.C., Sanches, A.C.C., Tonin, F.S., Fernandez-Llimos, F., Chierrito, D., & de Mello, J.C.P. (2023). Chemical constituents, anticancer and anti-Proliferative potential of Limonium species: A systematic review. Pharmaceuticals, 16(2), 293. https://doi.org/10.3390/ph16020293
Hatami, T., Emami, S.A., Miraghaee, S.S., & Mojarrab, M. (2014). Total phenolic contents and antioxidant activities of different extracts and fractions from the aerial parts of Artemisia biennis Willd. Iranian Journal of Pharmaceutical Research, 13(2), 551 559. https://doi.org/10.22037/ijpr.2014.1518
Kassymova, D., Zhusupova, G., Ogay, V., Zhussupova, A., Katragunta, K., Avula, B., & Khan, I.A. (2023). Phytochemical profiles and in vitro immunomodulatory activities of extracts obtained from Limonium gmelinii using different extraction methods. Plants, 12(23), 4019. https://doi.org/10.3390/plants12234019
Kerkatou, M., Redouane-Salah, A., León, F., Brouard, I., Mosset, P., Menad, A., & Benayache, F. (2016). Secondary Metabolites and Antioxidant Activity of Limonium duriusculum (de Girard) Kuntze Extracts. Asian Journal of Chemistry, 28(12), 2695 2700. https://doi.org/10.14233/ajchem.2016.20071
Khan, J., Deb, P.K., Priya, S., Medina, K.D., Devi, R., Walode, S.G., & Rudrapal, M. (2021). Dietary flavonoids: Cardioprotective potential with antioxidant effects and their pharmacokinetic, toxicological and therapeutic concerns. Molecules, 26(13), 4021. https://doi.org/10.3390/molecules26134021
Kong, J.M., Goh, N.K., Chia, L.S., & Chia, T.F. (2003). Recent advances in traditional plant drugs and orchids. Acta Pharmacologica Sinica, 24(1), 7-21.
Koutroumpa, K., Theodoridis, S., Warren, B.H., Jiménez, A., Celep, F., Doğan, M., Romeiras, M.M., Santos-Guerra, A., Fernández-Palacios, J.M., Caujapé-Castells, J., Moura, M., Menezes de Sequeira, M., & Conti, E. (2018). An expanded molecular phylogeny of Plumbaginaceae, with emphasis on Limonium (sea lavenders): Taxonomic implications and biogeographic considerations. Ecology and Evolution, 8(24), 12397–12424. https://doi.org/10.1002/ece3.4553
Mahasneh, A.M. (2002). Screening of some indigenous Qatari medicinal plants for antimicrobial activity. Phytotherapy Research: PTR, 16(8), 751–753. https://doi.org/10.1002/ptr.1037
Mariappan, G., Sundaraganesan, N., & Manoharan, S. (2012). The spectroscopic properties of anticancer drug apigenin investigated by using DFT calculations, FT-IR, FT-Raman and NMR analysis. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 95, 86–99. https://doi.org/10.1016/j.saa.2012.04.089
Medini, F., Bourgou, S., Lalancette, K.G., Snoussi, M., Mkadmini, K., Coté, I., Abdelly, C., Legault, J., & Ksouri, R. (2015). Phytochemical analysis, antioxidant, anti-inflammatory, and anticancer activities of the halophyte Limonium densiflorum extracts on human cell lines and murine macrophages. South African Journal of Botany, 99, 158 164. https://doi.org/10.1016/j.sajb.2015.04.007
Medini, F., Fellah, H., Ksouri, R., & Abdelly, C. (2014). Total phenolic, flavonoid and tannin contents and antioxidant and antimicrobial activities of organic extracts of shoots of the plant Limonium delicatulum. Journal of Taibah University for Science, 8(3), 216 224. https://doi.org/10.1016/j.jtusci.2014.01.003
Medini, F., Megdiche, W., Mshvildadze, V., Pichette, A., Legault, J., St-Gelais, A., & Ksouri, R. (2016). Antiviral-guided fractionation and isolation of phenolic compounds from Limonium densiflorum hydroalcoholic extract. Comptes Rendus Chimie, 19(6), 726 732. https://doi.org/10.1016/j.crci.2016.03.006
Murray, A.P., Rodriguez, S., Frontera, M.A., Tomas, M.A., & Mulet, M.C. (2004). Antioxidant metabolites from Limonium brasiliense (Boiss.) Kuntze. Zeitschrift für Naturforschung C, 59(7–8), 477–480.
Royal Botanic Gardens, Kew. (n.d.). Limonium otolepis (Schrenk) Kuntze. Plants of the World Online. Retrieved January 4, 2026, from https://powo.science.kew.org/taxon/urn:lsid:ipni.org:names:686902-1
Rudrapal, M., Khairnar, S. J., Khan, J., Dukhyil, A.B., Ansari, M.A., Alomary, M.N., Alshabrmi, F.M., Palai, S., Deb, P.K., & Devi, R. (2022). Dietary polyphenols and their role in oxidative stress-induced human diseases: insights into protective effects, antioxidant potentials and mechanism(s) of action. Frontiers in Pharmacology, 13, 806470. https://doi.org/10.3389/fphar.2022.806470
