Bioactive okra mucilage films with Aloysia citrodora extract: a natural approach to fish preservation under refrigeration
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Abstract
Background and Objectives: This study investigated the extraction of okra mucilage and its application in developing bioactive films enriched with Aloysia citrodora extract for food packaging. The effects of microwave-assisted extraction and UV treatment on film properties and preservation performance were evaluated.
Materials and Methods: Okra mucilage was extracted using conventional solvent and microwave-assisted methods. The phenolic profile of A. citrodora extract was analyzed by HPLC-DAD. Films containing up to 2% extract were prepared and subjected to UV treatment. Mechanical properties, thickness, water vapor permeability (WVP), antioxidant capacity (TPC and DPPH), antimicrobial activity, and preservation efficacy were assessed in refrigerated fish fillets.
Results: Microwave extraction slightly improved mucilage yield (8.7 vs. 8.3 g/100 g). Luteolin-7-O-diglucuronide and verbascoside were the dominant phenolics. UV-treated films with 2% extract showed higher tensile strength (45.71 MPa) and lower WVP (4.36 g mm/m² day kPa) compared to the controls. Antioxidant activity and antimicrobial inhibition against foodborne pathogens were significantly enhanced. Treated films delayed Pseudomonas aeruginosa growth in fish fillets.
Conclusion: Combining microwave extraction and UV treatment improved film functionality, highlighting their potential as natural active packaging materials.
1. Mehdipoor Damiri GR, Motamedzadegan A, Safari R, Shahidi SA, Ghorbani A. Evaluation of stability, physicochemical and antioxidant properties of extracted chlorophyll from Persian clover (Trifolium resupinatum L.). J Food Meas Charact 2021; 15: 327-340.
2. Salimbahrami SN, Ghorbani-HasanSaraei A, Tahermansouri H, Shahidi S-A. Synthesis, optimization via response surface methodology, and structural properties of carboxymethylcellulose/curcumin/graphene oxide biocomposite films/coatings for the shelf-life extension of shrimp. Int J Biol Macromol 2023; 253: 1-15.
3. Jorjani A, Najafian L, Farsi M, Shahidi SA. Effect of active films with free and nanoliposomal broccoli sprout protein hydrolysates on physicochemical, oxidative, microbial, and sensory properties of Cheddar cheese during cold storage. Int Dairy J 2025; 169: 1-14.
4. Shiryanpour S, Nouri L, Azizi M, Najafi A, Mohammadi Nafchi A. Active chitosan film containing single and double nanoemulsions of Oliveira decumbens Vent essential oil/anthocyanin of eggplant for chicken preservation. J Food Meas Charact 2025; 19: 7381-7403.
5. Choubaki S, Baghaei H, Mohammadi Nafchi A. Smart indicator for fish freshness monitoring: a chitosan/gelatin/zinc oxide nanoparticle composite incorporating pomegranate flower anthocyanin extract nanocapsules. Food Sci Nutr 2025; 13(9): e70944.
6. Ghorbani‐HasanSaraei A, Rafe A, Shahidi SA, Atashzar A. Microstructure and chemorheological behavior of whipped cream as affected by rice bran protein addition. Food Sci Nutr 2019; 7: 875-881.
7. Shafizadeh A, Golestan L, Ahmadi M, Darjani P, Ghorbani-HasanSaraei A. Encapsulation of Lactobacillus casei in alginate microcapsules: Improvement of the bacterial viability under simulated gastrointestinal conditions using flaxseed mucilage. J Food Meas Charact 2020; 14: 1901-1908.
8. Gao X, Liu J, Li B, Xie J. Antibacterial activity and antibacterial mechanism of lemon verbena essential oil. Molecules 2023; 28:3102.
9. Gholizadeh H, Ghorbani-HasanSaraei A, Tahermansouri H, Shahidi SA. The simultaneous adsorption and desorption of flavonoids from bitter orange peel by the carboxylated multi-walled carbon nanotubes. Carbon Lett 2019; 29: 273-279.
10. Sharif Nasirian V, Shahidi SA, Tahermansouri H, Chekin F. Application of graphene oxide in the adsorption and extraction of bioactive compounds from lemon peel. Food Sci Nutr 2021; 9: 3852-3862.
11. Azarashkan Z, Motamedzadegan A, Ghorbani-HasanSaraei A, Rahaiee S, Biparva P. Improvement of the stability and release of sulforaphane-enriched broccoli sprout extract nanoliposomes by co-encapsulation into basil seed gum. Food Bioproc Tech 2022; 15:1573-1587.
12. Nabet N, Gilbert-López B, Madani K, Herrero M, Ibáñez E, Mendiola JA. Optimization of microwave-assisted extraction recovery of bioactive compounds from Origanum glandulosum and Thymus fontanesii. Ind Crops Prod 2019; 129: 395-404.
13. Taghizadeh M, Mohammadifar MA, Sadeghi E, Rouhi M, Mohammadi R, Askari F, et al. Photosensitizer-induced cross-linking: A novel approach for improvement of physicochemical and structural properties of gelatin edible films. Food Res Int 2018; 112: 90-97.
14. Li C, Sheng L, Sun G, Wang L. The application of ultraviolet-induced photo-crosslinking in edible film preparation and its implication in food safety. Lwt 2020; 131: 109791.
15. Shahidi SA. Effect of solvent type on ultrasound-assisted extraction of antioxidant compounds from Ficaria kochii: Optimization by response surface methodology. Food Chem Toxicol 2022; 163: 112981.
16. Sprea RM, Fernandes LH, Pires TC, Calhelha RC, Rodrigues PJ, Amaral JS. Volatile compounds and biological activity of the essential oil of Aloysia citrodora Paláu: Comparison of hydrodistillation and microwave-assisted hydrodistillation. Molecules 2023; 28: 4528.
17. Kalkan E, Maskan M. Mucilage in okra: extraction, modelling, optimization and application. J Food Meas Charact 2023; 17:4812-4822.
18. Cruz D, Falé PL, Mourato A, Vaz PD, Serralheiro ML, Lino ARL. Preparation and physicochemical characterization of Ag nanoparticles biosynthesized by Lippia citriodora (Lemon Verbena). Colloids Surf B Biointerfaces 2010; 81: 67-73.
19. Lawal KG, Riaz A, Mostafa H, Stathopoulos C, Manikas I, Maqsood S. Development of carboxymethylcellulose based active and edible food packaging films using date seed components as reinforcing agent: physical, biological, and mechanical properties. Food Biophys 2023; 18: 497-509.
20. Pirnia M, Shirani K, Yazdi FT, Moratazavi SA, Mohebbi M. Characterization of antioxidant active biopolymer bilayer film based on gelatin-frankincense incorporated with ascorbic acid and Hyssopus officinalis essential oil. Food Chem X 2022; 14: 100300.
21. Hosseini MH, Razavi SH, Mousavi MA. Antimicrobial, physical and mechanical properties of chitosan‐based films incorporated with thyme, clove and cinnamon essential oils. J Food Process Preserv 2009; 33: 727-743.
22. Benbettaïeb N, Karbowiak T, Brachais C-H, Debeaufort F. Coupling tyrosol, quercetin or ferulic acid and electron beam irradiation to cross-link chitosan–gelatin films: A structure–function approach. Eur Polym J 2015; 67: 113-127.
23. Azmir J, Zaidul ISM, Rahman MM, Sharif K, Mohamed A, Sahena F, et al. Techniques for extraction of bioactive compounds from plant materials: A review. J Food Eng 2013; 117: 426-436.
24. Pesavento G, Calonico C, Bilia AR, Barnabei M, Calesini F, Addona R, et al. Antibacterial activity of Oregano, Rosmarinus and Thymus essential oils against Staphylococcus aureus and Listeria monocytogenes in beef meatballs. Food Control 2015; 54: 188-199.
25. Volpe M, Siano F, Paolucci M, Sacco A, Sorrentino A, Malinconico M, et al. Active edible coating effectiveness in shelf-life enhancement of trout (Oncorhynchusmykiss) fillets. LWT 2015; 60: 615-622.
26. Wang K, Li M, Wen X, Chen X, He Z, Ni Y. Optimization of ultrasound-assisted extraction of okra (Abelmoschus esculentus (L.) Moench) polysaccharides based on response surface methodology and antioxidant activity. Int J Biol Macromol 2018; 114: 1056-1063.
27. Strauss G, Gibson SM. Plant phenolics as cross-linkers of gelatin gels and gelatin-based coacervates for use as food ingredients. Food Hydrocoll 2004; 18: 81-89.
28. Lei Y, Wu H, Jiao C, Jiang Y, Liu R, Xiao D, et al. Investigation of the structural and physical properties, antioxidant and antimicrobial activity of pectin-konjac glucomannan composite edible films incorporated with tea polyphenol. Food Hydrocoll 2019; 94: 128-135.
29. Cui H, Cheng Q, Li C, Chen X, Lin L. Improving packing performance of lily polysaccharide based edible films via combining with sodium alginate and cold plasma treatment. Int J Biol Macromol 2022; 206: 750-758.
30. Bonilla J, Atarés L, Vargas M, Chiralt A. Effect of essential oils and homogenization conditions on properties of chitosan-based films. Food Hydrocoll 2012; 26: 9-16.
31. Jaradat N, Hawash M, Abualhasan MN, Qadi M, Ghanim M, Massarwy E, et al. Spectral characterization, antioxidant, antimicrobial, cytotoxic, and cyclooxygenase inhibitory activities of Aloysia citriodora essential oils collected from two Palestinian regions. BMC Complement Med Ther 2021; 21: 143.
32. Ojagh SM, Rezaei M, Razavi SH, Hosseini SM. Development and evaluation of a novel biodegradable film made from chitosan and cinnamon essential oil with low affinity toward water. Food Chem 2010; 122: 161-166.
33. Shafiee F, Moghadamnia AA, Shahandeh Z, Sadighian F, Khodadadi E. Evaluation of the antibacterial effects of aqueous and ethanolic leaf extracts of Aloysia citriodora (Lemon verbena) on Streptococcus mutans and Streptococcus sobrinus. Electron Physician 2016; 8: 3363-3368.
34. Shahidi F, Hossain A. Preservation of aquatic food using edible films and coatings containing essential oils: A review. Crit Rev Food Sci Nutr 2022; 62: 66-105.
2. Salimbahrami SN, Ghorbani-HasanSaraei A, Tahermansouri H, Shahidi S-A. Synthesis, optimization via response surface methodology, and structural properties of carboxymethylcellulose/curcumin/graphene oxide biocomposite films/coatings for the shelf-life extension of shrimp. Int J Biol Macromol 2023; 253: 1-15.
3. Jorjani A, Najafian L, Farsi M, Shahidi SA. Effect of active films with free and nanoliposomal broccoli sprout protein hydrolysates on physicochemical, oxidative, microbial, and sensory properties of Cheddar cheese during cold storage. Int Dairy J 2025; 169: 1-14.
4. Shiryanpour S, Nouri L, Azizi M, Najafi A, Mohammadi Nafchi A. Active chitosan film containing single and double nanoemulsions of Oliveira decumbens Vent essential oil/anthocyanin of eggplant for chicken preservation. J Food Meas Charact 2025; 19: 7381-7403.
5. Choubaki S, Baghaei H, Mohammadi Nafchi A. Smart indicator for fish freshness monitoring: a chitosan/gelatin/zinc oxide nanoparticle composite incorporating pomegranate flower anthocyanin extract nanocapsules. Food Sci Nutr 2025; 13(9): e70944.
6. Ghorbani‐HasanSaraei A, Rafe A, Shahidi SA, Atashzar A. Microstructure and chemorheological behavior of whipped cream as affected by rice bran protein addition. Food Sci Nutr 2019; 7: 875-881.
7. Shafizadeh A, Golestan L, Ahmadi M, Darjani P, Ghorbani-HasanSaraei A. Encapsulation of Lactobacillus casei in alginate microcapsules: Improvement of the bacterial viability under simulated gastrointestinal conditions using flaxseed mucilage. J Food Meas Charact 2020; 14: 1901-1908.
8. Gao X, Liu J, Li B, Xie J. Antibacterial activity and antibacterial mechanism of lemon verbena essential oil. Molecules 2023; 28:3102.
9. Gholizadeh H, Ghorbani-HasanSaraei A, Tahermansouri H, Shahidi SA. The simultaneous adsorption and desorption of flavonoids from bitter orange peel by the carboxylated multi-walled carbon nanotubes. Carbon Lett 2019; 29: 273-279.
10. Sharif Nasirian V, Shahidi SA, Tahermansouri H, Chekin F. Application of graphene oxide in the adsorption and extraction of bioactive compounds from lemon peel. Food Sci Nutr 2021; 9: 3852-3862.
11. Azarashkan Z, Motamedzadegan A, Ghorbani-HasanSaraei A, Rahaiee S, Biparva P. Improvement of the stability and release of sulforaphane-enriched broccoli sprout extract nanoliposomes by co-encapsulation into basil seed gum. Food Bioproc Tech 2022; 15:1573-1587.
12. Nabet N, Gilbert-López B, Madani K, Herrero M, Ibáñez E, Mendiola JA. Optimization of microwave-assisted extraction recovery of bioactive compounds from Origanum glandulosum and Thymus fontanesii. Ind Crops Prod 2019; 129: 395-404.
13. Taghizadeh M, Mohammadifar MA, Sadeghi E, Rouhi M, Mohammadi R, Askari F, et al. Photosensitizer-induced cross-linking: A novel approach for improvement of physicochemical and structural properties of gelatin edible films. Food Res Int 2018; 112: 90-97.
14. Li C, Sheng L, Sun G, Wang L. The application of ultraviolet-induced photo-crosslinking in edible film preparation and its implication in food safety. Lwt 2020; 131: 109791.
15. Shahidi SA. Effect of solvent type on ultrasound-assisted extraction of antioxidant compounds from Ficaria kochii: Optimization by response surface methodology. Food Chem Toxicol 2022; 163: 112981.
16. Sprea RM, Fernandes LH, Pires TC, Calhelha RC, Rodrigues PJ, Amaral JS. Volatile compounds and biological activity of the essential oil of Aloysia citrodora Paláu: Comparison of hydrodistillation and microwave-assisted hydrodistillation. Molecules 2023; 28: 4528.
17. Kalkan E, Maskan M. Mucilage in okra: extraction, modelling, optimization and application. J Food Meas Charact 2023; 17:4812-4822.
18. Cruz D, Falé PL, Mourato A, Vaz PD, Serralheiro ML, Lino ARL. Preparation and physicochemical characterization of Ag nanoparticles biosynthesized by Lippia citriodora (Lemon Verbena). Colloids Surf B Biointerfaces 2010; 81: 67-73.
19. Lawal KG, Riaz A, Mostafa H, Stathopoulos C, Manikas I, Maqsood S. Development of carboxymethylcellulose based active and edible food packaging films using date seed components as reinforcing agent: physical, biological, and mechanical properties. Food Biophys 2023; 18: 497-509.
20. Pirnia M, Shirani K, Yazdi FT, Moratazavi SA, Mohebbi M. Characterization of antioxidant active biopolymer bilayer film based on gelatin-frankincense incorporated with ascorbic acid and Hyssopus officinalis essential oil. Food Chem X 2022; 14: 100300.
21. Hosseini MH, Razavi SH, Mousavi MA. Antimicrobial, physical and mechanical properties of chitosan‐based films incorporated with thyme, clove and cinnamon essential oils. J Food Process Preserv 2009; 33: 727-743.
22. Benbettaïeb N, Karbowiak T, Brachais C-H, Debeaufort F. Coupling tyrosol, quercetin or ferulic acid and electron beam irradiation to cross-link chitosan–gelatin films: A structure–function approach. Eur Polym J 2015; 67: 113-127.
23. Azmir J, Zaidul ISM, Rahman MM, Sharif K, Mohamed A, Sahena F, et al. Techniques for extraction of bioactive compounds from plant materials: A review. J Food Eng 2013; 117: 426-436.
24. Pesavento G, Calonico C, Bilia AR, Barnabei M, Calesini F, Addona R, et al. Antibacterial activity of Oregano, Rosmarinus and Thymus essential oils against Staphylococcus aureus and Listeria monocytogenes in beef meatballs. Food Control 2015; 54: 188-199.
25. Volpe M, Siano F, Paolucci M, Sacco A, Sorrentino A, Malinconico M, et al. Active edible coating effectiveness in shelf-life enhancement of trout (Oncorhynchusmykiss) fillets. LWT 2015; 60: 615-622.
26. Wang K, Li M, Wen X, Chen X, He Z, Ni Y. Optimization of ultrasound-assisted extraction of okra (Abelmoschus esculentus (L.) Moench) polysaccharides based on response surface methodology and antioxidant activity. Int J Biol Macromol 2018; 114: 1056-1063.
27. Strauss G, Gibson SM. Plant phenolics as cross-linkers of gelatin gels and gelatin-based coacervates for use as food ingredients. Food Hydrocoll 2004; 18: 81-89.
28. Lei Y, Wu H, Jiao C, Jiang Y, Liu R, Xiao D, et al. Investigation of the structural and physical properties, antioxidant and antimicrobial activity of pectin-konjac glucomannan composite edible films incorporated with tea polyphenol. Food Hydrocoll 2019; 94: 128-135.
29. Cui H, Cheng Q, Li C, Chen X, Lin L. Improving packing performance of lily polysaccharide based edible films via combining with sodium alginate and cold plasma treatment. Int J Biol Macromol 2022; 206: 750-758.
30. Bonilla J, Atarés L, Vargas M, Chiralt A. Effect of essential oils and homogenization conditions on properties of chitosan-based films. Food Hydrocoll 2012; 26: 9-16.
31. Jaradat N, Hawash M, Abualhasan MN, Qadi M, Ghanim M, Massarwy E, et al. Spectral characterization, antioxidant, antimicrobial, cytotoxic, and cyclooxygenase inhibitory activities of Aloysia citriodora essential oils collected from two Palestinian regions. BMC Complement Med Ther 2021; 21: 143.
32. Ojagh SM, Rezaei M, Razavi SH, Hosseini SM. Development and evaluation of a novel biodegradable film made from chitosan and cinnamon essential oil with low affinity toward water. Food Chem 2010; 122: 161-166.
33. Shafiee F, Moghadamnia AA, Shahandeh Z, Sadighian F, Khodadadi E. Evaluation of the antibacterial effects of aqueous and ethanolic leaf extracts of Aloysia citriodora (Lemon verbena) on Streptococcus mutans and Streptococcus sobrinus. Electron Physician 2016; 8: 3363-3368.
34. Shahidi F, Hossain A. Preservation of aquatic food using edible films and coatings containing essential oils: A review. Crit Rev Food Sci Nutr 2022; 62: 66-105.
| Files | ||
| Issue | Vol 18 No 2 (2026) | |
| Section | Original Article(s) | |
| Keywords | ||
| Food preservation Antimicrobial agents Antioxidants Polysaccharides Plant extracts | ||
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How to Cite
1.
Daryabari SH, Ahmadi M, Fazeli F, Ariaii P. Bioactive okra mucilage films with Aloysia citrodora extract: a natural approach to fish preservation under refrigeration. Iran J Microbiol. 2026;18(2):285-294.
