Ultrasonic-Assisted Extraction of Propolis: Response Surface Optimization and Solvent-Dependent Bioactive Properties

Esen Bilge Biçer; Şeniz Karabıyıklı Çiçek; Ebru Yabaş

doi:10.35206/jan.1907468

EN

Ultrasonic-Assisted Extraction of Propolis: Response Surface Optimization and Solvent-Dependent Bioactive Properties

Abstract

Propolis is a resinous substance produced by honey bees (Apis mellifera) and is rich in biologically active compounds. Efficient extraction of these compounds depends largely on solvent selection and the optimization of extraction conditions. In this study, ultrasonic-assisted extraction (UAE) conditions from crude propolis were optimized using two solvents (dimethyl sulfoxide (DMSO) and ethanol) through a Box–Behnken experimental design based on response surface methodology (RSM). Response surface methodology (RSM) was employed to evaluate the effects of solvent concentration (0-100%), extraction temperature (30–50 °C), and extraction time (30-180 min) on extraction yield. The DMSO-based UAE model showed high statistical significance with an R² value of 0.9950 (F = 154.15, p <0.0001). The optimal extraction conditions were determined as 98.34% solvent concentration, 42.20°C extraction temperature, and 142.60 min extraction time, resulting in a yield of 54.17%. Similarly, the ethanol-based UAE model was also significant (F = 113.09, p <0.0001; R² = 0.9932), with optimal conditions of 99.19% solvent concentration, 45.26 °C extraction temperature, and 171.5 min extraction time, yielding 55.82%. In both solvent systems, solvent concentration was identified as the most influential parameter affecting extraction yield (p <0.0001). Bioactivity analyses revealed that DMSO extracts contained higher TPC (325.27 ± 0,05 mg GAE/g) and exhibited stronger antioxidant activity (DPPH SC50: 11.90 ± 1.6 µg/mL; ABTS SC50: 147.7 ± 0.4 µg/mL) compared with ethanol extracts. Conversely, ethanol extracts demonstrated stronger antimicrobial activity against Escherichia coli, Salmonella enterica serovar Typhimurium, Bacillus cereus, and Staphylococcus aureus, with consistent MIC values of 25 µg/mL. Overall, the findings indicate that both solvents provide high extraction efficiencies (~54-56%) under optimized UAE conditions. Furthermore, solvent selection plays a decisive role in determining the bioactivity profile. The results show that using DMSO in extraction is suitable for maximizing antioxidant properties, while using ethanol is more suitable for antimicrobial applications.

Keywords

Propolis, Ultrasonic-assisted Extraction, Response Surface Methodology, Antimicrobial Activity, Antioxidant Activity

Supporting Institution

Tokat Gaziosmanpaşa University Scientific Research Projects Coordination Unit

Project Number

2024-33

Ethical Statement

This study did not involve any human participants or animals. Therefore, ethics committee approval was not required.

Thanks

This study was supported by Tokat Gaziosmanpaşa University Scientific Research Projects Coordination Unit as a PhD thesis (Project Code: 2024-33).

References

Al-Ghamdi, A. A., Bayaqoob, N. I., Rushdi, A. I., Alattal, Y., Simoneit, B. R., El-Mubarak, A. H., and Al-Mutlaq, K. F. (2017). Chemical compositions and characteristics of organic compounds in propolis from Yemen. Saudi journal of biological sciences, 24(5), 1094-1103. https://doi.org/10.1016/j.sjbs.2016.12.012
Almuhayawi MS. (2020). Propolis as a novel antibacterial agent. Saudi J Biol Sci.,27(11):3079-3086. https://doi.org/10.1016/j.sjbs.2020.09.016
Bankova, V. S., de Castro, S. L., and Marcucci, M. C. (2000). Propolis: recent advances in chemistry and plant origin. Apidologie, 31(1), 3-15. https://doi.org/10.1051/apido:2000102
Bankova, V. (2005). Chemical diversity of propolis and the problem of standardization. Journal of ethnopharmacology, 100(1-2), 114-117. https://doi.org/10.1016/j.jep.2005.05.004
Bankova, V., Popova, M., and Trusheva, B. (2014). Propolis volatile compounds: chemical diversity and biological activity: a review. Chemistry Central Journal, 8(1), 28. https://doi.org/10.1186/1752-153X-8-28
Baş, D., and Boyacı, İ. H. (2007) Modeling and optimization I: Usability of response surface methodology. Journal of Food Engineering, 78(3), 836-845. https://doi.org/10.1016/j.jfoodeng.2005.11.024
Bezerra, M. A., Santelli, R. E., Oliveira, E. P., Villar, L. S., and Escaleira, L. A. (2008). Response surface methodology (RSM) as a tool for optimization in analytical chemistry. Talanta, 76(5), 965–977. https://doi.org/10.1016/j.talanta.2008.05.019
Bicer, E. B. (2024). Green Extraction Methods. Current Research from Science to Technology-Ii, Iksad Publications 19.
Blois, M. S. (1958). Antioxidant determinations by the use of a stable free radical. Nature, 181(4617), 1199-1200. https://doi.org/10.1038/1811199a0
Castaldo, S., and Capasso, F. (2002). Propolis, an old remedy used in modern medicine. Fitoterapia, 73(S1), S1–S6. https://doi.org/10.1016/s0367-326x(02)00185-5

