Green synthesis and characterization of silver nanoparticles from the aerial parts of Leucas lanata with enhanced antioxidant and antimicrobial activities

Green synthesis and characterization of silver nanoparticles from the aerial parts of Leucas lanata with enhanced antioxidant and antimicrobial activities | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Green synthesis and characterization of silver nanoparticles from the aerial parts of Leucas lanata with enhanced antioxidant and antimicrobial activities Ruchika Sharma, Sheetal Tyagi, Indra Rautela, Rakesh Kumar Bachheti, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7981923/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 11 You are reading this latest preprint version Abstract In the present study, a green, Phyto-assisted approach was employed to synthesize silver nanoparticles (AgNPs) using the aqueous extract of the aerial parts of Leucas lanata . The successful formation of AgNPs was evidenced by a characteristic surface plasmon resonance (SPR) band at 384 nm in the UV–Vis absorption spectrum. Elemental mapping and energy-dispersive X-ray (EDX) analysis further confirmed the presence of silver, showing a prominent peak at 2.7 keV. X-ray diffraction (XRD) patterns revealed distinct crystalline planes, which were consistent with the results of Selected area electron diffraction (SAED). High-resolution transmission electron microscopy (HRTEM) determined an average particle size of 25.37 nm. The nanoparticles exhibited a high zeta potential value, suggesting strong stability attributed to phytochemical-mediated capping, as corroborated by Fourier transform infrared spectroscopy (FT-IR). The biosynthesized L-AgNPs displayed potent antioxidant activity with an IC₅₀ value of 49 µg/mL in the DPPH assay. L-AgNPs exhibited a higher proportion of ABTS scavenging activity (12.26%) than the Leucas lanata extract at a lower concentration of 100 µg/mL, indicating enhanced antioxidant potential. Similarly, the FRAP value of L-AgNPs (21.56 ± 0.17 µM at 100 µg/mL) was significantly greater than that of the aqueous plant extract, further confirming their superior reducing capacity. Moreover, the silver nanoparticles demonstrated significant antimicrobial activity against Staphylococcus aureus, Escherichia coli, Salmonella abony, and Bacillus subtilis. The MIC value for Leucas lanata aqueous extract showed effects between 0.45 and 0.84 mg/mL, whereas L-AgNPs showed effects between 0.09 and 0.34 mg/mL across bacteria. To the best of our knowledge, this is the first report on the silver nanoparticle-synthesizing potential of Leucas lanata , highlighting its promising applications in biomedical and pharmaceutical fields. Silver nanoparticles Green-synthesis phytochemicals Capping agents zeta potential Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 1. Introduction Bacterial contamination is a significant cause of various issues in the food industry, medical devices, and water treatment. Both Gram-positive and Gram-negative bacteria are to blame for contamination and many human illnesses. Although many antibacterial agents are available on the market, they harm society due to their side effects, high cost, and toxicity [ 1 , 2 ]. All these obstacles have been largely overcome by the synthesis of metallic nanoparticles, which are found to be useful in various fields, including electronics, environmental remediation, the medical field, and agriculture (3–5). Metallic nanoparticles can be made from top to bottom and bottom to top. In the top-down method, metallic particles are converted into nanoparticles using various methods, including physical, chemical, and mechanical processes. In contrast, researchers employ physical or chemical vapor deposition, sol-gel, chemical reduction, hydrothermal, solvothermal, spray pyrolysis, laser ablation, and biomimetic techniques to create metallic nanoparticles utilizing the bottom-up strategy [ 6 – 9 ]. The most effective method among these is biomimetic, as it utilizes various microorganisms, enzymes, and plant extracts. In this method, readily available, non-toxic, and inexpensive materials are used to make metallic nanoparticles. Hence, this method is safe for both society and the environment [ 10 – 13 ]. Many silver nanoparticles have been made using the biomimetic method in the biomedical field. Most of these are prepared from plant extracts. Phytochemicals are present in the extracts of various plant parts, including root, stem, flower, fruit, seed, and leaf. Ag metal nanoparticles of different sizes are made by reducing silver ions [ 14 – 19 ]. In this research, AgNPs have been made from the extract of the Leucas lanata plant found in Karnaprayag, Uttarakhand, India, for the first time. Leucas lanata , also called woolly leucas, is a soft, densely woolly-haired perennial that grows between 700 and 300 meters above sea level in South India and the Himalayan Mountains. Known by its native name, Gumma or Biskapra, it is a member of the Lamiaceae family [ 20 , 21 ]. In Uttarakhand, people use an extract from the aerial part of the Leucas lanata plant to treat a variety of illnesses, including pertussis, stomach, and headaches [ 21 , 22 ]. While its paste treats wounds, many Uttarakhand residents use its leaves and petals, combined with cold water or milk, to treat illnesses such as colds, coughs, and diarrhea [ 20 ]. The Leucas lanata plant contains polyphenols, such as protocatechuic acid, caffeic acid, ferulic acid, gallic acid, and chlorogenic acid, according to HPLC studies (Fig. 1) [ 23 ]. Silver nanoparticles (AgNPs) synthesized through green methods have gained substantial attention due to their eco-friendly nature, cost-effectiveness, and enhanced biological activities. Various medicinal plants have been employed as reducing and stabilizing agents in the biosynthesis of AgNPs owing to the presence of bioactive phytochemicals [ 14 , 15 ]. Leucas lanata , a traditionally used medicinal herb, is known for its antibacterial, anti-inflammatory, and antioxidant properties [ 24 – 26 ]. Despite its ethnopharmacological significance and rich phytochemical profile, Leucas lanata remains underexplored in the context of nanobiotechnology. To date, no comprehensive scientific report is available on the use of the aerial parts of Leucas lanata for the green synthesis of AgNPs and their evaluation for antioxidant and antimicrobial activities. This study aims to bridge this knowledge gap by investigating the potential of Leucas lanata in the biosynthesis of AgNPs and assessing their biomedical relevance, thereby opening new perspectives in the field of plant-based nanomaterials. The objective of the current study was to produce, describe, and evaluate the biological activity of silver nanoparticles derived from aerial extract for application in the medical field, given the importance of Leucas lanata . Protocatechuic acid Caffeic acid Ferulic acid Gallic acid Chlorogenic acid Figure 1 Some polyphenol structures found in the Leucas lanata plant 2. Materials and Methods 2.1. Collection and identification of plant material The plant material was collected from Karnaprayag, Uttarakhand, India (30.2575° N, 79.2466° E) in October 2022. The collection of plant material was carried out in compliance with applicable local and national regulations, and no specific permits were required. The collection site was not located on protected or Forest land. The plant is collected from private land. A voucher specimen (No. 1203) was deposited at the Botanical Survey of India, Dehradun. The plant was identified as Leucas lanata (family Lamiaceae) by Dr. S. K. Singh, Scientist E and Head of the Department, Botanical Survey of India, Dehradun, India. The collection of plant material was carried out in accordance with all applicable local and national regulations. 2.2. Preparation of Leucas lanata plant extract Leucas lanata fresh aerial parts were cleaned twice or three times with tap water before being rinsed with distilled water to remove any dust or other visible particles from the leaves' surface. It took ten to twenty days in May for this aerial portion to dry at room temperature in the shade. After that, the dried plant was finely ground into a powder. Twenty grams of plant powder and 500 mL of water were heated to 60°C for 20 minutes to produce an aqueous extract of the leaves. After filtering the extract using Whatman filter paper No. 1, it was stored at 4°C for subsequent use [ 27 ]. 2.3. Green Synthesis of AgNPs To synthesize AgNPs, 1 mL of Leucas lanata extract was added individually to 7 mL, 8 mL, 9 mL, 10 mL, and 11 mL of aqueous solutions containing 1 mM (millimolar), 2 mM, 3 mM, 4 mM, and 5 mM AgNO3, respectively. The reaction mixture was centrifuged for 20 minutes at 4000 rpm and then dried at 60°C to produce the AgNPs, which exhibited a dark brown color at 5 mM and 1:10 AgNO₃ concentrations. 2.4. Characterization of AgNPs nanoparticle Using a UV-VIS spectrophotometer (JASCO V-650), the absorbance of AgNPs in ultrapure water was measured. Analysis of the spectra was done between 300 and 600 nm. The essential information regarding the synthesis of AgNPs is provided by the absorption peak at 400–500 nm [ 28 ]. AgNPs were found to have FT-IR (Fourier transform infrared) spectra between 4000 and 400 cm-1 (NICOLET 6700, Thermo Fisher Scientific, Germany). AgNPs' purity and percentage composition were examined using EDX (Energy Dispersive X-ray). The phase composition and structural analysis of the generated product were ascertained by utilizing X-ray beam diffraction (XRD, Bruker, D8 Advance, Germany) at room temperature with CuKα (λ = 1.5406 Å and step size = 0.02°) radiation in the scattering angle of 10–80°. The secondary electron mode of field-emission scanning electron microscopy (FE-SEM, Carl Zeiss, Ultra Plus) was used to examine the surface microstructure and grain size of the produced nanoparticles. Energy-dispersive X-ray spectroscopy (EDX), connected to and integrated with the FE-SEM equipment, was used to determine the chemical composition of the representative samples. The diameter of the produced AgNPs was measured, and their shape was determined using HETEM analysis. Ethanol was used to dissolve the material. A thin dispersion drop was put on a "staining mat." A copper grid coated with Lacey carbon was placed within the drop and coated side up. Following approximately 30 minutes of sonication, the grid was removed, allowed to air-dry for an additional 30 minutes, and then examined using a JEOL 2100 Transmission Electron Microscope. 