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

Nosiba A. Alsarayrah; Rafeezul Mohamed; Mohd Azwan Jaafar; Eshaifol A. Omar; Nosiba A. Alsarayrah; Rafeezul Mohamed; Mohd Azwan Jaafar; Eshaifol A. Omar

doi:10.3934/agrfood.2026010

AIMS Agriculture and Food

2026, Volume 11, Issue 1: 177-204. doi: 10.3934/agrfood.2026010

Previous Article Next Article

Research article

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

1.
Department of Toxicology, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, 13200 Kepala Batas, Pulau Pinang, Malaysia
2.
Department of Nutrition and Integrative Health, Faculty of Allied Health Sciences, Middle East University, Amman, 11831, Jordan
3.
Department of Biomedical Science, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Kepala Batas, Pulau Pinang, Malaysia
4.
Agri-Omics and Bioinformatics Programme (BN1), Biotechnology and Nanotechnology Research Centre, Malaysian Agricultural Research and Development Institute (MARDI) Headquarters, 43400 Serdang, Selangor, Malaysia

Received: 19 December 2025 Revised: 20 February 2026 Accepted: 10 March 2026 Published: 25 March 2026

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.
- propolis,
- stingless bees,
- antioxidant activity,
- phytochemicals,
- nutraceutical potential
Citation: Nosiba A. Alsarayrah, Rafeezul Mohamed, Mohd Azwan Jaafar, Eshaifol A. Omar. Comparative phytochemical and antioxidant characterization of propolis from different species of Malaysian stingless bees[J]. AIMS Agriculture and Food, 2026, 11(1): 177-204. doi: 10.3934/agrfood.2026010

Related Papers:

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.

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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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 doi: 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. doi: 10.1007/s12016-012-8322-2

Reader Comments

Your name:*

Email:*
© 2026 the Author(s), licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0)