Castro, M. L., Cury, J. A., Rosalen, P. L., Alencar, S. M., Ikegaki, M., Duarte, S., and Koo, H. (2007). Propolis from southeastern and northeastern of Brazil: the influence of seasonality in antibacterial activity and phenolic composition. Química Nova, 30, 1512-1516. https://doi.org/10.1590/S0100-40422007000700003
Chang, C. C., Yang, M. H., Wen, H. M., and Chern, J. C. (2002). Estimation of total flavonoid content in propolis by two complementary colometric methods. Journal of food and drug analysis, 10(3), 3. https://doi.org/10.38212/2224-6614.2748
Chemat, F., Rombaut, N., Sicaire, A. G., Meullemiestre, A., Fabiano-Tixier, A. S., and Abert-Vian, M. (2017). Ultrasound assisted extraction of food and natural products. Ultrasonics Sonochemistry, 34, 540–560. https://doi.org/10.1016/j.ultsonch.2016.06.035
Cottica, S. M., Sawaya, A. C. F., Eberlin, M. N., Franco, S. L., Zeoula, L. M., and Visentainer, J. V. (2011). Antioxidant activity and composition of propolis obtained by different methods of extraction. Journal of the Brazilian Chemical Society, 22(5), 929-935. https://doi.org/10.1590/S0103-50532011000500016
Cujic, N., Šavikin, K., Janković, T., Pljevljakušić, D., Stević, T., and Ibrić, S. (2016). Optimization of polyphenols extraction from dried chokeberry using response surface methodology. Journal of Applied Research on Medicinal and Aromatic Plants, 3(3), 88-97. https://doi.org/10.1016/j.foodchem.2015.08.008
Cunha, I. B. S., Sawaya, A. C. H. F., Caetano, F. M., Shimizu, M. T., Marcucci, M. C., Drezza, F. T., … Eberlin, M. N. (2004). Factors that influence the yield and composition of Brazilian propolis extracts. Journal of the Brazilian Chemical Society, 15(6), 964–970. https://doi.org/10.1590/S0103-50532004000600026
Cushnie, T. T., and Lamb, A. J. (2011). Recent advances in understanding the antibacterial properties of flavonoids. International journal of antimicrobial agents, 38(2), 99-107. https://doi.org/10.1016/j.ijantimicag.2011.02.014
Escriche, I., and Juan-Borrás, M. (2018). Standardizing the analysis of phenolic profile in propolis. Food Research International, 106, 834-841. https://doi.org/10.1016/j.foodres.2018.01.055
Galanakis, C. M. (2012). Recovery of high added-value components from food wastes: Conventional, emerging technologies and commercialized applications. Trends in Food Science and Technology, 26(2), 68-87. https://doi.org/10.1016/j.tifs.2012.03.003
Guan, X., and Yao, H. (2008). Optimization of viscozyme L-assisted extraction of oat bran protein using response surface methodology. Food Chemistry, 106(1), 345-351. https://doi.org/10.1016/j.foodchem.2007.05.041
Hasler, C. M. (2002). Functional foods: Benefits, concerns and challenges—A position paper from the American Council on Science and Health. The Journal of Nutrition, 132(12), 3772–3781. https://doi.org/10.1093/jn/132.12.3772
Herrera, M. C., and Luque de Castro, M. D. (2005). Ultrasound-assisted extraction of phenolic compounds from strawberries. Analytica Chimica Acta, 509(2), 217–222 https://doi.org/10.1016/j.chroma.2005.09.021
Huang, S., Zhang, C. P., Wang, K., Li, G. Q., & Hu, F. L. (2014). Recent advances in the chemical composition of propolis. Molecules, 19(12), 19610-19632. https://doi.org/10.3390/molecules191219610
Inui, S., Hatano, A., Yoshino, M., Hosoya, T., Shimamura, Y., Masuda, S., and Ahn, M. R. (2014). Identification of the phenolic compounds contributing to antibacterial activity in ethanol extracts of Brazilian red propolis. Journal of Agricultural and Food Chemistry, 62(3), 699–704. https://doi.org/10.1080/14786419.2014.898146
Karabıyıklı, Ş., and Öncül, N. (2016). Inhibitory effect of unripe grape products on foodborne pathogens. Journal of Food Processing and Preservation, 40(6), 1459–1465. https://doi.org/10.1111/jfpp.12731
Khuri, A. I., and Cornell, J. A. (1996). Response surfaces: designs and analyses. CRC press.
Knez, E., Kadac-Czapska, K., and Grembecka, M. (2025). Evaluation of Spectrophotometric Methods for Assessing Antioxidant Potential in Plant Food Samples—A Critical Approach. Applied Sciences, 15(11), 5925. https://doi.org/10.3390/app15115925
