Environmental Science and Pollution Research

, Volume 26, Issue 18, pp 18440–18450 | Cite as

Nanoencapsulated methyl salicylate as a biorational alternative of synthetic antifungal and aflatoxin B1 suppressive agents

  • Anupam Kujur
  • Amrita Yadav
  • Akshay Kumar
  • Prem Pratap Singh
  • Bhanu PrakashEmail author
Research Article


In view of the suspected negative impact of synthetic fungicides to the human health, nutritional quality, and non-targeted organisms, the use of plant-based antifungal agents has gained considerable interest to the agri-food industries. The aim of this study was to explore the antifungal and aflatoxin B1 (AFB1) inhibitory activity of chitosan (low molecular weight) encapsulated methyl salicylate. The nanoencapsulation of methyl salicylate (Ne-MS) has been characterized by SEM, FTIR, and XRD analysis. The encapsulation efficiency and loading capacity of Ne-MS ranged between 32–34% and 5–7% respectively. The minimum inhibitory concentration of Ne-MS (1.00 μL/mL) against the growth and aflatoxin B1 production by Aspergillus flavus was found to be lower than the free MS (1.50 μL/mL). Mode of action studies demonstrated that the Ne-MS cause a significant decrease in the ergosterol content, leakage of vital ions (Ca2+, Mg2+, and K+), utilization of different carbon source by the A. flavus. Further, the docking result showed ver1 and omt A gene of AFB1 biosynthesis are the possible molecular site of action of methyl salicylate. The in situ study revealed that Ne-MS had no significant negative impact on the organoleptic properties of the food system (maize) which strengthen its potential as a biorational alternative of synthetic fungicides.


Antifungal Aflatoxin B1 Methyl salicylate Mode of action Nanoencapsulation 



We are thankful to the Head, CAS in Botany, Banaras Hindu University, Varanasi, for instrumental facilities. We are also thankful to Indian Institute of Technology, Banaras Hindu University, Varanasi, for the SEM and XRD analyses.

Funding information

Financial support was by CSIR (JRF-Ref: 09/013(0706)/2017-EMR-I) and Science and Engineering Research Board (Scheme No. ECR/2016/000299) New Delhi, India.

Compliance with ethical standards

Conflict of interest

The authors declare that there are no conflicts of interest.


