Novel antimicrobial and edible coating films of chia seed/CMC hydrocolloids with betel leaf oil inclusion complexes for post-harvest fruit preservation: a case study on raspberry

Abstract

Betel leaf oil (BLO) is a natural essential oil composed of main active compounds such as chavibetol, eugenol, and alpha-pinene. These active compounds have major applications in promoting health benefits and antibacterial activity. However, despite its therapeutic promise, the practical application of BLO is hindered by several limitations, including its high volatility, hydrophobicity, and susceptibility to photodegradation. β-Cyclodextrin (β-CD) offers an effective strategy to improve solubility and bioactivity by forming inclusion complexes (ICs). This research integrates both computational and experimental methods to deliver detailed and comparative insights into the mechanistic dynamics of active compounds within the β-CD cavity, utilizing molecular docking and molecular dynamics simulations. The findings demonstrate that all ICs form spontaneously, and van der Waals forces are the major driving force. Comprehensive characterizations of ICs were performed using a suite of analytical techniques, confirming the successful formation and stability of the complexes. The findings indicated that ICs exhibited prolonged release over 12 h under various conditions and enhanced antibacterial efficacy against both Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria. Moreover, raspberries exhibited enhanced freshness after dip-coating with chia/CMC hydrocolloid incorporated with BLO/β-CD ICs, particularly the chia/CMC/1% BLO/β-CD ICs group. This treatment resulted in a significant prolongation of shelf life, reduced ethylene production, and a marked delay in decay. Additionally, bioactive natural coating films demonstrated significant potential as edible natural preservatives, providing a sustainable alternative to conventional chemical preservatives in food industry applications.

Keywords

Betel leaf oil / Inclusion complexes / Intermolecular interactions / Controlled release / Antibacterial activity

Author contributions

The authors confirm their contributions to the paper as follows: study conception and design: Supaphol P; data collection: Ngamplang P, Kornsuthisopon C, Chuysinuan P ; analysis and interpretation of results: Ngamplang P, Basharat G, Rungrotmongkol T, Mayer M, Bunchuay T, Laoviwat P, Pherkkhuntod C, Supaphol P; draft manuscript preparation: Ngamplang P, Choipang C, Mayer M, Supaphol P. All authors reviewed the results and approved the final version of the manuscript.

Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Acknowledgments

This research project is supported by the Second Century Fund (C2F), Chulalongkorn University (Batch 10, Track A), The Petroleum and Petrochemical College (PPC), Chulalongkorn University, Research Platform for Leading Education Grant: Article Review 2022, Chulalongkorn University: (Grant No. ReinUni_65_03_63_53), and Herbal Extracts-infused Advanced Wound Dressing Research Unit (Grant No. RU66_035_6300_001) that contributed to this research work.

Conflict of interest

The authors declare that they have no conflict of interest.

References

[1] Ullah R, Nadeem M, Khalique A, Imran M, Mehmood S, et al. 2015. Nutritional and therapeutic perspectives of Chia (Salvia hispanica L.): a review. Journal of Food Science and Technology 53:1750

[2] Mohd Ali N, Yeap SK, Ho WY, Beh BK, Tan SW, et al. 2012. The promising future of Chia, Salvia hispanica L. Journal of biomedicine & biotechnology 2012:171956

[3] Tavares KM, de Campos A, Luchesi BR, Resende AA, de Oliveira JE, et al. 2020. Effect of carboxymethyl cellulose concentration on mechanical and water vapor barrier properties of corn starch films. Carbohydrate Polymers 246:116521

[4] Singh T, Singh P, Pandey VK, Singh R, Dar AH. 2023. A literature review on bioactive properties of betel leaf (Piper betel L.) and its applications in food industry. Food Chemistry Advances 3:100536

[5] Naße KE, Heinen FS, Pawlowsky N, Schrimpf M, Monflier E, et al. 2024. The role of cyclodextrins in the acceleration of the reaction rate in a biphasic hydroformylations. Chemical Engineering Journal 497:154114

