Document Type : Original Article


1 Department of Pharmacognosy, Faculty of Pharmacy, University of Maiduguri, Maiduguri, Nigeria

2 Department of Pharmacognosy and Traditional Medicine, Faculty of Pharmacy Delta State University, Abraka, Nigeria

3 Department of Microbiology, Faculty of Natural Sciences, Kogi State University Anyigba, Nigeria

4 Department of Pharmaceutics and Pharmaceutical Microbiology, Faculty of Pharmacy, University of Maiduguri, Nigeria

5 Department of Biological Sciences, Faculty of Science, Taraba State University Jalingo, Jalingo, Nigeria

6 Department of Chemistry, Faculty of Physical Science, Ahmadu Bello University, Zaria, Nigeria


Background: Chitosan nanoparticle (chitosan-NPs) is a polymer obtained from the exoskeletons of crustaceans, and has been applied recently as a carrier for many drug agents. Multi-drug resistance has been the major set-back in the treatment of microbial infections globally.
Methods: Dibutyl phthalate (DBP) isolated from Melastomastrum capitatum leaves was encapsulated in chitosan-NPs and its antimicrobial activity was evaluated on selected multi-drug resistant pathogens. The isolated phthalate was characterized by FTIR, NMR and GC-MS. Chitosan-NPs encapsulated phthalate was prepared by ionic gelation of glutaraldehyde cross-linker. Antimicrobial activity of nano encapsulated drugs was carried by agar well diffusion at 0.5 µg/mL concentration. In vivo activity of nano encapsulated drugs were determined in thirty Swiss albino rats weighing 100-150g. Chitosan-NPs encapsulated treatment groups were administered at 0.5 µg/mL (i.p.) as compared with ciprofloxacin positive control group at 2.5 µg/mL.
Results: Chitosan-NPs encapsulated phthalate showed the strongest zones of inhibition against VRE ATCC 29212, MRSA NCTC 13435, Candida albicans ATCC 19231, and Clostridiodes difficile NCTC14385. Significant inhibition of bacterial growths was achieved by CSDBP encapsulated phthalate both in vitro and in vivo studies due to low concentrations in ALT, ALP, AST and creatinine, and high volume of WBC in rats. Non-Fickian drug release was observed by the formulations.
Conclusion: The study showed that chitosan-NPs mediated drug delivery exhibited strong antimicrobial activity with sustained release against multi-drug microbes in this study. This is promising, and can be employed as mediation for multi-drug resistant pathogens.

Graphical Abstract

Nanoencapsulation of Phthalate from Melastomastrum Capitatum (Fern.) in Chitosan-Nps as a Target Mediated Drug Delivery for Multi-Drug Resistant Pathogen


