Green Synthesis Characterization and Biological Assessment of Nickle based Microparticles using Justicia Adathoda L. Leaf Extract

Green Synthesis Characterization and Biological Assessment of Nickle based Microparticles using Justicia Adathoda L. 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Leaf Extract Waqas Ahmad, Aftab Alam, Noor Ul Islam, Wahab khan This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8368802/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Nickel-based oxide/hydroxide microparticles (NiO/Ni(OH)₂ MPs) were synthesized via a green way utilizing Justicia adathoda L. leaf extract, serving as novel reducing and stabilizing agent. Plants phytochemicals such as alkaloids, flavonoids, and phenolics facilitated the eco-friendly formations of NiO/Ni(OH) 2 MPs without toxic chemicals. Structure analysis through X-ray diffraction (XRD) confirmed the crystalline structure, while FTIR confirmed the involvement of functional groups in reduction and stabilizations. Morphological characterization by Scanning Electron Microscopy (SEM) revealed a granular dense structure microparticle of size about 1µm. Biological screening showed notable antibacterial and antifungal activities. NiO/Ni(OH) 2 MPs show 60.24% inhibition against Staphylococcus aureus and 70.24% inhibition against Pseudomonas aeruginosa, while antifungal activity reached 95% against Aspergillus fumigatus . Compared to standard pharmaceutical agents such as Ofloxacin and Miconazole, the NiO/Ni(OH) 2 MPs synthesized using Justicia adathoda L. leaf extract revealed potential antibacterial and antifungal activities. These results underline the value of Justicia adathoda as an eco-friendly and efficient biological resource for making functional NiO/Ni(OH) 2 MPs. The synthesized microparticles show strong applications in biomedical fields, including therapeutic formulations, antimicrobial coatings, and presenting a sustainable unconventional to chemically synthesized micromaterials. Nanoscience Green synthesis Ni based microparticle SEM FTIR XRD antimicrobial activity Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 INTODUCTION Green synthesis has become a major topic of interest in the field of Micro-nanotechnology (Shebl et al., 2019 ). Developments in micro-nanotechnology have led to the production of innovative materials, particularly ecofriendly micro-based-nanostructure synthetic procedures used to synthesize metal and metal oxide micromaterials of any shape and size (Fuku et al., 2020 ). Most physical, chemical, hybrid methods tend to be costly and detrimental due to the presence of harmful substances like solvents, reducing agents, and precursors (Shaik et al., 2016 ). In contrast, employing green techniques for the preparation of microparticles (MPs) is new economical, ecofriendly, and simpler to utensils. Consequently, such technique is increasingly utilized in the production of metallic based microparticles. A diverse span of natural deposit, inclusive of microorganisms, biomolecules, and plant extracts, was explored for green MP synthesis (F. A. Khan et al., 2016 ). Although numerous studies have been conducted on the green synthesis of MPs utilizing biomolecules and microbes, plant extracts are often favored because they are cost-effective, efficient, and do not require the complex processes involved in isolating and maintaining microbial cultures (M. Khan et al., 2022 ). Various plant species, their parts, and isolated compounds are effectively used to synthesize green MPs. Furthermore, the preparation of micromaterials can be made more efficiently by incorporating industrial and agricultural waste, in computation to being ecofriendly (Iftikhar et al., 2020 ). Phenolic chemicals can operate as capping agents in microparticles manufacturing, whereas biological molecules offer unique uses in micro-nanotechnology, making metal microparticles durable and trustworthy (Bar et al., 2009 ). Recent studies have proved the effectiveness of using micromaterial’s in different aspects of life, including environmental control, energy storage, solar cell, biomarking, bio probes, tissue engineering, cancer therapy, cancer diagnosis, and drug delivery (Mandal & Ganguly, 2011 ). The method of green synthesis consists in obtaining particles using safe materials as reducing agents and stabilizers, for example, products of biological origin. For example, the method can be carried out using extracts of plants or their individual parts, as well as microorganism (bacteria, yeast, fungi) (J. Singh et al., 2018 ). A plant contains various biochemicals and metabolites that can be used for the green synthesis of microparticles as they possess reducing and stabilizing properties. Green synthesis technologies are environmentally friendly, cost effective stable and nontoxic than traditional biological, physical, and chemical approaches (Mustapha et al., 2022 ). Biosynthesized metal and metal oxide MPs have rising applications in the medical sector including immunotherapy, dentistry, wound healing, diagnosis, regenerative medicines, bio toxicology and bio-sensing platform and its antiviral, fungicidal and bactericidal potentials (Pandit et al., 2022 ). Microparticles can be variable in dimension, shape, size and architectures(Machado et al., 2015 ). In the environmental field, the green synthesis of MPs has been used to control environmental pollution, such as with the degradation of organic dyes and chlorinated organic pollutants, heavy metal removal, in addition to the treatment of wastewater (Saif et al., 2016 ). Justicia adathoda L . also known as Malabar nut is a well-known and important medicinally important plant of family Acanthaceae growing in different Asian countries including Pakistan, Sirilanka, India and Nepal (Hossain & Hoq, 2016 ). Justicia adathoda L. has diverse applications in traditionally used Ayurvedic and Unani medicines. The local populace uses the plants' leaves to treat coughs, colds, and asthma. Justicia adathoda L . plant has several key properties, including antibacterial, antioxidant, antiulcer, antidiabetic, anti-allergic, anti-genotoxic, and anti-phlogiston. Adathoda vasica leaves contain phytochemicals including flavonoids, saponins, tannins, alkaloids and phenolic compounds. Justicia adathoda L . leaf extract contains the two most significant quinazoline alkaloids, vasicine and vasicione (Khadri et al., 2013 ). Phytochemicals are bioactive compounds naturally found in plants, and their composition varies greatly among different plant species and even within different parts of the same plant. The most common classes of phytochemicals include polyphenols (like flavonoids and tannins), terpenoids, alkaloids, carotenoids, and saponins. These compounds are involved in plant defense mechanisms and can also offer various health benefits to humans (Lydon & Duke, 1989). Antibacterial agents are utilized to inhibit the proliferation of harmful bacteria. They help prevent illness and eradicate microorganisms (Yao & Moellering, 2011 ). Numerous antibiotics are sourced from plants and various chemicals to combat diseases. Bacteria are the pathogens responsible for poisoning. For this assay, we used five different strains of pathogens (Balloux & van Dorp, 2017 ). A disc diffusion method was employed to assess bacterial activity. Bacteria are classified into two categories: Gram-negative and Gram-positive. Staphylococcus aureus (S. aureus) is typically recognized as a Gram-positive bacterium and is frequently used in antibacterial studies due to its significant potential to cause infections in humans. Escherichia coli (E. coli), a Gram-negative bacterium, is established in contaminated water and along with as a pathogenic microbe that leads to human illness, as well as serving as an indicator of water quality (Margesin, 2017). The antifungal test aims to eliminate microorganisms, especially fungi. Fungi pose significant risks and can lead to various illnesses. We utilize antifungal tests to identify infections caused by