HPLC-MS kaemferol and its glycosides identification

The article presents the results of kaempferol and its glycosides identification in plant extracts using the method of high-performance chromatography-mass spectrometry (HPLC-MS). The study is focused on the selection and optimization of chromatographic conditions for efficient separation and identification of these flavonoids with high biological activity, as well as at searching for new types of plant raw materials for the pharmaceutical industry of the Republic of Belarus. The use of the mass-spectrometric and spectrophotometric detection ensured high sensitivity and selectivity in the identification of kaempferol and its glycosides. Astragalin in an amount of more than 2.20 mg/g of absolutely dry raw material is contained in the extracts of Asclepias incarnate L., Geranium pratense L. and Glycyrrhiza uralensis Fisch. ex DC. Three glycosides of kaempferol were also identified in Glycyrrhiza uralensis Fisch. and Glycyrrhiza glabra L. extracts, and two glycosides – in the extracts of Borago officinalis L. The conducted study demonstrates the efficiency of the HPLC-MS method for analyzing flavonoids in multicomponent plant matrices and can be used for the quality control of plant raw materials and in pharmacognosy.

1. Riaz A., Rasul A., Hussain G., Zahoor M. K., Jabeen F., Subhani Z., Younis T., Ali M., Sarfraz I., Selamoglu Z. Astragalin: a bioactive phytochemical with potential therapeutic activities. Advances in Pharmacological and Pharmaceutical Sciences, 2018, vol. 108, p. 105769. https://doi.org/10.1155/2018/9794625

2. Ren J., Lu Y., Qian Y., Chen B., Wu T., Ji G. Recent progress regarding kaempferol for the treatment of various diseases (Review). Experimental and Therapeutic Medicine, 2019, vol. 18, pp. 2759–2776. https://doi.org/10.3892/etm.2019.7886

3. Qattan M. Y., Khan M. I., Alharbi S. H., Verma A. K., Al-Saeed F. A., Abduallah A. M., Al Areefy A. A. Therapeutic importance of kaempferol in the treatment of cancer through the modulation of cell signalling pathways. Molecules, 2022, vol. 27, no. 24, p. 8864. https://doi.org/10.3390/molecules27248864

4. De Morais E. F., de Oliveira L. Q. R., Farias Morais H. G., Souto Medeiros M. R., Freitas R. A., Rodini C. O., Coletta R. D. The anticancer potential of kaempferol: a systematic review based on in vitro studies. Cancers (Basel), 2024, vol. 16, no. 3, p. 585. https://doi.org/10.3390/cancers16030585.

5. Chen J., Zhong K., Qin S., Jing Y., Liu S., Li D., Peng C. Astragalin: a food-origin flavonoid with therapeutic effect for multiple diseases. Frontiers in Pharmacology, 2023, vol. 14, р. 1265960. https://doi.org/10.3389/fphar.2023.1265960

6. Wang Z., Lv J., Li X., Lin Q. The flavonoid astragalin shows anti‐tumor activity and inhibits PI3K/AKT signaling in gastric cancer. Chemical Biology & Drug Design, 2021, vol. 98, no. 5, pp. 779–786. https://doi.org/10.1111/cbdd.13933

7. Periferakis A., Periferakis K., Badarau I. A., Petran E. M., Popa D. C., Caruntu A., Costache R. S., Scheau C., Caruntu C., Costache D. O. Kaempferol: antimicrobial properties, sources, clinical, and traditional applications. International Journal of Molecular Sciences, 2022, vol. 23, no. 23, p. 15054. https://doi.org/10.3390/ijms232315054

8. Hossen M. J., Uddin M. B., Ahmed S. S. U., Yu Z. L., Cho J. Y. Kaempferol: review on natural sources and bioavailability. Biosynthesis, Food Sources and Therapeutic Uses. Nova Science Publishers, New York, NY, USA, 2016, pp. 101–150.

9. Calderón-Montaño J. M., Burgos-Morón E., Pérez-Guerrero C., López-Lázaro M. A review on the dietary flavonoid kaempferol. Mini-Reviews in Medicinal Chemistry, 2011, vol. 11, pp. 298–344.

10. Han Y. M., Koh J., Kim J. H., Lee J., Im J. P., Kim J. S. Astragalin inhibits nuclear factor-κB signaling in human colonic epithelial cells and attenuates experimental colitis in mice. Gut and Liver, 2021, vol. 15, no. 1, pp. 100–108. https://doi.org/10.5009/gnl19268

11. Guo Y., Wang T., Fu F. F., El-Kassaby Y. A., Wang G. Temporospatial flavonoids metabolism variation in Ginkgo biloba leaves. Frontiers in Genetics, 2020, vol. 11, p. 589326. https://doi.org/10.3389/fgene.2020.589326

12. Mehdi A., Al-ani Widad M. K., Raoof A. Isolation of astragalin from Iraqi Chenopodium album. Asian Journal of Pharmaceutical and Clinical Research, 2018, vol. 11, no. 12, pp. 530–535.

13. Adamtsevich N. Yu., Feskova А. V., Boltovskiy V. S., Titok V. V., Leontiev V. N. Isolation of flavonoids from Helichrysum arenarium flowers. Vesnіk Vіtsebskaga dzyarzhaўnaga ўnіversіteta = Vesnik of Vitebsk State University, 2021, no. 4, pp. 23–30 (in Russian).

14. Predicted LC-MS/MS Spectrum – 20V, Negative (HMDB0301694). The human metabolome database (HMDB). Available at: https://hmdb.ca/spectra/ms_ms/177781 (accessed 04 april 2025).

15. Jia S. S., Ma C. M., Li Y. H., Hao J. H. [Glycosides of phenolic acid and flavonoids from the leaves of Glycyrrhiza uralensis Ficsh]. Yao xue xue bao = Acta pharmaceutica Sinica, 1992, vol. 27, no. 6, pp. 441–444 (in Chinese).

16. Predicted LC-MS/MS Spectrum – 20V, Negative (HMDB0302426). The human metabolome database (HMDB). Available at: https://hmdb.ca/spectra/ms_ms/170734 (accessed 04 april 2025).

17. Fedoseeva G. M., Mirovich V. M., Goryachkina E. G., Perelomova M. V. Phytochemical analysis of plant materials containing flavonoids. Irkutsk, Irkutsk State Medical University, 2009. 67 p. (in Russian).