Development of Chitosan-Stabilized Pickering Emulsions Loaded with Supercritical Fluid Extraction of Honey Bee Pollen for Functional Food Applications
DOI:
https://doi.org/10.24925/turjaf.v14i2.519-525.8620Keywords:
Honey bee pollen , Pickering emulsion , Chitosan , Functional food , Supercritical fluid extractAbstract
Honey bee pollen (HBP) is a nutritionally rich natural product containing phenolics and antioxidants; however, its rigid outer wall limits the bioavailability of these bioactive compounds. Therefore advanced extraction and delivery systems are required to enhance stability and functional performance for food applications. This study aimed to encapsulate a supercritical fluid extract (SFE) of honey bee pollen within chitosan-stabilized Pickering emulsions in order to improve encapsulation efficiency and antioxidant activity. Honey bee pollen obtained from Bingöl, Türkiye, was extracted with supercritical CO₂. Pickering emulsions were prepared via ultrasound-assisted emulsification in the presence of chitosan as a stabilizer, with SFE-HBP at 5%, 10%, and 15% (w/v). The prepared emulsions were characterized for their microstructure, particle size, zeta potential, and encapsulation efficiency. Total phenolic content (TPC) and antioxidant activities were assessed using the Folin–Ciocalteu and CUPRAC assays, respectively. All formulations yielded stable, uniformly distributed Pickering emulsions with spherical droplets and no visible phase separation. Indeed, encapsulation efficiency increased with increasing SFE-HBP concentration, from 73.96% to 87.06%. The particle size ranges (7.22–12.44 µm) and zeta potential values (+21.73 to +28.24 mV) indicate good colloidal stability. For both TPC and CUPRAC antioxidant activities, a concentration-dependent increase was observed, with the highest values in the 15% SFE-HBP-containing emulsions. Collectively, these findings indicate that a chitosan-based Pickering emulsion is an effective delivery system for SFE-derived bioactive compounds from honey bee pollen. The method provides enhanced encapsulation efficiency, antioxidant capacity, and physicochemical stability, hence exhibiting great potential for functional food and nutraceutical applications.
References
Aguiar, J., Estevinho, B. N., & Santos, L. (2016). Microencapsulation of natural antioxidants for food application – The specific case of coffee antioxidants – A review. Trends in Food Science and Technology, 58, 21–39. https://doi.org/10.1016/j.tifs.2016.10.012
Alshallash, K. S., Abolaban, G., Elhamamsy, S. M., Zaghlool, A., Nasr, A., Nagib, A., El-Hakim, A. F. A., Zahra, A. A., Hamdy, A. E., & Taha, I. M. (2023). Bee Pollen as a Functional Product – Chemical Constituents and Nutritional Properties. Journal of Ecological Engineering, 24(2), 173–183. https://doi.org/10.12911/22998993/156611
Alshehri, K. M., & Abdella, E. M. (2023). Development of ternary nanoformulation comprising bee pollen-thymol oil extracts and chitosan nanoparticles for anti-inflammatory and anticancer applications. International Journal of Biological Macromolecules, 242(P1), 124584. https://doi.org/10.1016/j.ijbiomac.2023.124584
Apak, R., Güçlü, K., Özyürek, M., & Karademir, S. E. (2004). Novel total antioxidant capacity index for dietary polyphenols and vitamins C and E, using their cupric ion reducing capability in the presence of neocuproine: CUPRAC method. Journal of Agricultural and Food Chemistry, 52(26), 7970–7981. https://doi.org/10.1021/jf048741x
Aranaz, I., Alcántara, A. R., Civera, M. C., Arias, C., Elorza, B., Caballero, A. H., & Acosta, N. (2021). Chitosan: An overview of its properties and applications. Polymers, 13(19). https://doi.org/10.3390/polym13193256
