Encapsulation of Vitamins E and C by Whey Protein Concentrate– Chitosan via Spray Drying: Optimization, Physicochemical Characterization, and Release Rate Analysis

Document Type : Research Article-en

Authors

Pooneh Mojtahedi ¹

Mehdi Varidi ¹

Amir Mohammad Mortazavian ²

¹ Department of Food Science and Technology, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran

² Department of Food Science and Technology, Faculty of Nutrition Sciences and Food Technology, National Nutrition and Food Technology Research Institute, Shahid Beheshti University of Medical Sciences, Tehran, Iran

https://doi.org/10.22067/ifstrj.2026.96109.1497

Abstract

Increasing concerns about human health have increased demand for functional bioactive compounds, such as vitamins E and C. However, their inherent instability and susceptibility to environmental conditions have restricted their direct application in food systems. Encapsulation strategies offer an effective approach to protect these vitamins from degradation and enable controlled release. This study aimed to develop a stable encapsulation system for vitamins E and C using a whey protein concentrate–chitosan (WPC–CS) complex through spray drying, enabling simultaneous delivery of hydrophilic and lipophilic vitamins. Response surface methodology (RSM) was applied to optimize the formulation with the objective of maximizing encapsulation efficiency while minimizing moisture content and hygroscopicity. The optimal microcapsule exhibited encapsulation efficiencies of 93.83% and 93.6% for vitamins E and C, respectively, with a low moisture content (3.17%) and controlled hygroscopicity (13.18%). Release studies demonstrated a phase-dependent behavior: vitamin C showed negligible release in simulated mouth fluid (0.07% after 10 min) but a substantial cumulative release in simulated gastric fluid (85.74% after 120 min). In contrast, vitamin E exhibited a slower and sustained release profile, with 31.45% released after 120 min in simulated gastric fluid and no detectable release under mouth conditions. Scanning electron microscopy (SEM) analysis revealed that the microcapsules had a predominantly spherical morphology with a wrinkled surface, characteristic of protein–polysaccharide matrices formed during rapid drying. Fourier transform infrared spectroscopy (FTIR) confirmed successful encapsulation of both vitamins through interactions between WPC and CS. These findings indicate that WPC–CS complexes produced via spray drying provide an effective co-encapsulation and controlled-release system, enhancing vitamin stability and offering promising applications in functional foods and pharmaceutical formulations.

Keywords

Controlled Release

Microcapsules

Optimize

Spray Drier

Subjects

• Food Chemistry

---

Authors retain the copyright. This is an open access article distributed under Creative Commons Attribution 4.0 International License (CC BY 4.0)

