JOURNAL OF CHILEAN CHEMICAL SOCIETY

Vol 71 No 2 (2026): Journal of The Chilean Chemical Society - 2nd Special Issue in Tribute to Professor Mario Silva (In Memoriam)
Original Research Papers

CYCLOOXYGENASE-2 INHIBITORS AND ANTIOXIDANT COMPOUNDS IN BEE POLLEN IDENTIFIED USING HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY–EFFECT-DIRECTED ANALYSIS–MASS SPECTROMETRY

PDF
  • Siluana Katia Tischer Seraglio,
  • Esteban Narváez,
  • Luciano Valdemiro Gonzaga,
  • Mario Aranda,
  • Karem Henríquez-Aedo,
  • Ana Carolina Oliveira Costa
Siluana Katia Tischer Seraglio
Universidad del Bío-Bío
Published September 22, 2026
Keywords
  • Anti-inflammatory activity,
  • antioxidant activity,
  • bioactive compounds,
  • Brazilian bee pollen,
  • COX-2,
  • flavonoids
    • ...More
    Less
How to Cite
Seraglio, S. K. T., Narváez, E., Gonzaga, L. V., Aranda, M., Henríquez-Aedo, K., & Costa, A. C. O. (2026). CYCLOOXYGENASE-2 INHIBITORS AND ANTIOXIDANT COMPOUNDS IN BEE POLLEN IDENTIFIED USING HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY–EFFECT-DIRECTED ANALYSIS–MASS SPECTROMETRY. Journal of the Chilean Chemical Society, 71(2), 6558-6564. Retrieved from https://jcchems.com/index.php/JCCHEMS/article/view/3036

Copyright (c) 2026 SChQ

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

Abstract

Bee pollen has recognized nutritional and functional properties. This study analyzed Brazilian bee pollens for antioxidant compounds and cyclooxygenase-2 (COX-2) inhibitors using high-performance thin-layer chromatography–effect-directed analysis (HPTLC–EDA) and tentative identification via mass spectrometry (MS). Distinct HPTLC profiles were observed for each sample, reflecting compositional differences. Antioxidant activity was demonstrated by DPPH radical scavenging in multiple zones, and two COX-2 inhibitory zones were detected in one sample by HPTLC–EDA. These bioactivities were linked to phenolic compounds and MS/MS analysis allowed the tentative identification of seven flavonoids (isorhamnetin-3-O-rutinoside, pinobanksin-5-methyl-ether-3-O-acetate, quercetin-3-methyl-ether, quercetin-3-O-glucoside, kaempferol-3-O-rutinoside, isorhamnetin-O-acetyl-rutinoside, and quercetin-dimethyl-ether). Notably, quercetin-3-methyl-ether exhibited both antioxidant and COX-2 inhibitory activity, which is particularly relevant due to the constant search for COX inhibitors with no harmful cardiovascular effects. Therefore, bee pollen is a relevant matrix containing compounds with antioxidant and anti-inflammatory potential. This is the first report of isorhamnetin-O-acetyl-rutinoside and quercetin-dimethyl-ether in bee pollen.

 

