Antibacterial properties of biologically formed chitosan nanoparticles using aqueous leaf extract of Ocimum basilicum

Document Type: Research Paper


1 Department of Biotechnology, Imam Khomeini International University, Qazvin, Iran

2 Center of Biosciences Research, Imam Hossein comprehensive University, Tehran, Iran


Objective(s): Chitosan nanoparticles (CNPs) were prepared based on the ionic gelation of chitoan with anionic compounds of Ocimum basilicum leaf extract.
Materials and Methods: After addition of Ocimum basilicum leaf extract to chitosan solution, the physicochemical properties of the nanoparticles were determined by Field Emission Scanning Electron microscope (FESEM), Fourier Transform Infrared (FTIR) analysis, X-ray diffraction (XRD) Pattern, and Dynamic Light Scattering (DLS). The antibacterial activity of CNPs was evaluated by agar disc diffusion method.
Results: The synthesized nanoparticles were found to be nearly spherical shape with size in the range of 135-729 nm. FTIR analysis revealed the presence of polyphenolic; proteins and alkaloids compounds act as effective agents for converting chitosan to CNPs. Moreover, the synthesized nanoparticles showed potent antibacterial activity against Gram positive and Gram negative bacteria.
Conclusion: These results reveal that natural sources of materials such as plants could be used for preparation of CNPs instead of use of chemical substances.


1. Bulmer C, Margaritis A, Xenocostas A. Production and characterization of novel chitosan nanoparticles for controlled release of rHu-Erythropoietin. Biochem Eng J. 2012; 68: 61-69.

2. Patel J, Jivani N. Chitosan based nanoparticles in drug delivery. Int J Pharm Sci Nanotech. 2009; 2: 517-522.

3. Zhao L-M, Shi L-E, Zhang Z-L, Chen J-M, Shi D-D, Yang J, Tang Z-X. Preparation and application of chitosan nanoparticles and nanofibers. Braz J Chem Eng. 2011; 28: 353-362.

4. Kumar MNR. A review of chitin and chitosan applications. React Funct Polym. 2000; 46: 1-27.

5. Mogosanu GD, Grumezescu AM. Natural and synthetic polymers for wounds and burns dressing. Int J Pharm. 2014; 463: 127-136.

6. Su J, Zhang J, Liu L, Huang Y, Mason RP. Exploring feasibility of multicolored CdTe quantum dots for in vitro and in vivo fluorescent imaging. J Nanosci Nanotech. 2008; 8: 1174-1177.

7. Ajitha, B.; Ashok Kumar Reddy, Y.; Reddy, P.S. Biogenic nano-scale silver particles by Tephrosia purpurea leaf extract and their inborn antimicrobial activity. Spectrochim Acta Part A: Mol Biomol Spect. 2014, 121, 164-172.

8. Gilaki M. Biosynthesis of silver nanoparticles using plant extracts. J Biol Sci. 2010; 10: 465-467.

9. Bhumkar DR, Joshi HM, Sastry M, Pokharkar VB. Chitosan reduced gold nanoparticles as novel carriers for transmucosal delivery of insulin. Pharm Res. 2007; 24: 1415-1426.

10. Amidi M, Romeijn SG, Borchard G, Junginger HE, Hennink WE, Jiskoot W. Preparation and characterization of protein-loaded N-trimethyl chitosan nanoparticles as nasal delivery system. J Controlled Release. 2006; 111: 107-116.

11. Mansouri S, Cuie Y, Winnik F, Shi Q, Lavigne P, Benderdour M, Beaumont E, Fernandes JC. Characterization of folate-chitosan-DNA nanoparticles for gene therapy. Biomaterials. 2006; 27: 2060-1065.

12. Liu X, Hu Q, Fang Z, Zhang X, Zhang B. Magnetic chitosan nanocomposites: a useful recyclable tool for heavy metal ion removal. Langmuir. 2008; 25: 3-8.

13. Qi L, Xu Z, Jiang X, Hu C, Zou X. Preparation and antibacterial activity of chitosan nanoparticles. Carbohydr Res. 2004; 339: 2693-2700.

14. Cruz-Romero M, Murphy T, Morris M, Cummins E, Kerry J. Antimicrobial activity of chitosan, organic acids and nano-sized solubilisates for potential use in smart antimicrobially-active packaging for potential food applications. Food Control. 2013; 34: 393-397.