References

1. Agudelo-Chaparro, J., Ciro-Velásquez, H.J., Sepúlveda-Valencia, J.U., & Pérez-Monterroza, E.J. (2022). Microencapsulation of Lactobacillus rhamnosus ATCC 7469 by spray drying using maltodextrin, whey protein concentrate and trehalose. Food Science and Technology International, 28(6), 476-488. https://doi.org/10.1177/10820132211020621
2. Agustinisari, I., Mulia, K., & Nasikin, M. (2020). Physical and chemical properties of encapsulated eugenol using whey protein-maltodextrin mixed solution and chitosan as emulsifier and wall materials. AIP Conference Proceedings, 2255(1), 1-7. https://doi.org/10.1063/5.0013758
3. Anal, A.K., & Singh, H. (2007). Recent advances in microencapsulation of probiotics for industrial applications and targeted delivery. Trends in Food Science & Technology, 18(5), 240-251. https://doi.org/10.1016/j.tifs.2007.01.004
4. AOAC. (2005). Determination of moisture, ash, protein and fat. Official method of analysis of the association of analytical chemists. In: AOAC Washington DC.
5. Askari Vaselabadi, S., Gharibzahedi, S.M.T., Greiner, R., Vale, J.M., Ovenseri, A.C., Rashidinejad, A., & Roohinejad, S. (2025). Advancements in spray‐drying for the microencapsulation of fat‐soluble vitamins: Stability, bioavailability, and applications. Journal of Food Biochemistry, 2025(1), 9974476. https://doi.org/10.1155/jfbc/9974476
6. Asprea, M., Leto, I., Bergonzi, M.C., & Bilia, A.R. (2017). Thyme essential oil loaded in nanocochleates: Encapsulation efficiency, in vitro release study and antioxidant activity. LWT, 77, 497-502. https://doi.org/10.1016/j.lwt.2016.12.006
7. Barbosa, M., Borsarelli, C., & Mercadante, A. (2005). Light stability of spray-dried bixin encapsulated with different edible polysaccharide preparations. Food Research International, 38(8-9), 989-994. https://doi.org/10.1016/j.foodres.2005.02.018
8. Bazaria, B., & Kumar, P. (2016). Effect of whey protein concentrate as drying aid and drying parameters on physicochemical and functional properties of spray dried beetroot juice concentrate. Food Bioscience, 14, 21-27. https://doi.org/10.1016/j.fbio.2015.11.002
9. Budinčić, J.M., Petrović, L., Đekić, L., Fraj, J., Bučko, S., Katona, J., & Spasojević, L. (2021). Study of vitamin E microencapsulation and controlled release from chitosan/sodium lauryl ether sulfate microcapsules. Carbohydrate polymers, 251, 116988. https://doi.org/10.1016/J.CARBPOL.2020.116988
10. Carvalho-Silva, L.B.D. (2013). Physico-chemical properties of milk whey protein agglomerates for use in oral nutritional therapy Food and Nutrition Sciences, 4(9), 69-78. https://doi.org/10.4236/fns.2013.49a210
11. Chang, C., & Nickerson, M.T. (2018). Encapsulation of omega 3-6-9 fatty acids-rich oils using protein-based emulsions with spray drying. Journal of Food Science and Technology, 55(8), 2850-2861. https://doi.org/10.1007/s13197-018-3257-0
12. Chen, H., Mao, L., Hou, Z., Yuan, F., & Gao, Y. (2020). Roles of additional emulsifiers in the structures of emulsion gels and stability of vitamin E. Food Hydrocolloids, 99, 105372. https://doi.org/10.1016/j.foodhyd.2019.105372
13. Chen, X., McClements, D.J., Wang, J., Zou, L., Deng, S., Liu, W., Yan, C., Zhu, Y., Cheng, C., & Liu, C. (2018). Coencapsulation of (−)-Epigallocatechin-3-gallate and quercetin in particle-stabilized W/O/W emulsion gels: Controlled release and bioaccessibility. Journal of Agricultural and Food Chemistry, 66(14), 3691-3699. https://doi.org/10.1021/ACS.JAFC.7B05161
14. Chen, Y., Zhu,, Zhao, Y., Zhang, S., & Wang, W. (2022). Transcriptomics integrated with changes in cell wall material of chestnut (Castanea mollissima Blume) during storage provides a new insight into the “calcification” process. Foods, 11(8), 1136. https://doi.org/10.3390/foods11081136