Main Subjects

1. Poole K, Tetro K, Zhao Q, Neshat S, Heinrichs D E, Bianco N. (1996). Expression of the multidrug resistance operon mexA-mexB-oprM in Pseudomonas aeruginosa: mexR encodes a regulator of operon expression. Antimicrobial agents and chemotherapy, 40(9): 2021-2028. doi: 10.1128/AAC.40.9.2021.
2. World Health Organization. (2014). Antimicrobial resistance: global report on surveillance.
3. Gomez JE, Kaufmann-Malaga BB, Wivagg CN, Kim PB, Silvis MR, Renedo N, Ioerger TR, Ahmad R, Livny J, Fishbein S, Sacchettini JC, Carr SA, Hung DT. (2017). Ribosomal mutations promote the evolution of antibiotic resistance in a multidrug environment. Elife, 6: e20420. doi: 10.7554/eLife.20420.
4. Rolo J, Worning P, Boye Nielsen J, Sobral R, Bowden R, Bouchami O, Damborg P, Guardabassi L, Perreten V, Westh H, Tomasz A, de Lencastre H, Miragaia M. (2017). Evidence for the evolutionary steps leading to mecA-mediated β-lactam resistance in staphylococci. PLoS Genet., 13(4): e1006674. doi: 10.1371/journal.pgen.1006674.
5. Ziha-Zarifi I, Llanes C, Kohler T, Pechere JC, Plesiat P. (1999). In vivo emergence of multidrug-resistant mutants of Pseudomonas aeruginosa overexpressing the active efflux system MexA-MexB-OprM. Antimicrob Agents Chemother., 43 (2):287–291. doi:10.1128/AAC.43.2.287
6. Ward MJ, Goncheva M, Richardson E, McAdam PR, Raftis E, Kearns A, Daum RS, David MZ, Lauderdale TL, Edwards GF, Nimmo GR, Coombs GW, Huijsdens X, Woolhouse ME, Fitzgerald JR. (2016). Identification of source and sink populations for the emergence and global spread of the East-Asia clone of community-associated MRSA. Genome Biol., 26; 17(1):160. doi: 10.1186/s13059-016-1022-0.
7. Unemo M, Golparian D, Sánchez-Busó L, Grad Y, Jacobsson S, Ohnishi M, Lahra MM, Limnios A, Sikora AE, Wi T, Harris SR. (2016). The novel 2016 WHO Neisseria gonorrhoeae reference strains for global quality assurance of laboratory investigations: phenotypic, genetic and reference genome characterization. J Antimicrob Chemother., 71(11):3096-3108. doi: 10.1093/jac/dkw288.
8. Eyre DW, Sanderson ND, Lord E, Regisford-Reimmer N, Chau K, Barker L, Morgan M, Newnham R, Golparian D, Unemo M, Crook DW, Peto TE, Hughes G, Cole MJ, Fifer H, Edwards A, Andersson MI. (2018). Gonorrhoea treatment failure caused by a Neisseria gonorrhoeae strain with combined ceftriaxone and high-level azithromycin resistance, England, Euro Surveill, 23(27):1800323. doi: 10.2807/1560-7917.ES.2018.23.27.1800323.
9. Lee JYH, Monk IR, Gonçalves da Silva A, Seemann T, Chua KYL, Kearns A, Hill R, Woodford N, Bartels MD, Strommenger B, Laurent F, Dodémont M, Deplano A, Patel R, Larsen AR, Korman TM, Stinear TP, Howden BP. (2018). Global spread of three multidrug-resistant lineages of Staphylococcus epidermidis. Nat Microbiol., 3(10):1175-1185. doi: 10.1038/s41564-018-0230-7.
10. O’Neill GL, Murchan S, Gil-Setas A, Aucken HM. (2001). Identification and characterization of phage variants of a strain of epidemic methicillin-resistant Staphylococcus aureus (EMRSA-15). Journal of Clinical Microbiology, 39 (4):1540- 1548. doi:10.1128/JCM.39.4.1540-1548.2001.
11. WHO (2015). Global plan for antimicrobial resistance. Archivedfrom the original on 15 May 2015. Retrieved 9.
12. Dheer D, Gupta JPN, Shankar R. (2018). Tacrolimus: An updated review on delivering strategies for multifarious diseases. European Journal of Pharmaceutical Sciences 114: 217-227. doi: 10.1016/j.ejps.2017.12.017.
13. Cheba BA. (2011). Chitin and chitosan: marine biopolymers with unique properties and versatile applications. Global Journal of Biotechnology &. Biochemistry 6(3):149-153.
14. Badawy MEI, Rabea EI. (2011). A biopolymer chitosan and its derivatives as promising antimicrobial agents against plant pathogens and their applications in crop protection. International Journal of Carbohydrate Chemistry, 2011: 29.
15. Rosso AP, Martinelli M. (2020). Preparation and characterization of dendronized chitosan/gelatin-based nanogels. European Polymer Journal 124: 109506,
16. Molpeceres J, Aberturas MR, Guzman M. (2000). Biodegradable nanoparticles as a delivery system for cyclosporine: preparation and characterization. Journal of Microencapsulation 17(5) (2000) 599-614. doi: 10.1080/026520400417658.