2.5. Antioxidant properties Using the DPPH free radical scavenging assay technique, the antioxidant activity of the produced AgNPs was evaluated. In UV-VIS (Ultraviolet-Visible) spectroscopy, the deep violet DPPH solution in methanol exhibits absorption at 517 nm. An electron from an antioxidant causes it to lighten and turn yellow [ 29 , 30 ]. 1 mL of DPPH alcoholic solution was combined with an equivalent volume of Leucas lanata extract or L-AgNPs at varying concentrations, and the mixture was incubated for 30 minutes at room temperature. One common antioxidant is ascorbic acid. 1 mL of methanol was added to the reaction mixture in place of the extract or sample to obtain a control absorption. The modified FRAP method, as described by Benzie and Strain (1996), was used to perform the FRAP assay. The stock solution consisted of a 20 mM FeCl 3 · 6H 2 O solution in distilled water, 300 mM acetate buffer at pH 3.6, and a 10 mM TPTZ (2,4,6-tripyridyl-striazine) solution in 40 mM HCl. Next, FeCl 3 .6H 2 O (2.5 ml) was combined with acetate buffer (25 ml) and TPTZ (2.5 ml). Prior to use, the solution's temperature was increased to 37°C. Under dark conditions, 150 µL of plant extract and 2.85 mL of FRAP solution were left to react for 30 minutes. At 593 nm, the absorbance was measured. The standard curve, between 100 and 500 µg/mL FeSO 4 , was linear. The results were compared to ascorbic acid standards and expressed in µM Fe(II)/g dry mass. According to Ferreira-Santos et al. (2019), the extracts' capacity to scavenge ABTS radicals was evaluated. In 100 milliliters of distilled water, 0.360 grams of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt were dissolved to create a 7 mM ABTS stock solution. 0.066 g of the salt was dissolved in 100 mL of distilled water to create a 2.45 mM potassium persulfate (K₂S₂O₈) solution separately. The 2.45 mM K₂S₂O₈ and 7 mM ABTS solutions were mixed in equal quantities (10 mL each) to create the radical cation. The reaction mixture was allowed to sit at room temperature (25 ± 2°C) in the dark for 12 hours to achieve full radical production and absorbance stability. A UV-Vis spectrophotometer was used to confirm that the working solution had an absorbance of 0.700 ± 0.020 at 734 nm when the resultant concentrated ABTS⁺• solution was diluted 1:1 (v/v) with absolute ethanol. To perform the antioxidant capacity experiment, Leucas lanata extract or L-AgNPs were used at concentrations of 100, 200, 300, 400, and 500 µg/mL. For two hours, the reaction mixtures were allowed to sit at room temperature in the dark. Using spectrophotometry, a drop in absorbance at 734 nm was observed. Appropriate blanks and controls were utilized. 2.6. Antibacterial activity The antibacterial susceptibility of L-AgNPs against Salmonella abony , Bacillus subtilis , Staphylococcus aureus , and Escherichia coli was evaluated using the agar well diffusion method. A sterile glass spreader dipped in the bacterial culture's inoculum suspension was applied to the hardened nutrient agar plates, and the surface of the agar plates was then thoroughly cleaned. After loading the different concentrations of the produced L-AgNPs solution into the bored wells, the Petri plates were incubated for 24 hours at 37°C to examine the inhibition zone. The assay's standard was streptomycin at 10 µg/mL. The diameter of the inhibitory zone was measured in millimeters and compared to the control [ 31 ]. MIC (Minimum Inhibitor Concentration) Assay A tube dilution approach was used to determine the MIC. The investigated bacterial species were cultured for 24 hours and then diluted using the 0.5 McFarland standard in 10 mL of tryptic soy broth (TSB) to produce an inoculum of 108 CFU mL 1. Neem extracts at a range of strengths, from 5000 g/mL to 10.0 g/mL, were made in a series of culture tubes using DMSO. Each tube received 0.1 mL of the bacterial cell suspension, which was then incubated at 37°C for 24 hours. The development of the inoculum was ascertained by measuring the turbidity of the broth. The lowest extract concentration that inhibited the development of the test organism was known as the minimum inhibitory concentration, or MIC [ 32 , 33 ]. 3. Results and Discussion 3.1. UV-VIS (Ultraviolet-Visible) spectroscopy The reaction mixture's color shift from light brown to dark brown due to surface plasma resonances was the leading indicator of AgNP production. UV-VIS spectra of the colloidal solution of AgNPs were then taken, in which a peak at 384nm was observed for L-AgNPs, as shown in Fig. 2 . This is similar to the AgNPs mediated by the Fusarium exquisite, Fusarium solani, and Ficus carica extracts. Several biomolecules and functional groups found in Leucas lanata extract facilitate the bio-reduction and capping of synthetic L-AgNPs [ 34 , 35 ]. 3.2. X-ray beam diffraction (XRD) analysis X-ray diffraction was employed to ascertain the crystalline structure of the bio-synthesised L-AgNPs. Five peaks were found at 38.09°, 44.18°, 64.41°, 77.33°, and 81.44° in the XRD pattern as shown in Fig. 3, which is also represented by the SAED pattern. These peaks correspond to the [1 1 1], [2 0 0], [2 2 0], [3 1 1], and [2 2 2] planes, respectively. This result confirmed the cubic crystalline structure of L-AgNPs, whose basis was demonstrated in Joint Committee of Powder Diffraction Standards file numbers 040783 and 021098, as shown in different research. According to XRD, the size of synthesized L-AgNPs is estimated from the Debye-Scherrer formula, which ranges from 10nm to 75nm, while the average particle size is approximately 25.37nm, which is also vindicated by HRTEM result [ 36 – 40 ]. Figure 3 X-ray beam diffraction pattern of synthesized AgNPs 3.3. EDAX (Energy Dispersive X-ray) analysis EDAX was used to determine the chemical composition of the biomimetic synthesized L-AgNPs. The absorption peak for silver nanoparticles is generally believed to occur at 2.7 to 3 keV. In Fig. 4 , a peak was observed at 2.7 keV, which assures the presence of silver nanoparticles. Apart from this, signals for C and Cl were also obtained in EDX, indicating that the phytochemical present in the extract of Leucas lanata is responsible for the reduction of Ag + ions and the stability of AgNPs. The C-Cl group found in the biomolecule of Leucas lanata is the reason for the significantly lower level of Cl, which FTIR further verified. These findings are in good agreement with the published research [ 41 – 45 ]. The EDAX tool is also used for element mapping, which provides a digital photograph showing the distribution of elements in the specimen through the color dots. Figure 5 (a), (b), and (c) show the element mapping of C, Ag, and Cl, respectively, while Fig. 5 (d) represents the overlay element mapping of all elements presented with their homogenous distribution in the sample [ 46 ]. 3.4. FESEM (Field emission scanning electron microscopy) analysis FESEM assessed the surface morphology of L-AgNPs. The image of 100.00kx magnification is shown in Fig. 6 , according to which the shape of synthesized L-AgNPs was found to be spherical and rod-like. The aggregation in L-AgNPs is responsible for a rod-like shape; the same results are also visible from the HRTEM image of L-AgNPs [ 47 , 48 ]. 3.5. Fourier Transform Infrared analysis Fourier Transform Infrared (FTIR) analysis helps determine which functional groups are present in the biomolecules in Leucas lanata extract, which is essential for Ag + ion reduction, capping, and stability of AgNPs. Various sharp bands were obtained in FTIR of L-AgNPs between 3900cm − 1 and 532 cm − 1 (Fig. 7 ). The band near 3900 cm − 1 is due to N-H stretching, while the band at 3492 cm − 1 is due to O-H stretching vibration, indicating the presence of alcoholic and phenolic groups. The minor band observed in the region of 2917 cm − 1 is due to C-H stretching in an alkane. The band observed at 2376 cm − 1 corresponds to the ≡ C-C stretching vibration. The shared band at 1638 cm − 1 indicates the presence of a C = O or C = C stretching vibration in the aromatic ring. The bands found at 1382 cm − 1 and 1113 cm − 1 are due to the C-O, N-H stretching vibration of the amide linkage, while the bands found at 844 cm − 1 are due to the C-H banding of alkenes. The band at 532 cm − 1 is attributed to C-Cl or C-Br stretching vibrations, the presence of which has also been confirmed by EDAX. These bands indicate the presence of phytochemicals in the plant extract, which play a vital role in the formation process and act as capping and stabilizing agents [ 49 – 58 ]. 3.6. High-resolution transmission electron microscopy analysis High-resolution transmission electron microscopy (HRTEM) plays a crucial role in determining the morphology, including the size and shape of AgNPs. Figure 8 shows the particles of L-AgNPs at different magnifications. L-AgNPs have a spherical shape, but due to agglomeration, they also have rod-like morphology, which is also confirmed by the FESEM image (Fig. 8 a). For the average particle size, a total of 44 L-AgNPs were sampled to get the average particle size, for which the Image J software was used. The particle size of L-AgNPs ranges from 10 nm to 112 nm. However, particles with more than 50nm are very small, and the average particle size is 28.59nm, which is also verified by XRD (Fig. 8 e). In the SEAD pattern of synthesized L-AgNPs, eight spots are found that represent specific crystal planes. Figure 8 (e) shows eight concentric circles, out of which four are found to be [1 1 1], [2 0 0], [2 2 0], and [3 1 1] planes, which suggest the crystalline nature of L-AgNPs.The interplanar d spacing of 0.15nm for the [2 2 0] plane of L-AgNPs is shown in the figure. 7(f) [ 2 ] [ 59 , 60 ]. 