Arserim-Uçar, D. K. (2024). Engineered chitosan nanoparticles for encapsulation of thymol. The Journal of Food, 49, 1148–1161. https://doi.org/10.15237/gida.GD24055
Arserim-Uçar, D. K., & Çabuk, B. (2020). Emerging antibacterial and antifungal applications of nanomaterials on food products. Nanotoxicity: Prevention and Antibacterial Applications of Nanomaterials, 415–453. https://doi.org/10.1016/B978-0-12-819943-5.00027-0
Arserim-Uçar, D. K. (2020). Nanocontainers for food safety. Smart Nanocontainers, 105-117. https://doi.org/10.1016/B978-0-12-816770-0.00007-1
Aylanc, V., Falcão, S. I., & Vilas-Boas, M. (2023). Bee pollen and bee bread nutritional potential: Chemical composition and macronutrient digestibility under in vitro gastrointestinal system. Food Chemistry, 413, 135597. https://doi.org/https://doi.org/10.1016/j.foodchem.2023.135597
Baky, M. H., Abouelela, M. B., Wang, K., & Farag, M. A. (2023). Bee Pollen and Bread as a Super-Food: A Comparative Review of Their Metabolome Composition and Quality Assessment in the Context of Best Recovery Conditions. Molecules, 28(2), 715. https://doi.org/10.3390/molecules28020715
Baysal, G., Olcay, H. S., Keresteci, B., & Özpinar, H. (2022). The antioxidant and antibacterial properties of chitosan encapsulated with the bee pollen and the apple cider vinegar. Journal of Biomaterials Science, Polymer Edition, 33(8), 995–1011. https://doi.org/10.1080/09205063.2022.2031463
Beyki, M., Zhaveh, S., Khalili, S. T., Rahmani-Cherati, T., Abollahi, A., Bayat, M., Tabatabaei, M., & Mohsenifar, A. (2014). Encapsulation of Mentha piperita essential oils in chitosan-cinnamic acid nanogel with enhanced antimicrobial activity against Aspergillus flavus. Industrial Crops and Products, 54, 310–319. https://doi.org/10.1016/j.indcrop.2014.01.033
Campos, M. G. R., Bogdanov, S., de Almeida-Muradian, L. B., Szczesna, T., Mancebo, Y., Frigerio, C., & Ferreira, F. (2008). Pollen composition and standardisation of analytical methods. Journal of Apicultural Research, 47(2), 154–161. https://doi.org/10.1080/00218839.2008.11101443
Čeksterytė, V., Bliznikas, S., & Jaškūnė, K. (2023). The Composition of Fatty Acids in Bee Pollen, Royal Jelly, Buckthorn Oil and Their Mixtures with Pollen Preserved for Storage. Foods, 12(17), 3164. https://doi.org/10.3390/foods12173164
de Moura, S. C. S. R., Berling, C. L., Germer, S. P. M., Alvim, I. D., & Hubinger, M. D. (2018). Encapsulating anthocyanins from Hibiscus sabdariffa L. calyces by ionic gelation: Pigment stability during storage of microparticles. Food Chemistry, 241, 317–327. https://doi.org/https://doi.org/10.1016/j.foodchem.2017.08.095
de Souza Simões, L., Madalena, D. A., Pinheiro, A. C., Teixeira, J. A., Vicente, A. A., & Ramos, Ó. L. (2017). Micro- and nano bio-based delivery systems for food applications: In vitro behavior. Advances in Colloid and Interface Science, 243, 23–45. https://doi.org/10.1016/j.cis.2017.02.010
Denisow, B., & Denisow‐Pietrzyk, M. (2016). Biological and therapeutic properties of bee pollen: a review. Journal of the Science of Food and Agriculture, 96(13), 4303–4309. https://doi.org/10.1002/jsfa.7729
Dewettinck, K., & Huyghebaert, A. (1999). Fluidized bed coating in food technology. Trends in Food Science & Technology, 10(4–5), 163–168. https://doi.org/10.1016/S0924-2244(99)00041-2
Ecem Bayram, N. (2021). Vitamin, mineral, polyphenol, amino acid profile of bee pollen from Rhododendron ponticum (source of “mad honey”): nutritional and palynological approach. Journal of Food Measurement and Characterization, 15(3), 2659–2666. https://doi.org/10.1007/s11694-021-00854-5
