Stachys tibetica derived silver nanoparticles: A robust multifunctional material for enhanced biological activity and photocatalytic properties 

Stachys tibetica derived silver nanoparticles: A robust multifunctional material for enhanced biological activity and photocatalytic properties | 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 Stachys tibetica derived silver nanoparticles: A robust multifunctional material for enhanced biological activity and photocatalytic properties Naseer Ahmad Dar, Parvaiz A Dar, Mahpara Qadir, Wajaht Amin Shah This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5292367/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Stachys tibetica , a significant medicinal plant was collected from Ladakh (India) and subjected to extraction. The extract was titrated with AgNO 3 at 50ºC to obtain silver nanoparticles with plant extract acts as a reducing and capping agent. The eco-friendly synthetic procedure utilizes the phytochemicals in Stachys tibetica , ensuring a sustainable and non-toxic approach to nanoparticle production. The characterization of the silver nanoparticles was conducted using various techniques, including Fourier-transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), and dynamic light scattering (DLS). Field emission scanning electron microscopy (FE-SEM) was utilized to examine the surface morphology, revealing the irregular and rugged spherical shape of the synthesized nanoparticles. Stachys tibetica -derived silver nanoparticles (AgNPs) act as a versatile and robust multifunctional material with proficient bioactivity and catalytic properties. The synthesized silver nanoparticles displayed effective antioxidant and antimicrobial properties against Bacillus Cereus, Pseudomonas, Escherichia coli, and S.Aeureus . Additionally, the green nanoparticles degraded Rhodamine-B under sunlight irradiation within 1 hour at a rate constant of 0.066 min − 1 . Stachys tibetica silver nanoparticles FE-SEM XRD Rhodamine-b Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1. Introduction Nanotechnology an emerging multidisciplinary field having roots in basic sciences such as physics, chemistry and biology uses science to manipulate matter at molecular level and create nanoparticles of varying sizes, shapes and chemical compositions for their potential applications to human benefit [ 1 – 4 ]. Physical, chemical and environmentally friendly processes are used to create nanoparticles[ 5 , 6 ].In recent years, researchers have focused on developing efficient green chemistry procedures that use natural reducing capping and stabilizing chemicals to make metal nanoparticles with desired morphology and size. Scientists have been successful in increasing the efficacy of these materials by reducing particle size, because of their sub-microscopic size they have unique material characteristics and so synthesized nanoparticles are used in a various areas, such as engineering, medicine, environmental remediation and catalysis[ 7 ]. Environmental and toxicity concerns involved in some of synthetic methodologies involving the use of organic solvents, toxic reducing substances used for reduction of metals, etc have led for the demand of environmentally friendly, clean, reliable and biologically appropriate approaches. Nanoparticle synthesis from plant extracts and different types of plant biomass not only uses time conserving and environmentally friendly procedures but also these nanoparticles involve very simple and easy handling procedures[ 8 ]. Stachys is genus of 350–400 species of plants belonging to the family Lamiaceae, also called as mint family. Plants belonging to this family are found worldwide. Most members of this family are either perennial or annual herbs although some species of this family are shrubs or subshrubs[ 9 ]. Extensive phytochemical and pharmacological studies have been carried out on Stachys species, proving their ethnopharmacological benefits. Anti-inflammatory, antioxidant, analgesic, Reno protective, anxiolytic, and depressive activities are some of the prominent biological activities carried out on these species[ 10 ] [ 11 – 13 ]. The phytochemical content of these species is linked to a wide spectrum of medicinal benefits. As a result, the genus Stachys has garnered a lot of interest for the isolation and bioprospection of bioactive secondary metabolites of plant. In total, more than 200 compounds from this genus have been isolated, including terpenes (e.g., iridoids triterpenes, diterpenes,), polyphenols (e.g., lignans, flavone derivatives, phenylethanoid glycosides,), and phenolic acids etc [ 10 , 14 – 18 ]. Very efficient metallic nanoparticles like Iron, nickel, Copper, and Silver have been synthesized by researchers from the plants of the lamiaceae family eg. nanoparticles synthesized from Mentha Pulegium , exhibited potential antibacterial, cytotoxic, antioxidant and antifungal activities[ 19 – 21 ]. Besides being used as antibacterial agents nanoparticles also play the role of a catalyst for the degradation of various environmentally hazardous chemicals. Saliva officinalis mediated silver nanoparticles degrade Congo red dye very efficiently[ 22 ]. similarly, AgNPs from endostemon viscosus degrade methylene blue and crystal violet dyes[ 23 ]. Keeping these findings in view, the aim of our study is the green synthesis of silver nanoparticles from Stachys tibetica (locally known as yaktse), which is not reported in the literature yet. we were further motivated to carry out antibacterial, anti-oxidant and dye degradation properties of these silver nanoparticles. The sample plant for this study was collected from Minjhi, Ladakh, and Silver nanoparticles were synthesized from the extract of this plant using a 20 mM AgNO 3 solution. The bactericidal effect of these nanoparticles was checked against four bacterial strains namely Bacillus cereus, E.Coli, S.Aeureus , and Pseudomonas. The synthesized nanoparticles were further utilized as a catalyst in the degradation of environmentally hazardous dye Rhodamine-B with the use of a UV-Vis spectrophotometer. The absorbance spectrum of the solution was examined at various wavelengths. The absorbance value at 550-560nm depicted dye concentration throughout degradation. 2. Materials and Methods 2.1. Materials The ATR-FTIR Spectrophotometer (Agilent Technologies, USA) was used to study the structural properties of dried plant extract and silver nanoparticles at a constant temperature of 25°C. The wavenumber range was 500–4000 cm − 1 . FE-SEM was used to analyze the surface morphology of nanoparticles (FEI Quanta 200, USA) operated at 200V. XRD (Ultima-IV (Rigaku Corporation, Tokyo, Japan) uses Cu Kα radiation. The samples were scanned in a 2θ range of 10 − 80° at a scan rate of 5 min − 1 . 2.1.1. Collection of plant material The aerial parts of Stachys tibetica were collected from Minjhi region of Ladakh, india. The proper identification of material was done by curator Akhter Ahmed Malik of the Department of Taxonomy, Kashmir University. The voucher no [9062-(KASH)] has been deposited at the herbarium, Department of Botany, Kashmir University, Srinagar. 2.1.2. Extract preparation The collected material was shade-dried for ten days. 30 grams of ground plant material was soaked in distilled water and kept on standing for 5 days. The final solution was filtered through Whatman filter paper No.1 to obtain a uniform yellowish aqua extract of Stachys tibetica for further experimentation. 2.2. Methods 2.2.1. Preparation of silver Nanoparticle Stachtys tibetica extract-based nanoparticles were synthesized as per the green synthetic methodology reported for the synthesis of nanoparticles[ 24 ]. The aqueous extract of Stachys tibetica was dried on a water bath till the solution became deeply yellow. 20 mmol solution of AgNO 3 was filled in the burette and added to water-dissolved aqueous extract of Stachys tibetica dropwise the solution was continuously stirred on hotplate set at a temperature of 50 º C. Silver nitrate solution was continuously added till the colour of solution changes from yellowish to deep brown which marked the end point of reaction. The final solution was centrifuged at 8000 rpm for 15 minutes. The final nanoparticle was obtained by discarding the supernant. The silver nanoparticle obtained was washed with distilled water 3–4 times to obtain a pure mass of nano scale particles which was further dried in oven at 50 º C to obtain a powdery mass of nanoparticle. 