Investigation of phytochemical profile, cytotoxic and antioxidant activities of Michauxia campanuloides L’Hérit. Ex Aiton

Abstract

Michauxia campanuloides, a traditionally used plant in Southern Anatolia, was investigated for its bioactivity and phytochemical profiles. Methanolic extracts were first prepared from the roots and aerial parts of M. campanuloides, then each extract was suspended in water and partitioned with n-hexane, EtOAc and n-butanol, respectively. Methanolic extracts, subextracts and the remaining water extracts were evaluated for their in vitro cytotoxicity against colon (HCT116, SW480), breast (MDA-MB-231, HCC1937), and liver (HEP3B, HEPG2) cancer cell lines by MTS assay. The EtOAc subextract of roots showed the highest cytotoxic activity against all tested cancer cell lines (IC50 = 10–154.76 μg/mL). Antioxidant activities were assessed using ABTS, DPPH, FRAP, and CUPRAC assays. The EtOAc subextracts derived from both aerial parts and roots exhibited the highest antioxidant activity (353.40 ± 19.21 and 367.65 ± 16.38 mg TE/g in ABTS, respectively). HPTLC was used for phytochemical profiling of the extracts, while UPLC was used for the quantification of the investigated phenolic compounds, including rutin, chlorogenic acid, neochlorogenic acid, isoquercitrin, astragalin, caffeic acid, and quercetin. Astragalin (93.37 ± 0.11 mg/g) and isoquercitrin (51.22 ± 0.81 mg/g) were the main compounds of the EtOAc subextract. This is the first report on M. campanuloides evaluating its cytotoxic activity and quantification of the marker compounds.

Keywords

• Michauxia campanuloides
• cytotoxicity
• antioxidant
• HPTLC
• UPLC

References

1. [1] Al-Zein MS, Musselman LJ. Michauxia (Campanulaceae): A Western Asian genus honoring a North American pioneer botanist. Castanea. 2004;69:200-205.
2. [2] Ertekin AS, Aksoy A, Duran A, Yaprak A, Karavelioğulları AF, Kahraman A, A Checklist of the Flora of Turkey (Vascular Plants), first ed., ANG Vakfı/Nezahat Gökyiğit Botanik Bahçesi, Istanbul 2012.
3. [3] Özdemir E, Alpinar K. The wild edible plants of Western Nigde Aladaglar Mountains (Central Turkey). Istanbul J Pharm. 2013;41:66-74.
4. [4] Demirci Kayıran S, Ozhatay N. Wild plants used for medicinal purpose in Andırın. Turk J Pharm Sci. 2012;9:75-92.
5. [5] Mart S. Bahçe ve Hasanbeyli (Osmaniye) halkının kullandığı doğal bitkilerin etnobotanik yönden araştırılması. Adana: Çukurova University; 2006.
6. [6] Ebrahimabadi AH, Movahedpour MM, Batooli H, Ebrahimabadi EH, Mazoochi A, Mobarak Qamsari M. Volatile compounds analysis and antioxidant, antimicrobial and cytotoxic activities of Mindium laevigatum. Iran J Basic Med Sci. 2016;19:1337-1344. https://doi.org/10.22038/ijbms.2016.7921
7. [7] Güvenç A, Küpeli Akkol E, Hürkul MM, Süntar I, Keleş H. Wound healing and anti-inflammatory activities of the Michauxia L’Hérit (Campanulaceae) species native to Turkey. J Ethnopharmacol. 2012;139:401-408. http://doi.org/10.1016/j.jep.2011.11.024
8. [8] Koca RH, Hurkul MM, Kurt S, Koroglu A. The effect of Michauxia campanuloides L’Her. on some semen parameters in Wistar albino rats. Atatürk Univ J Vet Sci. 2020;15:138-144. https://doi.org/10.17094/ataunivbd.700491

