Valorization and Optimization of Agro-Food By-Products from Punica granatum Bark and Juglans regia Husk Combined with Syzygium aromaticum for Enhanced Antioxidant and Anti-Inflammatory Activities: A D-Optimal Mixture Design Approach

Background: The sustainable valorization of agri-food processing by-products has emerged as a scientifically promising strategy for the development of natural functional ingredients rich in bioactive compounds, contributing simultaneously to the circular bioeconomy and the formulation of health-promoting food and nutraceutical products.

Aims: The present study aimed to optimize a multi-component plant-derived formulation comprising Syzygium aromaticum (clove), Punica granatum bark (pomegranate), and Juglans regia husk (walnut) employing a D-optimal ABCD mixture design.

Materials and Methods: A ten-run D-optimal ABCD mixture design was employed to optimize the proportions of three plant-derived powders — Syzygium aromaticum (clove), Punica granatum bark, and Juglans regia husk — with TPC, TFC, and DPPH radical scavenging activity as response variables. The optimized formulation was characterized by LC-MS/MS for phenolic compound profiling. Antioxidant capacity was assessed using DPPH, ABTS, FRAP, and ferrous ion chelating assays. Anti-inflammatory activity was evaluated by BSA denaturation inhibition, and cytotoxicity by hemolytic activity assay on human erythrocytes.

Results: The optimal formulation comprised 58% pomegranate bark, 30% clove and 12% walnut husk. LC-MS/MS revealed that cloves was the richest source of acid (7,947.87 ± 255.68 µg/g) and quinic acid (5,235.29 ± 79.53 µg/g), pomegranate bark was characterized by catechin, epicatechin, and citric acid, and walnut husk by catechin, quercetin, and citric acid. The optimized mixture revealed a more enriched phenolic profile than the individual extracts, with substantial levels of rutin, kaempferol, naringenin, catechin, and epicatechin concentrations, suggesting synergistic phytochemical interactions. IC₅₀ values for DPPH, ABTS, ferrous iron chelating and BSA denaturation inhibition were 50.97 ± 0.31, 94.84 ± 1.34, 71.15 ± 0.72, and 4.33 ± 0.15 µg/mL, respectively; EC₅₀ for ferric reducing power was 170.83 ± 0.58 µg/ml. Hemolytic activity was negligible (13.38%) at the highest concentration tested (5 mg/mL), confirming erythrocyte biocompatibility.

Conclusions: The present study demonstrates that D-optimal mixture design constitutes an effective methodological strategy for the optimization of multi-component plant formulations, enabling the identification of synergistic combinations with enhanced phytochemical composition and biological activities.

Keywords

Syzygium aromaticum Punica granatum Bark Juglans regia Husk Phytochemical Interactions D-Optimal Mixture Design Antioxidant Activity Anti-Inflammatory Activity
  • Melissa Takka
    University of Béjaïa
  • Sofiane Dairi
    University of Jijel; University of Béjaïa
  • Sabrina Zeghichi-Hamri
    University of Béjaïa
  • Omar Aoun
    University Mohamed Boudiaf of M'sila
  • Nabil Kadri
    University of Bouira
  • Nabila Brahmi-Chendouh
    University of Béjaïa
  • Baya Ladjouze
    University of Béjaïa
  • Ouardia Bouallag
    University of Béjaïa
  • University of Béjaïa
  • Noelia Duarte
    University of Lisbon
  • Khodir Madani
    University of Béjaïa
  • Lila Boulekbache
    University of Béjaïa

How to Cite

Takka, M., Dairi, S., Zeghichi-Hamri, S., Aoun, O., Kadri, N., Brahmi-Chendouh, N., Ladjouze, B., Bouallag, O., Adel, K., Duarte, N., Madani, K., & Boulekbache, L. (2026). Valorization and Optimization of Agro-Food By-Products from Punica granatum Bark and Juglans regia Husk Combined with Syzygium aromaticum for Enhanced Antioxidant and Anti-Inflammatory Activities: A D-Optimal Mixture Design Approach. The North African Journal of Food and Nutrition Research, 9(SI), S98-S114. https://doi.org/10.51745/najfnr.9.SI.S98-S114

