Design, optimization, and antimicrobial assessment of Callicarpa longifolia-derived nanoparticles using quality by design (QbD) approach
DOI:
https://doi.org/10.69857/joapr.v13i4.1322Keywords:
Callicarpa longifolia-derived nanoparticles, Box-Behnken Design, Isolated extract fraction, Drug release, Antibacterial studyAbstract
Background: The purpose of this study is to focus on the design, optimization, and antimicrobial evaluation of ethanolic leaf extracts of Callicarpa longifolia-derived nanoparticles using the Quality by Design (QbD) technique. Methodology: Critical formulation parameters were optimized using a Box-Behnken Design. The optimized nanoparticles are characterized using Dynamic Light Scattering (DLS), Zeta Potential analysis, and Scanning Electron Microscopy (SEM), confirming their nanoscale size. Stability studies were conducted under various ICH recommendations. The antimicrobial activity of the isolated fraction extract and isolated fraction extract nanoparticles was assessed against Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria using the agar well diffusion method. Results and Discussion: By BBD, the optimized herbal nanoparticles have a particle size of 281.00 nm and an entrapment efficiency 88%. After characterization, the results of the optimized nanoparticles' particle size (349.3 nm), zeta potentials (-23.7 mV), % EE (86.25%), and spherical shape are confirmed by SEM. The % cumulative drug release of optimized nanoparticles is 86.12±0.79. Kinetic release model regression values of the optimized nanoparticles' R² values in different model kinetic releases are zero order (0.929), first order (0.971), Higuchi kinetic release (0.994), Korsmeyer kinetic release (0.994), and Hixon Crowell (0.978). Results revealed that the nanoparticle formulation exhibited significant antimicrobial efficacy. Conclusion: All things considered, the study shows how the QbD methodology may be successfully applied to create a stable and efficient nanoparticle system made from an isolated extract of C. longifolia, which has encouraging potential as a substitute antibacterial agent.
Downloads
References
Ansari P, Reberio AD, Ansari NJ, Kumar S, Khan JT, Chowdhury S, Abd El-Mordy FM, Hannan JMA, Flatt PR, Abdel-Wahab YHA, Seidel V. Therapeutic Potential of Medicinal Plants and Their Phytoconstituents in Diabetes, Cancer, Infections, Cardiovascular Diseases, Inflammation and Gastrointestinal Disorders. Biomedicines, 13(2), 454 (2025) https://doi.org/10.3390/biomedicines13020454
Bhoi A, Dwivedi SD, Singh D, Singh MR, Keshavkant S. Worldwide health scenario from the perspective of herbal medicine research. In Phytopharmaceuticals and herbal drugs. Academic Press,13-32 (2023) https://doi.org/10.1016/B978-0-323-99125-4.00014-7
Oyedepo TA, Palai S. Herbal remedies, toxicity, and regulations. Preparation of Phytopharmaceuticals for the Management of Disorders. Academic Press, 89-127 (2021) https://doi.org/10.1016/B978-0-12-820284-5.00014-9
Hassen G, Belete G, Carrera KG, Iriowen RO, Araya H, Alemu T, Solomon N, Bam DS, Nicola SM, Araya ME, Debele T, Zouetr M, Jain N. Clinical Implications of Herbal Supplements in Conventional Medical Practice: A US Perspective. Cureus, 14(7), e26893 (2022) http://dx.doi.org/10.7759/cureus.26893
