Author(s): Rozaini Mohd Zohdi, Nurul Najihah Yaacob, Nur Azyan Mohd Hasif, Muhammad Amirul Adli, Monporn Payaban, Richard Johari James, Fahimee Jaapar


DOI: 10.52711/0974-360X.2024.00158   

Address: Rozaini Mohd Zohdi1,2*, Nurul Najihah Yaacob3, Nur Azyan Mohd Hasif3, Muhammad Amirul Adli1,2, Monporn Payaban4, Richard Johari James1,5, Fahimee Jaapar6
1Atta-ur-Rahman Institute for Natural Product Discovery, Universiti Teknologi MARA Selangor Branch, Puncak Alam Campus, 42300 Puncak Alam, Selangor, Malaysia.
2Faculty of Pharmacy, Universiti Teknologi MARA, Selangor Branch, Puncak Alam Campus, 42300 Puncak Alam, Selangor, Malaysia.
3Faculty of Applied Sciences, Universiti Teknologi MARA, Selangor Branch, 40450 Shah Alam, Selangor, Malaysia.
4Faculty of Science, Rangsit University, Muang-Ake, Paholyothin Rd., PathumThani 12000, Thailand.
5Integrative Pharmacogenomics Institute (iPromise), Universiti Teknologi MARA Selangor Branch, Puncak Alam Campus, 42300 Puncak Alam, Selangor, Malaysia.
6Agrobiodiversity and Environmental Research Centre, MARDI Headquarter, 43400, Serdang, Selangor, Malaysia.
*Corresponding Author

Published In:   Volume - 17,      Issue - 3,     Year - 2024

Stingless bee propolis is known to contain a variety of bioactive compounds, including phenolic and flavonoids, which have been linked to its antibacterial properties. Nevertheless, the phytochemical compositions of stingless bee propolis are significantly influenced by a complex interplay of multiple factors such as geographical origin, floral source, and bee species. This study aimed to assess the physicochemical properties, phytochemical contents, and antibacterial activity of propolis from different stingless bee species found in the same environment and ecological system. The propolis samples obtained from Heterotrigona itama, Geniotrigona thoracica and Tetrigona apicalis were subjected to physicochemical analysis to determine the pH, moisture, lipid, resin, and wax contents. The total phenolic content (TPC) and total flavonoid content (TFC) were measured by Folin-Ciocalteu colorimetric and aluminium chloride methods, respectively. The antibacterial activity was determined using the agar well diffusion method against four Gram-positive bacteria, including Bacillus cereus, Micrococcus luteus, Streptococcus mutans, and Staphylococcus aureus. The physicochemical analysis of the propolis samples yielded the following results: moisture (12.17-16.45%), lipid (2.95-9.48%), resin (39.00-51.00%), wax (26.50-37.00%), and pH (5.07-5.61). Results revealed that propolis produced by G. thoracica displayed significantly higher moisture (16.45±0.38%), and resin contents (51.00±1.41%), as well as significantly lower percentage of lipid (2.95±0.12%), and wax content (26.50±0.71%). Similarly, G. thoracica propolis extract exhibited significantly higher TPC (200.70±0.06mg/mL GAE) and TFC (141.60±3.63mg/mL QE) values compared to the other propolis samples. Additionally, G. thoracica propolis extract was significantly active against B. cereus and M. luteus with inhibition zones of 15.00 and 16.00, respectively, and minimum inhibition concentration (MIC) of 390.63µg/mL. A strong correlation was found between resin content, TPC and antibacterial activity of propolis. This study indicated that the presence of high resin content in propolis resulted in a high concentration of phenolic compounds, which contributed significantly to its antibacterial activities. Furthermore, the research highlighted the species-dependent effect of propolis on its physicochemical characteristics, phytochemical composition, and antibacterial properties. The observed antibacterial efficacy of G. thoracica propolis suggested that the propolis extract held promise as an alternative treatment option against bacterial infections. Further research is warranted to fully elucidate the specific phenolic compounds that could contribute to its antibacterial properties.