  1. Arulmozhi V, Pandian K, Mirunalini S (2013) Ellagic acid encapsulated chitosan nanoparticles for drug delivery system in human oral cancer cell line (KB). Colloids Surf B Biointerfaces 110:313–320CrossRefGoogle Scholar
  2. Beyki M, Zhaveh S, Khalili ST, 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. Ind Crops Prod 54:310–319CrossRefGoogle Scholar
  3. Brent KJ, Hollomon DW (1998) Fungicide Resistance: The Assessment of Risk. Monograph no. 2. Frac, Global Crop Protection Federation, Brussels, pp 1–48Google Scholar
  4. Burt S (2004) Essential oils: their antibacterial properties and potential applications in foods—a review. Int J Food Microbiol 94:223–253Google Scholar
  5. Chen XG, Lee CM, Park HJ (2003) O/W emulsification for the self-aggregation and nanoparticle formation of linoleic acid modified chitosan in the aqueous system. J Agric Food Chem 51:3135–3139CrossRefGoogle Scholar
  6. Christofoli M, Costa EC, Bicalho KU, de Cássia Domingues V, Peixoto MF, Alves CC, Araújo WL, de Melo Cazal C (2015) Insecticidal effect of nanoencapsulated essential oils from Zanthoxylum rhoifolium (Rutaceae) in Bemisia tabaci populations. Ind Crop Prod 70:301–308CrossRefGoogle Scholar
  7. Coates J (2000) Interpretation of infrared spectra, a practical approach. Encyclopedia of analytical chemistry, vol 12, pp 10815–10837Google Scholar
  8. Cota-Arriola O, Onofre Cortez-Rocha M, Burgos-Hernández A, Marina Ezquerra-Brauer J, Plascencia-Jatomea M (2013) Controlled release matrices and micro/nanoparticles of chitosan with antimicrobial potential: development of new strategies for microbial control in agriculture. J Sci Food Agric 93:1525–1536CrossRefGoogle Scholar
  9. Darmadji P, Izumimoto M (1994) Effect of chitosan in meat preservation. Meat Sci 38:243–254CrossRefGoogle Scholar
  10. Diab MA, El-Sonbati AZ, Bader DMD (2011) Thermal stability and degradation of chitosan modified by benzophenone. Spectrochim Acta A 79:1057–1062CrossRefGoogle Scholar
  11. Donsì F, Annunziata M, Sessa M, Ferrari G (2011) Nanoencapsulation of essential oils to enhance their antimicrobial activity in foods. LWT - Food Sci Technol 44:1908–1914CrossRefGoogle Scholar
  12. Esmaeili A, Asgari A (2015) In vitro release and biological activities of Carum copticum essential oil (CEO) loaded chitosan nanoparticles. Int J Biol Macromol 81:283–290CrossRefGoogle Scholar
  13. Han XB, Zhao J, Cao JM, Zhang CS (2019) Essential oil of Chrysanthemum indicum L.: potential biocontrol agent against plant pathogen Phytophthora nicotianae. Environ Sci Pollut Res 26:7013–7023Google Scholar
  14. Helal GA, Sarhan MM, Abu Shahla ANK, Abou El-Khair EK (2007) Effects of Cymbopogon citratus L. essential oil on the growth: Morphogenesis and aflatoxin production of Aspergillus flavus ML2-strain. J Basic Microbiol 47:5–15CrossRefGoogle Scholar
  15. Hosseini SF, Zandi M, Rezaei M, Farahmandghavi F (2013) Two-step method for encapsulation of oregano essential oil in chitosan nanoparticles: preparation, characterization and in vitro release study. Carbohydr Polym 95:50–56CrossRefGoogle Scholar
  16. International Agency for Research on Cancer (IARC) (2002) Some Naturally Occurring Substances: Food Items and Constituents, Heterocyclic Aromatic Amines and Mycotoxins, vol. 82 of IARC Monographs on the Evaluation of Carcinogenic Risks to Humans. World Health Organization, International Agency for Research on Cancer, LyonGoogle Scholar
  17. Jingou J, Shilei H, Weiqi L, Danjun W, Tengfei W, Yi X (2011) Preparation, characterization of hydrophilic and hydrophobic drug in combine loaded chitosan/cyclodextrin nanoparticles and in vitro release study. Colloids Surf B Biointerfaces 83:103–107CrossRefGoogle Scholar
  18. Juliano C, Marchetti M, Campagna P, Usai M (2018) Antimicrobial activity and chemical composition of essential oil from Helichrysum microphyllum Cambess. subsp. tyrrhenicum Bacch., Brullo & Giusso collected in South-West Sardinia. Saudi J Biol Sci.
  19. Kaur R, Kaur S (2014) Role of polymers in drug delivery. J Drug Deliv Ther 4:32–36Google Scholar
  20. Keawchaoon L, Yoksan R (2011) Preparation, characterization and in vitro release study of carvacrol-loaded chitosan nanoparticles. Colloids Surf B Biointerfaces 84:163–171CrossRefGoogle Scholar
  21. Kiran S, Kujur A, Prakash B (2016) Assessment of preservative potential of Cinnamomum zeylanicum Blume essential oil against food borne molds, aflatoxin B1 synthesis, its functional properties and mode of action. Innov Food Sci Emerg Technol 37:184–191CrossRefGoogle Scholar
  22. Kujur A, Kiran S, Dubey NK, Prakash B (2017) Microencapsulation of Gaultheria procumbens essential oil using chitosan-cinnamic acid microgel: improvement of antimicrobial activity, stability and mode of action. LWT - Food Sci Technol 86:132–138CrossRefGoogle Scholar
  23. Le Grand F, George G, Akoka S (2005) Natural abundance 2H-ERETIC-NMR authentication of the origin of methyl salicylate. J Agric Food Chem 53:5125–5129CrossRefGoogle Scholar