Abbou, A., Kadri, N., Debbache, N., Dairi, S., Remini, H., Belbahi, A., Adel, K., Dahmoune, F., & Madani, K. (2019). Effect of precipitation solvent on some biological activities of polysaccharides from Pinus halepensis Mill. seeds. International Journal of Biological Macromolecules, *141*, 663–670. https://doi.org/10.1016/j.ijbiomac.2019.08.266 DOI: https://doi.org/10.1016/j.ijbiomac.2019.08.266

Alam, M. N., Bristi, N. J., & Rafiquzzaman, M. (2013). Review on in vivo and in vitro methods evaluation of antioxidant activity. Saudi Pharmaceutical Journal, 21(2), 143–152. https://doi.org/10.1016/j.jsps.2012.05.002 DOI: https://doi.org/10.1016/j.jsps.2012.05.002

Allane, T., & Benamara, S. (2010). Activité antioxydantes de quelques fruits communs et sauvages d'Algérie. Phytothérapie, 8(3), 171–175. https://doi.org/10.1007/s10298-010-0553-x DOI: https://doi.org/10.1007/s10298-010-0553-x

Alonso-Carrillo, N., de los Ángeles Aguilar-Santamaría, M., Vernon-Carter, E. J., Jiménez-Alvarado, R., Cruz-Sosa, F., & Román-Guerrero, A. (2017). Industrial Crops and Products, 103, 213–221. https://doi.org/10.1016/j.indcrop.2017.04.002 DOI: https://doi.org/10.1016/j.indcrop.2017.04.002

Andjelković, M., Van Camp, J., De Meulenaer, B., Depaemelaere, G., Socaciu, C., Verloo, M., & Verhe, R. (2006). Iron-chelation properties of phenolic acids bearing catechol and galloyl groups. Food Chemistry, 98(1), 23–31. https://doi.org/10.1016/j.foodchem.2005.05.044 DOI: https://doi.org/10.1016/j.foodchem.2005.05.044

Bassolé, I. H. N., & Juliani, H. R. (2012). Essential oils in combination and their antimicrobial properties. Molecules, 17(4), 3989–4006. https://doi.org/10.3390/molecules17043989 DOI: https://doi.org/10.3390/molecules17043989

Bernal-Mercado, A. T., Vazquez-Armenta, F. J., Tapia-Rodriguez, M. R., Islas-Osuna, M. A., Mata-Haro, V., Gonzalez-Aguilar, G. A., & Ayala-Zavala, J. F. (2018). Comparison of single and combined use of catechin, protocatechuic, and vanillic acids as antioxidant and antibacterial agents against uropathogenic Escherichia coli at planktonic and biofilm levels. Molecules, 23(11), 2813–2825. https://doi.org/10.3390/molecules23112813 DOI: https://doi.org/10.3390/molecules23112813

Brahmi, F., Khodir, M., Mohamed, C., & Pierre, D. (2017). Chemical composition and biological activities of Mentha species. In Aromatic and Medicinal Plants: Back to Nature (pp. 47–79). IntechOpen. https://doi.org/10.5772/67291 DOI: https://doi.org/10.5772/67291

Brown, J., & Mackey, H. (1968). Inhibition of heat-induced denaturation of serum proteins by mixtures of nonsteroidal anti-inflammatory agents and amino acids. Proceedings of the Society for Experimental Biology and Medicine, 128(1), 225–228. https://doi.org/10.3181/00379727-128-32984 DOI: https://doi.org/10.3181/00379727-128-32984

Cornell, J. A. (2002). Experiments with Mixtures: Designs, Models, and the Analysis of Mixture Data. Wiley. DOI: https://doi.org/10.1002/9781118204221

Similar Articles

1-10 of 121

You may also start an advanced similarity search for this article.

Most read articles by the same author(s)

1 2 > >> 

Downloads 124

-

Views 497

-

Country (Top 10)