Cite this article:
Rozaini Mohd Zohdi, Nurul Najihah Yaacob, Nur Azyan Mohd Hasif, Muhammad Amirul Adli, Monporn Payaban, Richard Johari James, Fahimee Jaapar. Comparative study of different Malaysian Stingless bee propolis: Physicochemical characterization, Phytochemical contents and Antibacterial activity. Research Journal of Pharmacy and Technology. 2024; 17(3):1021-8. doi: 10.52711/0974-360X.2024.00158

Rozaini Mohd Zohdi, Nurul Najihah Yaacob, Nur Azyan Mohd Hasif, Muhammad Amirul Adli, Monporn Payaban, Richard Johari James, Fahimee Jaapar. Comparative study of different Malaysian Stingless bee propolis: Physicochemical characterization, Phytochemical contents and Antibacterial activity. Research Journal of Pharmacy and Technology. 2024; 17(3):1021-8. doi: 10.52711/0974-360X.2024.00158   Available on:

1.    Ng, W. J., Sit, N. W., Ooi, P. A. C., Ee, K. Y., and Lim, T. M. Botanical origin differentiation of Malaysian stingless bee honey produced by Heterotrigona itama and Geniotrigona thoracica using chemometrics. Molecules. 2021; 26(24): 7628.
2.    Bueno, F. G. B., Kendall, L., Alves, D. A., Tamara, M. L., Heard, T., Latty, T., and Gloag, R. Stingless bee floral visitation in the global tropics and subtropics. Global Ecology and Conservation. 2023; e02454.
3.    Mustafa, M. Z., Yaacob, N. S., and Sulaiman, S. A. Reinventing the honey industry: Opportunities of the stingless bee. Malaysian Journal of Medical Sciences. 2018; 25(4): 1-5.
4.    Nafi, N. M., Zin, N. B. M., Pauzi, N., Khadar, A. S. A., Anisava, A. R., Badiazaman, A. A. M., and Mohd, K. S. Cytotoxicity, antioxidant and phytochemical screening of propolis extracts from four different Malaysian stingless bee species. Malaysian Journal of Fundamental and Applied Sciences. 2019; 15(2-1): 307-312.
5.    Samsudin, S. F., Mamat, M. R., and Hazmi, I. R. Taxonomic study on selected species of stingless bee (Hymenoptera: Apidae: Meliponini) in Peninsular Malaysia. Serangga. 2018; 23(2): 203-258.
6.    Burgett, M., Sangjaroen, P., Yavilat, J., and Chuttong, B. First report of hovering guard bees of the paleotropical stingless bee Tetrigona apicalis (Hymenoptera: Apidae: Meliponini). Apidologie. 2020; 51: 88-93.
7.    Pangestika, N. W., Atmowidi, T., and Kahono, S. Pollen load and flower constancy of three species of stingless bees (Hymenoptera, Apidae, Meliponinae). Tropical Life Sciences Research. 2017; 28(2): 179.
8.    Zohdi, R. M., Adli, M. A., Mohsin, H. F., Mokhtar, S. M., Low, A. L. M., Junaidi, A. H. A., and Jahrudin, D. H. J. GC-MS analysis and antibacterial activity of ethanolic and water extracts of Malaysian Heterotrigona itama propolis against selected human pathogenic bacteria. Malaysian Applied Biology. 2023; 52(2): 77-84.
9.    Menyiy, N. E., Al-Wali, N., Ghouizi, A. E., El-Guendouz, S., Salom, K., and Lyoussi, B. Potential therapeutic effect of Moroccan propolis in hyperglycemia, dyslipidemia, and hepatorenal dysfunction in diabetic rats. Iraninan Journal of Basic Medical Sciences. 2019; 22(11): 1331-1339. doi: 10.22038/ijbms.2019.33549.8004.