9. [9] Hürkul MM. Türkiye’de Yetişen Michauxia L’Herit. (Campanulaceae) türleri üzerinde Farmasötik Botanik yönünden araştırmalar. Ankara: Ankara University; 2011.
10. [10] Kurt S, Koca RH, Hürkul MM, Seker U, Köroğlu A. The antioxidant effect of Michauxia campanuloides on rat ovaries. J Hellenic Vet Med Soc. 2021;72:3163-3170. https://doi.org/10.12681/jhvms.28510
11. [11] Masoum S, Ghasemi-Estarki H, Seifi H, Ebrahimabadi EH, Parastar H. Analysis of the volatile chemical constituents in Mindium laevigatum by gas chromatography-mass spectrometry and correlative chemometric resolution methods. Microchem J. 2013;106:276-281. https://doi.org/10.1016/j.microc.2012.08.004
12. [12] Gulcin I. Antioxidants and antioxidant methods: An updated overview. Arch Toxicol. 2006;94:651-715. https://doi.org/10.1007/s00204-020-02689-3
13. [13] Khazir J, Mir BA, Pilcher L, Riley DL. Role of plants in anticancer drug discovery. Phytochem Lett. 2014;7:173-181. https://doi.org/10.1016/j.phytol.2013.11.010
14. [14] Folin O, Ciocalteu V. On tyrosine and tryptophane determinations in proteins. J Biol Chem. 1927;73:627-648.
15. [15] Wolsky RG, Salatino A. Analysis of propolis: Some parameters and procedures for chemical quality control. J Apic Res. 1998;37:99-105.
16. [16] Guzelmeric E, Reis R, Sen NB, Celik C, Özhan Y, Petrikaite V, Sipahi H, Ayfın A, Yeşilada E. Insights into the anti-inflammatory, analgesic, and anticancer potentials of the standardized extracts from three Cistus L. species. J Herb Med. 2023;41:100724. https://doi.org/10.1016/j.hermed.2023.100724
17. [17] Öztürk C, Şen NB, Güzelmeriç E, Kirmizibekmez H. Secondary metabolites from Scabiosa atropurpurea and their antioxidant and xanthine oxidase inhibitory activities. J Res Pharm. 2024;28:79-88. http://dx.doi.org/10.29228/jrp.676
18. [18] Cory AH, Owen TC, Barltrop JA, Cory JG. Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture. Cancer Commun. 1991;3:207-212. https://doi.org/10.3727/095535491820873191
19. [19] Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med. 1999;26:1231-1237. https://doi.org/10.1016/s0891-5849(98)00315-3
20. [20] Blois MS. Antioxidant determinations by the use of a stable free radical. Nature. 1958;181:1199-1200. https://doi.org/10.1038/1811199a0
21. [21] Benzie IFF, Strain JJ. The Ferric Reducing Ability of Plasma (FRAP) as a measure of "Antioxidant Power": The FRAP Assay. Anal Biochem. 1996;239:70-76. https://doi.org/10.1006/abio.1996.0292
22. [22] Apak R, Güçlü K, Özyürek M, Karademir SE. Novel total antioxidant capacity index for dietary polyphenols and vitamins C and E, using their cupric ion reducing capability in the presence of neocuproine: CUPRAC method. J Agric Food Chem. 2004;52:7970-7981. https://doi.org/10.1021/jf048741x
23. [23] Güzelmeriç E, Çelik C, Şen NB, Oçkun MA, Yeşilada E. Quali/quantitative research on herbal supplements containing black elder (Sambucus nigra L.) fruits. J Res Pharm. 2021;25:238-248. http://dx.doi.org/10.29228/jrp.14
24. [24] Reich E, Schibli A, High-Performance Thin-Layer Chromatography for the Analysis of Medicinal Plants, Georg Thieme Verlag KG, Stuttgart 2007. https://doi.org/10.1055/b-0034-65188
25. [25] ICH Guideline. Validation of analytical procedures: text and methodology Q2 (R1). Geneva, Switzerland: International Conference on Harmonization; 2005.