[6] Dhiman P, Bhatia M. 2020. Pharmaceutical applications of cyclodextrins and their derivatives. Journal of Inclusion Phenomena and Macrocyclic Chemistry 98:171-86

[7] Hejazi L, Mahboubi-Rabbani M, Mahdavi V, Alemi M, Khanniri E, et al. 2024. A critical review on sodium benzoate from health effects to analytical methods. Results in Chemistry 11:101798

[8] Kotronia M, Kavetsou E, Loupassaki S, Kikionis S, Vouyiouka S, et al. 2017. Encapsulation of oregano (Origanum onites L.) essential oil in β cyclodextrin ( β-CD): synthesis and characterization of the inclusion complexes. Bioengineering 4:73

[9] Jiang L, Yang J, Wang Q, Ren L, Zhou J. 2019. Physicochemical properties of catechin/ β-cyclodextrin inclusion complex obtained via coprecipitation. CyTA - Journal of Food 17:544-51

[10] Belhachemi A, Maatoug M, Canela-Garayoa R. 2022. GC-MS and GC-FID analyses of the essential oil of Eucalyptus camaldulensis grown under greenhouses differentiated by the LDPE cover-films. Industrial Crops and Products 178:114606

[11] Elizei PS, Krasaaekoopt W. 2014. Microencapsulation of litsea cubeba essential oil in β-cyclodextrin using paste and Co-precipitation methods. Kasetsart Journal - Natural Science 48:893-907

[12] Higuchi T, Connors KA. 1965. Phase Solubility Techniques. Advanced Analytical Chemistry of Instrumentation 4:117-212

[13] Saokham P, Muankaew C, Jansook P, Loftsson T. 2018. Solubility of Cyclodextrins and Drug/Cyclodextrin Complexes. Molecules 23:1161

[14] George S, Vasudevan D. 2012. Studies on the preparation, characterization, and solubility of 2-HP- β-cyclodextrin-meclizine HCl inclusion complexes. Journal of Young Pharmacists 4:220-27

[15] Chang C, Song M, Ma M, Song J, Cao F, et al. 2023. Preparation, characterization and molecular dynamics simulation of rutin-cyclodextrin inclusion complexes. Molecules 28:955

[16] Yazdani M, Tavakoli O, Khoobi M, Wu YS, Faramarzi MA, et al. 2022. Beta-carotene/cyclodextrin-based inclusion complex: improved loading, solubility, stability, and cytotoxicity. Journal of Inclusion Phenomena and Macrocyclic Chemistry 102:55-64

[17] Paiva-Santos AC, Ferreira L, Peixoto D, Silva F, Soares MJ, et al. 2022. Cyclodextrins as an encapsulation molecular strategy for volatile organic compounds- Pharmaceutical applications. Colloids and Surfaces B: Biointerfaces 218:112758

[18] Wen P, Zhu DH, Feng K, Liu FJ, Lou WY, et al. 2016. Fabrication of electrospun polylactic acid nanofilm incorporating cinnamon essential oil /β cyclodextrin inclusion complex for antimicrobial packaging. Food Chemistry 196:996-1004

[19] Ngamplang P, Choipang C, Chaiarwut S, Chuysinuan P, Supaphol P. 2025. Development of poly (vinyl alcohol)/carboxymethyl chitosan hydrogels loaded with encapsulated tea tree oil for wound care application. Journal of Drug Delivery Science and Technology 107:106867

[20] Marques CS, Carvalho SG, Bertoli LD, Villanova JCO, Pinheiro PF, et al. 2019. β-Cyclodextrin inclusion complexes with essential oils: obtention, characterization, antimicrobial activity and potential application for food preservative sachets. Food Research International 119:499-509

[21] Xiao Z, Yu P, Sun P, Kang Y, Niu Y, et al. 2024. Inclusion complexes of β cyclodextrin with isomeric ester aroma compounds: Preparation, characterization, mechanism study, and controlled release. Carbohydrate Polymers 333:121977