10.    Pereira, F. A. N., Barboza, J. R., Vasconcelos, C. C., Lopes, A. J. O., and Ribeiro, M. N. D. S. Use of stingless bee propolis and geopropolis against cancer - A literature review of preclinical studies. Pharmaceuticals. 2021; 14(11): 1161.
11.    Gupta, R., Rai, N., and Shetty, S. S. Herbal irrigants: underutilized option in dentistry. Research Journal of Pharmacy and Technology. 2019; 12(10): 5098-5100. doi: 10.5958/0974-360X.2019.00883.7.
12.    Jose, J., and Palanivelu, A. Antimicrobial efficacy of a novel root canal irrigant made from coconut and bromelain extract against biofilm forming pathogens – An in vitro evaluation. Research Journal of Pharmacy and Technology. 2021; 14(10): 5222-5226. doi: 10.52711/0974-360X.2021.00909.
13.    Handayani, B., R., M. S. M. A., Ridwan, R. D., and Aljunaid, M. The effect of Apis mellifera propolis on RUNX-2 and ALP during remodelling of orthodontic tooth movement. Research Journal of Pharmacy and Technology. 2021; 14(5): 2363-2366. doi: 10.52711/0974-360X.2021.00417.
14.    Campos, J. F., Dos Santos, H. F., Bonamigo, T., de Campos Domingues, N. L., de Picoli Souza, K., and Dos Santos, E. L. Stingless bee propolis: New insights for anticancer drugs. Oxidative Medicine and Cellular Longevity. 2021;1-18.
15.    Sharma, N., Sanadhya, S. Nagarajappa, R., Ramesh, G., and Naik, D. Antifungal activity of propolis, fluconazole and chlorhexidine against oral Candida albicans – A comparative in-vitro study. Research Journal of Pharmacy and Technology. 2022; 15(8): 3589-3594. doi: 10.52711/0974-360X.2022.00601.
16.    Mohiuddin, I., Kumar, T. R., Zargar, M. I., Wani, S. U. D., Mahdi, W. A., Alshehri, S., Alam, P., and Shakeel, F. GC-MS analysis, phytochemical screening, and antibacterial activity of Cerana indica propolis from Kashmir region. Separations. 2022; 9(11): 363.
17.    Kasote, D., Bankova, V., and Viljoen, A. M. Propolis: Chemical diversity and challenges in quality control. Phytochemistry Reviews. 2022; 21(6): 1887-1911.
18.    Jiang, X., Tian, J., Zheng, Y., Zhang, Y., Wu, Y., Zhang, C., and Hu, F. A new propolis type from Changbai mountains in North-east China: Chemical composition, botanical origin and biological activity. Molecules. 2019; 24(7): 1369.
19.    Kapare, H. S., and Sathiyanarayanan, L. Nutritional and therapeutic potential of propolis: A Review. Research Journal of Pharmacy and Technology. 2020; 13(7): 3545-3549. doi: 10.5958/0974-360X.2020.00627.7.
20.    Popova, M., Trusheva, B., and Bankova, V. Propolis of stingless bees: A phytochemist's guide through the jungle of tropical biodiversity. Phytomedicine. 2021; 86: 153098. doi: 10.1016/j.phymed.2019.153098.
21.    Adli, M. A., Zohdi, R. M., Othman, N. ’Aqilah, Amin, N. S. M., Mukhtar, S. M., Eshak, Z., Hazmi, I. R., and Jahrudin, D. H. J. Determination of antioxidant activity, total phenolic and flavonoid contents of Malaysian stingless bee propolis extracts. Journal of Sustainability Science and Management. 2022; 17(12): 133–144. doi: 10.46754/jssm.2022.12.012.
22.    AOAC, (2000). Official Methods of Analysis, Association of Official Analytical Chemists, Washington, DC, USA, 17th edition.
