Comparative phytochemical and antioxidant characterization of propolis from different species of Malaysian stingless bees

Abstract

Propolis derived from stingless bees is recognized for its rich bioactive composition and potential health-promoting properties. This study evaluated the phytochemical profile and antioxidant capacity of propolis extracts obtained from three Malaysian stingless bee species: Geniotrigona thoracica, Heterotrigona itama, and Tetrigona binghami. Gas chromatography–mass spectrometry (GC–MS) analysis identified diverse bioactive constituents and revealed marked compositional variations among the species examined. Antioxidant activities were assessed using DPPH, ABTS, and FRAP assays. All extracts exhibited strong antioxidant potential, as reflected by favorable IC₅₀/EC₅₀ values in the DPPH assay and consistent performance in the ABTS and FRAP assays. Significant correlations between antioxidant capacity and total phenolic and flavonoid contents indicated that phenolic compounds are the principal contributors to the observed effects. Overall, these findings demonstrate that Malaysian stingless bee propolis represents a valuable source of natural antioxidants and bioactive phytochemicals, underscoring its potential as a health-promoting and nutraceutical agent.

Keywords

propolis / stingless bees / antioxidant activity / phytochemicals / nutraceutical potential

References

[1] Salleh SNAS, Hanapiah NAM, Johari WLW, et al. (2021) Analysis of bioactive compounds and chemical composition of Malaysian stingless bee propolis water extracts. Saudi J Biol Sci 28: 6705-6710. https://doi.org/10.1016/j.sjbs.2021.07.049

[2] Silva-Beltrán NP, Umsza-Guez MA, Ramos Rodrigues DM, et al. (2021) Comparison of the biological potential and chemical composition of Brazilian and Mexican propolis. Appl Sci 11: 11417. https://doi.org/10.3390/app112311417

[3] Ahmed R, Tanvir E, Hossen MS, et al. (2017) Antioxidant properties and cardioprotective mechanism of Malaysian propolis in rats. Evidence-Based Complementary Altern Med 2017: 5370545. https://doi.org/10.1155/2017/5370545

[4] Ramadhan R, Kusuma IW, Egra S, et al. (2020) Diversity and honey properties of stingless bees from meliponiculture in East and North Kalimantan, Indonesia. Biodiversitas J Biol Diversity 21: No.10. https://doi.org/10.13057/biodiv/d211021

[5] Ismail NF, Maulidiani M, Omar S, et al. (2021) Classification of stingless bee honey based on species, dehumidification process and geographical origins using physicochemical and ATR-FTIR chemometric approach. J Food Compos Anal 104: 104126. https://doi.org/10.1016/j.jfca.2021.104126

[6] Ahangari Z, Naseri M, Vatandoost F (2018) Propolis: Chemical composition and its applications in endodontics. Iran Endod J 13: 285.

[7] Özer ED (2020) Propolis and potential use in food products. Turk J Agric-Food Sci Technol 8: 1139-1144. https://doi.org/10.24925/turjaf.v8i5.1139-1144.3324

[8] Abdullah NA, Ja'afar F, Yasin HM, et al. (2019) Physicochemical analyses, antioxidant, antibacterial, and toxicity of propolis particles produced by stingless bee Heterotrigona itama found in Brunei Darussalam. Heliyon 5: e02476. https://doi.org/10.1016/j.heliyon.2019.e02476

[9] Hossain S, Yousaf M, Liu Y, et al. (2022) An overview of the evidence and mechanism of drug-herb interactions between propolis and pharmaceutical drugs. Front Pharmacol 13: 876183. https://doi.org/10.3389/fphar.2022.876183

[10] Alsarayrah NA, Mohamed R, Omar EA (2025) Stingless bee propolis: a comprehensive review of chemical constituents and health efficacy. Nat Prod Bioprospect 15: 61. https://doi.org/10.1007/s13659-025-00545-4

[11] Mihai CM, Mărghitaș LA, Dezmirean DS, et al. (2011) Correlation between polyphenolic profile and antioxidant activity of propolis from Transylvania. Sci Papers Anim Sci Biotechnol 44: 100-100.