3036.png

PDF

References

  1. Qiao J, Zhang Y, Haubruge E, Wang K, El-Seedi HR, Dong J, et al. New insights into bee pollen: Nutrients, phytochemicals, functions and wall-disruption. Food Research International 2024;178:113934. https://doi.org/10.1016/j.foodres.2024.113934.
  2. Chantarudee A, Phuwapraisirisan P, Kimura K, Okuyama M, Mori H, Kimura A, et al. Chemical constituents and free radical scavenging activity of corn pollen collected from Apis mellifera hives compared to floral corn pollen at Nan, Thailand. BMC Complementary Medicine and Therapies 2012;12:45. https://doi.org/10.1186/1472-6882-12-45.
  3. Pascoal A, Rodrigues S, Teixeira A, Feás X, Estevinho LM. Biological activities of commercial bee pollens: Antimicrobial, antimutagenic, antioxidant and anti-inflammatory. Food and Chemical Toxicology 2014;63:233–9. https://doi.org/10.1016/j.fct.2013.11.010.
  4. Saliba ASMC, Sartori AG de O, Rosalen PL, Lazarini JG, do Amaral JEPG, da Luz CFP, et al. Bee pollen from bracatinga (Mimosa scabrella): Effects of gastrointestinal digestion and epithelial transport in vitro on phenolic profile and bioactivities. Food Research International 2024;196:115142. https://doi.org/10.1016/j.foodres.2024.115142.
  5. Zhang H, Zhu X, Huang Q, Zhang L, Liu X, Liu R, et al. Antioxidant and anti-inflammatory activities of rape bee pollen after fermentation and their correlation with chemical components by ultra-performance liquid chromatography-quadrupole time of flight mass spectrometry-based untargeted metabolomics. Food Chemistry 2023;409:135342. https://doi.org/10.1016/j.foodchem.2022.135342.
  6. Sen NB, Guzelmeric E, Vovk I, Glavnik V, Kırmızıbekmez H, Yesilada E. Phytochemical and bioactivity studies on Hedera helix L. (ivy) flower pollen and ivy bee pollen. Antioxidants 2023;12:1394. https://doi.org/10.3390/antiox12071394.
  7. Kostić AŽ, Milinčić DD, Gašić UM, Nedić N, Stanojević SP, Tešić ŽLj, et al. Polyphenolic profile and antioxidant properties of bee-collected pollen from sunflower (Helianthus annuus L.) plant. LWT 2019;112:108244. https://doi.org/10.1016/j.lwt.2019.06.011.
  8. Qiao J, Feng Z, Zhang Y, Xiao X, Dong J, Haubruge E, et al. Phenolamide and flavonoid glycoside profiles of 20 types of monofloral bee pollen. Food Chemistry 2023;405:134800. https://doi.org/10.1016/j.foodchem.2022.134800.
  9. El Ghouizi A, El Menyiy N, Falcão SI, Vilas-Boas M, Lyoussi B. Chemical composition, antioxidant activity, and diuretic effect of Moroccan fresh bee pollen in rats. Veterinary World 2020;13:1251–61. https://doi.org/10.14202/vetworld.2020.1251-1261.
  10. De-Melo AAM, Estevinho LM, Moreira MM, Delerue-Matos C, Freitas A da S de, Barth OM, et al. Phenolic profile by HPLC-MS, biological potential, and nutritional value of a promising food: Monofloral bee pollen. Journal of Food Biochemistry 2018;42:e12536. https://doi.org/10.1111/jfbc.12536.
  11. Zeb A. Concept, mechanism, and applications of phenolic antioxidants in foods. Journal of Food Biochemistry 2020;44:e13394. https://doi.org/10.1111/jfbc.13394.
  12. Lang Y, Gao N, Zang Z, Meng X, Lin Y, Yang S, et al. Classification and antioxidant assays of polyphenols: A review. Journal of Future Foods 2024;4:193–204. https://doi.org/10.1016/j.jfutfo.2023.07.002.
  13. Pisoschi AM, Iordache F, Stanca L, Cimpeanu C, Furnaris F, Geicu OI, et al. Comprehensive and critical view on the anti-inflammatory and immunomodulatory role of natural phenolic antioxidants. European Journal of Medicinal Chemistry 2024;265:116075. https://doi.org/10.1016/j.ejmech.2023.116075.
  14. Liu W, Cui X, Zhong Y, Ma R, Liu B, Xia Y. Phenolic metabolites as therapeutic in inflammation and neoplasms: Molecular pathways explaining their efficacy. Pharmacological Research 2023;193:106812. https://doi.org/10.1016/j.phrs.2023.106812.