15. Choi, K.-O., Ryu, J., Kwak, H.-S., & Ko, S. (2010). Spray-dried conjugated linoleic acid encapsulated with Maillard reaction products of whey proteins and maltodextrin. Food Science and Biotechnology, 19(4), 957-965. https://doi.org/10.1007/S10068-010-0134-7
16. Choi, Y.-R., & Chang, Y.H. (2018). Microencapsulation of gallic acid through the complex of whey protein concentrate-pectic polysaccharide extracted from Ulmus davidiana. Food Hydrocolloids, 85, 222-228. https://doi.org/10.1016/j.foodhyd.2018.07.022.
17. Comunian, T.A., Thomazini, M., Alves, A.J.G., de Matos Junior, F.E., de Carvalho Balieiro, J.C., & Favaro-Trindade, C.S. (2013). Microencapsulation of ascorbic acid by complex coacervation: Protection and controlled release. Food Research International, 52(1), 373-379. https://doi.org/10.1016/j.foodres.2013.03.028
18. De Queiroz, J.L.C., Costa, R.O.D.A., Matias, L.L.R., De Medeiros, A.F., Gomes, A.F.T., Pais, T.D.S., Passos, T.S., Maciel, B.L.L., Dos Santos, E.A., & Morais, A.H.D.A. (2018). Chitosan-whey protein nanoparticles improve encapsulation efficiency and stability of a trypsin inhibitor isolated from Tamarindus indica Food Hydrocolloids, 84, 247-256. https://doi.org/10.1016/j.foodhyd.2018.06.010
19. Delaporte, A., Duchemin, B., Grisel, M., & Gore, E. (2024). Impact of wall material-to-active ratio in the stability of spray-dried ascorbic acid using maltodextrin and gum arabic. Molecules, 29(15), 3587. https://doi.org/10.3390/molecules29153587
20. Desai, K., & Park, H. (2005). Encapsulation of vitamin C in tripolyphosphate cross-linked chitosan microspheres by spray drying. Journal of Microencapsulation, 22(2), 179-192. https://doi.org/10.1080/02652040400026533
21. Desai, K., & Park, H.J. (2006). Effect of manufacturing parameters on the characteristics of vitamin C encapsulated tripolyphosphate-chitosan microspheres prepared by spray-drying. Journal of Microencapsulation, 23(1), 91-103. https://doi.org/10.1080/02652040500435436
22. Díaz-Montes, E. (2023). Wall materials for encapsulating bioactive compounds via spray-drying: A review. Polymers, 15(12), 2659. https://doi.org/10.3390/polym15122659
23. Dickinson, E., Radford, S.J., & Golding, M. (2003). Stability and rheology of emulsions containing sodium caseinate: combined effects of ionic calcium and non-ionic surfactant. Food Hydrocolloids, 17(2), 211-220. https://doi.org/10.1016/S0268-005X(02)00055-3
24. Fernandes, R.V.D.B., Borges, S.V., Botrel, D.A., & de Oliveira, C.R. (2014). Physical and chemical properties of encapsulated rosemary essential oil by spray drying using whey protein–inulin blends as carriers. International Journal of Food Science and Technology, 49(6), 1522-1529. https://doi.org/10.1111/ijfs.12449
25. Frascareli, E., Silva, V., Tonon, R., & Hubinger, M. (2012). Effect of process conditions on the microencapsulation of coffee oil by spray drying. Food and Bioproducts Processing, 90(3), 413-424. https://doi.org/10.1016/j.fbp.2011.12.002
26. Guldiken, B., Linke, A., Capanoglu, E., Boyacioglu, D., Kohlus, R., Weiss, J., & Gibis, M. (2019). Formation and characterization of spray dried coated and uncoated liposomes with encapsulated black carrot extract. Journal of Food Engineering, 246, 42-50. https://doi.org/10.1016/j.jfoodeng.2018.10.025
27. Hinnenkamp, C., Reineccius, G., & Ismail, B.P. (2021). Efficient encapsulation of fish oil: Capitalizing on the unique inherent characteristics of whey cream and hydrolyzed whey protein. Journal of Dairy Science, 104(6), 6472-6486. https://doi.org/10.3168/jds.2020-19880
28. Hu, Y., He, C., Jiang, C., Liao, Y., Xiong, H., & Zhao, Q. (2020). Complexation with whey protein fibrils and chitosan: A potential vehicle for curcumin with improved aqueous dispersion stability and enhanced antioxidant activity. Food Hydrocolloids, 104, 105729. https://doi.org/10.1016/j.foodhyd.2020.105729
29. Katouzian, I., & Jafari, S.M. (2016). Nano-encapsulation as a promising approach for targeted delivery and controlled release of vitamins. Trends in Food Science & Technology, 53, 34-48. https://doi.org/10.1016/j.tifs.2016.05.002
30. Klaypradit, W., & Huang, Y.-W. (2008). Fish oil encapsulation with chitosan using ultrasonic atomizer. LWT-Food Science and Technology, 41(6), 1133-1139. https://doi.org/10.1016/j.lwt.2007.06.014
31. Lan, Y., Ohm, J.-B., Chen, B., & Rao, J. (2021). Microencapsulation of hemp seed oil by pea protein isolate− sugar beet pectin complex coacervation: Influence of coacervation pH and wall/core ratio. Food Hydrocolloids, 113, 106423. https://doi.org/10.1016/j.foodhyd.2020.106423
32. Lekshmi, R.K., Rahima, M., Chatterjee, N., Tejpal, C., Anas, K., Vishnu, K., Sarika, K., Asha, K., Anandan, R., & Suseela, M. (2019). Chitosan–Whey protein as efficient delivery system for squalene: Characterization and functional food application. International Journal of Biological Macromolecules, 135, 855-863. https://doi.org/10.1016/j.ijbiomac.2019.05.153
33. Li, K.-Y., Zhou, Y., Huang, G.-Q., Li, X.-D., & Xiao, J.-X. (2022). Preparation of powdered oil by spray drying the Pickering emulsion stabilized by ovalbumin–gum Arabic polyelectrolyte complex. Food Chemistry, 391, 133223. https://doi.org/10.1016/J.FOODCHEM.2022.133223
34. Liu, Q., Jing, Y., Han, C., Zhang, H., & Tian, Y. (2019). Encapsulation of curcumin in zein/caseinate/sodium alginate nanoparticles with improved physicochemical and controlled release properties. Food Hydrocolloids, 93, 432-442. https://doi.org/10.1016/j.foodhyd.2019.02.003
35. Luo, Y., Zhang, B., Whent, M., Yu, L.L., & Wang, Q. (2011). Preparation and characterization of zein/chitosan complex for encapsulation of α-tocopherol, and its in vitro controlled release study. Colloids and Surfaces B: Biointerfaces, 85(2), 145-152. https://doi.org/10.1016/J.COLSURFB.2011.02.020
36. Mohammadi, N., Ehsani, M.R., & Bakhoda, H. (2018). Design and evaluation of the release characteristics of caffeine-loaded microcapsules in a medicated chewing gum formulation. Food Biophysics, 13(3), 240-249. https://doi.org/10.1007/s11483-018-9530-y
37. Mojtahedi, P., Mortazavian, S.A.M., & Varidi, M. (2023). Encapsulation of vitamins E and C in whey protein concentrate-chitosan emulation: Physicochemical properties. Management Strategies and Engineering Sciences, 5(4), 136-146. https://msesj.com/index.php/mses/article/view/183
38. Muley, A.B., & Singhal, R.S. (2020). Extension of postharvest shelf life of strawberries (Fragaria ananassa) using a coating of chitosan-whey protein isolate conjugate. Food Chemistry, 329, 12-72. https://org/10.1016/j.foodchem.2020.127213
39. Oliveira, N.L., Espinal-Ruiz, M., Neves, I.C.O., Silva, S.H., de Resende, J.V., & Rogers, M.A. (2023). Evaluation of α-tocopherol microencapsulation stability with either coconut oil or canola oil cores in Greek yogurt and butter. Food Chemistry Advances, 2, 100277. https://doi.org/10.1016/j.focha.2023.100277
40. Pang, Y., Qin, A., Lin, X., Yang, L., Wang, Q., Wang, Z., Shan, Z., Li, S., Wang, J., & Fan, S. (2017). Biodegradable and biocompatible high elastic chitosan scaffold is cell-friendly both in vitro and in vivo. Oncotarget, 8(22), 35583. https://doi:18632/oncotarget.14709
41. Pateiro, M., Gómez, B., Munekata, P.E., Barba, F.J., Putnik, P., Kovačević, D.B., & Lorenzo, J.M. (2021). Nanoencapsulation of promising bioactive compounds to improve their absorption, stability, functionality and the appearance of the final food products. Molecules, 26(6), 1547. https://doi.org/10.3390/molecules26061547
42. Pinto, J.T., Faulhammer, E., Dieplinger, J., Dekner, M., Makert, C., Nieder, M., & Paudel, A. (2021). Progress in spray-drying of protein pharmaceuticals: Literature analysis of trends in formulation and process attributes. Drying Technology, 39(11), 1415-1446. https://doi.org/10.1080/07373937.2021.1903032