Ruiz-Riaguas, A., Zengin, G., Sinan, K. I., Salazar-Mendías, C., & Llorent-Martínez, E.J. (2020). Phenolic profile, antioxidant activity, and enzyme inhibitory properties of Limonium delicatulum (Girard) Kuntze and Limonium quesadense Erben. Journal of Chemistry, 2020, 1016208. https://doi.org/10.1155/2020/1016208
Saidana, D., Boussaada, O., Ayed, F., Mahjoub, M.A., Mighri, Z., & Helal, A.N. (2013). The in vitro free radical-scavenging and antifungal activities of the medicinal herb Limonium echioides L. growing wild in Tunisia. Journal of Food Processing and Preservation, 37(5), 533–540. https://doi.org/10.1111/j.1745-4549.2012.00673.x
Salehi, B., Venditti, A., Sharifi-Rad, M., Kręgiel, D., Sharifi-Rad, J., Durazzo, A., Lucarini, M., Santini, A., Souto, E.B., Novellino, E., Antolak, H., Azzini, E., Setzer, W.N., & Martins, N. (2019). The Therapeutic Potential of Apigenin. International Journal of Molecular Sciences, 20(6), 1305. https://doi.org/10.3390/ijms20061305
Sami, M., Sezen Karaoğlan, E., Hancı, H., and Kazaz, C. (2025). Some bioactivities of Limonium meyeri extracts and their separated compounds. BioResources, 20(1), 2171 2182. https://doi.org/10.15376/biores.20.1.2171-2182
Selamoglu, Z., Sevindik, M., Bal, C., Ozaltun, B., Sen, İ., & Pasdaran, A. (2020). Antioxidant, antimicrobial and DNA protection activities of phenolic content of Tricholoma virgatum (Fr.) P. Kumm. Biointerface Research in Applied Chemistry, 10(3), 5500 5506, https://doi.org/10.33263/BRIAC103.500506
Sennikov A.N. (Ed.) (2019). Flora of Uzbekistan, (Vol. 3). Ma’naviyat.
Siddiquee, R., Mahmood, T., Ansari, V.A., Ahsan, F., Bano, S., & Ahmad, S. (2025). Apigenin unveiled: An encyclopedic review of its preclinical and clinical insights. Discover Plants, 2, Article 11. https://doi.org/10.1007/s44372-024-00039-6
Singleton, V.L., Orthofer, R., & Lamuela-Raventós, R.M. (1999). Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. In L. Packer (Ed.), Methods in enzymology (Vol. 299, pp. 152–178). Academic Press. https://doi.org/10.1016/S0076-6879(99)99017-1
Sultan, M.H., Bedair, R., Ragab, O.G., Abd-ELShafy, E., Mahfouz, A.Y., & Daigham, G.E. (2024). Biological activities and ecological aspects of Limonium pruinosum (L.) collected from Wadi Hof Eastern Desert, Egypt, as a promising attempt for potential medical applications. Biomass Conversion and Biorefinery, 14, 23887–23907. https://doi.org/10.1007/s13399-023-04385-5
Sureda, A., Tejada, S., Khan, U.M., & Selamoglu, Z. (2023). An overview of the biological function of curcumin in the processes of oxidative stress, inflammation, nervous system, and lipid levels. Central Asian Journal of Medical and Pharmaceutical Sciences Innovation, 3(1), 1–11.
Tang, X.-H., Yan, L.-F., Gao, J., Yang, X.-L., Xu, Y.-X., Ge, H.-Y., & Yang, H.-D. (2012). Antitumor and immunomodulatory activity of polysaccharides from the root of Limonium sinense Kuntze. International Journal of Biological Macromolecules, 51(5), 1134 1139. https://doi.org/10.1016/j.ijbiomac.2012.08.037
Urmonov, D., & Ibragimov, A. (2025). Phytochemical and elemental profiling of Limonium otolepis growing in the Fergana Region, Uzbekistan: HPLC and ICP-OES analysis. Khimiya Rastitel’nogo Syr’ja, (2), 184-189. https://doi.org/10.14258/jcprm.20250215697
Van Loo, P., De Bruyn, A., & Buděšínský, M. (1986). Reinvestigation of the structural assignment of signals in the 1H and 13C NMR spectra of the flavone apigenin. Magnetic Resonance in Chemistry, 24, 879–882. https://doi.org/10.1002/mrc.1260241007
Whiting, P., Savchenko, T., Sarker, S.D., Rees, H.H., & Dinan, L. (1998). Phytoecdysteroids in the genus Limonium (Plumbaginaceae). Biochemical Systematics and Ecology, 26, 695–698.
Zhusupova, G.E., Artamonova, N.A., Abilov, Z.A., & Orazbaeva, Zh.K. (2006). Lipophilic pigments and fatty acids from the aerial parts of certain plant species of the genus Limonium. Chemistry of Natural Compounds, 42, 512–514. https://doi.org/10.1007/s10600-006-0200-9

Details

Primary Language

English

Subjects

Natural Products and Bioactive Compounds

Journal Section

Research Article

Authors

Dadahon Urmonov ^*
0000-0001-9693-3646
Uzbekistan

Ikbol Zhalolov This is me
0009-0003-3530-5699
Uzbekistan

Uchkun Ishimov This is me
0009-0001-9021-6782
Uzbekistan

Dilmurodjon Turdaliev This is me
0009-0008-0334-8279
Uzbekistan

Shakhnoz Azimova
0000-0003-1803-7728
Uzbekistan

Alidjan Ibragimov This is me
0000-0002-6204-1145
Uzbekistan

Early Pub Date

September 17, 2026

Publication Date

-

Submission Date

October 31, 2025

Acceptance Date

July 8, 2026

Published in Issue

Year 2026 Number: Advanced Online Publication

IZ

https://izlik.org/JA75RR96HC

Cite

RIS / Bibtex

APA

Urmonov, D., Zhalolov, I., Ishimov, U., Turdaliev, D., Azimova, S., & Ibragimov, A. (2026). Total phenolic content, antimicrobial activity, and LC-MS analysis of Limonium otolepis with isolation of apigenin. International Journal of Secondary Metabolite, Advanced Online Publication. https://izlik.org/JA75RR96HC

AMA

1.Urmonov D, Zhalolov I, Ishimov U, Turdaliev D, Azimova S, Ibragimov A. Total phenolic content, antimicrobial activity, and LC-MS analysis of Limonium otolepis with isolation of apigenin. Int. J. Sec. Metabolite. 2026;(Advanced Online Publication). https://izlik.org/JA75RR96HC

Chicago

Urmonov, Dadahon, Ikbol Zhalolov, Uchkun Ishimov, Dilmurodjon Turdaliev, Shakhnoz Azimova, and Alidjan Ibragimov. 2026. “Total Phenolic Content, Antimicrobial Activity, and LC-MS Analysis of Limonium Otolepis With Isolation of Apigenin”. International Journal of Secondary Metabolite, no. Advanced Online Publication. https://izlik.org/JA75RR96HC.

EndNote

Urmonov D, Zhalolov I, Ishimov U, Turdaliev D, Azimova S, Ibragimov A (September 1, 2026) Total phenolic content, antimicrobial activity, and LC-MS analysis of Limonium otolepis with isolation of apigenin. International Journal of Secondary Metabolite Advanced Online Publication

IEEE

[1]D. Urmonov, I. Zhalolov, U. Ishimov, D. Turdaliev, S. Azimova, and A. Ibragimov, “Total phenolic content, antimicrobial activity, and LC-MS analysis of Limonium otolepis with isolation of apigenin”, Int. J. Sec. Metabolite, no. Advanced Online Publication, Sept. 2026, [Online]. Available: https://izlik.org/JA75RR96HC

ISNAD

Urmonov, Dadahon - Zhalolov, Ikbol - Ishimov, Uchkun - Turdaliev, Dilmurodjon - Azimova, Shakhnoz - Ibragimov, Alidjan. “Total Phenolic Content, Antimicrobial Activity, and LC-MS Analysis of Limonium Otolepis With Isolation of Apigenin”. International Journal of Secondary Metabolite. Advanced Online Publication (September 1, 2026). https://izlik.org/JA75RR96HC.

JAMA

1.Urmonov D, Zhalolov I, Ishimov U, Turdaliev D, Azimova S, Ibragimov A. Total phenolic content, antimicrobial activity, and LC-MS analysis of Limonium otolepis with isolation of apigenin. Int. J. Sec. Metabolite. 2026. Available at https://izlik.org/JA75RR96HC.

MLA

Urmonov, Dadahon, et al. “Total Phenolic Content, Antimicrobial Activity, and LC-MS Analysis of Limonium Otolepis With Isolation of Apigenin”. International Journal of Secondary Metabolite, no. Advanced Online Publication, Sept. 2026, https://izlik.org/JA75RR96HC.

Vancouver

1.Dadahon Urmonov, Ikbol Zhalolov, Uchkun Ishimov, Dilmurodjon Turdaliev, Shakhnoz Azimova, Alidjan Ibragimov. Total phenolic content, antimicrobial activity, and LC-MS analysis of Limonium otolepis with isolation of apigenin. Int. J. Sec. Metabolite [Internet]. 2026 Sep. 1;(Advanced Online Publication). Available from: https://izlik.org/JA75RR96HC