17. Aminabhavi TM, Dharupaneedi SP, More AU. (2017). The role of nanotechnology & chitosan-based biomaterials for tissue engineering and therapeutic delivery. Chitosan Based Biomaterials, 2:1-29. doi:10.1016/B978-0-08-100228-5.00001-8.
18. Adhikari HS, Yadav PN. (2018). Anticancer activity of chitosan, chitosan derivatives, and their mechanism of action. International Journal of Biomaterials, vol. 2018: 29. doi:
19. Saeidnia S. (2014). Phthalates, Encyclopaedia of toxicology 3rd Edition.
20. Occupational Safety and Health Administration (OSHA), Phthalic acid. Available online at:
21. Lotha G (2013). Phthalic acid chemical compound safety. Britannica.
22. Hiraiwa Y, Morinaka A, Fukushima T, Kudo T. (2009). Metallo-β-lactamase inhibitory activity of phthalic acid derivatives. Bioorganic and Medicinal Chemistry Letters, 19 (17):5162-5165. doi:10.1016/j.bmcl.2009.07.018.
23. Cassano R, De-Amicis F, Servidio C, Curcio F, Trombino S. (2020). Preparation, characterization and in vitro evaluation of resveratrol-loaded nano-spheres potentially useful for human breast carcinoma. Journal of Drug Delivery Science and Technology, 57:101748,
24. Sasidharan S, Chen Y, Saravanan D, Sundram KM, Latha LY. (2011). Extraction, isolation and characterization of bioactive compounds from plants' extracts. Afr J. Tradit. Complement Altern Med., 8(1): 1-10, PMID: 22238476.
25.  Saraswathi SV, Saravanan D, Santhakumar K. (2017). Isolation of quercetin from the methanolic extract of Lagerstroemia speciosa by HPLC technique, its cytotoxicity against MCF-7 cells and photocatalytic activity. J. Photochem Photobiol, B 171:20- 26. doi: 10.1016/j.jphotobiol.2017.04.031.
26. Alfarra HY, Muhammd NO. (2014). HPLC separation and isolation of asiaticoside from Centella asiatica and its biotransformation by A. niger. Int. J. Pharm. Med. & Bio. Sc. 3(3):1-8.
27. Gini TG, Jothi GJ. (2018). Column chromatography and HPLC analysis of phenolic compounds in the fractions of Salvinia molesta Mitchell. Egyptian Journal of Basic and Applied Sciences, 5(3):197-203.
28. Ukwubile CA, Ahmed A, Katsayal UA,  Ya'u J, Mejida S. (2019). GC-MS analysis of bioactive compounds from Melastomastrum capitatum (Vahl) Fern. leaf methanol extract: An anticancer plant, Scientific African 3(2019) e00059. doi:
29. Lazaridou M, Christodoulou E, Nerantzaki M, Kostoglou M, Lambropoulou DA, Katsarou A, Pantopoulos K, Bikiaris DN. (2020). Formulation and In-vitro Characterization of Chitosan-Nanoparticles Loaded with the Iron Chelator Deferoxamine Mesylate (DFO). Pharmaceutics, 12(3) :238. doi: 10.3390/pharmaceutics12030238.
30. Naskar S, Sharma S, Kuotsu K. (2019). Chitosan-based nanoparticles: An overview of biomedical applications and its preparation. Journal of Drug Delivery Science and Technology, 49:66-81.
31. Shariatinia Z. (2020). Biopolymeric Nanocomposites in Drug Delivery. In: Nayak A, Hasnain M. (Eds) Advanced Biopolymeric Systems for Drug Delivery. Advances in Material Research and Tech. Springer, Cham. doi:10.1007/978-3-030- 46923-8-10.
32. Ramachandran R, Shanmughave P. (2010). Preparation and characterization of biopolymeric nanoparticles used in drug delivery. Indian Journal of Biochemistry & Biophysics, 47(2010) 56-59.
33. The European Committee on Antimicrobial Susceptibility Testing (EUCAST),
34. Mohammed FADL, Alaasar MM. (2018). In Vitro Susceptibility Testing of Silver and Zinc Nanoparticles on Different Fungal Species. The Medical Journal of Cairo University,86: 2639-2643.doi: 10.21608/mjcu.2018.58068.
35. Clinical and Laboratory Standards Institute (CLSI) (2012). Methods for antimicrobial susceptibility testing of anaerobic bacteria, Approved Standard 8th Edition CLSI document M11-A8. Wayne, PA: Clinical and Laboratory Standards Institute, 32 (5).
36. Andrews JM. (2001). Determination of minimum inhibitory concentrations. J. Antimicrob Chemother.48 (1):5-16. doi: 10.1093/jac/48.suppl_1.5.
 37. Escárcega-González CE, Garza-Cervantes JA,Vazquez-Rodríguez A, Montelongo-Peralta LZ, Treviño-Gonzalez MT, Barriga-Castro ED, Saucedo-Salazar EM, Morales RMC, Regalado-Soto DI, Treviño-González FM, Rosales JLC, Cruz RV, Morones-Ramirez JR. (2018).  In vivo antimicrobial activity of silver nanoparticles produced via a green chemistry synthesis using Acacia rigidula as a reducing and capping agent.  International Journal of Nanomedicine, 13: 2349-2363. doi: