Towards the chemical standardisation of Epilobium hirsutum leaves

Authors

DOI:

https://doi.org/10.15587/2519-4852.2026.349856

Keywords:

Epilobium hirsutum, стандартизація, фенольні сполуки, енотеїн B, флавоноїди, ВЕТШХ, ВЕРХ-ДАД

Abstract

Epilobium hirsutum L. is widely used in traditional European medicine to treat urological and inflammatory conditions. Despite its widespread use, no pharmacopoeial monographs on its plant material have been published to date. The aim of this study was to establish physicochemical parameters and identify marker phenolic compounds for the standardisation of plant leaves.

Method. Seven E. hirsutum leaf samples collected in Ukraine, Lithuania, and Poland for analysis. The following parameters were analysed: loss on drying, total ash, and acid-insoluble ash content according to the State Pharmacopoeia of Ukraine. The qualitative composition was analysed using high-performance thin-layer chromatography (HPTLC), and the quantitative analysis of total phenolic compounds, flavonoids, and specific marker compounds was performed using UV spectrophotometry and HPLC-DAD.

Results. As a result, it was established that the content of foreign impurities did not exceed 2% in the analysed samples, the drying loss was 6.4–8.1%, and the total ash was 4.0–5.7%. The HPTLC method was used to determine gallic acid, isoquercitrin, avicularin, guijaverin and hyperoside as key compounds for the plant quality control. Total phenolics ranged between 0.78 and 1.52 mg GAE/g dw, while total flavonoid content was 1.9–5.5 mg HE/g dw. The HPLC method showed that the dominant polyphenolics is oenothein B (39.9–65.7 mg/g), followed by oenothein A, gallic, chlorogenic, and ellagic acids, hyperoside, isoquercitrin, and quercetin. The present components can be proposed for the development of raw material standardisation parameters.

Conclusion. The obtained data confirm that E. hirsutum leaves meet requirements of the State Pharmacopeia of Ukraine and European Pharmacopoeia and can serve as the basis for developing a monograph. Oenothein B, hyperoside, and gallic acid can be proposed as identification markers; in addition, the total phenolic content can be assessed

Author Biographies

Kateryna Uminska, Zhytomyr Basic Pharmaceutical Professional College

PhD

Liudas Ivanauskas, Lithuanian University of Health Sciences

Doctor of Biomedical Sciences, Professor, Head of Department

Department of Analytical and Toxicological Chemistry

Laurynas Jarukas, Lithuanian University of Health Sciences

PhD

Department of Analytical and Toxicological Chemistry

Victoriya Georgiyants, National University of Pharmacy

Doctor of Pharmaceutical Sciences, Professor, Head of Department

Department of Pharmaceutical Chemistry

Olha Mykhailenko, National University of Pharmacy; Kiel University; UCL School of Pharmacy

Doctor of Pharmaceutical Sciences, Associate Professor

Department of Pharmaceutical Chemistry

Humboldt Fellow

Visiting Researcher

References

  1. Indrayanto, G. (2024). Regulation and standardization of herbal drugs: Current status, limitation, challenge’s and future prospective. Profiles of Drug Substances, Excipients and Related Methodology, 153–199. https://doi.org/10.1016/bs.podrm.2023.11.003
  2. Aware, C. B., Patil, D. N., Suryawanshi, S. S., Mali, P. R., Rane, M. R., Gurav, R. G. et al. (2022). Natural bioactive products as promising therapeutics: A review of natural product-based drug development. South African Journal of Botany, 151, 512–528. https://doi.org/10.1016/j.sajb.2022.05.028
  3. Herbs of commerce (1992). American Herbal Products Association. Available at: https://law.resource.org/pub/us/cfr/ibr/001/ahpa.herbs.1992.pdf
  4. Guideline on quality of herbal medicinal products2/traditional herbal medicinal products. (2022). European Medicinal Agency. Available at: https://www.ema.europa.eu/en/quality-herbal-medicinal-products-traditional-herbal-medicinal-products-scientific-guideline
  5. Dmytrenko, O., Golembiovska, O., Pashuk, V., Zgonnyk, S. (2025). Problems of classification, safety assessment and risk management of medical devices with biologically active substances. ScienceRise: Pharmaceutical Science, 2 (54), 86–104. https://doi.org/10.15587/2519-4852.2025.328031
  6. Ak, G., Zengin, G., Mahomoodally, M. F., Llorent-Martínez, E., Orlando, G., Chiavaroli, A. et al. (2021). Shedding Light into the Connection between Chemical Components and Biological Effects of Extracts from Epilobium hirsutum: Is It a Potent Source of Bioactive Agents from Natural Treasure? Antioxidants, 10 (9), 1389. https://doi.org/10.3390/antiox10091389
  7. Stolarczyk, M., Naruszewicz, M., Kiss, A. K. (2013). Extracts from Epilobium sp. herbs induce apoptosis in human hormone-dependent prostate cancer cells by activating the mitochondrial pathway. Journal of Pharmacy and Pharmacology, 65 (7), 1044–1054. https://doi.org/10.1111/jphp.12063
  8. Assessment report on Epilobium angustifolium L. and/or Epilobium parviflorum Schreb., herba (2016). European Medicines Agency, 20. Available at: https://www.scribd.com/document/524645489/epilobium-parviflorum-schreb-herba-en-1?language_settings_changed=English
  9. Epilobii herba (willow herb) (2024). ESCOP, 1–16. Available at: https://www.escop.com/downloads/epilobii-herba-willow-herb/
  10. Barakat, H. H., Hussein, S. A. M., Marzouk, M. S., Merfort, I., Linscheid, M., Nawwar, M. A. M. (1997). Polyphenolic metabolites of Epilobium hirsutum. Phytochemistry, 46 (5), 935–941. https://doi.org/10.1016/s0031-9422(97)00370-1
  11. Karakurt, S., Semiz, A., Celik, G., Gencler-Ozkan, A. M., Sen, A., Adali, O. (2016). Contribution of ellagic acid on the antioxidant potential of medicinal plant Epilobium hirsutum. Nutrition and Cancer, 68 (1), 173–183. https://doi.org/10.1080/01635581.2016.1115092
  12. Kiss, A. K., Bazylko, A., Filipek, A., Granica, S., Jaszewska, E., Kiarszys, U. et al. (2011). Oenothein B’s contribution to the anti-inflammatory and antioxidant activity of Epilobium sp. Phytomedicine, 18 (7), 557–560. https://doi.org/10.1016/j.phymed.2010.10.016
  13. Mohammadi Bazargani, M., Falahati-Anbaran, M., Rohloff, J. (2021). Comparative Analyses of Phytochemical Variation Within and Between Congeneric Species of Willow Herb, Epilobium hirsutum and E. parviflorum: Contribution of Environmental Factors. Frontiers in Plant Science, 11. https://doi.org/10.3389/fpls.2020.595190
  14. Sheikh, N. A., Desai, T. R., Patel, R. D. (2016). Pharmacognostic Evaluation of Epilobium hirsutum Linn. Pharmacognosy Journal, 8 (3), 226–229. https://doi.org/10.5530/pj.2016.3.8
  15. Uminska, K., Korinek, M., Ivanauskas, L., El‐Shazly, M., Georgiyants, V., Chen, Y.et al. (2025). Assessment of the Phenolics Content in Epilobium angustifolium and Epilobium hirsutum Extracts and Their Pharmacological Activity. Archiv Der Pharmazie, 358 (5). https://doi.org/10.1002/ardp.202400765