  15. Desai SJ, Prickril B, Rasooly A. Mechanisms of phytonutrient modulation of cyclooxygenase-2 (COX-2) and inflammation related to cancer. Nutrition and Cancer 2018;70:350–75. https://doi.org/10.1080/01635581.2018.1446091.
  16. Reguigui A, Baglyas M, Cselőtey A, Romdhane M, Móricz ÁM. High-performance thin-layer chromatography combined with effect-directed assays revealed bioactivity profiles of Salvia aegyptiaca, S. verbenaca, and S. officinalis. Journal of Chromatography B 2025;1261:124653. https://doi.org/10.1016/j.jchromb.2025.124653.
  17. Galarce-Bustos O, Pavón-Pérez J, Henríquez-Aedo K, Aranda M. An improved method for a fast screening of α-glucosidase inhibitors in cherimoya fruit (Annona cherimola Mill.) applying effect-directed analysis via high-performance thin-layer chromatography-bioassay-mass spectrometry. Journal of Chromatography A 2019;1608:460415. https://doi.org/10.1016/j.chroma.2019.460415.
  18. Shi X, Xi X, Tang X, Chen Y. Quantitation of common mycotoxins (aflatoxin B1 and deoxynivalenol) in cereals by high performance thin layer chromatography with smartphone image capture and ImageJ analysis. Journal of Future Foods 2026;6:48–54. https://doi.org/10.1016/j.jfutfo.2024.08.011.
  19. López K, Espinoza-Bello A, Carrasco J, Peña-Farfal C, Aranda M, Henríquez-Aedo K. Multivariate optimization of microwave-assisted enzyme digestion of α-casein for generation of bioactive peptides. Journal of the Chilean Chemical Society 2023;68:5963–8. https://doi.org/10.4067/s0717-97072023000305963.
  20. Zhang Y, Yu J, Lai S, Song J, Wu X, Wang D, et al. Rapid determination of histamine level in seafood using read-out strips based on high-performance thin layer chromatography modified with self-visualization nanomaterials. Food Control 2021;122:107816. https://doi.org/10.1016/j.foodcont.2020.107816.
  21. Morlock GE. High-performance thin-layer chromatography combined with effect-directed assays and high-resolution mass spectrometry as an emerging hyphenated technology: A tutorial review. Analytica Chimica Acta 2021;1180:338644. https://doi.org/10.1016/j.aca.2021.338644.
  22. Cabezudo I, Salazar MO, Ramallo IA, Furlan RLE. Effect-directed analysis in food by thin-layer chromatography assays. Food Chemistry 2022;390:132937. https://doi.org/10.1016/j.foodchem.2022.132937.
  23. Sharma B, Islam A, Sharma A. HPTLC-MS: An advance approach in herbal drugs using fingerprint spectra and mass spectroscopy. Traditional Medicine Research 2023;8:10. https://doi.org/10.53388/TMR20220428001.
  24. Sen NB, Vovk I, Kırmızıbekmez H, Guzelmeric E. Phytochemical and bioactivity evaluation of bee pollen and androecia of Castanea, Salix, and Quercus species. Antioxidants 2024;14:40. https://doi.org/10.3390/antiox14010040.
  25. Ju Z, Li M, Xu J, Howell DC, Li Z, Chen F-E. Recent development on COX-2 inhibitors as promising anti-inflammatory agents: The past 10 years. Acta Pharmaceutica Sinica B 2022;12:2790–807. https://doi.org/10.1016/j.apsb.2022.01.002.
  26. Carrasco-Sandoval J, Rebolledo P, Peterssen-Fonseca D, Fischer S, Wilckens R, Aranda M, et al. A fast and selective method to determine phenolic compounds in quinoa (Chenopodium quinoa Will) seeds applying ultrasound-assisted extraction and high-performance liquid chromatography. Chemical Papers 2021;75:431–8. https://doi.org/10.1007/s11696-020-01313-z.
  27. Alimoglu G, Guzelmeric E, Yuksel PI, Celik C, Deniz I, Yesilada E. Monofloral and polyfloral bee pollens: Comparative evaluation of their phenolics and bioactivity profiles. LWT 2021;142:110973. https://doi.org/10.1016/j.lwt.2021.110973.