fungi. These organisms can damage fruits, vegetables, and plants (Leyva Salas et al., 2017 ). Numerous plants are recognized for their antifungal properties. Unsanitary food and environments weaken the immune system, facilitating the spread of fungal infections. Antifungal medications are necessary to treat such infections (Das et al., 2013 ). Metallic nickel based microparticles are those that are used as the primary raw material without combining them with other metals, compounds or elements. The term nickel microparticle used in this review also refers to metallic nickel microparticle (Jaji et al., 2020 ). The successful production of Ni based MPs employing various plant extracts, as well as their enhanced biological potential, call for additional research into other medicinally essential plants for future enhancement (Hussain et al., 2023 ). Ni based MPs have gained much attention due to their unique magnetic, chemical, and physical properties as well as their potential applications in various technological fields such as catalysis (Bibi et al., 2017 a), battery manufacture (Cheng et al., 2020 ), incorporation in textile (Jiao et al., 2019 ), and optical switches (Reena Mary et al., 2011 ). The current study aimed to synthesis Ni based MPs using Justicia adathoda L. leaf extract. The produced (NiO/Ni(OH)) 2 MPs) were examined by using modern analytical techniques, including X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM). Within inclusion, the antibacterial and antifungal potentials of the prepared Nickel based microparticles were measured. METHODOLOGY Chemicals and other materials The chemicals used were Distilled water and Nickel chloride. These chemicals were of analytical grade. Compounds with great clarity were purchased from Sigma-Aldrich and used without any further purification. Double distilled water was used throughout all the involved experiments as a solvent. The raw materials included NiCl, Ni (NO 3 )2-6H 2 O (98%, Sigma-Aldrich), salvia plant extract that was collected from the natural habitats and methylene blue (MB; 98%, Sigma-Aldrich) dye. Synthesis of the plant extract The leaves of Justicia adathoda L . were harvested in Khyber Pakhtunkhwa's Tehsil Timergara District Dir Lower. Following collection, the plant was identified and authenticated by the Flora of Pakistan. The collected plant leaves have been clean with filter water, shadow dried and grind within fine talc by utilizing pistil and motor. Approximately ten grams of powder were taken and 200 mL of distilled were added in a glass container and placed for 48 hours. The sample was then filtered through Whatman filter paper and leaf extract was obtained in a sterilized conical flask (Ullah et al., 2025 ). Green synthesis of Ni based MPs Ni based microparticles (NiMPs) were synthesized following the standard plant-mediated green reduction method (Sharma et al., 2009 ). The extract from Justicia adathoda L . was utilized as a bio reductant in the production of nickel based microparticles. To form a NiCl solution (2M Nickel chloride solution), 0.95g of nickel chloride was dissolved in 150 ml of distilled water. Initially, the Nickel chloride solution was stirred at room temperature for one minute before incorporating the Justicia adathoda L . leaf extract. The mixture was then agitated for one hour. Following the addition of the leaf extract, the solution transitioned from colorless to a brownish-black hue, indicating the formation of Ni based MPs. Subsequently, the samples underwent centrifugation for 40 min at 14,000 rpm at 60℃.The resulting remnant from the centrifugation be desiccated at standard temperature 60℃ prior to characterization and biological assessment. Characterization of the synthesized Ni based MPs Characterization of microparticles stands as the most important factor in comprehending and controlling the synthesis and application of microparticles. This vital factor can be achieved by utilizing a variety of procedures including X-ray diffraction (XRD, D8-Advance Bruker) (G et al., 2024). NiO/Ni(OH) 2 MPs was characterized by using X-ray powder diffraction using X-ray diffractometer [Model: Pert Pro (PAN Analytical). The XRD spectrum was taken over 2θ range of 5 ◦ -80 ℴ (Salem & Fouda, 2021). Fourier transform infrared (FT-IR) analysis, using IR Prestige-21 Shimadzu 400–4000 cm−1 . Fourier transform infrared (FT-IR) analysis, using IR Prestige-21 Shimadzu, was implemented to characterize the functional groups on the surface of the Ni MP. The scanned spectra were in the range 400–4000 cm − 1 at a resolution of 4 cm − 1 (Mohamed et al., 2017 a). UV-Vis analysis was performed using perkin Elmer UV Win lab 7.3.0.340 /220126 to study the optical properties as well as band gap of microparticles (Behzadi et al., 2015 ), field electron scanning electron microscopy (FESEM)/ energy-dispersive X-ray spectroscopy (EDX, Tescan Brno-Mira3 Lmu), The purity of Ni MPs was confirmed by the means of EDX, which is capable of accurately measuring Ni along with displaying the Ni peaks without any impurities (Sadeghi et al., 2016 ). Elemental analysis was carried out by using EDX analysis to analyze the % amount of carbon, nickel and oxygen (Silva et al., 2001 ). SEM was carried out to analyze the surface morphology of Ni based microparticles. The morphology of the prepared sample of Ni-MP was examined using a Scanning Electron Microscope SEM analysis (JEOL jsm-6480 LV) (Bouremana et al., 2015 ). Antimicrobial Assay The antimicrobial activity of the studied MPs was evaluated against 5 bacterial strains (Escherichia coli ATCC 25922, B. subtilis, Staphylococcus aureus, P. auriginosa, and Salmonella Typhi) , belonging to Gram-positive and Gram-negative bacteria, and 7 yeast strains, Trichophyton rubrum, Candida albicans, Aspergillus niger, Microsporum canis, Fusarium lini, Candida glabirata, Aspergillus fumigatus. Were isolated from leaves extract of plant products (Ed-Dra et al., 2020 ). Before use, bacterial strains were revivified with subcultures in tryptone soya agar (TSA, Biokar, Beauvais, France) at 37℃ for 24 h, while yeast was revivified with subcultures in Sabouraud dextrose agar with chloramphenicol (Biokar, Beauvais, France) at 25℃ for 48 h. Then, a microbial suspension equivalent to 0.5 McFarland (108 cfu/mL) was prepared in sterile physiological water (0.9% NiCL) and inoculated by swabbing on plates containing Mueller–Hinton agar (Biokar, Beauvais, France) (Ed-Dra et al., 2020 ). On the surface of each plate, 10 µL of each MP solution at a concentration of 10 µg/mL was dropped on 6 mm diameter filter paper discs (Whatman Grade 4 Qualitative Filter Papers, Merck, Germany. Sterile distilled water (10 µL) was used as a negative control, while gentamicin (30 µg) was used as a positive control for bacteria and 10 µL of Canaflucan (fluconazole) Win® capsule 150 mg with a concentration of 0.6 µg/mL was used as a positive control for the yeast strain. The used plates were incubated at 37 ℃ for 18–24 h for bacteria and 25℃ for 44–48℃ for yeast. After incubation, the inhibition diameter was measured in millimeters (disk included). The antimicrobial activity was classified into three levels based on the produced inhibitory diameter: weak (inhibition zone ≤ 12.0 mm), intermediate/moderate (12.1 mm ≤ inhibition zone ≤ 20.0 mm), and strong (inhibition zone ≥ 20.1 mm). Statistical Analysis Origin software and Microsoft Excel were used for the generation of figures and statistical analysis. In this study, the experiments were carried out in triplicate, and the difference between groups was determined by using the student t-test (P < 0.05). RESULTS AND DISCUSSION NiO/Ni(OH) 2 MPs Preparation and its Corroboration by SEM Spectroscopy The SEM image of the resulting microparticles, as shown in the Fig. 1 , displayed a crystalline, and slightly uneven shape, with a diameter 1 µm. Scanning electron microscopy became employed to scrutinize the physical dimensions with morphological characteristics of the biosynthesized Ni based microparticles (Ni et al., 2013 ). The image shows that the microparticles have a highly crystalline as well as highly agglomerate and appear as cluster of microparticles with a size range of 1µm. The agglomeration of microparticles may be due to the exposure of microparticles to large volume of heat during calcination step (Saiganesh et al. 2020) or because of high surface tension of the ultrafine microparticles and high surface energy (kamiya et al. 2018) (Ying et al., 2022 b). The SEM result is compatible with the result of (Pooyandeh et al. 2020) who study the morphology of Ni based microparticles which are prepared by the sol-gel method using two different precursors nickel chloride hexahydrate and nickel nitrate hexahydrate. Our results are accurately similar to other studies in literature where it was revealed that Ni based MPs have important microparticles in other previous SEM analysis) (Ying et al., 2022 a). XRD Analysis The XRD pattern was employed to characterize the structural features, crystalline information, and phase purity of the precursor NiCl.6H2O and the synthesized Ni based MPs. The results match well with the standard cards of pure NiCl2.6H2O JCPDS card (No. 025-10440) (Wegner et al., 2017 ).. To determine the known phase of the Ni based MPs, an X-ray diffraction analysis was conducted. The XRD pattern of green produced Ni based microparticles has been displaced. The spectra revealed five distinct independent peaks at 2Ɵ = 10 ℴ , 24 ℴ , 27 ℴ , 29 ℴ and 32 ℴ corresponding to intensity planes (1900), (1200), (1200), (1700) and (1000) respectively. The highest intensity for the peak observed was at 2θ value 29°. The diffraction peak data calm coordinate with the FCC fabric report from the Joint Committee on Powder Diffraction Standards (JCPDS). Sprinkling small unexplained peaks were also seen, which could be due to bioorganic piece from the Justicia adathoda L. leaf extract crystallizing on the nickel surface (Chandra et al., 2014 ). FTIR Analysis FT-IR spectroscopy was used to study the functional groups present on the raw materials, NiCl, NiNO3 and the product Ni based MPs. The FT-IR of the raw materials and products are summarized in figure (Mohamed et al., 2017 b). FTIR spectroscopy in the spectral range of 400 cm − 1 to 4000 cm − 1 was performed to know about the possible involvement of different functional groups of plant extract in the reduction process of the Nickel based microparticles. The FTIR profile of NiMPs displayed vibration at 3309.75 cm 1 for-OH groups, at 2943.78 and 2831.77 cm 1 indicating the C-H stretching, while other multiple peaks at 1559.94, 1559.94, 1449.04,1415.19,1114.62 and 1021.53 cm − 1 were related to C = C and C-O stretching for aromatic ring and polyphenols. The peak at 616.36 cm − 1 displayed information about NiO in NiNO3. The presence of Ni̶ (OH)2 and NiO in the FT-IR spectra indirectly indicates the synthesis of Ni based MPs. Our results are meticulously similar to other studies in literature where it was revealed that Ni based MPs have important microparticles and related with other previous FTIR analysis (Li et al., 2021 ). The FTIR spectra displays functional groups exist in diverse locations. Figure shows displaced FTIR spectra of Ni based MPs and control leaf extract. Justicia adathoda L. leaf extract exhibits similar transmittance peaks at 3309 and 616.36 cm-1. The extensive and secure bands at 3309 cm-1 were caused by the leaf extract's bounded hydroxyl (-OH) or amine group (-NH). The C-H group can be examined by the peak screen at 2943cm-1. The tensile vibration of the C _ O, C = C group caused the peaks at 1559.94 cm-1, 1449.04 cm-1, 1415.19 cm-1,1114.62 cm-1,1021.53 cm-1, and Ni based peak at 616.36 cm-1 (Thirumagal & Jeyakumari, 2020 ). Antibacterial activities of Justicia adathoda L. leaf extract and NiO/(OH) 2 MPs We used the disc diffusion method to examine the antibacterial activity of Ni based microparticles biosynthesized from Justicia adathoda L . leaf extract. Six exertions in all were used to examine the biological activity. Three are gram negative (Escherichia. Coli), (Salmonella. Typhi) and (P. Aeruginosa) and three are gram positive (S. Aureus), (M. Letus) and (B. Subtilis). The finding was described in table. The consequences opposed five exertions were conflict with those prevail utilize streptomycin as the positive control. A region of hindering was seen after a day. Vernier callipers were used to measure the inhibition zone (Luzala et al., 2022 ). The concentration of Ni based MPs increased in tandem with the inhibition of bacterial growth. The sample's inhibition of gram-negative bacteria is 90.2%. 84.25% inhibition against gram-positive S. aureus bacteria and E. coli (Luzala et al., 2022 ). A previous study found that biosynthetic Ni based MPs derived from solanum nigrum leaf extract showed a correlation between the particle's size and antibacterial activity, significantly more antibacterial action found in smaller particles. The precise mechanism by which Ni based MPs inhibit bacterial growth is unknown, based on studies in the literature (Dikshit et al., 2021 ). Sondi and Sondi, though, confirmed that the antibacterial activity of Ni based MPs against gram-negative bacteria depended on the Ni based MPs concentration and was directly associated to the establishment of pits in bacterial cell walls. Our results are meticulously similar to other studies in literature where it was revealed that Ni based MPs have important antibacterial activities against the verified bacterial strains (Balogun et al., 2024 ). Table 1 Percent inhibition of NiO/(OH) 2 microparticles S.no Bacterial species Percentage inhibition of sample Percentage inhibition of drug) ofloxacin) 1 E. Coli 55.3% 90.2% 2 B. subtilis No inhibition 85.21% 3 S. aureus 60.24% 84.25% 4 P. auriginosa 70.24% 92.61% 5 S. typhi No inhibition 88.31% Antifungal Activity Justicia adathoda L. leaf extract was utilize in antifungal research to assess the antifungal efficiency of biologically formed Ni based MPs against numerous fungus strains (P. Singh et al., 2020). The experiment was run, Table comprises the outcomes. Seven fungi such as Candia albicans, Microsprum canis, Trichophyton rubrum, Fusarium lini. Aspergillus fumigatus and Candida glabirata were used. In this experiment, diverse fungal strains were exposed to the antifungal possessions of green produced Ni based microparticles from Justicia adathoda L. leaf extract. The results showed hopeful antifungal activities, Aspergillus was strongly inhibited by Ni based MPs, which resulted in 95%. This indicates that Ni based MPs have a great deal of potential for managing this pathogenic fungus. 85% inhibition was shown by Trichophyton rubrum and 82% Candida albicans , while the remaining fungal species exabits i-e Candida glabitus, Aspergillus niger, Fusarium lini 75% inhibition (Álvarez-Chimal et al., 2022). Overall, the result demonstrates the antifungal properties of Ni based microparticles mediated from Justicia adathoda L. leaf extract and exhibit significant potential in fighting Candida albicans, Trichophyton rubrum, Aspergillus Niger, Fusarium lini, Microsporum canis, Aspergillus fumigatus, and Candida Glabirata Important repressive effects were also seen when associated to the other three strains. Micanazole and Amphotericin-B were utilized as encouraging control and distilled water was used as an adverse control (Mohapatra et al., 2024 ). A previous examination detected the antifungal features of Ni based microparticles made with extract from Camellia sinensis , or green tea (Bibi et al., 2017 b). Table 2 Percent inhibition of NiO/(OH) 2 MPs S.no Fungal species Linear growth in sample in mm Percentage Inhibition Drug Sample Control 1. Trichophyton rubrum 15mm 100mm 85% Miconazole 2. Candida albicans 18mm 100mm 82% Miconazole 3. Aspergillus niger 20mm 100mm 80% Amphotericin-B 4. Microsporum canis 25mm 100mm 75% Miconazole 5. Fusarium lini 40mm 100mm 60% Miconazole 6. Candida glabirata 35mm 100mm 65% Miconazole 7. Aspergillus fumigatus 5mm 100mm 95% Miconazole CONCLUSION Recent study has focused on the green production of MPs utilize plant extracts. Plant metabolites promote the environmentally benign production of microparticles. Plant-produced MPs have been used in numerous medicinal and industrial submissions. In the recent study, Ni based MPs were formed biologically with Justicia adathoda L . extract as a caping agent, which is a maintainable and biologically friendly method that is a cheap alternate to other traditional procedures. The Ni based MPs and leaf extract derived from Justicia adathoda L. were also tested for antibacterial and antifungal activities. The aqueous extract of Justicia Aadathoda L. was shown to be less effective than Ni based MPs in terms of antibacterial and antifungal properties. The supplied plant extracts and Ni based MPs remained tested for MIC and MBC. Both the Ni based MPs, and the extract revealed probable antibacterial and antifungal properties when tested against the DPPH and ABTS radicals. The current study concluded that Justicia adathoda L .-based NiO/Ni(OH) 2 MPs should be used as alternate treatments due to their antibacterial and antifungal capabilities. Though, additional vivo testing determination is required to expand their applications to biotic systems. References Balloux, F., & van Dorp, L. (2017). Q&A: What are pathogens, and what have they done to and for us?. BMC biology , 15 , 1-6. DOIhttps://doi.org/10.1186/s12915-017-0433-z Balogun, S. A., Abolarinwa, T. O., Adesanya, F. A., Ateba, C. N., & Fayemi, O. E. (2024). Spectroscopic and antibacterial activities of cobalt and nickel nanoparticles: a comparative analysis. Journal of Analytical Science and Technology , 15 (1), 33. https://doi.org/10.1186/s40543-024-00446-0 Bar, H., Bhui, D. 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Chemistry–A European Journal , 23 (26), 6330-6340. https://doi.org/10.1002/chem.201605251 Yao, J. D., & Moellering Jr, R. C. (2011). Antibacterial agents. Manual of clinical microbiology , 1041-1081. https://doi.org/10.1128/9781555816728.ch65. Ying, S., Guan, Z., Ofoegbu, P. C., Clubb, P., Rico, C., He, F., & Hong, J. (2022). Green synthesis of nanoparticles: Current developments and limitations. Environmental Technology & Innovation , 26 , 102336. https://doi.org/10.1016/j.eti.2022.102336 Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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microparticles\u003c/em\u003e\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-8368802/v1/084268ff9abbcc697b5e2df8.jpg"},{"id":98441531,"identity":"075fb768-3518-4b8a-b741-3edc5917efd8","added_by":"auto","created_at":"2025-12-17 17:05:34","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":28358,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eAnalysis of NiO/Ni(OH)\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u003cem\u003e microparticles using XRD spectroscopy\u003c/em\u003e\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-8368802/v1/35ae81ed88d817af5aa0aec3.jpg"},{"id":98440599,"identity":"b9c2b892-6d3e-4b04-ae81-2b7998efe211","added_by":"auto","created_at":"2025-12-17 17:04:05","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":49902,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eAnalysis of NiO/Ni(OH)\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u003cem\u003e microparticles\u003c/em\u003e\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-8368802/v1/66f3abaf481777f64b516934.jpg"},{"id":98400991,"identity":"4ab90a8f-f25a-43c2-8600-1dd518c6e331","added_by":"auto","created_at":"2025-12-17 11:35:56","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":47333,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eAnti-bacterial activity of NiO/(OH)\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u003cem\u003e microoparticle\u003c/em\u003e\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-8368802/v1/11ef56837dd2d4304292d248.jpg"},{"id":98400998,"identity":"ff848c12-0dde-4c5b-870b-4dd278c48a44","added_by":"auto","created_at":"2025-12-17 11:35:56","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":50679,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eAntifungal activity of NiO/Ni(OH)\u003c/em\u003e\u003csub\u003e\u003cem\u003e2 \u003c/em\u003e\u003c/sub\u003e\u003cem\u003eMPs\u003c/em\u003e\u003c/p\u003e","description":"","filename":"5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-8368802/v1/76c328bd02970932c30b82aa.jpg"},{"id":98775009,"identity":"61bd2834-5397-4e7f-a556-4498cdb69d29","added_by":"auto","created_at":"2025-12-22 12:17:54","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":958393,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8368802/v1/5f274404-cc0f-4ba8-bbcf-332bf657c279.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003e\u003cstrong\u003eGreen Synthesis Characterization and Biological Assessment of Nickle based Microparticles using\u003c/strong\u003e\u003cem\u003e\u003cstrong\u003e Justicia Adathoda L\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e. Leaf Extract\u003c/strong\u003e\u003c/p\u003e","fulltext":[{"header":"INTODUCTION","content":"\u003cp\u003eGreen synthesis has become a major topic of interest in the field of Micro-nanotechnology (Shebl et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Developments in micro-nanotechnology have led to the production of innovative materials, particularly ecofriendly micro-based-nanostructure synthetic procedures used to synthesize metal and metal oxide micromaterials of any shape and size (Fuku et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Most physical, chemical, hybrid methods tend to be costly and detrimental due to the presence of harmful substances like solvents, reducing agents, and precursors (Shaik et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). In contrast, employing green techniques for the preparation of microparticles (MPs) is new economical, ecofriendly, and simpler to utensils. Consequently, such technique is increasingly utilized in the production of metallic based microparticles. A diverse span of natural deposit, inclusive of microorganisms, biomolecules, and plant extracts, was explored for green MP synthesis (F. A. Khan et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Although numerous studies have been conducted on the green synthesis of MPs utilizing biomolecules and microbes, plant extracts are often favored because they are cost-effective, efficient, and do not require the complex processes involved in isolating and maintaining microbial cultures (M. Khan et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Various plant species, their parts, and isolated compounds are effectively used to synthesize green MPs. Furthermore, the preparation of micromaterials can be made more efficiently by incorporating industrial and agricultural waste, in computation to being ecofriendly (Iftikhar et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Phenolic chemicals can operate as capping agents in microparticles manufacturing, whereas biological molecules offer unique uses in micro-nanotechnology, making metal microparticles durable and trustworthy (Bar et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Recent studies have proved the effectiveness of using micromaterial\u0026rsquo;s in different aspects of life, including environmental control, energy storage, solar cell, biomarking, bio probes, tissue engineering, cancer therapy, cancer diagnosis, and drug delivery (Mandal \u0026amp; Ganguly, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). The method of green synthesis consists in obtaining particles using safe materials as reducing agents and stabilizers, for example, products of biological origin. For example, the method can be carried out using extracts of plants or their individual parts, as well as microorganism (bacteria, yeast, fungi) (J. Singh et al., \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). A plant contains various biochemicals and metabolites that can be used for the green synthesis of microparticles as they possess reducing and stabilizing properties. Green synthesis technologies are environmentally friendly, cost effective stable and nontoxic than traditional biological, physical, and chemical approaches (Mustapha et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Biosynthesized metal and metal oxide MPs have rising applications in the medical sector including immunotherapy, dentistry, wound healing, diagnosis, regenerative medicines, bio toxicology and bio-sensing platform and its antiviral, fungicidal and bactericidal potentials (Pandit et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Microparticles can be variable in dimension, shape, size and architectures(Machado et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). In the environmental field, the green synthesis of MPs has been used to control environmental pollution, such as with the degradation of organic dyes and chlorinated organic pollutants, heavy metal removal, in addition to the treatment of wastewater (Saif et al., \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). \u003cem\u003eJusticia adathoda L\u003c/em\u003e. also known as Malabar nut is a well-known and important medicinally important plant of family Acanthaceae growing in different Asian countries including Pakistan, Sirilanka, India and Nepal (Hossain \u0026amp; Hoq, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). \u003cem\u003eJusticia adathoda L.\u003c/em\u003e has diverse applications in traditionally used Ayurvedic and Unani medicines. The local populace uses the plants' leaves to treat coughs, colds, and asthma. \u003cem\u003eJusticia adathoda L\u003c/em\u003e. plant has several key properties, including antibacterial, antioxidant, antiulcer, antidiabetic, anti-allergic, anti-genotoxic, and anti-phlogiston. Adathoda vasica leaves contain phytochemicals including flavonoids, saponins, tannins, alkaloids and phenolic compounds. \u003cem\u003eJusticia adathoda L\u003c/em\u003e. leaf extract contains the two most significant quinazoline alkaloids, vasicine and vasicione (Khadri et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Phytochemicals are bioactive compounds naturally found in plants, and their composition varies greatly among different plant species and even within different parts of the same plant. The most common classes of phytochemicals include polyphenols (like flavonoids and tannins), terpenoids, alkaloids, carotenoids, and saponins. These compounds are involved in plant defense mechanisms and can also offer various health benefits to humans (Lydon \u0026amp; Duke, 1989). Antibacterial agents are utilized to inhibit the proliferation of harmful bacteria. They help prevent illness and eradicate microorganisms (Yao \u0026amp; Moellering, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Numerous antibiotics are sourced from plants and various chemicals to combat diseases. Bacteria are the pathogens responsible for poisoning. For this assay, we used five different strains of pathogens (Balloux \u0026amp; van Dorp, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). A disc diffusion method was employed to assess bacterial activity. Bacteria are classified into two categories: Gram-negative and Gram-positive. Staphylococcus aureus (S. aureus) is typically recognized as a Gram-positive bacterium and is frequently used in antibacterial studies due to its significant potential to cause infections in humans. Escherichia coli (E. coli), a Gram-negative bacterium, is established in contaminated water and along with as a pathogenic microbe that leads to human illness, as well as serving as an indicator of water quality (Margesin, 2017). The antifungal test aims to eliminate microorganisms, especially fungi. Fungi pose significant risks and can lead to various illnesses. We utilize antifungal tests to identify infections caused by fungi. These organisms can damage fruits, vegetables, and plants (Leyva Salas et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Numerous plants are recognized for their antifungal properties. Unsanitary food and environments weaken the immune system, facilitating the spread of fungal infections. Antifungal medications are necessary to treat such infections (Das et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Metallic nickel based microparticles are those that are used as the primary raw material without combining them with other metals, compounds or elements. The term nickel microparticle used in this review also refers to metallic nickel microparticle (Jaji et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The successful production of Ni based MPs employing various plant extracts, as well as their enhanced biological potential, call for additional research into other medicinally essential plants for future enhancement (Hussain et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Ni based MPs have gained much attention due to their unique magnetic, chemical, and physical properties as well as their potential applications in various technological fields such as catalysis (Bibi et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2017\u003c/span\u003ea), battery manufacture (Cheng et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), incorporation in textile (Jiao et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), and optical switches (Reena Mary et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). The current study aimed to synthesis Ni based MPs using \u003cem\u003eJusticia adathoda L.\u003c/em\u003e leaf extract. The produced (NiO/Ni(OH))\u003csub\u003e2\u003c/sub\u003e MPs) were examined by using modern analytical techniques, including X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM). Within inclusion, the antibacterial and antifungal potentials of the prepared Nickel based microparticles were measured.\u003c/p\u003e"},{"header":"METHODOLOGY","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eChemicals and other materials\u003c/h2\u003e \u003cp\u003eThe chemicals used were Distilled water and Nickel chloride. These chemicals were of analytical grade. Compounds with great clarity were purchased from Sigma-Aldrich and used without any further purification. Double distilled water was used throughout all the involved experiments as a solvent. The raw materials included NiCl, Ni (NO\u003csub\u003e3\u003c/sub\u003e)2-6H\u003csub\u003e2\u003c/sub\u003eO (98%, Sigma-Aldrich), salvia plant extract that was collected from the natural habitats and methylene blue (MB; 98%, Sigma-Aldrich) dye.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eSynthesis of the plant extract\u003c/h3\u003e\n\u003cp\u003eThe leaves of \u003cem\u003eJusticia adathoda L\u003c/em\u003e. were harvested in Khyber Pakhtunkhwa's Tehsil Timergara District Dir Lower. Following collection, the plant was identified and authenticated by the Flora of Pakistan. The collected plant leaves have been clean with filter water, shadow dried and grind within fine talc by utilizing pistil and motor. Approximately ten grams of powder were taken and 200 mL of distilled were added in a glass container and placed for 48 hours. The sample was then filtered through Whatman filter paper and leaf extract was obtained in a sterilized conical flask (Ullah et al., \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2025\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003eGreen synthesis of Ni based MPs\u003c/h3\u003e\n\u003cp\u003eNi based microparticles (NiMPs) were synthesized following the standard plant-mediated green reduction method (Sharma et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). The extract from \u003cem\u003eJusticia adathoda L\u003c/em\u003e. was utilized as a bio reductant in the production of nickel based microparticles. To form a NiCl solution (2M Nickel chloride solution), 0.95g of nickel chloride was dissolved in 150 ml of distilled water. Initially, the Nickel chloride solution was stirred at room temperature for one minute before incorporating the \u003cem\u003eJusticia adathoda L\u003c/em\u003e. leaf extract. The mixture was then agitated for one hour. Following the addition of the leaf extract, the solution transitioned from colorless to a brownish-black hue, indicating the formation of Ni based MPs. Subsequently, the samples underwent centrifugation for 40 min at 14,000 rpm at 60℃.The resulting remnant from the centrifugation be desiccated at standard temperature 60℃ prior to characterization and biological assessment.\u003c/p\u003e\n\u003ch3\u003eCharacterization of the synthesized Ni based MPs\u003c/h3\u003e\n\u003cp\u003eCharacterization of microparticles stands as the most important factor in comprehending and controlling the synthesis and application of microparticles. This vital factor can be achieved by utilizing a variety of procedures including X-ray diffraction (XRD, D8-Advance Bruker) (G et al., 2024). NiO/Ni(OH)\u003csub\u003e2\u003c/sub\u003e MPs was characterized by using X-ray powder diffraction using X-ray diffractometer [Model: Pert Pro (PAN Analytical). The XRD spectrum was taken over 2θ range of 5\u003csup\u003e◦\u003c/sup\u003e-80\u003csup\u003eℴ\u003c/sup\u003e (Salem \u0026amp; Fouda, 2021). Fourier transform infrared (FT-IR) analysis, using IR Prestige-21 Shimadzu 400\u0026ndash;4000 \u003csup\u003ecm\u0026minus;1\u003c/sup\u003e. Fourier transform infrared (FT-IR) analysis, using IR Prestige-21 Shimadzu, was implemented to characterize the functional groups on the surface of the Ni MP. The scanned spectra were in the range 400\u0026ndash;4000\u003csup\u003ecm\u0026thinsp;\u0026minus;\u0026thinsp;1\u003c/sup\u003e at a resolution of 4\u003csup\u003ecm\u0026thinsp;\u0026minus;\u0026thinsp;1\u003c/sup\u003e (Mohamed et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2017\u003c/span\u003ea). UV-Vis analysis was performed using perkin Elmer UV Win lab 7.3.0.340 /220126 to study the optical properties as well as band gap of microparticles (Behzadi et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), field electron scanning electron microscopy (FESEM)/ energy-dispersive X-ray spectroscopy (EDX, Tescan Brno-Mira3 Lmu), The purity of Ni MPs was confirmed by the means of EDX, which is capable of accurately measuring Ni along with displaying the Ni peaks without any impurities (Sadeghi et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Elemental analysis was carried out by using EDX analysis to analyze the % amount of carbon, nickel and oxygen (Silva et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). SEM was carried out to analyze the surface morphology of Ni based microparticles. The morphology of the prepared sample of Ni-MP was examined using a Scanning Electron Microscope SEM analysis (JEOL jsm-6480 LV) (Bouremana et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003eAntimicrobial Assay\u003c/h3\u003e\n\u003cp\u003eThe antimicrobial activity of the studied MPs was evaluated against 5 bacterial strains \u003cem\u003e(Escherichia coli ATCC 25922, B. subtilis, Staphylococcus aureus, P. auriginosa, and Salmonella Typhi)\u003c/em\u003e, belonging to Gram-positive and Gram-negative bacteria, and 7 yeast strains, \u003cem\u003eTrichophyton rubrum, Candida albicans, Aspergillus niger, Microsporum canis, Fusarium lini, Candida glabirata, Aspergillus fumigatus.\u003c/em\u003e Were isolated from leaves extract of plant products (Ed-Dra et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Before use, bacterial strains were revivified with subcultures in tryptone soya agar (TSA, Biokar, Beauvais, France) at 37℃ for 24 h, while yeast was revivified with subcultures in Sabouraud dextrose agar with chloramphenicol (Biokar, Beauvais, France) at 25℃ for 48 h. Then, a microbial suspension equivalent to 0.5 McFarland (108 cfu/mL) was prepared in sterile physiological water (0.9% NiCL) and inoculated by swabbing on plates containing Mueller\u0026ndash;Hinton agar (Biokar, Beauvais, France) (Ed-Dra et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). On the surface of each plate, 10 \u0026micro;L of each MP solution at a concentration of 10 \u0026micro;g/mL was dropped on 6 mm diameter filter paper discs (Whatman Grade 4 Qualitative Filter Papers, Merck, Germany. Sterile distilled water (10 \u0026micro;L) was used as a negative control, while gentamicin (30 \u0026micro;g) was used as a positive control for bacteria and 10 \u0026micro;L of Canaflucan (fluconazole) Win\u0026reg; capsule 150 mg with a concentration of 0.6 \u0026micro;g/mL was used as a positive control for the yeast strain. The used plates were incubated at 37 ℃ for 18\u0026ndash;24 h for bacteria and 25℃ for 44\u0026ndash;48℃ for yeast. After incubation, the inhibition diameter was measured in millimeters (disk included). The antimicrobial activity was classified into three levels based on the produced inhibitory diameter: weak (inhibition zone\u0026thinsp;\u0026le;\u0026thinsp;12.0 mm), intermediate/moderate (12.1 mm\u0026thinsp;\u0026le;\u0026thinsp;inhibition zone\u0026thinsp;\u0026le;\u0026thinsp;20.0 mm), and strong (inhibition zone\u0026thinsp;\u0026ge;\u0026thinsp;20.1 mm).\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eStatistical Analysis\u003c/h2\u003e \u003cp\u003eOrigin software and Microsoft Excel were used for the generation of figures and statistical analysis. In this study, the experiments were carried out in triplicate, and the difference between groups was determined by using the student t-test (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS AND DISCUSSION","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eNiO/Ni(OH)\u003csub\u003e2\u003c/sub\u003e MPs Preparation and its Corroboration by SEM Spectroscopy\u003c/h2\u003e \u003cp\u003eThe SEM image of the resulting microparticles, as shown in the Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, displayed a crystalline, and slightly uneven shape, with a diameter 1 \u0026micro;m. Scanning electron microscopy became employed to scrutinize the physical dimensions with morphological characteristics of the biosynthesized Ni based microparticles (Ni et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). The image shows that the microparticles have a highly crystalline as well as highly agglomerate and appear as cluster of microparticles with a size range of 1\u0026micro;m. The agglomeration of microparticles may be due to the exposure of microparticles to large volume of heat during calcination step (Saiganesh et al. 2020) or because of high surface tension of the ultrafine microparticles and high surface energy (kamiya et al. 2018) (Ying et al., \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2022\u003c/span\u003eb). The SEM result is compatible with the result of (Pooyandeh et al. 2020) who study the morphology of Ni based microparticles which are prepared by the sol-gel method using two different precursors nickel chloride hexahydrate and nickel nitrate hexahydrate. Our results are accurately similar to other studies in literature where it was revealed that Ni based MPs have important microparticles in other previous SEM analysis) (Ying et al., \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2022\u003c/span\u003ea).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eXRD Analysis\u003c/h2\u003e \u003cp\u003eThe XRD pattern was employed to characterize the structural features, crystalline information, and phase purity of the precursor NiCl.6H2O and the synthesized Ni based MPs. The results match well with the standard cards of pure NiCl2.6H2O JCPDS card (No. 025-10440) (Wegner et al., \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).. To determine the known phase of the Ni based MPs, an X-ray diffraction analysis was conducted. The XRD pattern of green produced Ni based microparticles has been displaced. The spectra revealed five distinct independent peaks at 2Ɵ = 10\u003csup\u003eℴ\u003c/sup\u003e, 24\u003csup\u003eℴ\u003c/sup\u003e, 27\u003csup\u003eℴ\u003c/sup\u003e, 29\u003csup\u003eℴ\u003c/sup\u003e and 32\u003csup\u003eℴ\u003c/sup\u003e corresponding to intensity planes (1900), (1200), (1200), (1700) and (1000) respectively. The highest intensity for the peak observed was at 2θ value 29\u0026deg;. The diffraction peak data calm coordinate with the FCC fabric report from the Joint Committee on Powder Diffraction Standards (JCPDS). Sprinkling small unexplained peaks were also seen, which could be due to bioorganic piece from the \u003cem\u003eJusticia adathoda L.\u003c/em\u003e leaf extract crystallizing on the nickel surface (Chandra et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eFTIR Analysis\u003c/h2\u003e \u003cp\u003eFT-IR spectroscopy was used to study the functional groups present on the raw materials, NiCl, NiNO3 and the product Ni based MPs. The FT-IR of the raw materials and products are summarized in figure (Mohamed et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2017\u003c/span\u003eb). FTIR spectroscopy in the spectral range of 400 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e to 4000 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e was performed to know about the possible involvement of different functional groups of plant extract in the reduction process of the Nickel based microparticles. The FTIR profile of NiMPs displayed vibration at 3309.75 cm 1 for-OH groups, at 2943.78 and 2831.77 cm 1 indicating the C-H stretching, while other multiple peaks at 1559.94, 1559.94, 1449.04,1415.19,1114.62 and 1021.53 cm\u0026thinsp;\u0026minus;\u0026thinsp;1 were related to C\u0026thinsp;=\u0026thinsp;C and C-O stretching for aromatic ring and polyphenols. The peak at 616.36 cm\u0026thinsp;\u0026minus;\u0026thinsp;1 displayed information about NiO in NiNO3. The presence of Ni̶ (OH)2 and NiO in the FT-IR spectra indirectly indicates the synthesis of Ni based MPs. Our results are meticulously similar to other studies in literature where it was revealed that Ni based MPs have important microparticles and related with other previous FTIR analysis (Li et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The FTIR spectra displays functional groups exist in diverse locations. Figure shows displaced FTIR spectra of Ni based MPs and control leaf extract. \u003cem\u003eJusticia adathoda L.\u003c/em\u003e leaf extract exhibits similar transmittance peaks at 3309 and 616.36 cm-1. The extensive and secure bands at 3309 cm-1 were caused by the leaf extract's bounded hydroxyl (-OH) or amine group (-NH). The C-H group can be examined by the peak screen at 2943cm-1. The tensile vibration of the C\u003csup\u003e_\u003c/sup\u003eO, C\u0026thinsp;=\u0026thinsp;C group caused the peaks at 1559.94 cm-1, 1449.04 cm-1, 1415.19 cm-1,1114.62 cm-1,1021.53 cm-1, and Ni based peak at 616.36 cm-1 (Thirumagal \u0026amp; Jeyakumari, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eAntibacterial activities of\u003c/b\u003e \u003cb\u003eJusticia adathoda L.\u003c/b\u003e \u003cb\u003eleaf extract and NiO/(OH)\u003c/b\u003e\u003csub\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sub\u003e \u003cb\u003eMPs\u003c/b\u003e\u003c/p\u003e \u003cp\u003eWe used the disc diffusion method to examine the antibacterial activity of Ni based microparticles biosynthesized from \u003cem\u003eJusticia adathoda L\u003c/em\u003e. leaf extract. Six exertions in all were used to examine the biological activity. Three are gram negative \u003cem\u003e(Escherichia. Coli), (Salmonella. Typhi) and (P. Aeruginosa)\u003c/em\u003e and three are gram positive \u003cem\u003e(S. Aureus), (M. Letus) and (B. Subtilis).\u003c/em\u003e The finding was described in table. The consequences opposed five exertions were conflict with those prevail utilize streptomycin as the positive control. A region of hindering was seen after a day. Vernier callipers were used to measure the inhibition zone (Luzala et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The concentration of Ni based MPs increased in tandem with the inhibition of bacterial growth. The sample's inhibition of gram-negative bacteria is 90.2%. 84.25% inhibition against gram-positive S. aureus bacteria and E. coli (Luzala et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). A previous study found that biosynthetic Ni based MPs derived from solanum nigrum leaf extract showed a correlation between the particle's size and antibacterial activity, significantly more antibacterial action found in smaller particles. The precise mechanism by which Ni based MPs inhibit bacterial growth is unknown, based on studies in the literature (Dikshit et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Sondi and Sondi, though, confirmed that the antibacterial activity of Ni based MPs against gram-negative bacteria depended on the Ni based MPs concentration and was directly associated to the establishment of pits in bacterial cell walls. Our results are meticulously similar to other studies in literature where it was revealed that Ni based MPs have important antibacterial activities against the verified bacterial strains (Balogun et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003e\u003cem\u003ePercent inhibition of NiO/(OH)\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e \u003cem\u003emicroparticles\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eS.no\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eBacterial species\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePercentage inhibition of sample\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePercentage inhibition of drug) ofloxacin)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eE. Coli\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e55.3%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e90.2%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eB. subtilis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNo inhibition\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e85.21%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eS. aureus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e60.24%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e84.25%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eP. auriginosa\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e70.24%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e92.61%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eS. typhi\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNo inhibition\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e88.31%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eAntifungal Activity\u003c/h2\u003e \u003cp\u003e \u003cem\u003eJusticia adathoda L.\u003c/em\u003e leaf extract was utilize in antifungal research to assess the antifungal efficiency of biologically formed Ni based MPs against numerous fungus strains (P. Singh et al., 2020). The experiment was run, Table comprises the outcomes. Seven fungi such as \u003cem\u003eCandia albicans, Microsprum canis, Trichophyton rubrum, Fusarium lini. Aspergillus fumigatus\u003c/em\u003e and \u003cem\u003eCandida glabirata\u003c/em\u003e were used. In this experiment, diverse fungal strains were exposed to the antifungal possessions of green produced Ni based microparticles from \u003cem\u003eJusticia adathoda L.\u003c/em\u003e leaf extract. The results showed hopeful antifungal activities, \u003cem\u003eAspergillus\u003c/em\u003e was strongly inhibited by Ni based MPs, which resulted in 95%. This indicates that Ni based MPs have a great deal of potential for managing this pathogenic fungus. 85% inhibition was shown by \u003cem\u003eTrichophyton rubrum\u003c/em\u003e and 82% \u003cem\u003eCandida albicans\u003c/em\u003e, while the remaining fungal species exabits i-e \u003cem\u003eCandida glabitus, Aspergillus niger, Fusarium lini\u003c/em\u003e 75% inhibition (\u0026Aacute;lvarez-Chimal et al., 2022). Overall, the result demonstrates the antifungal properties of Ni based microparticles mediated from \u003cem\u003eJusticia adathoda L.\u003c/em\u003e leaf extract and exhibit significant potential in fighting \u003cem\u003eCandida albicans, Trichophyton rubrum, Aspergillus Niger, Fusarium lini, Microsporum canis, Aspergillus fumigatus, and Candida Glabirata\u003c/em\u003e Important repressive effects were also seen when associated to the other three strains. Micanazole and Amphotericin-B were utilized as encouraging control and distilled water was used as an adverse control (Mohapatra et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). A previous examination detected the antifungal features of Ni based microparticles made with extract from \u003cem\u003eCamellia sinensis\u003c/em\u003e, or green tea (Bibi et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2017\u003c/span\u003eb).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003e\u003cem\u003ePercent inhibition of NiO/(OH)\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e \u003cem\u003eMPs\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eS.no\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eFungal species\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eLinear growth in sample in mm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePercentage\u003c/p\u003e \u003cp\u003eInhibition\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eDrug\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSample\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eTrichophyton rubrum\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e85%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMiconazole\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCandida albicans\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e82%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMiconazole\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eAspergillus niger\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e20mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e80%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAmphotericin-B\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eMicrosporum canis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e25mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e75%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMiconazole\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eFusarium lini\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e40mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e60%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMiconazole\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCandida glabirata\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e35mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e65%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMiconazole\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eAspergillus fumigatus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100mm\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e95%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMiconazole\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eRecent study has focused on the green production of MPs utilize plant extracts. Plant metabolites promote the environmentally benign production of microparticles. Plant-produced MPs have been used in numerous medicinal and industrial submissions. In the recent study, Ni based MPs were formed biologically with \u003cem\u003eJusticia adathoda L\u003c/em\u003e. extract as a caping agent, which is a maintainable and biologically friendly method that is a cheap alternate to other traditional procedures. The Ni based MPs and leaf extract derived from \u003cem\u003eJusticia adathoda L.\u003c/em\u003e were also tested for antibacterial and antifungal activities. The aqueous extract of \u003cem\u003eJusticia Aadathoda L.\u003c/em\u003e was shown to be less effective than Ni based MPs in terms of antibacterial and antifungal properties. The supplied plant extracts and Ni based MPs remained tested for MIC and MBC. Both the Ni based MPs, and the extract revealed probable antibacterial and antifungal properties when tested against the DPPH and ABTS radicals. The current study concluded that \u003cem\u003eJusticia adathoda L\u003c/em\u003e.-based NiO/Ni(OH)\u003csub\u003e2\u003c/sub\u003e MPs should be used as alternate treatments due to their antibacterial and antifungal capabilities. Though, additional vivo testing determination is required to expand their applications to biotic systems.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eBalloux, F., \u0026amp; van Dorp, L. (2017). Q\u0026amp;A: What are pathogens, and what have they done to and for us?. \u003cem\u003eBMC biology\u003c/em\u003e, \u003cem\u003e15\u003c/em\u003e, 1-6. DOIhttps://doi.org/10.1186/s12915-017-0433-z\u003c/li\u003e\n\u003cli\u003eBalogun, S. A., Abolarinwa, T. O., Adesanya, F. A., Ateba, C. N., \u0026amp; Fayemi, O. E. (2024). 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Green synthesis of nanoparticles: Current developments and limitations. \u003cem\u003eEnvironmental Technology \u0026amp; Innovation\u003c/em\u003e, \u003cem\u003e26\u003c/em\u003e, 102336. https://doi.org/10.1016/j.eti.2022.102336\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"Nazarbayev University","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Green synthesis, Ni based microparticle, SEM, FTIR, XRD, antimicrobial activity","lastPublishedDoi":"10.21203/rs.3.rs-8368802/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8368802/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eNickel-based oxide/hydroxide microparticles (NiO/Ni(OH)₂ MPs) were synthesized via a green way utilizing \u003cem\u003eJusticia adathoda L.\u003c/em\u003e leaf extract, serving as novel reducing and stabilizing agent. Plants phytochemicals such as alkaloids, flavonoids, and phenolics facilitated the eco-friendly formations of NiO/Ni(OH)\u003csub\u003e2\u003c/sub\u003e MPs without toxic chemicals. Structure analysis through X-ray diffraction (XRD) confirmed the crystalline structure, while FTIR confirmed the involvement of functional groups in reduction and stabilizations. Morphological characterization by Scanning Electron Microscopy (SEM) revealed a granular dense structure microparticle of size about 1µm. Biological screening showed notable antibacterial and antifungal activities. NiO/Ni(OH)\u003csub\u003e2\u003c/sub\u003e MPs show 60.24% inhibition against \u003cem\u003eStaphylococcus aureus\u003c/em\u003e and 70.24% inhibition against \u003cem\u003ePseudomonas aeruginosa,\u003c/em\u003e while antifungal activity reached 95% against \u003cem\u003eAspergillus fumigatus\u003c/em\u003e. Compared to standard pharmaceutical agents such as \u003cem\u003eOfloxacin\u003c/em\u003e and \u003cem\u003eMiconazole,\u003c/em\u003e the NiO/Ni(OH)\u003csub\u003e2\u003c/sub\u003e MPs synthesized using \u003cem\u003eJusticia adathoda L. \u003c/em\u003eleaf extract revealed potential antibacterial and antifungal activities. These results underline the value of \u003cem\u003eJusticia adathoda\u003c/em\u003e as an eco-friendly and efficient biological resource for making functional NiO/Ni(OH)\u003csub\u003e2\u003c/sub\u003e MPs. The synthesized microparticles show strong applications in biomedical fields, including therapeutic formulations, antimicrobial coatings, and presenting a sustainable unconventional to chemically synthesized micromaterials.\u003c/p\u003e","manuscriptTitle":"Green Synthesis Characterization and Biological Assessment of Nickle based Microparticles using Justicia Adathoda L. Leaf Extract","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-17 11:35:48","doi":"10.21203/rs.3.rs-8368802/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"a85bd2ab-1027-4385-a591-82096cf80697","owner":[],"postedDate":"December 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":59700704,"name":"Nanoscience"}],"tags":[],"updatedAt":"2025-12-17T11:35:48+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-17 11:35:48","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8368802","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8368802","identity":"rs-8368802","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}