3.7. Dynamic Light Scattering (DLS) The size distribution and zeta potential of L-AgNPs were ascertained by DLS analysis. The zeta potential can be used to assess the stability of AgNPs. The zeta potential gives information about the charge of the phytochemicals acting as capping reagents. The higher the charge on the surface, the greater the repulsion force existing between the synthesized L-AgNPs, due to which the aggregation of L-AgNPs will decrease and the dispersity of nanoparticles will increase in the medium. The zeta potential value of synthesized L-AgNPs was − 25.4mV, as shown in Fig. 9 (a). According to previous research, a voltage of 30 mV is considered the most stable range for nanosuspensions. It also confirms that synthesized L-AgNPs are relatively stable and have a good colloidal nature. In contrast, Fig. 9 (b) shows the particle size distribution of L-AgNPs, which shows that particles are of a higher particle size of more than 100 nm as compared to the particle size suggested by XRD and HRTEM. The difference is due to the sample preparation procedure. In the DLS method, the size of L-AgNPs is obtained as the hydrodynamic diameter, which is why the particle size predicted by DLS is higher than predicted by XRD and HRTEM [ 58 – 63 ]. 3.8. Antioxidant activity The antioxidant activity was determined using the DPPH assay. In this case, ascorbic acid was taken as a control. The antioxidant activity of ascorbic acid, Leucas lanata plant extract, and silver nanoparticles made from Leucas lanata is completely dose-dependent towards free radicals like DPPH, as shown in Fig. 10 (a). The scavenging activity of DPPH of plant extract was lower than that of synthesized L-AgNPs and almost equal to that of ascorbic acid at a high concentration of 500µg/ml. The %RAS values for the plant extract and L-AgNPs at 100 µg/ml were 38% and 45%, respectively. For a maximum concentration of 500 µg/ml, the %RAS values were 81% and 93%, respectively. The IC 50 value for the plant extract of Leucas lanata was 154 µg/ml, whereas for L-AgNPs, it was found to be 49 µg/ml, as shown in Fig. 10 (b), which is significantly lower compared to the Leucas lanata plant extract. This result is also confirmed by results obtained for silver nanoparticles mediated by Cucumis prophetarum [ 64 – 66 ]. The aqueous extract of Leucas lanata and L-AgNPs demonstrated their ABTS radical scavenging activity in Fig. 10 (c). When compared to aqueous plant extract, L-AgNPs exhibit a larger proportion of ABTS scavenging activity. At lower concentrations of 100 µg/ml, L-AgNPs exhibit 12.26% inhibition, whereas at higher concentrations of 500 µg/ml, they exhibit 59.92% inhibition. Nonetheless, the plant's aqueous extract exhibits 8.75% at lower concentrations of 100 µg/ml and 53.17% at higher concentrations of 500 µg/ml. Figure 10 (d) showed the FRAP assay of L-AgNPs and the aqueous extract of Leucas lanata . L-AgNPs have a higher FRAP than aqueous plant extract. L-AgNPs show 21.56 ± 0.17 µM at a lower concentration of 100 µg/ml and 83.31 ± 0.15 µM at higher values of 500 µg/ml. However, at a lower concentration of 100 µg/ml, the aqueous solution of the plant extract shows 14.29 ± 0.11, whereas at a higher concentration of 500 µg/ml, it shows 69.54 ± 0.21. The current findings showed that, in addition to silver nanoparticles exhibited potent antioxidant activity compared with aqueous plant extract of Leucas lanata , as previously supported by the previous study [ 67 – 69 ] Fig: 10(a): DPPH (2,2-Diphenyl-1-picrylhydrazyl) assay analysis for Antioxidant activity Figure 10 (b): DPPH (2,2-Diphenyl-1-picrylhydrazyl) Free radical scavenging Activity AgNPs and IC 50 values 3.9. Antibacterial Activity The antibacterial activity of, was tested using two gram-positive and two gram-negative pathogens: Salmonella ebony , Bacillus subtilis, Escherichia coli , and Staphylococcus aureus as positive and negative controls, respectively, streptomycin and All Bacteria culture diluted 10 8 CFU/ml and spread on agar plate for incubation 24 hr at 37ºC to measure the inhibition zones. DMSO were employed as observed Fig. 11 and Fig. 12 . The activity of the produced L-AgNPs against gram-positive and gram-negative bacteria was nearly identical. Silver nanoparticles derived from the peel of Citus sinensis and Acacia leucophloea also showed comparable outcomes [ 70 , 71 ]. The table displays the inhibition zone at various concentrations. Compared to L-AgNPs, the inhibitory zone of the plant extract is smaller. At the lowest concentration of L-AgNPs, Bacillus subtilis exhibited the highest inhibitory zone, measuring 10 mm (Table 1 ). When bacteria and silver ions interact, it inhibits essential processes such as DNA replication, respiratory enzyme function, and ATP synthesis, ultimately leading to cell death [ 72 – 73 ]. Table 1 Inhibition Zone Diameter(mm) for different Bacteria Concentration Staphylococcus aureus(P) Escherichia coli(Q) Salmonella abony(R) Bacillus subtilis (S) Leucas lanata Plant extract L-AgNPs Leucas lanata Plant extract L-AgNPs Leucas lanata Plant extract L-AgNPs Leucas lanata Plant extract L-AgNPs 2000µg/mL (A) 6 mm 20mm 10 mm 25mm 10 mm 17mm 16 mm 20mm 1000µg/mL (B) 4 mm 10 mm 6 mm 19mm 4mm 12mm 6 mm 15mm 500µg/mL (C) --------- 5 mm -------- 11mm -------- 8mm ------- 12mm 250µg/mL (D) -------- ------- -------- 6mm ------- ------ --------- 10mm Streptomycin µg/mL(F) Positive Control 20 mm 23 mm 24 mm 29mm 20 mm 20mm 20mm 23mm DMSO (Dimethyl sulfoxide) (E) (Negative Control) -------- ------- -------- ------- -------- ------- -------- ------- MIC (Minimum Inhibitory Concentration) against Bacteria Figure 13 shows that the aqueous extract of Leucas lanata exhibited effects between 0.45 and 0.84 mg/mL, whereas L-AgNPs showed effects between 0.09 and 0.34 mg/mL across the bacteria. With a minimum inhibitory concentration (MIC) of 0.84 mg/mL, the plant extract in aqueous solution showed the most potent effects against Salmonella abony . Bacillus subtilis was inhibited with a minimum inhibitory concentration (MIC) of 0.76 mg/mL, whereas E.coli showed the strongest action with the lowest MIC value of 0.45 mg/mL. MIC 0.68 mg/ml is moderate for Staphylococcus aureus . With a minimum inhibitory concentration (MIC) of 0.09 mg/mL, L-AgNPs exhibited the most notable efficacy against E. coli . The MIC value for the second activity against Staphylococcus aureus was 0.16 mg/mL. Bacillus subtilis and Salmonella abony , on the other hand, exhibit 0.34 and 0.24 mg/ml, respectively. Consequently, L-AgNPs exhibit a MIC value that is noticeably higher than that of the plant's aqueous extract. 4. Limitations and Future Scope of the Study The study has some limitations that warrant further investigation. The exact mechanism by which phytochemicals interact to stabilize and cap the silver nanoparticles was not fully elucidated, leaving a gap in understanding the underlying processes. The antimicrobial activity was evaluated against a limited number of bacterial strains; expanding the investigation to include a broader range of microorganisms, such as fungi and drug-resistant strains, would provide more comprehensive insights into the efficacy of the nanoparticles. Additionally, the environmental impact of Leucas lanata -based silver nanoparticles, including their biodegradability and long-term stability within ecological systems, remains unexplored. The absence of in vivo studies to validate the in vitro antioxidant and antimicrobial efficacy represents another limitation, as such studies are critical for practical and clinical applications. Addressing these limitations in future research would significantly enhance the relevance and applicability of the study. 5. Conclusion Several biological entities have recently been investigated for their ability to mediate the green synthesis of silver nanoparticles (AgNPs). In this study, Leucas lanata extract, prepared from its aerial parts, was employed as a natural reducing and capping agent for the synthesis of AgNPs. Characterization techniques, such as UV-VIS spectroscopy, confirmed nanoparticle formation with a distinct absorption peak at 384 nm, while EDX analysis verified the elemental composition of the silver. Morphological assessment using HRTEM and structural evaluation through XRD indicated that the L-AgNPs were predominantly spherical with an average particle diameter of under 29 nm. The antioxidant capacity of the synthesized nanoparticles was evaluated using the DPPH, ABTS, and FRAP radical scavenging methods, which suggest an enhanced antioxidant efficiency of L-AgNPs. The antibacterial efficacy of L-AgNPs was tested against two Gram-positive (Staphylococcus aureus, Bacillus subtilis) and two Gram-negative (Escherichia coli, Salmonella abony) strains. L-AgNPs exhibit a MIC value that is noticeably higher than that of the plant's aqueous extract. Results demonstrated consistent inhibitory zones across all four bacterial species, with a slightly higher zone of inhibition observed against Bacillus subtilis . These findings highlight the effectiveness of Leucas lanata as a novel, eco-friendly source for the synthesis of bio-functional silver nanoparticles with promising antimicrobial and antioxidant properties. Abbreviations SPR Surface plasmon resonance AgNPs Silver Nanoparticles, UV-VIS Ultraviolet- Visible EDX Energy Dispersive X-ray XRD X-ray beam diffraction, FTIR Fourier Transform Infrared FESEM Field emission scanning electron microscopy HRTEM High-resolution transmission electron microscopy DPPH 2,2-Diphenyl-1-picrylhydrazyl mM millimolar JCPDS Joint Committee on Powder Diffraction Standards SAED Selected area electron diffraction DMSO Dimethyl sulfoxide and L-AgNPs:Silver nanoparticles mediated by Leucas lanata extract ℃ Degree Celsius mm millimetre gm gram:FRAP:Ferric Reducing Antioxidant Power TPTZ:2,4,6-tripyridyl-striazine ABTS 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) MIC Minimum Inhibitory Concentration CFU Colony-Forming Unit Declarations Author Contributions Statement : "R.S., S.T., and I. R. wrote the main manuscript text, and R.K.B. A.B. and L.A.W. prepared different tables and figures. All authors reviewed the manuscript." Funding Statement: This research received no specific grant from any funding agency in any sectors. Data Availability: All relevant data are within the manuscript Declarations Ethics, consent to participate, and consent to publish Not applicable. Ethics declaration Not applicable. Clinical trial number Not applicable. Competing interests The authors declare no competing interests. References Ameen F, Srinivasan P, Selvankumar T, Kamala-Kannan S, Al Nadhari S, Almansob A, Govarthanan M (2019) Phytosynthesis of silver nanoparticles using Mangifera indica flower extract as bioreductant and their broad-spectrum antibacterial activity. Bioorg Chem 88:102970. https://doi.org/10.1016/j.bioorg.2019.102970 Rana A, Kumari A, Chaudhary AK, Srivastava R, Kamil D, Vashishtha P, Sharma SN (2023) An investigation of antimicrobial activity for plant pathogens by green-synthesized silver nanoparticles using Azadirachta indica and Mangifera indica. 