  28. Ogbole OO, Ndabai NC, Akinleye TE, Attah AF. Evaluation of peptide-rich root extracts of Calliandria portoriscensis (Jacq.) Benth (Mimosaceae) for in vitro antimicrobial activity and brine shrimp lethality. BMC Complementary Medicine and Therapies 2020;20:30. https://doi.org/10.1186/s12906-020-2836-6.
  29. Pedan V, Weber C, Do T, Fischer N, Reich E, Rohn S. HPTLC fingerprint profile analysis of cocoa proanthocyanidins depending on origin and genotype. Food Chemistry 2018;267:277–87. https://doi.org/10.1016/j.foodchem.2017.08.109.
  30. Ayoola-Oresanya IO, Sonibare MA, Gueye B, Paliwal R, Abberton MT, Morlock GE. Effect-directed profiling and identification of bioactive metabolites from field, in vitro-grown and acclimatized Musa spp. accessions using high-performance thin-layer chromatography-mass spectrometry. Journal of Chromatography A 2020;1616:460774. https://doi.org/10.1016/j.chroma.2019.460774.
  31. Oyarzún P, Carrasco J, Peterssen D, Tereucan G, Aranda M, Henríquez-Aedo K. A high throughput method for detection of cyclooxygenase-2 enzyme inhibitors by effect-directed analysis applying high performance thin layer chromatography-bioassay-mass spectrometry. Journal of Chromatography A 2023;1711:464426. https://doi.org/10.1016/j.chroma.2023.464426.
  32. Thakur M, Nanda V. Screening of Indian bee pollen based on antioxidant properties and polyphenolic composition using UHPLC-DAD-MS/MS: A multivariate analysis and ANN based approach. Food Research International 2021;140:110041. https://doi.org/10.1016/j.foodres.2020.110041.
  33. Acito M, Varfaj I, Brighenti V, Cengiz EC, Rondini T, Fatigoni C, et al. A novel black poplar propolis extract with promising health-promoting properties: Focus on its chemical composition, antioxidant, anti-inflammatory, and anti-genotoxic activities. Food & Function 2024;15:4983–99. https://doi.org/10.1039/D3FO05059A.
  34. Falcão SI, Vale N, Gomes P, Domingues MRM, Freire C, Cardoso SM, et al. Phenolic profiling of portuguese propolis by LC–MS spectrometry: Uncommon propolis rich in flavonoid glycosides. Phytochemical Analysis 2013;24:309–18. https://doi.org/10.1002/pca.2412.
  35. Brugnerotto P, Dortzbach D, Blainski É, Falkenberg D de B, Vieira VF, Vieira H, et al. Identificação de plantas com potencial apícola provenientes de sistemas de monitoramento de colmeias no estado de Santa Catarina, Brasil. Research, Society and Development 2021;10:1–16. https://doi.org/10.33448/rsd-v10i7.16254.
  36. Mosić M, Trifković J, Vovk I, Gašić U, Tešić Ž, Šikoparija B, et al. Phenolic composition influences the health-promoting potential of bee-pollen. Biomolecules 2019;9:783. https://doi.org/10.3390/biom9120783.
  37. Kumar K, Debnath P, Singh S, Kumar N. An overview of plant phenolics and their involvement in abiotic stress tolerance. Stresses 2023;3:570–85. https://doi.org/10.3390/stresses3030040.
  38. Lin D, Xiao M, Zhao J, Li Z, Xing B, Li X, et al. An overview of plant phenolic compounds and their importance in human nutrition and management of type 2 diabetes. Molecules 2016;21:1374. https://doi.org/10.3390/molecules21101374.
  39. Shahidi F, Zhong Y. Measurement of antioxidant activity. Journal of Functional Foods 2015;18:757–81. https://doi.org/10.1016/j.jff.2015.01.047.
  40. Moita E, Gil-Izquierdo A, Sousa C, Ferreres F, Silva LR, Valentão P, et al. Integrated analysis of COX-2 and iNOS derived inflammatory mediators in LPS-stimulated RAW macrophages pre-exposed to Echium plantagineum L. bee pollen extract. PLoS One 2013;8:e59131. https://doi.org/10.1371/journal.pone.0059131.
  41. Maruyama H, Sakamoto T, Araki Y, Hara H. Anti-inflammatory effect of bee pollen ethanol extract from Cistus sp. of Spanish on carrageenan-induced rat hind paw edema. BMC Complementary Medicine and Therapies 2010;10:30. https://doi.org/10.1186/1472-6882-10-30.