El Ghouizi, A., Bakour, M., Laaroussi, H., Ousaaid, D., El Menyiy, N., Hano, C., & Lyoussi, B. (2023). Bee Pollen as Functional Food: Insights into Its Composition and Therapeutic Properties. Antioxidants, 12(3), 557. https://doi.org/10.3390/antiox12030557
El-Kholy, W. M., Soliman, T. N., & Darwish, A. M. G. (2019). Evaluation of date palm pollen (Phoenix dactylifera L.) encapsulation, impact on the nutritional and functional properties of fortified yoghurt. PLOS ONE, 14(10), e0222789. https://doi.org/10.1371/journal.pone.0222789
Erol, E. (2024). Quantitative Analysis of Bioaccessible Phenolic Compounds in Aegean Bee Bread Using LC-HRMS Coupled with a Human Digestive System Model. Chemistry and Biodiversity, 21(3). https://doi.org/10.1002/cbdv.202301497
Fameau, A.-L., Arnould, A., & Saint-Jalmes, A. (2014). Responsive self-assemblies based on fatty acids. Current Opinion in Colloid & Interface Science, 19(5), 471–479. https://doi.org/10.1016/j.cocis.2014.08.005
Gardana, C., Del Bo, C., Quicazán, M. C., Corrrea, A. R., & Simonetti, P. (2018). Nutrients, phytochemicals and botanical origin of commercial bee pollen from different geographical areas. Journal of Food Composition and Analysis, 73, 29–38. https://doi.org/10.1016/j.jfca.2018.07.009
Harman, C. L. G., Patel, M. A., Guldin, S., & Davies, G. (2019). ScienceDirect Recent developments in Pickering emulsions for biomedical applications. Current Opinion in Colloid & Interface Science, 39, 173–189. https://doi.org/10.1016/j.cocis.2019.01.017
Hemmingsen, L. M., Boracchia, L., Hagen, N. E., Guareschi, F., Sonvico, F., Škalko-basnet, N., Myrseth, L., Boracchia, L., & Hagen, N. E. (2025). Lecithin-chitosan nanoparticles for co-delivery of curcumin and β-caryophyllene — potential applications in chronic wound care. Journal of Biomaterials Science, Polymer Edition, 1–25. https://doi.org/10.1080/09205063.2025.2582737
Hızır-Kadı, İ., Gültekin-Özgüven, M., Altin, G., Demircan, E., & Özçelik, B. (2020). Liposomal nanodelivery systems generated from proliposomes for pollen extract with improved solubility and in vitro bioaccessibility. Heliyon, 6(9), e05030. https://doi.org/10.1016/j.heliyon.2020.e05030
Hsieh, W. C., Chang, C. P., & Gao, Y. L. (2006). Controlled release properties of Chitosan encapsulated volatile Citronella Oil microcapsules by thermal treatments. Colloids and Surfaces B: Biointerfaces, 53(2), 209–214. https://doi.org/10.1016/j.colsurfb.2006.09.008
Iber, B. T., Kasan, N. A., Torsabo, D., & Omuwa, J. W. (2022). A review of various sources of chitin and chitosan in nature. Journal of Renewable Materials, 10(4), 1097–1123. https://doi.org/10.32604/JRM.2022.018142
Kim, S. B., Jo, Y. H., Liu, Q., Ahn, J. H., Hong, I. P., Han, S. M., Hwang, B. Y., & Lee, M. K. (2015). Optimization of extraction condition of bee pollen using response surface methodology: Correlation between anti-melanogenesis, antioxidant activity, and phenolic content. Molecules, 20(11), 19764–19774. https://doi.org/10.3390/molecules201119656
Klinger, C., Żółtowska-Aksamitowska, S., Wysokowski, M., Tsurkan, M. V., Galli, R., Petrenko, I., Machałowski, T., Ereskovsky, A., Martinović, R., Muzychka, L., Smolii, O. B., Bechmann, N., Ivanenko, V., Schupp, P. J., Jesionowski, T., Giovine, M., Joseph, Y., Bornstein, S. R., Voronkina, A., & Ehrlich, H. (2019). Express Method for Isolation of Ready-to-Use 3D Chitin Scaffolds from Aplysina archeri (Aplysineidae: Verongiida) Demosponge. Marine Drugs, 17(2), 131. https://doi.org/10.3390/md17020131
Klojdová, I., Milota, T., Smetanová, J., & Stathopoulos, C. (2023). Encapsulation: A Strategy to Deliver Therapeutics and Bioactive Compounds? Pharmaceuticals, 16(3), 1–19. https://doi.org/10.3390/ph16030362
Komosinska-Vassev, K., Olczyk, P., Kaźmierczak, J., Mencner, L., & Olczyk, K. (2015). Bee pollen: Chemical composition and therapeutic application. Evidence-Based Complementary and Alternative Medicine, 2015. https://doi.org/10.1155/2015/297425
Köppler, K., Vorwohl, G., & Koeniger, N. (2007). Comparison of pollen spectra collected by four different subspecies of the honey bee Apis mellifera. Apidologie, 38(4), 341–353. https://doi.org/10.1051/apido:2007020
Malagnini, V., Cappellari, A., Marini, L., Zanotelli, L., Zorer, R., Angeli, G., Ioriatti, C., & Fontana, P. (2022). Seasonality and Landscape Composition Drive the Diversity of Pollen Collected by Managed Honey Bees. Frontiers in Sustainable Food Systems, 6, 1–9. https://doi.org/10.3389/fsufs.2022.865368
Maqsoudlou, A., Sadeghi Mahoonak, A., Mohebodini, H., & Koushki, V. (2020). Stability and structural properties of bee pollen protein hydrolysate microencapsulated using maltodextrin and whey protein concentrate. Heliyon, 6(5), e03731. https://doi.org/10.1016/j.heliyon.2020.e03731
Mcclements, D. J. (2012). Advances in fabrication of emulsions with enhanced functionality using structural design principles. Current Opinion in Colloid & Interface Science, 17(5), 235–245. https://doi.org/10.1016/j.cocis.2012.06.002
Munin, A., & Edwards-Lévy, F. (2011). Encapsulation of Natural Polyphenolic Compounds; a Review. Pharmaceutics, 3(4), 793–829. https://doi.org/10.3390/pharmaceutics3040793
Mwangi, W. W., Ho, K. W., Tey, B. T., & Chan, E. S. (2016). Effects of environmental factors on the physical stability of pickering-emulsions stabilized by chitosan particles. Food Hydrocolloids, 60, 543–550. https://doi.org/10.1016/j.foodhyd.2016.04.023
Nedovic, V., Kalusevic, A., Manojlovic, V., Levic, S., & Bugarski, B. (2011). An overview of encapsulation technologies for food applications. Procedia Food Science, 1, 1806–1815. https://doi.org/10.1016/j.profoo.2011.09.265
Ntohogian, S., Gavriliadou, V., Christodoulou, E., Nanaki, S., Lykidou, S., Naidis, P., Mischopoulou, L., Barmpalexis, P., Nikolaidis, N., & Bikiaris, D. N. (2018). Chitosan nanoparticles with encapsulated natural and UF-purified annatto and saffron for the preparation of UV protective cosmetic emulsions. Molecules, 23(9), 2107.
Prđun, S., Svečnjak, L., Valentić, M., Marijanović, Z., & Jerković, I. (2021). Characterization of bee pollen: Physico-chemical properties, headspace composition and FTIR spectral profiles. Foods, 10(9), 2103. https://doi.org/10.3390/foods10092103
Sagona, S., Pozzo, L., Peiretti, P. G., Biondi, C., Giusti, M., Gabriele, M., Pucci, L., & Felicioli, A. (2017). Palynological origin, chemical composition, lipid peroxidation and fatty acid profile of organic Tuscanian bee-pollen. Journal of Apicultural Research, 56(2), 136–143. https://doi.org/10.1080/00218839.2017.1287995
Sarkar, A., & Dickinson, E. (2020). Sustainable food-grade Pickering emulsions stabilized by plant-based particles. Current Opinion in Colloid & Interface Science, 49, 69–81. https://doi.org/10.1016/j.cocis.2020.04.004
Schröder, A., Sprakel, J., Schroën, K., Spaen, J. N., & Berton-Carabin, C. C. (2018). Coalescence stability of Pickering emulsions produced with lipid particles: A microfluidic study. Journal of Food Engineering, 234, 63–72. https://doi.org/10.1016/j.jfoodeng.2018.04.007
Sharkawy, A., Barreiro, M. F., & Rodrigues, A. E. (2020). Chitosan-based Pickering emulsions and their applications: A review. Carbohydrate Polymers, 250, 116885. https://doi.org/10.1016/j.carbpol.2020.116885
Slinkard, K., & Singleton, V. (1977). Total phenol analysis: automation and comparison with manual methods. American Journal of Enology and Viticulture, 28(1), 49–55. https://doi.org/10.5344/ajev.1977.28.1.49
Song, T., Xiong, Z., Shi, T., Yuan, L., & Gao, R. (2022). Effect of glutamic acid on the preparation and characterization of Pickering emulsions stabilized by zein. Food Chemistry, 366, 130598. https://doi.org/10.1016/j.foodchem.2021.130598
Sotelo-Boyás, M. E., Correa-Pacheco, Z. N., Bautista-Baños, S., & Corona-Rangel, M. L. (2017). Physicochemical characterization of chitosan nanoparticles and nanocapsules incorporated with lime essential oil and their antibacterial activity against food-borne pathogens. LWT, 77, 15–20. https://doi.org/https://doi.org/10.1016/j.lwt.2016.11.022
Tackenberg, M. W., Krauss, R., Schuchmann, H. P., & Kleinebudde, P. (2015). Encapsulation of orange terpenes investigating a plasticisation extrusion process. Journal of Microencapsulation, 32(4), 408–417. https://doi.org/10.3109/02652048.2015.1035686
Thakur, M., Pant, K., Naik, R. R., & Nanda, V. (2020). Optimization of spray drying operating conditions for production of functional milk powder encapsulating bee pollen. Drying Technology, 39(6), 1–14. https://doi.org/10.1080/07373937.2020.1720225
Tiwari, M., Basavaraj, M. G., & Dugyala, V. R. (2024). Pickering Emulsions: Role of Particle Wettability and Adhesive Force on Droplet Bridging. Langmuir, 40(50), 26474–26486. https://doi.org/10.1021/acs.langmuir.4c03241
Velderrain-Rodríguez, G. R., Acevedo-Fani, A., González-Aguilar, G. A., & Martín-Belloso, O. (2019). Encapsulation and stability of a phenolic-rich extract from mango peel within water-in-oil-in-water emulsions. Journal of Functional Foods, 56, 65–73. https://doi.org/10.1016/j.jff.2019.02.045
Xu, X., Sun, L., Dong, J., & Zhang, H. (2009). Breaking the cells of rape bee pollen and consecutive extraction of functional oil with supercritical carbon dioxide. Innovative Food Science & Emerging Technologies, 10(1), 42–46. https://doi.org/10.1016/j.ifset.2008.08.004
Xu, Y., Yan, X., Zheng, H., Li, J., Wu, X., Xu, J., Zhen, Z., & Du, C. (2024). The application of encapsulation technology in the food Industry: Classifications, recent Advances, and perspectives. Food Chemistry: X, 21, 101240. https://doi.org/10.1016/j.fochx.2024.101240
Xue, X., Zhao, J., Chen, L., Zhou, J., Yue, B., Li, Y., Wu, L., & Liu, F. (2012). Analysis of coenzyme Q10 in bee pollen using online cleanup by accelerated solvent extraction and high performance liquid chromatography. Food Chemistry, 133(2), 573–578. https://doi.org/10.1016/j.foodchem.2011.12.085
Yang, K., Wu, D., Ye, X., Liu, D., Chen, J., & Sun, P. (2013). Characterization of Chemical Composition of Bee Pollen in China. Journal of Agricultural and Food Chemistry, 61(3), 708–718. https://doi.org/10.1021/jf304056b
Zhang, M., Li, X., Zhou, L., Chen, W., & Marchioni, E. (2023). Protein-Based High Internal Phase Pickering Emulsions: A Review of Their Fabrication, Composition and Future Perspectives in the Food Industry. Foods. https://doi.org/10.3390/foods12030482
Zhaveh, S., Mohsenifar, A., Beiki, M., Khalili, S. T., Abdollahi, A., Rahmani-Cherati, T., & Tabatabaei, M. (2015). Encapsulation of Cuminum cyminum essential oils in chitosan-caffeic acid nanogel with enhanced antimicrobial activity against Aspergillus flavus. Industrial Crops and Products, 69, 251–256. https://doi.org/10.1016/j.indcrop.2015.02.028
Zuluaga, C. M., Serrato, J. C., & Quicazan, M. C. (2015). Bee-pollen structure modification by physical and biotechnological processing: Influence on the availability of nutrients and bioactive compounds. Chemical Engineering Transactions, 43, 79–84. https://doi.org/10.3303/CET1543014
Downloads
Published
Issue
Section
License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.