[22] Aman A, Ali S, Mahalapbutr P, Krusong K, Wolschann P, et al. 2023. Enhancing solubility and stability of sorafenib through cyclodextrinbased inclusion complexation: in silico and in vitro studies. RSC Advances 13:27244-54

[23] Kirschner KN, Yongye AB, Tschampel SM, González-Outeiriño J, Daniels CR, et al. 2008. GLYCAM06: a generalizable biomolecular force field. Carbohydrates. Journal of Computational Chemistry 29:622-55

[24] Case DA, Aktulga HM, Belfon K, Ben-Shalom I, Berryman J, et al. 2022. AMBER 2022: Reference Manual. University of California, San Francisco. doi: 10.13140/RG.2.2.31337.77924

[25] Sangsanoh P, Chaiarwut S, Choipang C, Niyompanich J, Suwantong O, et al. 2023. Cannabidiol /β-cyclodextrin inclusion complex-loaded poly(vinyl alcohol) semi-solid hydrogels for potential use in wound management. Journal of Polymers and the Environment 31:3982-97

[26] Gupta RK, Guha P, Srivastav PP. 2023. Phytochemical and biological studies of betel leaf (Piper betle L.): Review on paradigm and its potential benefits in human health. Acta Ecologica Sinica 43:721-32

[27] Sahu AK, Kar SS, Kumari P, Dey SK. 2022. An overview of Betel vine (Piper Betle L.): nutritional, pharmacological and economical promising natural reservoir. Advances in Horticultural Science 36:63-80

[28] Shah SK, Garg G, Jhade D, Patel N. 2016. Piper betle: phytochemical, pharmacological and nutritional value in health management. International Journal of Pharmaceutical Sciences Review and Research 38:181-89

[29] Kringel DH, Antunes MD, Klein B, Crizel RL, Wagner R, et al. 2017. Production, Characterization, and Stability of Orange or Eucalyptus Essential Oil/ β-Cyclodextrin Inclusion Complex. Journal of Food Science 82:2598-605

[30] Jagdale SC, Gawali VU, Kuchekar BS, Chabukswar AR. 2011. Formulation and in vitro evaluation of taste-masked oro-dispersible dosage form of diltiazem hydrochloride. Brazilian Journal of Pharmaceutical Sciences 47:907-16

[31] Loftsson T, Hreinsdóttir D, Másson M. 2005. Evaluation of cyclodextrin solubilization of drugs. International journal of pharmaceutics 302:18-28

[32] Herrera A, Rodríguez FJ, Bruna JE, Abarca RL, Galotto MJ, et al. 2019. Antifungal and physicochemical properties of inclusion complexes based on β-cyclodextrin and essential oil derivatives. Food Research International 121:127-35

[33] Zhumakova S, Ten A, Zharkynbek T, Yu V, Seilkhanov T, et al. 2022. NMR study of the inclusion complexes of β-cyclodextrin with diphenhydramine, clonidine and tolperisone. SN Applied Sciences 4:10

[34] Wang J, Cao Y, Sun B, Wang C. 2011. Physicochemical and release characterisation of garlic oil- β-cyclodextrin inclusion complexes. Food Chemistry 127:1680-85

[35] Silva I, Feitosa E, Santos M, Silva R, Rocha M, et al. 2020. Theoretical and experimental investigations on inclusion complex β-cyclodextrin and sulcatone: a cardiovascular activity evaluation. Journal of the Brazilian Chemical Society 31:1064-77

[36] Shi C, Zhou A, Fang D, Lu T, Wang J, et al. 2021. Oregano essential oil /β cyclodextrin inclusion compound polylactic acid/polycaprolactone electrospun nanofibers for active food packaging. SSRN Electronic Journal 445:136746

[37] Borghetti GS, Lula IS, Sinisterra RD, Bassani VL. 2009. Quercetin/betacyclodextrin solid complexes prepared in aqueous solution followed by spray-drying or by physical mixture. AAPS PharmSciTech 10:235-42