15. Shukla SK, Mishra AK, Arotiba OA, Mamba BB. Chitosan-based nanomaterials: A state-of-the-art review. Int J Biological Macromoles. 2013; 59: 46-58.

16. Kavitha K, Baker S, Rakshith D, Kavitha H, Harini B, Satish S. Plants as green source towards synthesis of nanoparticles. Int Res J Biological Sci. 2013; 2: 66-76.

17. Agnihotri SA, Mallikarjuna NN, Aminabhavi TM. Recent advances on chitosan-based micro-and nanoparticles in drug delivery. J Controlled Release. 2004; 100: 5-28.

18. Calvo P, Remunan Lopez C, Vila Jato J, Alonso M. Novel hydrophilic chitosan polyethylene oxide nanoparticles as protein carriers. J Appl Polym Sci. 1997; 63: 125-132.

19. Dyer A, Hinchcliffe M, Watts P, Castile J, Jabbal-Gill I,Nankervis R, Smith A, Illum L. Nasal delivery of insulin using novel chitosan based formulations: a comparative study in two animal models between simple chitosan formulations and chitosan nanoparticles. Pharm Res. 2002; 19: 998-1008.

20. Yang S-J, Lin F-H, Tsai H-M, Lin C-F, Chin H-C, Wong J-M, Shieh M-J. Alginate-folic acid-modified chitosan nanoparticles for photodynamic detection of intestinal neoplasms. Biomaterials. 2011; 32: 2174-2182.

21. Prasad M, Jayalakshmi K, Rindhe G. Antibacterial activity of Ocimum species and their phytochemical and antioxidant potential. Int J Microbio Res. 2012; 4: 302.

22. Paschapur MS, Patil M, Kumar R, Patil SR. Evaluation of aqueous extract of leaves of Ocimum kilimandscharicum on wound healing activity in albino wistar rats. Int J PharmTech Res. 2009; 1: 544-550.

23. Pang X, Zhitomirsky I. Electrodeposition of composite hydroxyapatite–chitosan films. Mater Chem Phys. 2005; 94: 245-251.

24. Calvo P, Remunan-Lopez C, Vila-Jato J, Alonso M. Development of positively charged colloidal drug carriers: chitosan-coated polyester nanocapsules and submicron-emulsions. Colloid Polym Sci. 1997; 275: 46-53.

25. Meisner D, Pringle J, Mezei M. Liposomal ophthalmic drug delivery. III. Pharmacodynamic and biodisposition studies of atropine. Int J Pharm. 1989; 55: 105-113.

26. Meisner D, Pringle J, Mezei M. Liposomal pulmonary drug delivery I. In vivo disposition of atropine base in solution and liposomal form following endotracheal instillation to the rabbit lung. J Microencapsulation. 1989; 6:3 79-87.

27. Jancy, M.E.; Inbathamizh, L. Green synthesis and characterization of nano silver using leaf extract of Morinda pubescens. Asian J Pharm Clin Res. 2012; 5:159-162.

28. Christaki, E.; Bonos, E.; Giannenas, I.; Florou-Paneri, P. Aromatic plants as a source of bioactive compounds. Agriculture. 2012; 2: 228-243.

29. Banerjee T, Mitra S, Singh AK, Sharma RK, Maitra A. Preparation, characterization and biodistribution of ultrafine chitosan nanoparticles. Int J Pharm. 2002; 243: 93-105.

30. Bodnar M, Hartmann JF, Borbely J. Preparation and characterization of chitosan-based nanoparticles. Biomacromolecules. 2005; 6: 2521-2527.

31. Kweka EJ, Nkya HM, Lyaruu L, Kimaro EE, Mwang’onde BJ, Mahande AM. Efficacy of Ocimum kilimandscharicum plant extracts after four years of storage against Anopheles gambiae ss. J Cell Animal Biol. 2009; 3: 171-174.

32. Safari J, Azizi F, Sadeghi M. Chitosan nanoparticles as a green and renewable catalyst in the synthesis of 1, 4-dihydropyridine under solvent-free conditions. New J Chem. 2015; 39: 1905-1909.

33. Jha AK, Prasad K, Prasad K, Kulkarni A. Plant system: nature’s nanofactory. Coll Sur B: Biointerfaces. 2009; 73: 219-223.

34. Roller S, Covill N. The antifungal properties of chitosan in laboratory media and apple juice. Int J Food Microbiol. 1999; 47: 67-77.