43. Pudjiastuti, P., Wafiroh, S., Hendradi, E., Darmokoesoemo, H., Harsini, M., Fauzi, M.A.R.D., Nahar, L., & Sarker, S.D. (2020). Disintegration, in vitro dissolution, and drug release kinetics profiles of k-carrageenan-based nutraceutical hard-shell capsules containing salicylamide. Open Chemistry, 18(1), 226-231. https://doi.org/10.1515/chem-2020-0028
44. Rabkin, B., Tirosh, O., & Kanner, J. (2022). Reactivity of vitamin E as an antioxidant in red meat and the stomach medium. Journal of Agricultural and Food Chemistry, 70(38), 12172-12179. https://doi.org/10.1021/ACS.JAFC.2C03674
45. Rot, T., Kovačević, D., Habschied, K., & Mastanjević, K. (2025). N-nitrosamines in meat products: formation, detection and regulatory challenges. Processes, 13(5), 1555. https://doi.org/10.3390/pr13051555
46. Schmitt, J.M., Baumann, J.M., & Morgen, M.M. (2022). Predicting spray dried dispersion particle size via machine learning regression methods: Schmitt, Baumann and Morgen. Pharmaceutical Research, 39(12), 3223-3239. https://doi.org/10.1007/s11095-022-03370-3
47. Somchue, W., Sermsri, W., Shiowatana, J., & Siripinyanond, A. (2009). Encapsulation of α-tocopherol in protein-based delivery particles. Food Research International, 42(8), 909-914. https://doi.org/10.1016/J.FOODRES.2009.04.021
48. Stabrauskiene, J., Pudziuvelyte, L., & Bernatoniene, J. (2024). Optimizing encapsulation: Comparative analysis of spray-drying and freeze-drying for sustainable recovery of bioactive compounds from Citrus x paradisi peels. Pharmaceuticals, 17(5), 596. https://doi.org/10.3390/ph17050596
49. Suwannasang, S., Zhong, Q., Thumthanaruk, B., Uttapap, D., Puttanlek, C., Vatanyoopaisarn, S., & Rungsardthong, V. (2022). Optimization of wall material composition for production of spray-dried Sacha inchi oil microcapsules with desirable physicochemical properties. Food and Bioprocess Technology, 15(11), 2499-2514. https://doi.org/10.1007/s11947-022-02893-2
50. Tan, Y., Li, R., Liu, C., Mundo, J.M., Zhou, H., Liu, J., & McClements, D.J. (2020). Chitosan reduces vitamin D bioaccessibility in food emulsions by binding to mixed micelles. Food & Function, 11(1), 187-199. https://doi.org/10.1039/C9FO02164G
51. Wang, F., & Mutukumira, A.N. (2022). Microencapsulation of Limosilactobacillus reuteri DPC16 by spray drying using different encapsulation wall materials. Journal of Food Processing and Preservation, 46(10), e16880. https://doi.org/10.1111/jfpp.16880
52. Wang, L., Gao, Y., Li, J., Subirade, M., Song, Y., & Liang, L. (2016). Effect of resveratrol or ascorbic acid on the stability of α-tocopherol in O/W emulsions stabilized by whey protein isolate: Simultaneous encapsulation of the vitamin and the protective antioxidant. Food Chemistry, 196, 466-474. https://doi.org/10.1016/j.foodchem.2015.09.071
53. Wang, S., Ye, X., Sun, Y., Liang, J., Yue, P., & Gao, X. (2021). Nanocomplexes derived from chitosan and whey protein isolate enhance the thermal stability and slow the release of anthocyanins in simulated digestion and prepared instant coffee. Food Chemistry, 336, 127707. https://doi.org/10.1016/j.foodchem.2020.127707
54. Xu, W., Lv, K., Mu, W., Zhou, S., & Yang, Y. (2021). Encapsulation of α-tocopherol in whey protein isolate/chitosan particles using oil-in-water emulsion with optimal stability and bioaccessibility. LWT, 148, 111724. https://doi.org/10.1016/j.lwt.2021.111724
55. Xu, W., Tang, Y., Yang, Y., Wang, G., & Zhou, S. (2020). Establishment of a stable complex formed from whey protein isolate and chitosan and its stability under environmental stresses. International Journal of Biological Macromolecules, 165, 2823-2833. https://doi.org/10.1016/j.ijbiomac.2020.10.130
56. Yousefi, S., Rajaei, P., Nateghi, L., Nodeh, H.R., & Rashidi, L. (2023). Encapsulation of sesamol and retinol using alginate and chitosan-coated W/O/W multiple emulsions containing Tween 80 and Span 80. International Journal of Biological Macromolecules, 242, 124766. https://doi.org/10.1016/j.ijbiomac.2023.124766