Kumova, U. (2002). Önemli bir arı ürünü: Propolis. Uludag Bee Journal, 2(2), 10-24. https://izlik.org/JA86YC62MR
Maran, J. P., Manikandan, S., Thirugnanasambandham, K., Nivetha, C. V., & Dinesh, R. (2013). Box–Behnken design based statistical modeling for ultrasound-assisted extraction of corn silk polysaccharide. Carbohydrate polymers, 92(1), 604-611. https://doi.org/10.1016/j.carbpol.2012.09.020
Marcucci, M. C. (1995). Propolis: Chemical composition, biological properties and therapeutic activity. Apidologie, 26(2), 83–99. https://doi.org/10.1051/apido:19950202
McDonnell, G., and Russell, A. D. (1999). Antiseptics and disinfectants: activity, action, and resistance. Clinical microbiology reviews, 12(1), 147-179. https://doi.org/10.1128/cmr.12.1.147
Montgomery, D. C. (2017). Design and analysis of experiments (9th ed.). Wiley.
Myers, R. H., Montgomery, D. C., and Anderson-Cook, C. M. (2016). Response surface methodology: Process and product optimization using designed experiments. John Wiley and Sons.
Oroian, M., Dranca, F., and Ursachi, F. (2020). Comparative evaluation of maceration, microwave and ultrasonic-assisted extraction of phenolic compounds from propolis. Journal of food science and technology, 57(1), 70-78. https://doi.org/10.1007/s13197-019-04031-x
Permukaan, K. G. B. (2021). Optimization of extraction temperature and time on phenolic compounds and antioxidant activity of Malaysian propolis Trigona spp. aqueous extract using “response surface methodology. Malaysian Journal of Analytical Sciences, 25(4), 649-660.
Picot-Allain, M. C. N., Ramasawmy, B., and Emmambux, M. N. (2022). Extraction, characterisation, and application of pectin from tropical and sub-tropical fruits: a review. Food Reviews International, 38(3), 282-312. https://doi.org/10.1080/87559129.2020.1733008
Przybyłek, I., and Karpiński, T. M. (2019). Antibacterial properties of propolis. Molecules, 24(11), 2047. https://doi.org/10.3390/molecules24112047
Roberfroid, M. B. (2002). Global view on functional foods: European perspectives. British Journal of Nutrition, 88(S2), S133–S138. https://doi.org/10.1079/BJN2002677
Silva, J. F. M., de Souza, M. C., Matta, S. R., de Andrade, M. R., and Vidal, F. V. N. (2006). Correlation analysis between phenolic levels of Brazilian propolis extracts and their antimicrobial and antioxidant activities. Food chemistry, 99(3), 431-435. https://doi.org/10.1016/j.foodchem.2005.07.055
Sforcin, J. M. (2007). Propolis and the immune system: a review. Journal of ethnopharmacology, 113(1), 1-14. https://doi.org/10.1016/j.jep.2007.05.012
Soquetta, M. B., Terra, L. M., and Bastos, C. P. (2018). Green technologies for the extraction of bioactive compounds from fruits and vegetables. CyTA-Journal of Food, 16(1), 400-412. https://doi.org/10.1080/19476337.2017.1411978
Speciale, A., Costanzo, R., Puglisi, S., Musumeci, R., and Cacciola, F. (2006). Antibacterial activity of propolis and its chemical composition. Food Chemistry, 95(4), 580–587. https://doi.org/10.1179/joc.2006.18.2.164
Thamnopoulos, I. A. I., Michailidis, G. F., Fletouris, D. J., Badeka, A. V., Kontominas, M. G., and Angelidis, A. S. (2018). Inhibitory activity of propolis against Listeria monocytogenes in milk stored under refrigeration. Food Microbiology, 73, 168–176. https://doi.org/10.1016/j.fm.2018.01.021
Trusheva, B., Trunkova, D., and Bankova, V. (2007). Different extraction methods of biologically active components from propolis: A preliminary study. Chemistry Central Journal, 1, 13. https://doi.org/10.1186/1752-153X-1-13
Wiegand, I., Hilpert, K., and Hancock, R. E. (2008). Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nature protocols, 3(2), 163-175. https://doi.org/10.1038/nprot.2007.521
Zhang, Q. W., Lin, L. G., and Ye, W. C. (2018). Techniques for extraction and isolation of natural products: a comprehensive review. Chinese Medicine, 13(1), 1-26. https://doi.org/10.1186/s13020-018-0177-x