  16. Vlase, A.-M., Toiu, A., Tomuță, I., Vlase, L., Muntean, D., Casian, T. et al. (2022). Epilobium Species: From Optimization of the Extraction Process to Evaluation of Biological Properties. Antioxidants, 12 (1), 91. https://doi.org/10.3390/antiox12010091
  17. Vlase, A.-M., Toiu, A., Gligor, O., Muntean, D., Casian, T., Vlase, L. et al. (2024). Investigation of Epilobium hirsutum L. Optimized Extract’s Anti-Inflammatory and Antitumor Potential. Plants, 13 (2), 198. https://doi.org/10.3390/plants13020198
  18. Mykhailenko, O., Jalil, B., Uminska, K., Ivanauskas, L., Gudžinskas, Z., Heinrich, M. (2025). The phenology of Epilobium hirsutum L.: assessing marker compounds variability of a pharmaceutically important plant remedy. Frontiers in Pharmacology, 16. https://doi.org/10.3389/fphar.2025.1602819
  19. Derzhavna farmakopeia Ukrainy (2015). Kharkiv: Derzhavne pidpryiemstvo «Ukrainskyi naukovyi farmakopeinyi tsentr yakosti likarskykh zasobiv», 1128. Available at: https://sphu.org/wp-content/uploads/2024/09/content_1t_2-izd.pdf
  20. Vilkickyte, G., Petrikaite, V., Marksa, M., Ivanauskas, L., Jakstas, V., Raudone, L. (2023). Fractionation and Characterization of Triterpenoids from Vaccinium vitis-idaea L. Cuticular Waxes and Their Potential as Anticancer Agents. Antioxidants, 12 (2), 465. https://doi.org/10.3390/antiox12020465
  21. Hawthorn leaf and flower (Crataegi folium cum flore) (2008). European Pharmacopoeia. Available at: http://www.uspbpep.com/ep60/hawthorn%20leaf%20and%20flower%201432e.pdf
  22. Ivanauskas, L., Uminska, K., Gudžinskas, Z., Heinrich, M., Georgiyants, V., Kozurak, A. et al. (2024). Phenological Variations in the Content of Polyphenols and Triterpenoids in Epilobium angustifolium Herb Originating from Ukraine. Plants, 13 (1), 120. https://doi.org/10.3390/plants13010120
  23. Wagner, H., Bladt, S. (Eds.) (1996). Plant Drug Analysis. Flavonoid drugs including Ginkgo biloba and Echinaceae species. Berlin: Springer, 195–245. https://doi.org/10.1007/978-3-642-00574-9
  24. Harborne, J. B. (1998). Phytochemical methods. A guide to modern technique of plant analysis. London: Chapman and Hill, 285. Available at: https://link.springer.com/book/9780412572609
  25. Bobkova, І., Burianova, V., Honcharuk, S., Dunaievska, O., Uminska, K., Khranivska, V. (2022). Anatomical investigation of sheets Chamenerion angustifolium (L.) SCOP. Fitoterapia, 1, 77–81. https://doi.org/10.33617/2522-9680-2022-1-77
  26. Granica, S., Piwowarski, J. P., Czerwińska, M. E., Kiss, A. K. (2014). Phytochemistry, pharmacology and traditional uses of different Epilobium species (Onagraceae): A review. Journal of Ethnopharmacology, 156, 316–346. https://doi.org/10.1016/j.jep.2014.08.036
  27. Remmel, I., Vares, L., Toom, L., Matto, V., Raal, A. (2012). Phenolic Compounds in Five Epilobium Species Collected from Estonia. Natural Product Communications, 7 (10). https://doi.org/10.1177/1934578x1200701017
  28. Jürgenson, S., Matto, V., Raal, A. (2012). Vegetational variation of phenolic compounds inEpilobium angustifolium. Natural Product Research, 26 (20), 1951–1953. https://doi.org/10.1080/14786419.2011.643310
  29. Everette, J. D., Bryant, Q. M., Green, A. M., Abbey, Y. A., Wangila, G. W., Walker, R. B. (2010). Thorough Study of Reactivity of Various Compound Classes toward the Folin−Ciocalteu Reagent. Journal of Agricultural and Food Chemistry, 58 (14), 8139–8144. https://doi.org/10.1021/jf1005935
Towards the chemical standardisation of Epilobium hirsutum leaves

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Published

2026-01-19

How to Cite

Uminska, K., Ivanauskas, L., Jarukas, L., Georgiyants, V., & Mykhailenko, O. (2026). Towards the chemical standardisation of Epilobium hirsutum leaves. ScienceRise: Pharmaceutical Science. https://doi.org/10.15587/2519-4852.2026.349856

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2026: Article in press

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Pharmaceutical Science

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Copyright (c) 2026 Kateryna Uminska, Liudas Ivanauskas, Laurynas Jarukas, Victoriya Georgiyants, Olha Mykhailenko

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