[12] Asgharpour F, Moghadamnia AA, Kazemi S, et al. (2020) Applying GC-MS analysis to identify chemical composition of Iranian propolis prepared with different solvent and evaluation of its biological activity. Caspian J Int Med 11: 191. https://doi.org/10.22088/cjim.11.2.191

[13] Harif Fadzilah N, Jaapar MF, Jajuli R, et al. (2017) Total phenolic content, total flavonoid and antioxidant activity of ethanolic bee pollen extracts from three species of Malaysian stingless bee. J Apic Res 56: 130-135. https://doi.org/10.1080/00218839.2017.1287996

[14] Hatano A, Nonaka T, Yoshino M, et al. (2012) Antioxidant activity and phenolic constituents of red propolis from Shandong, China. Food Sci Technol Res 18: 577-584. https://doi.org/10.3136/fstr.18.577

[15] Ahmadinejad F, Geir Møller S, Hashemzadeh-Chaleshtori M, et al. (2017) Molecular mechanisms behind free radical scavengers function against oxidative stress. Antioxidants 6: 51. https://doi.org/10.3390/antiox6030051

[16] Tai A,Sawano T,Yazama F, et al. (2011) Evaluation of antioxidant activity of vanillin by using multiple antioxidant assays. Biochimica et Biophysica Acta (BBA)-General Subjects 1810: 170-177. https://doi.org/10.1016/j.bbagen.2010.11.004

[17] Eltawaty SI, Suliman MB, El-Hddad S (2023) Chemical composition, and antibacterial and antifungal activities of crude extracts from Pistacia lentiscus L. fruit. Tropical J Nat Prod Res 7: 4049-4054. https://doi.org/10.26538/tjnpr/v7i9.30

[18] Bakkali F, Averbeck S, Averbeck D, et al. (2008) Biological effects of essential oils-A review. Food Chem Toxicol 46: 446-475. https://doi.org/10.1016/j.fct.2007.09.106

[19] Gertsch J,Leonti M,Raduner S, et al. (2008) Beta-caryophyllene is a dietary cannabinoid. Proc Nat Acad Sci 105: 9099-9104. https://doi.org/10.1073/pnas.0803601105

[20] Aljaafari MN, Alkhoori MA, Hag-Ali M, et al. (2022) Contribution of aldehydes and their derivatives to antimicrobial and immunomodulatory activities. Molecules 27: 3589. https://doi.org/10.3390/molecules27113589

[21] Aparna V, Dileep KV, Mandal PK, et al. (2012) Anti-inflammatory property of n-hexadecanoic acid: Structural evidence and kinetic assessment. Chem Biol Drug Des 80: 434-439. https://doi.org/10.1111/j.1747-0285.2012.01418.x

[22] Ilozue NM, Okoye P-A, Ekpunobi UE (2024) Phytochemical evaluation, GC-MS profiling and antimicrobial activity of two herbal mixtures marketed in Anambra state. South Asian Res J Nat Prod 7: 184-196. http://archive.go4subs.com/id/eprint/1937

[23] Saleem M (2009) Lupeol, a novel anti-inflammatory and anti-cancer dietary triterpene. Cancer Lett 285: 109-115. https://doi.org/10.1016/j.canlet.2009.04.033

[24] Rakariyatham K, Zhou D, Rakariyatham N, et al. (2020) Sapindaceae (Dimocarpus longan and Nephelium lappaceum) seed and peel by-products: Potential sources for phenolic compounds and use as functional ingredients in food and health applications. J Functional Foods 67: 103846. https://doi.org/10.1016/j.jff.2020.103846

[25] Rubab M, Chelliah R, Saravanakumar K, et al. (2020) Bioactive potential of 2-Methoxy-4-vinylphenol and benzofuran from Brassica oleracea L. var. capitate f, rubra (red cabbage) on oxidative and microbiological stability of beef meat. Foods 9: 568. https://doi.org/10.3390/foods9050568

[26] Mostofa MG, Reza AA, Khan Z, et al. (2024) Apoptosis-inducing anti-proliferative and quantitative phytochemical profiling with in silico study of antioxidant-rich Leea aequata L. leaves. Heliyon 10: e23400. https://doi.org/10.1016/j.heliyon.2023.e23400

[27] Adli MA, Zohdi RM, Othman NA, et al. (2022) Determination of antioxidant activity, total phenolic and flavonoid contents of Malaysian stingless bee propolis extracts. J Sustainability Sci Manage 17: 132-143. https://doi.org/10.46754/jssm.2022.12.012

[28] Mokhtar SU (2019) Comparison of total phenolic and flavonoids contents in Malaysian propolis extract with two different extraction solvents. Int J Eng Technol Sci 6: No.2. https://doi.org/10.15282/ijets.v6i2.2577

[29] Yıldırım HK (2022) Assessment of propolis treated by different extraction methods. Braz Arch Biol Technol 65: e22210251. https://doi.org/10.1590/1678-4324-2022210251

[30] Awang N, Ali Na, Majid F, et al. (2018) Total flavonoids and phenolic contents of sticky and hard propolis from 10 species of Indo-Malayan stingless bees. Malays J Anal Sci 22: 877-884. https://doi.org/10.17576/mjas-2018-2205-15

[31] Zohdi RM, Yaacob NN, Hasif NAM, et al. (2024) Comparative study of different Malaysian Stingless bee propolis: Physicochemical characterization, Phytochemical contents and Antibacterial activity. Res J Pharm Technol 17: 1021-1028. https://doi.org/10.52711/0974-360x.2024.00158

[32] Campos JF, Santos UPd, Rocha PdSd, et al. (2015) Antimicrobial, antioxidant, anti-inflammatory, and cytotoxic activities of propolis from the stingless bee Tetragonisca fiebrigi (Jatai). Evidence-Based Complementary Altern Med 2015: 296186. https://doi.org/10.1155/2015/296186

[33] Ferreira LMdMC,Souza PDQd,Pereira RR, et al. (2024) Preliminary study on the chemical and biological properties of propolis extract from stingless bees from the northern region of Brazil. Processes 12: 700. https://doi.org/10.3390/pr12040700

[34] Badiazaman AM, Zin NBM, Annisava AR, et al. (2019) Phytochemical screening and antioxidant properties of stingless bee Geniotrigona thoracica propolis. Malays J Fundam Appl Sci 15: 330-335. https://doi.org/10.11113/mjfas.v15n2-1.1557

[35] Fabris S, Bertelle M, Astafyeva O, et al. (2013) Antioxidant properties and chemical composition relationship of Europeans and Brazilians propolis. Pharmacol Pharm 4: 46-51. https://doi.org/10.4236/pp.2013.41006

[36] Kumazawa S, Hamasaka T, Nakayama T (2004) Antioxidant activity of propolis of various geographic origins. Food Chem 84: 329-339. https://doi.org/10.1016/s0308-8146(03)00216-4

[37] Idris L, Adli MA, Yaacop NN, et al. (2023) Phytochemical screening and antioxidant activities of Geniotrigona thoracica propolis extracts derived from different locations in Malaysia. Malays J Fundam Appl Sci 19: 1023-1032. https://doi.org/10.11113/mjfas.v19n6.3128

[38] Zohdi RM, Adli MA, Miskan AM, et al. (2025) Chemical profile, physicochemical properties, and antioxidant activity of Malaysian propolis: Insights from honeybee and stingless bee. J Sci Math Lett 13: 1-9. https://doi.org/10.37134/jsml.vol13.1.1.2025

[39] Kurek-Górecka A, Keskin Ş, Bobis O, et al. (2022) Comparison of the antioxidant activity of propolis samples from different geographical regions. Plants 11: 1203. https://doi.org/10.3390/plants11091203

[40] Olszowy-Tomczyk M (2021) How to express the antioxidant properties of substances properly? Chem Pap 75: 6157-6167. https://doi.org/10.1007/s11696-021-01799-1.

[41] Bankova V, Popova M, Trusheva B (2014) Propolis volatile compounds: Chemical diversity and biological activity: A review. Chem Cent J 8: 28. https://doi.org/10.1016/B978-0-444-62650-1.00007-6

[42] Isla MI, Nieva Moreno MI, Zampini C, et al. (2013) Argentine propolis: Flavonoid and chalcone content and its relation with the functional properties. Nova Science Publishers, Hauppauge, New York, 161-170. http://hdl.handle.net/11336/147944

[43] Miguel MG (2013) Chemical and biological properties of propolis from the western countries of the Mediterranean basin and Portugal. Int J Pharm Pharm Sci 5: 403-409. http://hdl.handle.net/10400.1/6333

[44] Chan GC-F, Cheung K-W, Sze DM-Y (2013) The immunomodulatory and anticancer properties of propolis. Clin Rev Allergy Immunol 44: 262-273. https://doi.org/10.1007/s12016-012-8322-2.