Details

Primary Language

English

Subjects

Food Engineering

Journal Section

Research Article

Authors

Esen Bilge Biçer ^*
0000-0003-0500-8303
Türkiye

Şeniz Karabıyıklı Çiçek
0000-0001-9287-9400
Türkiye

Ebru Yabaş
0000-0001-7163-3057
Türkiye

Publication Date

May 12, 2026

Submission Date

March 11, 2026

Acceptance Date

May 11, 2026

Published in Issue

Year 2026 Volume: 9 Number: 1

DOI

https://doi.org/10.35206/jan.1907468

IZ

https://izlik.org/JA42NR35GA

APA

Biçer, E. B., Karabıyıklı Çiçek, Ş., & Yabaş, E. (2026). Ultrasonic-Assisted Extraction of Propolis: Response Surface Optimization and Solvent-Dependent Bioactive Properties. Journal of Apitherapy and Nature, 9(1), 76-94. https://doi.org/10.35206/jan.1907468

AMA

1.Biçer EB, Karabıyıklı Çiçek Ş, Yabaş E. Ultrasonic-Assisted Extraction of Propolis: Response Surface Optimization and Solvent-Dependent Bioactive Properties. J.Apit.Nat. 2026;9(1):76-94. doi:10.35206/jan.1907468

Chicago

Biçer, Esen Bilge, Şeniz Karabıyıklı Çiçek, and Ebru Yabaş. 2026. “Ultrasonic-Assisted Extraction of Propolis: Response Surface Optimization and Solvent-Dependent Bioactive Properties”. Journal of Apitherapy and Nature 9 (1): 76-94. https://doi.org/10.35206/jan.1907468.

EndNote

Biçer EB, Karabıyıklı Çiçek Ş, Yabaş E (May 1, 2026) Ultrasonic-Assisted Extraction of Propolis: Response Surface Optimization and Solvent-Dependent Bioactive Properties. Journal of Apitherapy and Nature 9 1 76–94.

IEEE

[1]E. B. Biçer, Ş. Karabıyıklı Çiçek, and E. Yabaş, “Ultrasonic-Assisted Extraction of Propolis: Response Surface Optimization and Solvent-Dependent Bioactive Properties”, J.Apit.Nat., vol. 9, no. 1, pp. 76–94, May 2026, doi: 10.35206/jan.1907468.

ISNAD

Biçer, Esen Bilge - Karabıyıklı Çiçek, Şeniz - Yabaş, Ebru. “Ultrasonic-Assisted Extraction of Propolis: Response Surface Optimization and Solvent-Dependent Bioactive Properties”. Journal of Apitherapy and Nature 9/1 (May 1, 2026): 76-94. https://doi.org/10.35206/jan.1907468.

JAMA

1.Biçer EB, Karabıyıklı Çiçek Ş, Yabaş E. Ultrasonic-Assisted Extraction of Propolis: Response Surface Optimization and Solvent-Dependent Bioactive Properties. J.Apit.Nat. 2026;9:76–94.

MLA

Biçer, Esen Bilge, et al. “Ultrasonic-Assisted Extraction of Propolis: Response Surface Optimization and Solvent-Dependent Bioactive Properties”. Journal of Apitherapy and Nature, vol. 9, no. 1, May 2026, pp. 76-94, doi:10.35206/jan.1907468.

Vancouver

1.Esen Bilge Biçer, Şeniz Karabıyıklı Çiçek, Ebru Yabaş. Ultrasonic-Assisted Extraction of Propolis: Response Surface Optimization and Solvent-Dependent Bioactive Properties. J.Apit.Nat. 2026 May 1;9(1):76-94. doi:10.35206/jan.1907468

e-ISSN: 2667-4734 Publication Model: Continuous Publication Citation Indexes: TR Dizin

Abstract

Ultrasonic-Assisted Extraction of Propolis: Response Surface Optimization and Solvent-Dependent Bioactive Properties

Abstract

Keywords

Supporting Institution

Project Number

Ethical Statement

Thanks

References

Details

Primary Language

Subjects

Journal Section

Authors

Publication Date

Submission Date

Acceptance Date

Published in Issue

DOI

IZ

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