  42. Cavallero A, Vidotto F, Sbrana C, Peres Fabbri L, Petroni G, Gabriele M. Antioxidant-rich polyfloral bee pollen exerts antimicrobial activity and anti-inflammatory effect in A549 lung epithelial cells by modulating the NF-κB pathway. Foods 2025;14:802. https://doi.org/10.3390/foods14050802.
  43. Ghafouri-Fard S, Shoorei H, Khanbabapour Sasi A, Taheri M, Ayatollahi SA. The impact of the phytotherapeutic agent quercetin on expression of genes and activity of signaling pathways. Biomedicine & Pharmacotherapy 2021;141:111847. https://doi.org/10.1016/j.biopha.2021.111847.
  44. Lesjak M, Beara I, Simin N, Pintać D, Majkić T, Bekvalac K, et al. Antioxidant and anti-inflammatory activities of quercetin and its derivatives. Journal of Functional Foods 2018;40:68–75. https://doi.org/10.1016/j.jff.2017.10.047.
  45. Skalski B, Lis B, Pecio Ł, Kontek B, Olas B, Żuchowski J, et al. Isorhamnetin and its new derivatives isolated from sea buckthorn berries prevent H2O2/Fe – Induced oxidative stress and changes in hemostasis. Food and Chemical Toxicology 2019;125:614–20. https://doi.org/10.1016/j.fct.2019.02.014.
  46. Abdel Bar FM, Alonazi R, Elekhnawy E, Samra RM, Alqarni MH, Badreldin H, et al. HPLC-PDA and in vivo anti-inflammatory potential of isorhamnetin-3-O-β-D-glucoside from Zygophyllum simplex L. Journal of Ethnopharmacology 2025;338:119089. https://doi.org/10.1016/j.jep.2024.119089.
  47. Lim HJ, Prajapati R, Seong SH, Jung HA, Choi JS. Antioxidant and antineuroinflammatory mechanisms of kaempferol-3-O-β-D-glucuronate on lipopolysaccharide-stimulated BV2 microglial cells through the Nrf2/HO-1 signaling cascade and MAPK/NF-κB pathway. ACS Omega 2023;8:6538–49. https://doi.org/10.1021/acsomega.2c06916.
  48. Elangovan B. A review on pharmacological studies of natural flavanone: pinobanksin. 3 Biotech 2024;14:111. https://doi.org/10.1007/s13205-023-03904-5.
  49. Wang Y, Tang C, Zhang H. Hepatoprotective effects of kaempferol 3-O-rutinoside and kaempferol 3-O-glucoside from Carthamus tinctorius L. on CCl4-induced oxidative liver injury in mice. Journal of Food and Drug Analysis 2015;23:310–7. https://doi.org/10.1016/j.jfda.2014.10.002.
  50. Zheng Y-Z, Deng G, Liang Q, Chen D-F, Guo R, Lai R-C. Antioxidant activity of quercetin and its glucosides from propolis: A theoretical study. Scientific Reports 2017;7:7543. https://doi.org/10.1038/s41598-017-08024-8.
  51. Dok-Go H, Lee KH, Kim HJ, Lee EH, Lee J, Song YS, et al. Neuroprotective effects of antioxidative flavonoids, quercetin, (+)-dihydroquercetin and quercetin 3-methyl ether, isolated from Opuntia ficus-indica var. saboten. Brain Research 2003;965:130–6. https://doi.org/10.1016/S0006-8993(02)04150-1.
  52. Matias A, Nunes SL, Poejo J, Mecha E, Serra AT, Madeira PJA, et al. Antioxidant and anti-inflammatory activity of a flavonoid-rich concentrate recovered from Opuntia ficus-indica juice. Food & Function 2014;5:3269–80. https://doi.org/10.1039/C4FO00071D.
  53. Wei B-L, Lu C-M, Tsao L-T, Wang J-P, Lin C-N. In vitro anti-inflammatory effects of quercetin 3-O-methyl ether and other constituents from Rhamnus species. Planta Medica 2001;67:745–7. https://doi.org/10.1055/s-2001-18339.
  54. Song J-H, Mun S-H, Mishra S, Kim S-R, Yang H, Choi SS, et al. Quercetin-3-methyl ether induces early apoptosis to overcome HRV1B immune evasion, suppress viral replication, and mitigate inflammatory pathogenesis. Biomolecules & Therapeutics (Seoul) 2025;33:388–98. https://doi.org/10.4062/biomolther.2024.204.

Copyright @2019 | Designed by: Open Journal Systems Chile Logo Open Journal Systems Chile Support OJS, training, DOI, Indexing, Hosting OJS

Code under GNU license: OJS PKP