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standard\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"10.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7981923/v1/b137c343da32153dcbd04867.jpg"},{"id":97684504,"identity":"69b47021-a86b-474e-9d4a-cf67032975d9","added_by":"auto","created_at":"2025-12-08 10:07:02","extension":"jpg","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":77885,"visible":true,"origin":"","legend":"\u003cp\u003eInhibition Zone of \u003cem\u003eLeucas lanata\u003c/em\u003e extract against different Bacteria (P-\u003cem\u003eStaphylococcus aureus\u003c/em\u003e; Q- \u003cem\u003eEscherichia coli\u003c/em\u003e; R- \u003cem\u003eSalmonella abony\u003c/em\u003e; S- \u003cem\u003eBacillus subtilis\u003c/em\u003e)\u003c/p\u003e","description":"","filename":"11.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7981923/v1/303bece0a19365befe954a49.jpg"},{"id":97684357,"identity":"5e75c8ec-bab3-447e-ac19-40067745c862","added_by":"auto","created_at":"2025-12-08 10:06:05","extension":"jpg","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":119501,"visible":true,"origin":"","legend":"\u003cp\u003eInhibition Zone of L- AgNPs against different Bacteria (P- \u003cem\u003eStaphylococcus aureus\u003c/em\u003e; Q- \u003cem\u003eEscherichia coli\u003c/em\u003e; R- \u003cem\u003eSalmonella abony\u003c/em\u003e; S- \u003cem\u003eBacillus subtilis\u003c/em\u003e)\u003c/p\u003e","description":"","filename":"12.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7981923/v1/151609f1dd004356ceaa59ce.jpg"},{"id":97684459,"identity":"bc3c071c-6181-45ef-90a9-a2e5c53ac6e0","added_by":"auto","created_at":"2025-12-08 10:06:40","extension":"jpg","order_by":13,"title":"Figure 13","display":"","copyAsset":false,"role":"figure","size":170558,"visible":true,"origin":"","legend":"\u003cp\u003eThe MIC of plant extract and L- AgNPs against different bacterial strains\u003c/p\u003e","description":"","filename":"13.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7981923/v1/ef7f68ff88428896a484a8c2.jpg"},{"id":97902455,"identity":"7eaaeea2-a947-4afb-81ee-8c855b549e6a","added_by":"auto","created_at":"2025-12-10 15:52:25","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2791189,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7981923/v1/feca03c9-7794-43aa-bc91-84048ba15b4f.pdf"},{"id":97684225,"identity":"9b1bdda8-922e-4edc-b51e-d2808ce90a58","added_by":"auto","created_at":"2025-12-08 10:05:30","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":998137,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7981923/v1/2610b478709bd5a66250480e.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Green synthesis and characterization of silver nanoparticles from the aerial parts of Leucas lanata with enhanced antioxidant and antimicrobial activities","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eBacterial contamination is a significant cause of various issues in the food industry, medical devices, and water treatment. Both Gram-positive and Gram-negative bacteria are to blame for contamination and many human illnesses. Although many antibacterial agents are available on the market, they harm society due to their side effects, high cost, and toxicity [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. All these obstacles have been largely overcome by the synthesis of metallic nanoparticles, which are found to be useful in various fields, including electronics, environmental remediation, the medical field, and agriculture (3\u0026ndash;5). Metallic nanoparticles can be made from top to bottom and bottom to top. In the top-down method, metallic particles are converted into nanoparticles using various methods, including physical, chemical, and mechanical processes. In contrast, researchers employ physical or chemical vapor deposition, sol-gel, chemical reduction, hydrothermal, solvothermal, spray pyrolysis, laser ablation, and biomimetic techniques to create metallic nanoparticles utilizing the bottom-up strategy [\u003cspan additionalcitationids=\"CR7 CR8\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. The most effective method among these is biomimetic, as it utilizes various microorganisms, enzymes, and plant extracts. In this method, readily available, non-toxic, and inexpensive materials are used to make metallic nanoparticles. Hence, this method is safe for both society and the environment [\u003cspan additionalcitationids=\"CR11 CR12\" citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Many silver nanoparticles have been made using the biomimetic method in the biomedical field. Most of these are prepared from plant extracts. Phytochemicals are present in the extracts of various plant parts, including root, stem, flower, fruit, seed, and leaf. Ag metal nanoparticles of different sizes are made by reducing silver ions [\u003cspan additionalcitationids=\"CR15 CR16 CR17 CR18\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn this research, AgNPs have been made from the extract of the \u003cem\u003eLeucas lanata\u003c/em\u003e plant found in Karnaprayag, Uttarakhand, India, for the first time. \u003cem\u003eLeucas lanata\u003c/em\u003e, also called woolly leucas, is a soft, densely woolly-haired perennial that grows between 700 and 300 meters above sea level in South India and the Himalayan Mountains. Known by its native name, Gumma or Biskapra, it is a member of the \u003cem\u003eLamiaceae family\u003c/em\u003e [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. In Uttarakhand, people use an extract from the aerial part of the \u003cem\u003eLeucas lanata\u003c/em\u003e plant to treat a variety of illnesses, including pertussis, stomach, and headaches [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. While its paste treats wounds, many Uttarakhand residents use its leaves and petals, combined with cold water or milk, to treat illnesses such as colds, coughs, and diarrhea [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. The \u003cem\u003eLeucas lanata\u003c/em\u003e plant contains polyphenols, such as protocatechuic acid, caffeic acid, ferulic acid, gallic acid, and chlorogenic acid, according to HPLC studies (Fig.\u0026nbsp;1) [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eSilver nanoparticles (AgNPs) synthesized through green methods have gained substantial attention due to their eco-friendly nature, cost-effectiveness, and enhanced biological activities. Various medicinal plants have been employed as reducing and stabilizing agents in the biosynthesis of AgNPs owing to the presence of bioactive phytochemicals [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. \u003cem\u003eLeucas lanata\u003c/em\u003e, a traditionally used medicinal herb, is known for its antibacterial, anti-inflammatory, and antioxidant properties [\u003cspan additionalcitationids=\"CR25\" citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Despite its ethnopharmacological significance and rich phytochemical profile, \u003cem\u003eLeucas lanata\u003c/em\u003e remains underexplored in the context of nanobiotechnology. To date, no comprehensive scientific report is available on the use of the aerial parts of \u003cem\u003eLeucas lanata\u003c/em\u003e for the green synthesis of AgNPs and their evaluation for antioxidant and antimicrobial activities. This study aims to bridge this knowledge gap by investigating the potential of \u003cem\u003eLeucas lanata\u003c/em\u003e in the biosynthesis of AgNPs and assessing their biomedical relevance, thereby opening new perspectives in the field of plant-based nanomaterials.\u003c/p\u003e\u003cp\u003eThe objective of the current study was to produce, describe, and evaluate the biological activity of silver nanoparticles derived from aerial extract for application in the medical field, given the importance of \u003cem\u003eLeucas lanata\u003c/em\u003e.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eProtocatechuic acid Caffeic acid Ferulic acid\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eGallic acid Chlorogenic acid\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eFigure\u0026nbsp;1\u003c/strong\u003e\u003cp\u003eSome polyphenol structures found in \u003cem\u003ethe Leucas lanata\u003c/em\u003e plant\u003c/p\u003e\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1. Collection and identification of plant material\u003c/h2\u003e\u003cp\u003eThe plant material was collected from Karnaprayag, Uttarakhand, India (30.2575\u0026deg; N, 79.2466\u0026deg; E) in October 2022. The collection of plant material was carried out in compliance with applicable local and national regulations, and no specific permits were required. The collection site was not located on protected or Forest land. The plant is collected from private land. A voucher specimen (No. 1203) was deposited at the Botanical Survey of India, Dehradun. The plant was identified as \u003cem\u003eLeucas lanata\u003c/em\u003e (family Lamiaceae) by Dr. S. K. Singh, Scientist E and Head of the Department, Botanical Survey of India, Dehradun, India. The collection of plant material was carried out in accordance with all applicable local and national regulations.