Javed MN, Dahiya ES, Ibrahim AM, Alam MS, Khan FA, Pottoo FH. Recent advancement in clinical application of nanotechnological approached targeted delivery of herbal drugs. Nanophytomedicine, Concept to Clinic, 151-72 (2020) https://doi.org/10.1007/978-981-15-4909-0_9
Taleuzzaman M, Jain P, Verma R, Iqbal Z, Mirza MA. Eugenol as a Potential Drug Candidate: A Review. Current Topics in Medicinal Chemistry, 21(20), 1804-1815 (2021) https://doi.org/10.2174/1568026621666210701141433
Wang CY, Chen LH, He HL, Liu Y, Wang Q. The complete chloroplast genome of Callicarpa longifolia Lamk. var. floccosa Schauer (Lamiaceae). Mitochondrial DNA Part B, 6(12), 3473-3474 (2021) https://doi.org/10.1080/23802359.2021.2002210
Tourabi M, Metouekel A, Ghouizi AE, Jeddi M, Nouioura G, Laaroussi H, Hosen ME, Benbrahim KF, Bourhia M, Salamatullah AM, Nafidi HA. Efficacy of various extracting solvents on phytochemical composition, and biological properties of Mentha longifolia L. leaf extracts. Scientific Reports, 13(1), 18028 (2023) https://doi.org/10.1038/s41598-023-45030-5
Yu M, Gouvinhas I, Rocha J, Barros AI. Phytochemical and antioxidant analysis of medicinal and food plants towards bioactive food and pharmaceutical resources. Scientific reports, 11(1), 10041 (2021) https://doi.org/10.1038/s41598-021-89437-4
Nghakliana F, Zosangzuali M, Lalremruati M, Lalmuansangi C. Ethnopharmacological Applications and Phytochemical Compounds of the Genus Callicarpa L. Bioactives and Pharmacology of Lamiaceae Apple Academic Press, 1, 343-366 (2023) https://doi.org/10.1201/9781003346142
Dewi MK, Chaerunisaa AY, Muhaimin M, Joni IM. Improved Activity of Herbal Medicines through Nanotechnology. Nanomaterials (Basel), 12(22), 4073 (2022) https://doi.org/10.3390/nano12224073
Awlqadr FH, Majeed KR, Altemimi AB, Hassan AM, Qadir SA, Saeed MN, Faraj AM, Salih TH, Abd Al‐Manhel AJ, Najm MA, Tsakali E. Nanotechnology-Based Herbal Medicine: Preparation, Synthesis, and Applications in Food and Medicine. Journal of Agriculture and Food Research, 19, 101661 (2025) https://doi.org/10.1016/j.jafr.2025.101661
Kaur J, Anwer MK, Sartaj A, Panda BP, Ali A, Zafar A, Kumar V, Gilani SJ, Kala C, Taleuzzaman M. ZnO nanoparticles of Rubia cordifolia extract formulation developed and optimized with QbD application, considering ex vivo skin permeation, antimicrobial and antioxidant properties. Molecules, 27(4), 1450 (2021) https://doi.org/10.3390/molecules27041450
Taleuzzaman M, Sartaj A, Kumar Gupta D, Gilani SJ, Mirza MA. Phytosomal gel of Manjistha extract (MJE) formulated and optimized with central composite design of Quality by Design (QbD). Journal of Dispersion Science and Technology, 44(2), 236-44 (2021) https://doi.org/10.1080/01932691.2021.1942036
Dewi MK, Chaerunisaa AY, Muhaimin M, Joni IM. Improved Activity of Herbal Medicines through Nanotechnology. Nanomaterials (Basel), 12(22), 4073 (2022) https://doi.org/10.3390/nano12224073
Lopalco A, Iacobazzi RM, Lopedota AA, Denora N. Recent Advances in Nanodrug Delivery Systems Production, Efficacy, Safety, and Toxicity. Methods Molecular Biology, 2834, 303-332 (2024) http://dx.doi.org/10.1007/978-1-0716-4003-6_15
Sharma D, Gupta A, Rawat R, Sharma S, Yadav JS, Saxena A. Exploring nano formulation drug delivery of herbal actives for enhanced therapeutic efficacy: A comprehensive Review. Intelligent Pharmacy, 3(1), 26-34 (2024) https://doi.org/10.1016/j.ipha.2024.07.004
Huang L, Luo S, Tong S, Lv Z, Wu J. The development of nanocarriers for natural products. Wiley Interdisciplinary Reviews: Nanomedicine and Nanobiotechnology, 16(3), e1967 (2024) https://doi.org/10.1002/wnan.1967
Rombolà L, Scuteri D, Marilisa S, Watanabe C, Morrone LA, Bagetta G, Corasaniti MT. Pharmacokinetic Interactions between Herbal Medicines and Drugs: Their Mechanisms and Clinical Relevance. Life (Basel), 10(7), 106 (2020) https://doi.org/10.3390/life10070106
Abdul Rahim R, Jayusman PA, Muhammad N, Ahmad F, Mokhtar N, Naina Mohamed I, Mohamed N, Shuid AN. Recent Advances in Nanoencapsulation Systems Using PLGA of Bioactive Phenolics for Protection against Chronic Diseases. Int J Environ Res Public Health. 16(24), 4962 (2019) https://doi.org/10.3390/ijerph16244962
Roshan Z, Haddadi‐Asl V, Ahmadi H, Moussaei M. Curcumin‐encapsulated poly (lactic‐co‐glycolic acid) nanoparticles: a comparison of drug release kinetics from particles prepared via electrospray and nanoprecipitation. Macromolecular Materials and Engineering. 309 (7):2400040, (2024) https://doi.org/10.1002/mame.202400040
Awlqadr FH, Majeed KR, Altemimi AB, Hassan AM, Qadir SA, Saeed MN, Faraj AM, Salih TH, Abd Al‐Manhel AJ, Najm MA, Tsakali E. Nanotechnology-based herbal medicine: Preparation, synthesis, and applications in food and medicine. Journal of Agriculture and Food Research. 31:101661, (2025), https://doi.org/10.1016/j.jafr.2025.101661
Zagalo DM, Sousa J, Simões S. Quality by design (QbD) approach in marketing authorization procedures of non-biological complex drugs: a critical evaluation. European Journal of Pharmaceutics and Biopharmaceutics, 178, 1-24 (2022) https://doi.org/10.1016/j.ejpb.2022.07.014
Cunha S, Costa CP, Moreira JN, Lobo JM, Silva AC. Using the quality by design (QbD) approach to optimize formulations of lipid nanoparticles and nano emulsions: A review. Nanomedicine: Nanotechnology, Biology and Medicine, 28, 102206 (2020) https://doi.org/10.1016/j.nano.2020.102206
Beg S, Haneef J, Rahman M, Peraman R, Taleuzzaman M, Almalki WH. Introduction to analytical quality by design. InHandbook of Analytical Quality by Design 2021 Jan 1 (pp. 1-14). Academic Press. https://doi.org/10.1016/B978-0-12-820332-3.00009-1
Alam MS, Srivastava N. Extraction, Phytochemical Characterization and Assessment of Antioxidant and Antimicrobial Properties of Callicarpa Longifolia. Biochemical and Cellular Archives, 25(1), 777-786 (2025) https://doi.org/10.51470/bca.2025.25.1.777
Alam MS, Shrivastava N. Exploring key compounds in callicarpa longifolia: a study on isolation and identification. Journal of Applied Pharmaceutical Research, 13(1), 75-85 (2025) https://doi.org/10.69857/joapr.v13i1.751
Sindhu RK, Gupta R, Wadhera G, Kumar P. Modern Herbal Nanogels: Formulation, Delivery Methods, and Applications. Gels, 8(2), 97 (2022) https://doi.org/10.3390/gels8020097
Awlqadr FH, Majeed KR, Altemimi AB, Hassan AM, Qadir SA, Saeed MN, Faraj AM, Salih TH, Abd Al‐Manhel AJ, Najm MA, Tsakali E. Nanotechnology-Based Herbal Medicine: Preparation, Synthesis, and Applications in Food and Medicine. Journal of Agriculture and Food Research, 19, 101661 (2025) https://doi.org/10.1016/j.jafr.2025.101661
Sharaf NS, Shetta A, Elhalawani JE, Mamdouh W. Applying Box-Behnken Design for Formulation and Optimization of PLGA-Coffee Nanoparticles and Detecting Enhanced Antioxidant and Anticancer Activities. Polymers (Basel), 14(1), 144 (2021) https://doi.org/10.3390/polym14010144
Alam P, Shakeel F, Taleuzzaman M, Foudah AI, Alqarni MH, Aljarba TM, Alshehri S, Ghoneim MM. Box-Behnken Design (BBD) application for optimization of chromatographic conditions in RP-HPLC method development for the estimation of thymoquinone in nigella sativa seed powder. Processes, 10(6), 1082 (2022) https://doi.org/10.3390/pr10061082
Sisodiya S, Gautam S, Aneja D, Debnath M. Exploring the usage of Olive leaf and green synthesized Olive leaf metallic nanoparticles as soap supplement for the formulation and optimization of innovative germicidal herbal soap. Industrial Crops and Products, 219, 119012 (2024) https://doi.org/10.1016/j.indcrop.2024.119012
Luiz MT, Viegas JS, Abriata JP, Viegas F, de Carvalho Vicentini FT, Bentley MV, Chorilli M, Marchetti JM, Tapia-Blacido DR. Design of experiments (DoE) to develop and to optimize nanoparticles as drug delivery systems. European Journal of Pharmaceutics and Biopharmaceutics, 165, 127-48 (2021) https://doi.org/10.1016/j.ejpb.2021.05.011
Bala Chennaiah M, Kumar KD, Kumar BS, Tanneeru SR. Characterisation of zinc oxide nanoparticles–herbal synthesised coated with Ocimum tenuiflorum. Advances in Materials and Processing Technologies, 8(2), 466-77 (2021) https://doi.org/10.1080/2374068X.2021.1934642
Altammar KA. A review on nanoparticles: characteristics, synthesis, applications, and challenges. Frontiers in Microbiology, 14, 1155622 (2023) https://doi.org/10.3389/fmicb.2023.1155622
Taleuzzaman M, Ahmed MM, Chattopadhyay M. Particle size role, Importance and Strategy of HPLC Analysis-An update. Int Arch BioMed Clin Res., 2(2), (2016) http://dx.doi.org/10.21276/iabcr.2016.2.2.2
Kumari V, Tyagi P, Sangal A. In-Vitro kinetic release study of illicium verum (Chakraphool) polymeric nanoparticles. Materials Today: Proceedings, 60(1), 14-20 (2022) https://doi.org/10.1016/j.matpr.2021.11.014
Santadkha T, Skolpap W, Thitapakorn V. Diffusion Modeling and in vitro release kinetics studies of curcumin-loaded superparamagnetic nanomicelles in cancer drug delivery system. Journal of Pharmaceutical Sciences, 111(6), 1690-99 (2022) https://doi.org/10.1016/j.xphs.2021.11.015
Rahat I, Imam SS, Rizwanullah M, Alshehri S, Asif M, Kala C, Taleuzzaman M. Thymoquinone-entrapped chitosan-modified nanoparticles: formulation optimization to preclinical bioavailability assessments. Drug Delivery, 28(1), 973-84 (2021) https://doi.org/10.1080/10717544.2021.1927245
Santadkha T, Skolpap W, Thitapakorn V. Diffusion Modeling and in vitro release kinetics studies of curcumin-loaded superparamagnetic nanomicelles in cancer drug delivery system. Journal of Pharmaceutical Sciences, 111(6), 1690-99 (2022) https://doi.org/10.1016/j.xphs.2021.11.015
Weng J, Tong HHY, Chow SF. In Vitro Release Study of the Polymeric Drug Nanoparticles: Development and Validation of a Novel Method. Pharmaceutics, 12(8), 732 (2020) https://doi.org/10.3390/pharmaceutics12080732
Varghese RM, Aravind Kumar S, Rajeshkumar S. Antibacterial activity of herbal formulation against common oral pathogens. Bioinformation, 19(5), 663-669 (2023 https://doi.org/10.6026/97320630019663
Varghese RM, Kumar A, Shanmugam R. Antimicrobial activity of zinc oxide nanoparticles synthesized using Ocimum tenuiflorum and Ocimum gratissimum herbal formulation against oral pathogens. Cureus, 16(2), e53562 (2024) https://doi.org/10.7759/cureus.53562
Shanmugam R, Govindharaj S, Arunkumar P, Sanjana GS, Manigandan P, Sulochana G, Padmapriya A. Preparation of a herbal mouthwash with lemongrass and mint-mediated zinc oxide nanoparticles and evaluation of its antimicrobial and cytotoxic properties. Cureus, 16(2), e53671 (2024) https://doi.org/10.7759/cureus.53671
Hossain TJ. Methods for screening and evaluation of antimicrobial activity: A review of protocols, advantages, and limitations. European Journal of Microbiology and Immunology, 14(2), 97-115 (2024) https://doi.org/10.1556/1886.2024.00035
Mihailovic V, Sreckovic N, Nedic ZP, Dimitrijevic S, Matic M, Obradovic A, Selakovic D, Rosic G, Katanic Stankovic JS. Green synthesis of silver nanoparticles using Salvia verticillata and Filipendula ulmaria extracts: Optimization of synthesis, biological activities, and catalytic properties. Molecules, 28(2), 808 (2023) https://doi.org/10.3390/molecules28020808
Choi S, Kang BS, Choi G, Kang M, Park H, Kim N, Chang PS, Kwak MK, Jeong HE, Jung HS. Multichamber PLGA Microparticles with Enhanced Monodispersity and Encapsulation Efficiency Fabricated by a Batch‐Microfluidic Hybrid Approach. Advanced NanoBiomed Research, 3(10), 2300044 (2023) https://doi.org/10.1002/anbr.202300044
Li L, Li Z, Guo Y, Zhang K, Mi W, Liu J. Preparation of uniform-sized GeXIVA [1, 2]-loaded PLGA microspheres as long-effective release system with high encapsulation efficiency. Drug Delivery, 29(1), 2283-95 (2022) https://doi.org/10.1080/10717544.2022.2089297
Benavides Castillo L, Martinez Y. The concentration and type of emulsifier rules the oil/water and water/oil/water emulsion size distribution. Chemical Engineering Communications, 210(11), 2064-71 (2023) https://doi.org/10.1080/00986445.2023.2169680
Dong R, Wang F, Jing D, Liu Y, Bao Y. The Stirring Effect on the Crystal Morphology of p-Acetamidobenzoic Acid Solution Crystallization. Crystals, 15(3), 284 (2025) https://doi.org/10.3390/cryst15030284
Liaqat N, Jahan N, Khalil-Ur-Rahman, Anwar T, Qureshi H. Green synthesized silver nanoparticles: Optimization, characterization, antimicrobial activity, and cytotoxicity study by hemolysis assay. Frontiers in Chemistry, 10, 952006 (2022) https://doi.org/10.3389/fchem.2022.952006
Ryu S, Nam SH, Baek JS. Green Synthesis of Silver Nanoparticles (AgNPs) of Angelica Gigas Fabricated by Hot-Melt Extrusion Technology for Enhanced Antifungal Effects. Materials (Basel), 15(20), 7231 (2022) https://doi.org/10.3390/ma15207231
Akhtar K, Khan SA, Khan SB, Asiri AM. Scanning electron microscopy: Principle and applications in nanomaterials characterization. Springer International Publishing, 113-145 (2018) https://doi.org/10.1007/978-3-319-92955-2_4
Sarma SK, Dutta U, Bharali A, Kumar S, Baruah S, Sarma H, Laloo D, Sahu BP. Isolation of curcumin from Lakadong turmeric of Meghalaya and development of its PLGA-Cur-NS loaded nanogel for potential anti-inflammatory and cutaneous wound healing activity in Wistar rats. Future Journal of Pharmaceutical Sciences, 9(1), 85 (2023) https://doi.org/10.1186/s43094-023-00534-9
Manjunath AM, Priya S, Jyothi D. Mucoadhesive Chitosan-Coated PLGA Nanoparticles of Ashwagandha Extract for Colon-Targeted Delivery. Ind. J. Pharm. Edu. Res., 57(4), 971-82 (2023) http://dx.doi.org/10.5530/ijper.57.4.119
Dalli M, El Guerraf A, Azizi SE, Benataya K, Azghar A, Mi-Kyung J, Maleb A, Bonglee K, Gseyra N. Loaded n-hydroxyapatite/SSG 3D scaffolds as a drug delivery system of nigella sativa fractions for the management of local antibacterial infections. Nanomaterials, 12(5), 856 (2022) https://doi.org/10.3390/nano12050856
Salim SA, Abed T, Abdelazim EB, Eissa NG, Elsabahy M, Kamoun EA. Exploring the Therapeutic Potential of Plant Extract-Loaded Polymeric Nanoparticles for Advanced Wound Healing: A Comprehensive Review. Arabian Journal for Science and Engineering, 13, 1-9 (2025) https://doi.org/10.1007/s13369-025-10247-1
Vladár AE, Hodoroaba VD. Characterization of nanoparticles by scanning electron microscopy. Characterization of nanoparticles, 7-27 (2020) https://doi.org/10.1016/B978-0-12-814182-3.00002-X
Elumalai K, Srinivasan S, Shanmugam A. Review of the efficacy of nanoparticle-based drug delivery systems for cancer treatment. Biomedical Technology, 5, 109-22 (2024) https://doi.org/10.1016/j.bmt.2023.09.001
Khattab A, Nattouf A. Optimization of entrapment efficiency and release of clindamycin in microsponge based gel. Scientific Reports, 11(1), 23345 (2021) https://doi.org/10.1038/s41598-021-02826-7
Fahmy OM, Eissa RA, Mohamed HH, Eissa NG, Elsabahy M. Machine learning algorithms for prediction of entrapment efficiency in nanomaterials. Methods, 218, 133-40 (2023) https://doi.org/10.1016/j.ymeth.2023.08.008
Baltz N, Scherließ R. Entrapment efficiency methodology for lipid nanoparticles-a literature review. Open Nano, 100251 (2025) https://doi.org/10.1016/j.onano.2025.100251
Ewii UE, Attama AA, Olorunsola EO, Onugwu AL, Nwakpa FU, Anyiam C, Chijioke C, Ogbulie T. Nanoparticles for Drug Delivery: Insight Into In Vitro and In Vivo Drug Release from Nanomedicines. Nano TransMed, 100083 (2025) https://doi.org/10.1016/j.ntm.2025.100083
Yang L, Wu W, Yang J, Xu M. Nanoparticle-mediated delivery of herbal-derived natural products to modulate immunosenescence-induced drug resistance in cancer therapy: a comprehensive review. Frontiers in Oncology, 15, 1567896 (2025) https://doi.org/10.3389/fonc.2025.1567896
Mehta J, Pathania K, Pawar SV. Recent overview of nanotechnolog based approaches for targeted delivery of nutraceuticals. Sustainable Food Technology, (2025) https://doi.org/10.1039/D5FB00122F
Ezzariga N, Moukal A, Asdadi A, Lemkhente Z, Moustaoui F, Kaaya A, Aghrouch M. Evaluation of the Antimicrobial Activity of 20 Essential Oils and Their Combinations on Bacterial and Fungal Strains. Cureus, 17(2), e79499 (2025) https://doi.org/10.7759/cureus.79499
Nagore P, Ghotekar S, Mane K, Ghoti A, Bilal M, Roy A. Structural properties and antimicrobial activities of Polyalthia longifolia leaf extract-mediated CuO nanoparticles. BioNanoScience, 11(2), 579-89 (2021) https://doi.org/10.1007/s12668-021-00851-4
Tourabi M, Metouekel A, Ghouizi AE, Jeddi M, Nouioura G, Laaroussi H, Hosen ME, Benbrahim KF, Bourhia M, Salamatullah AM, Nafidi HA. Efficacy of various extracting solvents on phytochemical composition, and biological properties of Mentha longifolia L. leaf extracts. Scientific Reports, 13(1), 18028 (2023) https://doi.org/10.1038/s41598-023-45030-5
Published
How to Cite
Issue
Section
Copyright (c) 2025 Md. Shakeel Alam, Nidhi Srivastava
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