[38] Castro JC, Pante GC, de Souza DS, Pires TY, Miyoshi JH, et al. 2022. Molecular inclusion of Cymbopogon martinii essential oil with β-cyclodextrin as a strategy to stabilize and increase its bioactivity. Food Hydrocolloids for Health 2:100066

[39] Pinto LMA, Fraceto LF, Santana MHA, Pertinhez TA, Junior SO, et al. 2005. Physico-chemical characterization of benzocaine- β-cyclodextrin inclusion complexes. Journal of Pharmaceutical and Biomedical Analysis 39:956-63

[40] Prabu S, Swaminathan M, Sivakumar K, Rajamohan R. 2015. Preparation, characterization and molecular modeling studies of the inclusion complex of Caffeine with Beta-cyclodextrin. Journal of Molecular Structure 1099:616-24

[41] Menezes PP, Serafini MR, Quintans-Júnior LJ, Silva GF, Oliveira JF, et al. 2014. Inclusion complex of (-)-linalool and β-cyclodextrin. Journal of Thermal Analysis and Calorimetry 115:2429-37

[42] Ke Q, Zhu J, Su D, Pan F, Meng Q, et al. 2024. Preparation, physicochemical characterization, and computational studies of aldehyde aroma compounds/cyclodextrin inclusion complexes. Industrial Crops and Products 211:118245

[43] Gholami R, Azizi K, Ganjali Koli M. 2024. Unveiling the dynamic and thermodynamic interactions of hydrocortisone with β-cyclodextrin and its methylated derivatives through insights from molecular dynamics simulations. Scientific Reports 14:12495

[44] Fereidounpour P, Steinmann C, Larsen KL. 2024. Prediction of the free energy of binding for cyclodextrin-steroid complexes: phase solubility and molecular dynamics studies. Journal of Inclusion Phenomena and Macrocyclic Chemistry 104:535-46

[45] Middelburg J. 2024. The Gibbs free energy. Cham: Springer. pp. 35-46 doi: 10.1007/978-3-031-53407-2_4

[46] Pina LTS, Rabelo TK, Borges LP, Gonçalves VSS, Silva AS, et al. 2024. Antihyperalgesic effect of γ-terpinene complexed in β-cyclodextrin on neuropathic pain model induced by tumor cells. International journal of pharmaceutics 662:124538

[47] de Freitas CAB, Costa CHS, da Costa KS, da Paz SPA, Silva JRA, et al. 2022. Assessment of host-guest molecular encapsulation of eugenol using β cyclodextrin. Frontiers in Chemistry 10:1061624

[48] Sharma D, Dhingra S, Banerjee A, Saha S, Bhattacharyya J, et al. 2022. Designing suture-proof cell-attachable copolymer-mediated and curcumin- β-cyclodextrin inclusion complex loaded aliphatic polyesterbased electrospun antibacterial constructs. International Journal of Biological Macromolecules 216:397-413

[49] Jamnongkan T, Kaewpirom S. 2010. Potassium release kinetics and water retention of controlled-release fertilizers based on chitosan hydrogels. Journal of Polymers and the Environment 18:413-21

[50] Rostamitabar M, Abdelgawad AM, Jockenhoevel S, Ghazanfari S. 2021. Drug-eluting medical textiles: from fiber production and textile fabrication to drug loading and delivery. Macromolecular Bioscience 21:e2100021

[51] Riva SC, Opara UO, Fawole OA. 2020. Recent developments on postharvest application of edible coatings on stone fruit: a review. Scientia Horticulturae 262:109074

[52] Ali S, Ullah MA, Nawaz A, Naz S, Shah AA, et al. 2022. Carboxymethyl cellulose coating regulates cell wall polysaccharides disassembly and delays ripening of harvested banana fruit. Postharvest Biology and Technology 191:111978

[53] Valero D, Serrano M. 2013. Growth and ripening stage at harvest modulates postharvest quality and bioactive compounds with antioxidant activity. Stewart Postharvest Review 9( 3):1-8