35. Choi B-K, Kim K-Y, Yoo Y-J, Oh S-J, Choi J-H, Kim C-Y. In vitro antimicrobial activity of a chitooligosaccharide mixture against Actinobacillusactinomycetemcomitans and Streptococcus mutans. Int J antimicrobial agents. 2001; 18: 553-557.

36. Muzzarelli R, Tarsi R, Filippini O, Giovanetti E, Biagini G, Varaldo P. Antimicrobial properties of N-carboxybutyl chitosan. Antimicrob Agents Chemother. 1990; 34: 2019-23.

37. De Campos AM, Sánchez A, Alonso MaJ. Chitosan nanoparticles: a new vehicle for the improvement of the delivery of drugs to the ocular surface. Application to cyclosporin A. Int J Pharm. 2001; 224: 159-168.

38. Xu Y, Du Y. Effect of molecular structure of chitosan on protein delivery properties of chitosan nanoparticles. Int J Pharm. 2003; 250: 215-226.

39. Perumalla A, Hettiarachchy NS. Green tea and grape seed extracts—Potential applications in food safety and quality. Food Res Int. 2011; 44: 827-839.

40. Cowan MM. Plant products as antimicrobial agents. Clinical Microbiol Rev. 1999; 12: 564-582.

41. Chattopadhyay D, Inamdar M. Studies on synthesis, characterization and viscosity behaviour of nano chitosan. Res J Eng Sci. 2012; 1: 9-15.

42. Brack H, Tirmizi S, Risen W. A spectroscopic and viscometric study of the metal ion-induced gelation of the biopolymer chitosan. Polymer. 1997; 38: 2351-2362.

43. Berger J, Reist M, Mayer JM, Felt O, Gurny R. Structure and interactions in chitosan hydrogels formed by complexation or aggregation for biomedical applications. Eur J Pharm Biopharm. 2004; 57: 35-52.

44. Berger J, Reist M, Mayer JM, Felt O, Peppas N, Gurny R. Structure and interactions in covalently and ionically crosslinked chitosan hydrogels for biomedical applications. Eur J Pharm Biopharm. 2004; 57: 19-34.

45. Noble L, Gray AI, Sadiq L, Uchegbu IF. A non-covalently cross-linked chitosan based hydrogel. Int J Pharm. 1999; 192: 173-182.

46. Shu X, Zhu K. A novel approach to prepare tripolyphosphate/chitosan complex beads for controlled release drug delivery. Int J Pharm. 2000; 201: 51-58.

47. Boonyo W, Junginger HE, Waranuch N, Polnok A, Pitaksuteepong T. Preparation and characterization of particles from chitosan with different molecular weights and their trimethyl chitosan derivatives for nasal immunization. J Met Mater Miner. 2008; 18: 59-6.

48. Huang K-S, Sheu Y-R, Chao I-C. Preparation and properties of nanochitosan. Polym-Plast Tech Eng. 2009; 48: 1239-1243.

49. Heras A, Rodriguez N, Ramos V, Agullo E. N-methylene phosphonic chitosan: a novel soluble derivative. Carbohydr Polym. 2001; 44: 1-8.

50. Holme KR, Perlin AS. Chitosan N-sulfate. A water-soluble polyelectrolyte. Carbohydr Res. 1997; 302: 7-12.

51. Welsh ER, Schauer CL, Qadri SB, Price RR. Chitosan cross-linking with a water-soluble, blocked diisocyanate. 1. Solid state. Biomacromolecules. 2002; 3: 1370-1374.

52. Sugimoto M, Morimoto M, Sashiwa H, Saimoto H, Shigemasa Y. Preparation and characterization of water-soluble chitin and chitosan derivatives. Carbohydr Polym. 1998; 36: 49-59.

53. Xie W, Xu P, Wang W, Liu Q. Preparation and antibacterial activity of a water-soluble chitosan derivative. Carbohydr Polym. 2002; 50: 35-40.

54. Avadi M, Sadeghi A, Tahzibi A, Bayati K, Pouladzadeh M, Zohuriaan-Mehr M, Rafiee-Tehrani M. Diethylmethyl chitosan as an antimicrobial agent: Synthesis, characterization and antibacterial effects. Eur Polym J. 2004; 40: 1355-13561.

55. Mitra A, Dey B. Chitosan microspheres in novel drug delivery systems. Ind J Pharm Sci. 2011; 73: 355.