  24. Lee KY, Jo WH, Kwon IC, Kim YH, Jeong SY (1998) Physicochemical characteristics of self-aggregates of hydrophobically modified chitosans. Langmuir 14:2329–2332CrossRefGoogle Scholar
  25. Liu J, Sun L, Zhang N, Zhang J, Guo J, Li C, Rajput SA, Qi D (2016) Effects of nutrients in substrates of different grains on aflatoxin B1 production by Aspergillus flavus. Biomed Res Int.
  26. Merino N, Berdejo D, Bento R, Salman H, Lanz M, Maggi F, Sanchez-Gomez S, Garcia-Gonzaloa D, Pagan R (2019) Antimicrobial efficacy of Thymbra capitata (L.) Cav. essential oil loaded in self-assembled zein nanoparticles in combination with heat. Ind Crop Prod 133:98–104CrossRefGoogle Scholar
  27. Moosavy MH, Basti AA, Misaghi A, Salehi TZ, Abbasifar R, Mousavi HAE, Noori N (2008) Effect of Zataria multiflora Boiss. essential oil and nisin on Salmonella typhimurium and Staphylococcus aureus in a food model system and on the bacterial cell membranes. Food Res Int 41:1050–1057CrossRefGoogle Scholar
  28. Nitta SK, Numata K (2013) Biopolymer-based nanoparticles for drug/gene delivery and tissue engineering. Int J Mol Sci 14:1629e1654CrossRefGoogle Scholar
  29. Pinto NDOF, Rodrigues THS, Pereira RDCA, eSilva LMA, Cáceres CA, de Azeredo HMC, Muniz CR, de Brito ES, Canuto KM (2016) Production and physico-chemical characterization of nanocapsules of the essential oil from Lippia sidoides Cham. Ind Crops Prod 86:279–288CrossRefGoogle Scholar
  30. Prakash B, Kiran S (2016) Essential oils: a traditionally realized natural resource for food preservation. Curr Sci 110:1890Google Scholar
  31. Prakash B, Kujur A, Yadav A, Kumar A, Singh PP, Dubey NK (2018) Nanoencapsulation: an efficient technology to boost the antimicrobial potential of plant essential oils in food system. Food Control 1(89):1–1CrossRefGoogle Scholar
  32. Qiu C, Chang R, Yang J, Ge S, Xiong L, Zhao M, Li M, Sun Q (2017) Preparation and characterization of essential oil-loaded starch nanoparticles formed by short glucan chains. Food Chem 221:1426–1433CrossRefGoogle Scholar
  33. Singh MP (2009) Application of Biolog FF MicroPlate for substrate utilization and metabolite profiling of closely related fungi. J Microbiol Methods 77(1):102–108CrossRefGoogle Scholar
  34. Srivastava AK, Singh D, Roy BK (2017) Structural interactions of curcumin biotransformed molecules with the N-terminal residues of cytotoxic-associated gene a protein provide insights into suppression of oncogenic activities. Interdiscip Sci Comput Life Sci 9:116–129CrossRefGoogle Scholar
  35. Tchameni SN, Mbiakeu SN, Sameza ML, Jazet PMD, Tchoumbougnang F (2018) Using Citrus aurantifolia essential oil for the potential biocontrol of Colocasia esculenta (taro) leaf blight caused by Phytophthora colocasiae. Environ Sci Pollut Res 25:29929–29935CrossRefGoogle Scholar
  36. Tian J, Huang B, Luo X, Zeng H, Ban X, He J, Wang Y (2012) The control of Aspergillus flavus with Cinnamomum jensenianum Hand.-Mazz essential oil and its potential use as a food preservative. Food Chem 130:520–527CrossRefGoogle Scholar
  37. U.S. Code of Federal Regulations (2016) Title 21, TITLE 21efood and drugs Part 182.Section.¼182.20. (Accessed 14 November 2016)
  38. Vaseeharan B, Ramasamy P, Chen JC (2010) Antibacterial activity of silver nanoparticles (AgNps) synthesized by tea leaf extracts against pathogenic Vibrio harveyi and its protective efficacy on juvenile Feneropenaeus indicus. Lett Appl Microbiol 50:352–356CrossRefGoogle Scholar
  39. Wang C (2006) The use of essential oils as natural preservatives for berry fruits. HortSci 41:1042–1043CrossRefGoogle Scholar
  40. Wang H, Wang J, Li L, Hsiang T, Wang M, Shang S, Yu Z (2016) Metabolic activities of five botryticides against Botrytis cinerea examined using the Biolog FF MicroPlate. Sci Rep 6:31025CrossRefGoogle Scholar
  41. Yu J (2012) Current understanding on aflatoxin biosynthesis and future perspective in reducing aflatoxin contamination. Toxins 4(11):1024–1057CrossRefGoogle Scholar
  42. Yu J, Chang PK, Ehrlich KC, Cary JW, Bhatnagar D, Cleveland TE, Gary A, Linz JE, Woloshuk CP, Bennett JW (2004) Clustered pathway genes in aflatoxin biosynthesis. Appl Environ Microbiol 70:1253–1262CrossRefGoogle Scholar
  43. Zamora-Mora V, Fernández-Gutiérrez M, González-Gómez Á, Sanz B, San Román J, Goya GF, Hernández R, Mijangos C (2017) Chitosan nanoparticles for combined drug delivery and magnetic hyperthermia: from preparation to in vitro studies. Carbohydr Polym 157:361–370CrossRefGoogle Scholar
  44. Zhaveh S, Mohsenifar A, Beiki M, Khalili ST, 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. Ind Crop Prod 69:251–256CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Anupam Kujur
    • 1
  • Amrita Yadav
    • 1
  • Akshay Kumar
    • 1
  • Prem Pratap Singh
    • 1
  • Bhanu Prakash
    • 1
    Email author
  1. 1.Centre of Advanced Study in Botany, Institute of ScienceBanaras Hindu UniversityVaranasiIndia

Personalised recommendations