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2. Preparation of \u003cem\u003eLeucas lanata\u003c/em\u003e plant extract\u003c/h2\u003e\u003cp\u003e\u003cem\u003eLeucas lanata\u003c/em\u003e fresh aerial parts were cleaned twice or three times with tap water before being rinsed with distilled water to remove any dust or other visible particles from the leaves' surface. It took ten to twenty days in May for this aerial portion to dry at room temperature in the shade. After that, the dried plant was finely ground into a powder. Twenty grams of plant powder and 500 mL of water were heated to 60\u0026deg;C for 20 minutes to produce an aqueous extract of the leaves. After filtering the extract using Whatman filter paper No. 1, it was stored at 4\u0026deg;C for subsequent use [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e2.3. Green Synthesis of AgNPs\u003c/h2\u003e\u003cp\u003eTo synthesize AgNPs, 1 mL of \u003cem\u003eLeucas lanata\u003c/em\u003e extract was added individually to 7 mL, 8 mL, 9 mL, 10 mL, and 11 mL of aqueous solutions containing 1 mM (millimolar), 2 mM, 3 mM, 4 mM, and 5 mM AgNO3, respectively. The reaction mixture was centrifuged for 20 minutes at 4000 rpm and then dried at 60\u0026deg;C to produce the AgNPs, which exhibited a dark brown color at 5 mM and 1:10 AgNO₃ concentrations.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003e2.4. Characterization of AgNPs nanoparticle\u003c/h2\u003e\u003cp\u003eUsing a UV-VIS spectrophotometer (JASCO V-650), the absorbance of AgNPs in ultrapure water was measured. Analysis of the spectra was done between 300 and 600 nm. The essential information regarding the synthesis of AgNPs is provided by the absorption peak at 400\u0026ndash;500 nm [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. AgNPs were found to have FT-IR (Fourier transform infrared) spectra between 4000 and 400 cm-1 (NICOLET 6700, Thermo Fisher Scientific, Germany). AgNPs' purity and percentage composition were examined using EDX (Energy Dispersive X-ray). The phase composition and structural analysis of the generated product were ascertained by utilizing X-ray beam diffraction (XRD, Bruker, D8 Advance, Germany) at room temperature with CuKα (λ\u0026thinsp;=\u0026thinsp;1.5406 \u0026Aring; and step size\u0026thinsp;=\u0026thinsp;0.02\u0026deg;) radiation in the scattering angle of 10\u0026ndash;80\u0026deg;. The secondary electron mode of field-emission scanning electron microscopy (FE-SEM, Carl Zeiss, Ultra Plus) was used to examine the surface microstructure and grain size of the produced nanoparticles. Energy-dispersive X-ray spectroscopy (EDX), connected to and integrated with the FE-SEM equipment, was used to determine the chemical composition of the representative samples. The diameter of the produced AgNPs was measured, and their shape was determined using HETEM analysis. Ethanol was used to dissolve the material. A thin dispersion drop was put on a \"staining mat.\" A copper grid coated with Lacey carbon was placed within the drop and coated side up. Following approximately 30 minutes of sonication, the grid was removed, allowed to air-dry for an additional 30 minutes, and then examined using a JEOL 2100 Transmission Electron Microscope.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003e2.5. Antioxidant properties\u003c/h2\u003e\u003cp\u003eUsing the DPPH free radical scavenging assay technique, the antioxidant activity of the produced AgNPs was evaluated. In UV-VIS (Ultraviolet-Visible) spectroscopy, the deep violet DPPH solution in methanol exhibits absorption at 517 nm. An electron from an antioxidant causes it to lighten and turn yellow [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. 1 mL of DPPH alcoholic solution was combined with an equivalent volume of \u003cem\u003eLeucas lanata\u003c/em\u003e extract or L-AgNPs at varying concentrations, and the mixture was incubated for 30 minutes at room temperature. One common antioxidant is ascorbic acid. 1 mL of methanol was added to the reaction mixture in place of the extract or sample to obtain a control absorption.\u003c/p\u003e\u003cp\u003eThe modified FRAP method, as described by Benzie and Strain (1996), was used to perform the FRAP assay. The stock solution consisted of a 20 mM FeCl\u003csub\u003e3\u003c/sub\u003e \u0026middot; 6H\u003csub\u003e2\u003c/sub\u003eO solution in distilled water, 300 mM acetate buffer at pH 3.6, and a 10 mM TPTZ (2,4,6-tripyridyl-striazine) solution in 40 mM HCl. Next, FeCl\u003csub\u003e3\u003c/sub\u003e.6H\u003csub\u003e2\u003c/sub\u003eO (2.5 ml) was combined with acetate buffer (25 ml) and TPTZ (2.5 ml). Prior to use, the solution's temperature was increased to 37\u0026deg;C. Under dark conditions, 150 \u0026micro;L of plant extract and 2.85 mL of FRAP solution were left to react for 30 minutes. At 593 nm, the absorbance was measured. The standard curve, between 100 and 500 \u0026micro;g/mL FeSO\u003csub\u003e4\u003c/sub\u003e, was linear. The results were compared to ascorbic acid standards and expressed in \u0026micro;M Fe(II)/g dry mass.\u003c/p\u003e\u003cp\u003eAccording to Ferreira-Santos et al. (2019), the extracts' capacity to scavenge ABTS radicals was evaluated. In 100 milliliters of distilled water, 0.360 grams of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt were dissolved to create a 7 mM ABTS stock solution. 0.066 g of the salt was dissolved in 100 mL of distilled water to create a 2.45 mM potassium persulfate (K₂S₂O₈) solution separately. The 2.45 mM K₂S₂O₈ and 7 mM ABTS solutions were mixed in equal quantities (10 mL each) to create the radical cation. The reaction mixture was allowed to sit at room temperature (25\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C) in the dark for 12 hours to achieve full radical production and absorbance stability.\u003c/p\u003e\u003cp\u003eA UV-Vis spectrophotometer was used to confirm that the working solution had an absorbance of 0.700\u0026thinsp;\u0026plusmn;\u0026thinsp;0.020 at 734 nm when the resultant concentrated ABTS⁺\u0026bull; solution was diluted 1:1 (v/v) with absolute ethanol. To perform the antioxidant capacity experiment, \u003cem\u003eLeucas lanata\u003c/em\u003e extract or L-AgNPs were used at concentrations of 100, 200, 300, 400, and 500 \u0026micro;g/mL. For two hours, the reaction mixtures were allowed to sit at room temperature in the dark. Using spectrophotometry, a drop in absorbance at 734 nm was observed. Appropriate blanks and controls were utilized.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003e2.6. Antibacterial activity\u003c/h2\u003e\u003cp\u003eThe antibacterial susceptibility of L-AgNPs against \u003cem\u003eSalmonella abony\u003c/em\u003e, \u003cem\u003eBacillus subtilis\u003c/em\u003e, \u003cem\u003eStaphylococcus aureus\u003c/em\u003e, and \u003cem\u003eEscherichia coli\u003c/em\u003e was evaluated using the agar well diffusion method. A sterile glass spreader dipped in the bacterial culture's inoculum suspension was applied to the hardened nutrient agar plates, and the surface of the agar plates was then thoroughly cleaned. After loading the different concentrations of the produced L-AgNPs solution into the bored wells, the Petri plates were incubated for 24 hours at 37\u0026deg;C to examine the inhibition zone. The assay's standard was streptomycin at 10 \u0026micro;g/mL. The diameter of the inhibitory zone was measured in millimeters and compared to the control [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003cb\u003eMIC (Minimum Inhibitor Concentration) Assay\u003c/b\u003e\u003c/p\u003e\u003cp\u003eA tube dilution approach was used to determine the MIC. The investigated bacterial species were cultured for 24 hours and then diluted using the 0.5 McFarland standard in 10 mL of tryptic soy broth (TSB) to produce an inoculum of 108 CFU mL 1. Neem extracts at a range of strengths, from 5000 g/mL to 10.0 g/mL, were made in a series of culture tubes using DMSO. Each tube received 0.1 mL of the bacterial cell suspension, which was then incubated at 37\u0026deg;C for 24 hours. The development of the inoculum was ascertained by measuring the turbidity of the broth. The lowest extract concentration that inhibited the development of the test organism was known as the minimum inhibitory concentration, or MIC [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e"},{"header":"3. Results and Discussion","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\u003ch2\u003e3.1. UV-VIS (Ultraviolet-Visible) spectroscopy\u003c/h2\u003e\u003cp\u003eThe reaction mixture's color shift from light brown to dark brown due to surface plasma resonances was the leading indicator of AgNP production. UV-VIS spectra of the colloidal solution of AgNPs were then taken, in which a peak at 384nm was observed for L-AgNPs, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003e. This is similar to the AgNPs mediated by the \u003cem\u003eFusarium exquisite, Fusarium solani, and Ficus carica\u003c/em\u003e extracts. Several biomolecules and functional groups found in \u003cem\u003eLeucas lanata\u003c/em\u003e extract facilitate the bio-reduction and capping of synthetic L-AgNPs [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003e3.2. X-ray beam diffraction (XRD) analysis\u003c/h2\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eX-ray diffraction was employed to ascertain the crystalline structure of the bio-synthesised L-AgNPs. Five peaks were found at 38.09\u0026deg;, 44.18\u0026deg;, 64.41\u0026deg;, 77.33\u0026deg;, and 81.44\u0026deg; in the XRD pattern as shown in Fig.\u0026nbsp;3, which is also represented by the SAED pattern. These peaks correspond to the [1 1 1], [2 0 0], [2 2 0], [3 1 1], and [2 2 2] planes, respectively. This result confirmed the cubic crystalline structure of L-AgNPs, whose basis was demonstrated in Joint Committee of Powder Diffraction Standards file numbers 040783 and 021098, as shown in different research. According to XRD, the size of synthesized L-AgNPs is estimated from the Debye-Scherrer formula, which ranges from 10nm to 75nm, while the average particle size is approximately 25.37nm, which is also vindicated by HRTEM result [\u003cspan additionalcitationids=\"CR37 CR38 CR39\" citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eFigure\u0026nbsp;3\u003c/strong\u003e\u003cp\u003eX-ray beam diffraction pattern of synthesized AgNPs\u003c/p\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003e\u003cb\u003e3.3. EDAX (Energy Dispersive X-ray) analysis\u003c/b\u003e\u003c/h2\u003e\u003cp\u003eEDAX was used to determine the chemical composition of the biomimetic synthesized L-AgNPs. The absorption peak for silver nanoparticles is generally believed to occur at 2.7 to 3 keV. In Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e4\u003c/span\u003e, a peak was observed at 2.7 keV, which assures the presence of silver nanoparticles. Apart from this, signals for C and Cl were also obtained in EDX, indicating that the phytochemical present in the extract of \u003cem\u003eLeucas lanata\u003c/em\u003e is responsible for the reduction of Ag\u003csup\u003e+\u003c/sup\u003e ions and the stability of AgNPs. The C-Cl group found in the biomolecule of \u003cem\u003eLeucas lanata\u003c/em\u003e is the reason for the significantly lower level of Cl, which FTIR further verified. These findings are in good agreement with the published research [\u003cspan additionalcitationids=\"CR42 CR43 CR44\" citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe EDAX tool is also used for element mapping, which provides a digital photograph showing the distribution of elements in the specimen through the color dots. Figure\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e5\u003c/span\u003e (a), (b), and (c) show the element mapping of C, Ag, and Cl, respectively, while Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e5\u003c/span\u003e(d) represents the overlay element mapping of all elements presented with their homogenous distribution in the sample [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003e3.4. FESEM (Field emission scanning electron microscopy) analysis\u003c/h2\u003e\u003cp\u003eFESEM assessed the surface morphology of L-AgNPs. The image of 100.00kx magnification is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e6\u003c/span\u003e, according to which the shape of synthesized L-AgNPs was found to be spherical and rod-like. The aggregation in L-AgNPs is responsible for a rod-like shape; the same results are also visible from the HRTEM image of L-AgNPs [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003ch2\u003e3.5. Fourier Transform Infrared analysis\u003c/h2\u003e\u003cp\u003eFourier Transform Infrared (FTIR) analysis helps determine which functional groups are present in the biomolecules in \u003cem\u003eLeucas lanata\u003c/em\u003e extract, which is essential for Ag\u003csup\u003e+\u003c/sup\u003e ion reduction, capping, and stability of AgNPs. Various sharp bands were obtained in FTIR of L-AgNPs between 3900cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and 532 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e7\u003c/span\u003e). The band near 3900 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e is due to N-H stretching, while the band at 3492 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e is due to O-H stretching vibration, indicating the presence of alcoholic and phenolic groups. The minor band observed in the region of 2917 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e is due to C-H stretching in an alkane. The band observed at 2376 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e corresponds to the \u0026equiv;\u0026thinsp;C-C stretching vibration. The shared band at 1638 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e indicates the presence of a C\u0026thinsp;=\u0026thinsp;O or C\u0026thinsp;=\u0026thinsp;C stretching vibration in the aromatic ring. The bands found at 1382 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and 1113 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e are due to the C-O, N-H stretching vibration of the amide linkage, while the bands found at 844 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e are due to the C-H banding of alkenes. The band at 532 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e is attributed to C-Cl or C-Br stretching vibrations, the presence of which has also been confirmed by EDAX. These bands indicate the presence of phytochemicals in the plant extract, which play a vital role in the formation process and act as capping and stabilizing agents [\u003cspan additionalcitationids=\"CR50 CR51 CR52 CR53 CR54 CR55 CR56 CR57\" citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\u003ch2\u003e3.6. High-resolution transmission electron microscopy analysis\u003c/h2\u003e\u003cp\u003eHigh-resolution transmission electron microscopy (HRTEM) plays a crucial role in determining the morphology, including the size and shape of AgNPs. Figure\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e8\u003c/span\u003e shows the particles of L-AgNPs at different magnifications. L-AgNPs have a spherical shape, but due to agglomeration, they also have rod-like morphology, which is also confirmed by the FESEM image (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e8\u003c/span\u003ea). For the average particle size, a total of 44 L-AgNPs were sampled to get the average particle size, for which the Image J software was used. The particle size of L-AgNPs ranges from 10 nm to 112 nm. However, particles with more than 50nm are very small, and the average particle size is 28.59nm, which is also verified by XRD (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e8\u003c/span\u003ee). In the SEAD pattern of synthesized L-AgNPs, eight spots are found that represent specific crystal planes. Figure\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e8\u003c/span\u003e(e) shows eight concentric circles, out of which four are found to be [1 1 1], [2 0 0], [2 2 0], and [3 1 1] planes, which suggest the crystalline nature of L-AgNPs.The interplanar d spacing of 0.15nm for the [2 2 0] plane of L-AgNPs is shown in the figure. 7(f) [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e] [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e, \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\u003ch2\u003e3.7. Dynamic Light Scattering (DLS)\u003c/h2\u003e\u003cp\u003eThe size distribution and zeta potential of L-AgNPs were ascertained by DLS analysis. The zeta potential can be used to assess the stability of AgNPs. The zeta potential gives information about the charge of the phytochemicals acting as capping reagents. The higher the charge on the surface, the greater the repulsion force existing between the synthesized L-AgNPs, due to which the aggregation of L-AgNPs will decrease and the dispersity of nanoparticles will increase in the medium. The zeta potential value of synthesized L-AgNPs was \u0026minus;\u0026thinsp;25.4mV, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e9\u003c/span\u003e (a). According to previous research, a voltage of 30 mV is considered the most stable range for nanosuspensions. It also confirms that synthesized L-AgNPs are relatively stable and have a good colloidal nature. In contrast, Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e9\u003c/span\u003e(b) shows the particle size distribution of L-AgNPs, which shows that particles are of a higher particle size of more than 100 nm as compared to the particle size suggested by XRD and HRTEM. The difference is due to the sample preparation procedure. In the DLS method, the size of L-AgNPs is obtained as the hydrodynamic diameter, which is why the particle size predicted by DLS is higher than predicted by XRD and HRTEM [\u003cspan additionalcitationids=\"CR59 CR60 CR61 CR62\" citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\u003ch2\u003e3.8. Antioxidant activity\u003c/h2\u003e\u003cp\u003eThe antioxidant activity was determined using the DPPH assay. In this case, ascorbic acid was taken as a control. The antioxidant activity of ascorbic acid, \u003cem\u003eLeucas lanata\u003c/em\u003e plant extract, and silver nanoparticles made from \u003cem\u003eLeucas lanata\u003c/em\u003e is completely dose-dependent towards free radicals like DPPH, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e10\u003c/span\u003e(a). The scavenging activity of DPPH of plant extract was lower than that of synthesized L-AgNPs and almost equal to that of ascorbic acid at a high concentration of 500\u0026micro;g/ml. The %RAS values for the plant extract and L-AgNPs at 100 \u0026micro;g/ml were 38% and 45%, respectively. For a maximum concentration of 500 \u0026micro;g/ml, the %RAS values were 81% and 93%, respectively. The IC\u003csub\u003e50\u003c/sub\u003e value for the plant extract of \u003cem\u003eLeucas lanata\u003c/em\u003e was 154 \u0026micro;g/ml, whereas for L-AgNPs, it was found to be 49 \u0026micro;g/ml, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e10\u003c/span\u003e(b), which is significantly lower compared to the \u003cem\u003eLeucas lanata\u003c/em\u003e plant extract. This result is also confirmed by results obtained for silver nanoparticles mediated by \u003cem\u003eCucumis prophetarum\u003c/em\u003e [\u003cspan additionalcitationids=\"CR65\" citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e]. \u003cem\u003eThe\u003c/em\u003e aqueous extract of Leucas lanata and L-AgNPs demonstrated their ABTS radical scavenging activity in Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e10\u003c/span\u003e(c). When compared to aqueous plant extract, L-AgNPs exhibit a larger proportion of ABTS scavenging activity. At lower concentrations of 100 \u0026micro;g/ml, L-AgNPs exhibit 12.26% inhibition, whereas at higher concentrations of 500 \u0026micro;g/ml, they exhibit 59.92% inhibition. Nonetheless, the plant's aqueous extract exhibits 8.75% at lower concentrations of 100 \u0026micro;g/ml and 53.17% at higher concentrations of 500 \u0026micro;g/ml. Figure\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e10\u003c/span\u003e(d) showed the FRAP assay of L-AgNPs and the aqueous extract of \u003cem\u003eLeucas lanata\u003c/em\u003e. L-AgNPs have a higher FRAP than aqueous plant extract. L-AgNPs show 21.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17 \u0026micro;M at a lower concentration of 100 \u0026micro;g/ml and 83.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15 \u0026micro;M at higher values of 500 \u0026micro;g/ml. However, at a lower concentration of 100 \u0026micro;g/ml, the aqueous solution of the plant extract shows 14.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11, whereas at a higher concentration of 500 \u0026micro;g/ml, it shows 69.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21. The current findings showed that, in addition to silver nanoparticles exhibited potent antioxidant activity compared with aqueous plant extract of \u003cem\u003eLeucas lanata\u003c/em\u003e, as previously supported by the previous study [\u003cspan additionalcitationids=\"CR68\" citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e]\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eFig: 10(a): DPPH (2,2-Diphenyl-1-picrylhydrazyl) assay analysis for Antioxidant activity\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eFigure\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e10\u003c/span\u003e(b): DPPH (2,2-Diphenyl-1-picrylhydrazyl) Free radical scavenging Activity AgNPs and IC\u003csub\u003e50\u003c/sub\u003e values\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\u003ch2\u003e3.9. Antibacterial Activity\u003c/h2\u003e\u003cp\u003eThe antibacterial activity of, was tested using two gram-positive and two gram-negative pathogens: \u003cem\u003eSalmonella ebony\u003c/em\u003e, \u003cem\u003eBacillus subtilis, Escherichia coli\u003c/em\u003e, and \u003cem\u003eStaphylococcus aureus\u003c/em\u003e as positive and negative controls, respectively, streptomycin and All Bacteria culture diluted 10\u003csup\u003e8\u003c/sup\u003e CFU/ml and spread on agar plate for incubation 24 hr at 37\u0026ordm;C to measure the inhibition zones. DMSO were employed as observed Fig.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e11\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e12\u003c/span\u003e. The activity of the produced L-AgNPs against gram-positive and gram-negative bacteria was nearly identical. Silver nanoparticles derived from the peel of \u003cem\u003eCitus sinensis\u003c/em\u003e and \u003cem\u003eAcacia leucophloea\u003c/em\u003e also showed comparable outcomes [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e, \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e]. The table displays the inhibition zone at various concentrations. Compared to L-AgNPs, the inhibitory zone of the plant extract is smaller. At the lowest concentration of L-AgNPs, \u003cem\u003eBacillus subtilis\u003c/em\u003e exhibited the highest inhibitory zone, measuring 10 mm (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). When bacteria and silver ions interact, it inhibits essential processes such as DNA replication, respiratory enzyme function, and ATP synthesis, ultimately leading to cell death [\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e73\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\u003eInhibition Zone Diameter(mm) for different Bacteria\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"9\"\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\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eConcentration\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e\u003cp\u003e\u003cem\u003eStaphylococcus aureus(P)\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e\u003cp\u003e\u003cem\u003eEscherichia coli(Q)\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e\u003cp\u003e\u003cem\u003eSalmonella abony(R)\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e\u003cp\u003e\u003cem\u003eBacillus subtilis (S)\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cem\u003eLeucas lanata\u003c/em\u003e Plant extract\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eL-AgNPs\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eLeucas lanata\u003c/em\u003e Plant extract\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eL-AgNPs\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cem\u003eLeucas lanata\u003c/em\u003e Plant extract\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eL-AgNPs\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cem\u003eLeucas lanata\u003c/em\u003e Plant extract\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eL-AgNPs\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e2000\u0026micro;g/mL (A)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6 mm\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\u003e10 mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e25mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e10 mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e17mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e16 mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e20mm\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1000\u0026micro;g/mL (B)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4 mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e10 mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6 mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e19mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e4mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e12mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e6 mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e15mm\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e500\u0026micro;g/mL (C)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e---------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5 mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e--------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e11mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e--------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e8mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e12mm\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e250\u0026micro;g/mL (D)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e--------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e--------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e---------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e10mm\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eStreptomycin \u0026micro;g/mL(F)\u003c/p\u003e\u003cp\u003ePositive Control\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e20 mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e23 mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e24 mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e29mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e20 mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e20mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e20mm\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e23mm\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDMSO (Dimethyl sulfoxide) (E)\u003c/p\u003e\u003cp\u003e(Negative Control)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e--------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e--------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e--------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e--------\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-------\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\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eMIC (Minimum Inhibitory Concentration) against Bacteria\u003c/b\u003e\u003c/p\u003e\u003cp\u003eFigure \u003cspan refid=\"Fig12\" class=\"InternalRef\"\u003e13\u003c/span\u003e shows that the aqueous extract of Leucas lanata exhibited effects between 0.45 and 0.84 mg/mL, whereas L-AgNPs showed effects between 0.09 and 0.34 mg/mL across the bacteria. With a minimum inhibitory concentration (MIC) of 0.84 mg/mL, the plant extract in aqueous solution showed the most potent effects against \u003cem\u003eSalmonella abony\u003c/em\u003e. \u003cem\u003eBacillus subtilis\u003c/em\u003e was inhibited with a minimum inhibitory concentration (MIC) of 0.76 mg/mL, whereas \u003cem\u003eE.coli\u003c/em\u003e showed the strongest action with the lowest MIC value of 0.45 mg/mL. MIC 0.68 mg/ml is moderate for \u003cem\u003eStaphylococcus aureus\u003c/em\u003e. With a minimum inhibitory concentration (MIC) of 0.09 mg/mL, L-AgNPs exhibited the most notable efficacy against \u003cem\u003eE. coli\u003c/em\u003e. The MIC value for the second activity against \u003cem\u003eStaphylococcus aureus\u003c/em\u003e was 0.16 mg/mL. \u003cem\u003eBacillus subtilis\u003c/em\u003e and \u003cem\u003eSalmonella abony\u003c/em\u003e, on the other hand, exhibit 0.34 and 0.24 mg/ml, respectively. Consequently, L-AgNPs exhibit a MIC value that is noticeably higher than that of the plant's aqueous extract.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e"},{"header":"4. Limitations and Future Scope of the Study","content":"\u003cp\u003eThe study has some limitations that warrant further investigation. The exact mechanism by which phytochemicals interact to stabilize and cap the silver nanoparticles was not fully elucidated, leaving a gap in understanding the underlying processes. The antimicrobial activity was evaluated against a limited number of bacterial strains; expanding the investigation to include a broader range of microorganisms, such as fungi and drug-resistant strains, would provide more comprehensive insights into the efficacy of the nanoparticles. Additionally, the environmental impact of \u003cem\u003eLeucas lanata\u003c/em\u003e-based silver nanoparticles, including their biodegradability and long-term stability within ecological systems, remains unexplored. The absence of in vivo studies to validate the in vitro antioxidant and antimicrobial efficacy represents another limitation, as such studies are critical for practical and clinical applications. Addressing these limitations in future research would significantly enhance the relevance and applicability of the study.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eSeveral biological entities have recently been investigated for their ability to mediate the green synthesis of silver nanoparticles (AgNPs). In this study, \u003cem\u003eLeucas lanata\u003c/em\u003e extract, prepared from its aerial parts, was employed as a natural reducing and capping agent for the synthesis of AgNPs. Characterization techniques, such as UV-VIS spectroscopy, confirmed nanoparticle formation with a distinct absorption peak at 384 nm, while EDX analysis verified the elemental composition of the silver. Morphological assessment using HRTEM and structural evaluation through XRD indicated that the L-AgNPs were predominantly spherical with an average particle diameter of under 29 nm. The antioxidant capacity of the synthesized nanoparticles was evaluated using the DPPH, ABTS, and FRAP radical scavenging methods, which suggest an enhanced antioxidant efficiency of L-AgNPs. The antibacterial efficacy of L-AgNPs was tested against two Gram-positive \u003cem\u003e(Staphylococcus aureus, Bacillus subtilis)\u003c/em\u003e and two Gram-negative \u003cem\u003e(Escherichia coli, Salmonella abony)\u003c/em\u003e strains. L-AgNPs exhibit a MIC value that is noticeably higher than that of the plant's aqueous extract. Results demonstrated consistent inhibitory zones across all four bacterial species, with a slightly higher zone of inhibition observed against \u003cem\u003eBacillus subtilis\u003c/em\u003e. These findings highlight the effectiveness of \u003cem\u003eLeucas lanata\u003c/em\u003e as a novel, eco-friendly source for the synthesis of bio-functional silver nanoparticles with promising antimicrobial and antioxidant properties.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eSPR\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eSurface plasmon resonance\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eAgNPs\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eSilver Nanoparticles,\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eUV-VIS\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eUltraviolet- Visible\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eEDX\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eEnergy Dispersive X-ray\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eXRD\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eX-ray beam diffraction,\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eFTIR\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eFourier Transform Infrared\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eFESEM\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eField emission scanning electron microscopy\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eHRTEM\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eHigh-resolution transmission electron microscopy\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eDPPH\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003e2,2-Diphenyl-1-picrylhydrazyl\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003emM\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003emillimolar\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eJCPDS\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eJoint Committee on Powder Diffraction Standards\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eSAED\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eSelected area electron diffraction\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eDMSO\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eDimethyl sulfoxide and L-AgNPs:Silver nanoparticles mediated by \u003cem\u003eLeucas lanata\u003c/em\u003e extract\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003e℃\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eDegree Celsius\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003emm\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003emillimetre\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003egm\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003egram:FRAP:Ferric Reducing Antioxidant Power TPTZ:2,4,6-tripyridyl-striazine\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eABTS\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003e2,2\u0026prime;-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eMIC\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eMinimum Inhibitory Concentration\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eCFU\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eColony-Forming Unit\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor Contributions Statement\u003c/strong\u003e:\u0026nbsp;\"R.S., S.T., and I. R. wrote the main manuscript text, and R.K.B. A.B. and L.A.W. prepared different tables and figures. All authors reviewed the manuscript.\"\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding Statement:\u003c/strong\u003e This research received no specific grant from any funding agency in any sectors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability:\u003c/strong\u003e All relevant data are within the manuscript\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclarations\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics, consent to participate, and consent to publish\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical trial number\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAmeen F, Srinivasan P, Selvankumar T, Kamala-Kannan S, Al Nadhari S, Almansob A, Govarthanan M (2019) Phytosynthesis of silver nanoparticles using Mangifera indica flower extract as bioreductant and their broad-spectrum antibacterial activity. 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Molecules 26(16):5057. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/molecules26165057\u003c/span\u003e\u003cspan address=\"10.3390/molecules26165057\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"discover-nano","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"narl","sideBox":"Learn more about [Discover Nano](https://www.springer.com/journal/11671)","snPcode":"11671","submissionUrl":"https://submission.nature.com/new-submission/11671/3","title":"Discover Nano","twitterHandle":"@SpringerOpen","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Silver nanoparticles, Green-synthesis, phytochemicals, Capping agents, zeta potential","lastPublishedDoi":"10.21203/rs.3.rs-7981923/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7981923/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIn the present study, a green, Phyto-assisted approach was employed to synthesize silver nanoparticles (AgNPs) using the aqueous extract of the aerial parts of \u003cem\u003eLeucas lanata\u003c/em\u003e. The successful formation of AgNPs was evidenced by a characteristic surface plasmon resonance (SPR) band at 384 nm in the UV–Vis absorption spectrum. Elemental mapping and energy-dispersive X-ray (EDX) analysis further confirmed the presence of silver, showing a prominent peak at 2.7 keV. X-ray diffraction (XRD) patterns revealed distinct crystalline planes, which were consistent with the results of Selected area electron diffraction (SAED). High-resolution transmission electron microscopy (HRTEM) determined an average particle size of 25.37 nm. The nanoparticles exhibited a high zeta potential value, suggesting strong stability attributed to phytochemical-mediated capping, as corroborated by Fourier transform infrared spectroscopy (FT-IR). The biosynthesized L-AgNPs displayed potent antioxidant activity with an IC₅₀ value of 49 µg/mL in the DPPH assay. L-AgNPs exhibited a higher proportion of ABTS scavenging activity (12.26%) than the \u003cem\u003eLeucas lanata\u003c/em\u003e extract at a lower concentration of 100 µg/mL, indicating enhanced antioxidant potential. Similarly, the FRAP value of L-AgNPs (21.56 ± 0.17 µM at 100 µg/mL) was significantly greater than that of the aqueous plant extract, further confirming their superior reducing capacity. Moreover, the silver nanoparticles demonstrated significant antimicrobial activity against \u003cem\u003eStaphylococcus aureus, Escherichia coli, Salmonella abony, \u003c/em\u003eand\u003cem\u003e Bacillus subtilis.\u003c/em\u003e The MIC value for \u003cem\u003eLeucas lanata\u003c/em\u003e aqueous extract showed effects between 0.45 and 0.84 mg/mL, whereas L-AgNPs showed effects between 0.09 and 0.34 mg/mL across bacteria. To the best of our knowledge, this is the first report on the silver nanoparticle-synthesizing potential of \u003cem\u003eLeucas lanata\u003c/em\u003e, highlighting its promising applications in biomedical and pharmaceutical fields.\u003c/p\u003e","manuscriptTitle":"Green synthesis and characterization of silver nanoparticles from the aerial parts of Leucas lanata with enhanced antioxidant and antimicrobial activities","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-08 09:59:04","doi":"10.21203/rs.3.rs-7981923/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-12-31T11:13:08+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-30T22:43:15+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-13T03:55:22+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"88051114165748144368493383425065599737","date":"2025-12-05T12:51:42+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"62814343767496433033699897700979019042","date":"2025-12-04T10:54:16+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"122077986874405387677250891531209414019","date":"2025-12-04T08:05:50+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-12-04T05:00:50+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-12-02T17:50:53+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-11-28T11:46:38+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-11-28T11:38:01+00:00","index":"","fulltext":""},{"type":"submitted","content":"Discover Nano","date":"2025-11-28T11:29:03+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"discover-nano","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"narl","sideBox":"Learn more about [Discover Nano](https://www.springer.com/journal/11671)","snPcode":"11671","submissionUrl":"https://submission.nature.com/new-submission/11671/3","title":"Discover Nano","twitterHandle":"@SpringerOpen","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"74400df9-89c2-4733-89d2-30324e2107f0","owner":[],"postedDate":"December 8th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-04-27T21:08:25+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-08 09:59:04","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7981923","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7981923","identity":"rs-7981923","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}