23.    Touzani, S., Embaslat, W., Imtara, H., Kmail, A., Kadan, S., Zaid, H., ElArabi, I., Badiaa, L., and Saad, B. In vitro evaluation of the potential use of propolis as a multitarget therapeutic product: Physicochemical properties, chemical composition, and immunomodulatory, antibacterial, and anticancer properties. 2019; 4836378. doi: 10.1155/2019/4836378.
24.    Pratami, D. K., Mun’im, A., Sundowo, A., and Sahlan, M. Phytochemical profile and antioxidant activity of propolis ethanolic extract from Tetragonula bee. Pharmacognosy Journal. 2018; 10(1): 128–135. Doi: 10.5530/pj.2018.1.23.
25.    Farasat, M., Khavari-Nejad, R. A., Mohammad, S., Nabavi, B., and Namjooyan, F. Antioxidant activity, total phenolics and flavonoid contents of some edible green seaweeds from northern coasts of the Persian Gulf. Iranian Journal of Pharmaceutical Research. 2014;13(1).
26.    Clinical And Laboratory Standards Institute. M100 Performance Standards for Antimicrobial Susceptibility Testing (28th ed.). 2018.
27.    Sarker, S. D., Nahar, L., and Kumarasamy, Y. Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth, and its application in the in vitro antibacterial screening of phytochemicals. Methods. 2007; 42(4): 321–324. doi: 10.1016/j.ymeth.2007.01.006.
28.    Lavinas, F. C., Macedo, E. H. B. C., Sá, G. B. L., Amaral, A. C. F., Silva, J. R.A., Azevedo, M. M. B., Vieira, B. A., Domingos, T. F. S., Vermelho, A. B., Carneiro, C. S., and Rodrigues, I. A. Brazilian stingless bee propolis and geopropolis: promising sources of biologically active compounds. Revista Brasileira de Farmacognosia. 2019; 19(3). doi: 10.1016/j.bjp.2018.11.007.
29.    Zullkiflee, N., Taha, H., and Usman, A. Propolis: Its role and efficacy in human health and diseases. Molecules. 2022; 27(18): 6120.
30.    Abdullah, N. A., Ja’afar, F., Yasin, H. M., Taha, H., Petalcorin, M. I. R., Mamit, M. H., Kusrini, E., and Usman, A. Physicochemical analyses, antioxidant, antibacterial, and toxicity of propolis particles produced by stingless bee Heterotrigona itama found in Brunei Darussalam. Heliyon. 2019; 5(9). doi: 10.1016/j.heliyon.2019.e02476.
31.    Abdullah, N. A., Zullkiflee, N., Zaini, S. N. Z., Taha, H., Hashim, F., and Usman, A. Phytochemicals, mineral contents, antioxidants, and antimicrobial activities of propolis produced by Brunei stingless bees Geniotrigona thoracica, Heterotrigona itama, and Tetrigona binghami. Saudi Journal of Biological Sciences. 2020; 27(11): 2902–2911. doi: 10.1016/j.sjbs.2020.09.014.
32.    Wallace, H. M., and Lee, D. J. Resin-foraging by colonies of Trigona sapiens and T. hockingsi (Hymenoptera: Apidae, Meliponini) and consequent seed dispersal of Corymbia torelliana (Myrtaceae). Apidologie. 2010; 41: 428–435. doi: 10.1051/apido/2009074.
33.    Benedick, S., Gansau, J. A., and Ahmad, A. H. Foraging behaviour of Heterotrigona itama (Apidae: Meliponini) in residential areas. Pertanika Journal of Tropical Agricultural Science. 2021; 44(2): 485-502.
34.    Shanahan, M., and Spivak, M. Resin Use by Stingless Bees: A Review. Insects. 2021;12:719. doi: 10.3390/insects12080719.
35.    Dias, L. G., Pereira, A. P., and Estevinho, L. M. Comparative study of different Portuguese samples of propolis: Pollinic, sensorial, physicochemical, microbiological characterization and antibacterial activity. Food and Chemical Toxicology. 2012; 50(12): 4246-4253. doi: 10.1016/j.fct.2012.08.056.
36.    Menyiy, N. E., Bakour, M., El-Ghouizi, A., El-Guendouz, S., and Lyoussi, B. Influence of geographic origin and plant source on physicochemical properties, mineral content, and antioxidant and antibacterial activities of Moroccan propolis. International Journal of Food Science. 2021; 5570224.
37.    Ibrahim, N., Niza, N. F. S. M., Rodi, M. M. M., Zakaria, A. J., Ismail, Z., and Mohd, K. S. Chemical and biological analyses of malaysian stingless bee propolis extracts. Malaysian Journal of Analytical Science. 2016; 20(2): 413-422. doi: 10.17576/mjas-2016-2002-26.
38.    Jobir, M. D., and Belay, A. Comparative study of different Ethiopian propolis: In vivo wound healing, antioxidant, antibacterial, physicochemical properties and mineral profiles. Journal of Apitherapy. 2020; 7(2): 31-48. doi: 10.5455/ja.20200225125850.
39.    Aboulghazi, A., Touzani, S., Fadil, M., and Lyoussi, B. Physicochemical characterization and in vitro evaluation of the antioxidant and anticandidal activities of Moroccan propolis. Veterinary World. 2022; 15(2): 341-349. doi: 10.14202/vetworld.2022.341-349.
40.    Asem, N., Abdul Gapar, N. A., Abd Hapit, N. H., and Omar, E. A. Correlation between total phenolic and flavonoid contents with antioxidant activity of Malaysian stingless bee propolis extract. Journal of Apicultural Research. 2019; 59(4): 437–442. doi: 10.1080/00218839.2019.1684050.
41.    Awang, N., Ali, A., Adibah Abd Majid, F., Hamzah, S., and Bahri A. R. S. Total flavonoids and phenolic contents of sticky and hard propolis from 10 species of Indo-Malayan stingless bees. Malaysian Journal of Analytical Sciences. 2018; 22: 877–884. doi: 10.17576/mjas-2018-2205-15.
42.    Idris, L., Adli, M. A., Yaacop, N. N., and Zohdi, R. M. Phytochemical screening and antioxidant activities of Geniotrigona thoracica propolis extracts derived from different locations in Malaysia. Malaysian Journal of Fundamental and Applied Sciences. 2023; 19: 1023-1032.
43.    Touzani, S., Al-Waili, N., Menyiy, N. E., Filipic, B., Pereyra, A., Arabi, I. E., Al-Waili, W., and Lyoussi, B. Chemical analysis and antioxidant content of various propolis samples collected from different regions and their impact on antimicrobial activities. Asian Pacific Journal of Tropical Medicine. 2018; 11(7): 436-442. doi: 10.4103/1995-7645.237188.
44.    Luján, M. D. R. M., Reséndez, A. M., Barrón, G. S. G., Carrillo, J. L. R., and Inungaray, M. L. C. Antibacterial activity and phenolic content of propolis extracts obtained by different extraction methods. Nova Scientia. 2018; 10(20): 397-412.
45.    Sforcin, J.M. Biological properties and therapeutic applications of propolis. Phytotherapy Research. 2016;30(6):894-905.
46.    Almuhayawi, M.S. Propolis as a novel antibacterial agent. Saudi Journal of Biological Sciences. 2020; 27(11): 3079-3086.
47.    Kalia, P., Kumar, N. R., and Harjai, K. Preventive effect of honeybee propolis on Salmonella enterica serovar Typhimurium infected BALB/c mice: A hematological study. Research Journal of Pharmacy and Technology. 2020; 13(7): 3389-3393. doi: 10.5958/0974-360X.2020.00602.2.

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