INTRODUCTION: Helichrysum leucocephalum Boiss. (Asteraceae) is an endemic plant to Iran. No reports have studied the cytotoxicity of the plant. The current study aimed to evaluate the cytotoxicity of H. leucocephalum collected from Fars province (Iran) against MCF-7 and HDF cell lines using HPLC-based activity profiling and to annotate the active constituents by LC-ESIQTOF-MS/MS. METHODS: H. leucocephalum was collected from three locations in Fars province. The dried flowers and leaves were separately extracted by percolation using methanol. The crude extracts were fractionated by liquid-liquid partitioning with dichloromethane (DCM) and aqueous methanol. The cytotoxicity of the fractions was evaluated against MCF-7 and HDF cells by Alamarblue assay. HPLC-based activity profiling was used to track the active constituents. LC-MS dereplication strategy was used for the annotation of the compounds in the active time window. LC-MS data were preprocessed by MZmine 3.3.0 and submitted to multivariate analysis to compare the differences and similarities in the metabolites of the samples. RESULTS: The DCM fractions showed a dose-dependent cytotoxicity against the cancerous cells (IC50s, 9.8-105.1 μg/ml). In general, the metabolites of the flowers and their cytotoxicity were higher than the leaves. LCESIMS/MS analyses revealed that prenylated and geranylated α,β-unsaturated spiroketal phloroglucinols were among the active constituents. CONCLUSION: It can be concluded that H. leucocephalum is a rich source of phloroglucinol derivatives with cytotoxic activities. Further phytochemical analysis is needed to characterize the bioactive components.

Department of Analytical Chemistry Faculty of Science Charles University Czech Republic

Department of Pharmacognosy School of Pharmacy Mashhad University of Medical Sciences Mashhad Iran

Department of Pharmacology and Toxicology School of Pharmacy Mashhad University of Medical Sciences Mashhad Iran

Department of Traditional Pharmacy School of Pharmacy Mashhad University of Medical Sciences Mashhad Iran

Forest and Rangeland Research Department Khorasan Razavi Agricultural and Natural Resources Research and Education Center AREEO Mashhad Iran

Mass Spectrometry Laboratory Section of Chemistry Faculty of Science Charles University Prague Czech Republic

Medical Toxicology Research Center Pharmaceutical Technology Institute Mashhad University of Medical Sciences Mashhad Iran

Student Research Committee Mashhad University of Medical Sciences Mashhad Iran

Targeted Drug Delivery Research Center Pharmaceutical Technology Institute Mashhad University of Medical Sciences Mashhad Iran

Zobrazit více v PubMed

Süntar I., Akkol E.K., Keles H., Yesilada E., Sarker S.D. Exploration of the wound healing potential of Helichrysum graveolens (Bieb.) Sweet: isolation of apigenin as an active component. J. Ethnopharmacol. 2013;149(1):103–110. doi: 10.1016/j.jep.2013.06.006. PubMed DOI

Akaberi M., Sahebkar A., Azizi N., Emami S.A. Everlasting flowers: phytochemistry and pharmacology of the genus Helichrysum. Ind. Crops Prod. 2019;138 doi: 10.1016/j.indcrop.2019.111471. DOI

Ghahreman A. Iran University Press; Tehran: 1994. Iranian Choromophites (Botanical Systematic)

Mastelic J., Politeo O., Jerkovic I., Radosevic N. Composition and antimicrobial activity of Helichrysum italicum essential oil and its terpene and terpenoid fractions. Chem. Nat. Compd. 2005;41(1):35–40.

Czinner E., Hagymasi K., Blazovics A., Kery A., Szőke É., Lemberkovics E. The in vitro effect of Helichrysi flos on microsomal lipid peroxidation. J. Ethnopharmacol. 2001;77(1):31–35. doi: 10.1016/s0378-8741(01)00258-6. PubMed DOI

Mozaffarian V. No 59. Research Institute of Forests and Rangelands; Tehran: 2008. (Compositae: Anthemideae & Echinopeae—In Flora of Iran).

POWO . Faciltated by the Royal Botanic Gardens; Kew: 2022. Plants of the World Online. Published on the internet.

Jahromi M.A.F., Dehshahri S., Forouzandeh Samani S. Volatile Composition, Antimicrobial and free radical scavenging activities of essential oil and total extract of Helichrysum leucocephalum Boiss. Trends Pharmacol. Sci. 2017;3(3):193–200.

Motamedifar M., Nozari A., Azhdari Ghasrodashti E. In vitro investigation of hydro-alcoholic extract of Helichrysum leucocephalum on the inhibition of Streptococcus mutans growt. J. Dent. Biomater. 2017;4(4):484–488.

Akaberi M., Emami S.A., Vatani M., Tayarani-Najaran Z. Evaluation of antioxidant and anti-melanogenic activity of different extracts of aerial parts of N. sintenisii in murine melanoma B16F10 Cells, Iran. J. Pharm. Res. 2018;17(1):225. PubMed PMC

Akaberi M., Danton O., Tayarani-Najaran Z., Asili J., Iranshahi M., Emami S.A., Hamburger M. HPLC-based activity profiling for antiprotozoal compounds in the endemic Iranian medicinal plant Helichrysum oocephalum. J. Nat. Prod. 2019;82(4):958–969. doi: 10.1021/acs.jnatprod.8b01031. PubMed DOI

Pluskal T., Castillo S., Villar-Briones A., Orešič M. MZmine 2: modular framework for processing, visualizing, and analyzing mass spectrometry-based molecular profile data. BMC Bioinf. 2010;11(1):1–11. PubMed PMC

Bozicevic A., Dobrzyński M., Bie H., Gafner F., Garo E., Hamburger M. Automated comparative metabolite profiling of large LC-ESIMS data sets in an ACD/MS workbook suite add-in, and data clustering on a new open-source web platform FreeClust. Anal. Chem. 2017;89:12682–12689. doi: 10.1021/acs.analchem.7b02221. PubMed DOI

Akaberi M., Najaran Z.T., Azizi N., Emami S.A. Metabolite profiling and antiprotozoal activity of three endemic Iranian Helichrysum species. Ind. Crops Prod. 2021;174 doi: 10.1016/j.indcrop.2021.114196. DOI

Sagbo I.J., Otang-Mbeng W. Anti-proliferative and genotoxic activities of the Helichrysum petiolare Hilliard & BL Burtt. Sci. Pharm. 2020;88(4):49. doi: 10.3390/scipharm88040049. DOI

Lourens A., Van Vuuren S., Viljoen A., Davids H., Van Heerden F. Antimicrobial activity and in vitro cytotoxicity of selected South African Helichrysum species. South Afr. J. Bot. 2011;77(1):229–235. doi: 10.1016/j.sajb.2010.05.006. DOI

Akinfenwa A.O., Sagbo I.J., Makhaba M., Mabusela W.T., Hussein A.A. Helichrysum genus and compound activities in the management of diabetes mellitus. Plants. 2022;11(10):1386. doi: 10.3390/plants11101386. PubMed DOI PMC

Matić I.Z., Aljancić I., Vajs V., Jadranin M., Gligorijević N., Milosavljević S., Juranić Z.D. Cancer-suppressive potential of extracts of endemic plant Helichrysum zivojinii: effects on cell migration, invasion and angiogenesis. Nat. Prod. Commun. 2013;8(9):1291–1296. PubMed

Bigović D., Savikin K., Janković T., Menković N., Zdunić G., Stanojković T., Djurić Z. Antiradical and cytotoxic activity of different Helichrysum plicatum flower extracts. Nat. Prod. Commun. 2011;6(6):819–822. PubMed

Lawal O.A., Ogunwande I.A., Kasali A.A., Opoku A.R., Oyedeji A.O. Chemical composition, antibacterial and cytotoxic activities of essential oil from the leaves of Helichrysum odoratissimum grown in South Africa. J. Essent. Oil-Bear. Plants. 2015;18(1):236–241. doi: 10.1080/0972060X.2014.901618. DOI

Afoulous S., Ferhout H., Raoelison E.G., Valentin A., Moukarzel B., Couderc F., Bouajila J. Helichrysum gymnocephalum essential oil: chemical composition and cytotoxic, antimalarial and antioxidant activities, attribution of the activity origin by correlations. Molecules. 2011;16(10):8273–8291. doi: 10.3390/molecules16108273. PubMed DOI PMC

Potterat O., Hamburger M. Combined use of extract libraries and HPLC-based activity profiling for lead discovery: potential, challenges, and practical considerations. Planta Med. 2014;80(14):1171–1181. doi: 10.1055/s-0034-1382900. PubMed DOI

Potterat O., Hamburger M. Concepts and technologies for tracking bioactive compounds in natural product extracts: generation of libraries, and hyphenation of analytical processes with bioassays. Nat. Prod. Rep. 2013;30(4):546–564. doi: 10.1039/c3np20094a. PubMed DOI

Peron G., Pant D.R., Shrestha S.S., Rajbhandary S., Dall'Acqua S. An integrated LC-ESI-MS(n) and high resolution LC-ESI-QTOF approach for the identification of phloroglucinols from Nepalese Hypericum japonicum. Molecules. 2020;25(24) doi: 10.3390/molecules25245937. PubMed DOI PMC

Wang C.-J., Jiang Y.-Q., Liu D.-H., Yan X.-H., Ma S.-C. Characterization of phloroglucinol derivatives and diterpenes in Euphorbia ebracteolata Hayata by utilizing ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry. J. Pharm. Anal. 2013;3(4):292–297. doi: 10.1016/j.jpha.2013.01.002. PubMed DOI PMC

Sleno L., Daneshfar R., Eckert G.P., Müller W.E., Volmer D.A. Mass spectral characterization of phloroglucinol derivatives hyperforin and adhyperforin. Rapid Commun. Mass Spectrom. 2006;20(18):2641–2648. doi: 10.1002/rcm.2650. PubMed DOI