2.2.2. Antibacterial Activity A panel of four bacterial strains namely Staphylococcus aureus, Bacillus cereus, Pseudomonas and E.coli , obtained from Microbial type culture collection (MTCC), Institute of Microbial Technology, Chandigarh, India. These bacterial strains were cultivated on nutrient agar plates and kept on agar slants at 37ºC. Microorganism cell suspension in 0.9% NaCl was adjusted to 0.5 Mc Farland to obtain approximately 106 cfu/ml. 2.2.3. Antioxidant properties The free radical scavenging potential of AgNP, plant extract, and standard ascorbic acid was checked against DPPH(2,2-Diphenyl-1-Picrylhydrazyl) using a modified approach proposed by W brand Williams et al[ 25 ]. In different test tubes solutions with concentrations (20,40,60,80,100,120,140,160,180,200 µg/ml) of AgNP, plant extract, and ascorbic acid were prepared). To each solution, DPPH(1mM) prepared in methanol was added and mixed thoroughly. The solution obtained after the addition of DPPH was incubated in the dark for 30 minutes at room temperature before being measured for UV absorbance at 517nm with a UV-Vis spectrophotometer (perkin-Elmer Lambda 950:UK). The DPPH solution obtained without the addition of a sample was used as a control to measure radical scavenging activity. The Free radical scavenging activity of all samples was calculated using the formula; Radical scavenging activity = \(\:\frac{{AB}_{C-{AB}_{S}}}{{AB}_{C}}\times\:100\) Where AB C is the absorbance of methanol + DPPH, and AB S is the absorbance of sample solution + DPPH. 2.2.4. Degradation of rhodamine-B dye 10mg of Rhodamine-b dye were typically added to 1000 mL of double distilled water as a stock solution. A control of dye and NaBH 4 (0.06M) without the inclusion of silver nanoparticles was also kept. 5ml freshly prepared NaBH 4 solution was added to 20ml of rhodamine B solution and subsequently 5ml of nano catalyst of a definite concentration was added to it. Prior to irradiation, the reaction suspension was thoroughly mixed to ensure that the working solution was clearly balanced. Following that, the dispersion was exposed to visible light and continuously monitored. Aliquots of 2–3 mL solution were filtered and utilized to analyze the photocatalytic degradation of dye at particular time intervals. The absorbance spectrum of the solution was examined with a UV-Vis spectrophotometer at various wavelengths. The absorbance value at 550-560nm depicted dye concentration throughout degradation. 3. Results and Discussion Nanoparticles serve an effective scientific and research tools having an excellent catalytic efficiency due to their large surface area. Silver nanoparticles synthesized through green approach act as functional materials that have been explored for curbing of various modern biological, industrial and environmental challenges. These nanoparticles synthesized from the aqueous extract of Stachys tibetica were characterised using various techniques. Initial screening was carried out through FT-IR analysis, that revealed the presence of various functional groups. P-XRD revealed the crystallinity, and the morphology was screened using SEM analysis. The synthesised nanoparticles were explored in various directions ranging from bioactivity, antioxidant mitigation to water pollution remediation. The extensive discussions are addressed in various sections 3.1. FTIR Analysis of Synthesized Nanoparticle The FTIR (Fig. 1 a) spectra of produced AgNPs were examined to determine the existence of potential functional groups inside biomolecules that bind to the surface of silver for bioreduction as shown in the Fig. 1 (a). IR spectrum of impregnated silver nanoparticle shows absorption bands at 2975cm − 1 corresponding to N-H Stretching, 2150cm − 1 corresponding to C-N double bond stretching, 1609cm − 1 corresponding to N-H bending, 1380cm − 1 corresponding to N-O stretching whereas plant extract absorbs at 2945cm − 1 ,2041cm − 1 ,1609cm − 1 ,1392cm − 1 . 3.2. XRD Analysis The XRD pattern (Fig. 1 b) of silver nanoparticles shows strong peaks at 2Ɵ = 33.21º, 38.47º, 47.37º and 77º which correspond to the (1 1 1), (2 0 0), (2 2 0), (3 1 1), Bragg’s reflections of the face-centered cubic structure of silver. In addition, unassigned peaks that were weaker than those of silver could be related to the bioorganic chemicals forming on the surface of the AgNPs[ 26 ]. 3.3. SEM And EDS Analysis of synthesized nanoparticle Field-Emission Scanning electron Microscope (FE-SEM) was used to analyze the surface topography and morphology of S.Tibetica based AgNPs. It is apparent from the SEM monograps that the material has developed into regular nanospheres with a rugged surface which increases the surface area of the catalyst Fig. 2 (a and b). The existence of silver metal in the produced nanoparticles was confirmed using energy dispersive X-ray spectroscopy (EDS). As seen in Fig. 2 (C). A sharp peak at 3 KeV established the existence of elemental silver in the nanoparticles. The EDS examination of produced AgNPs revealed indications of elements like Carbon(C), Nitrogen(N), and Oxygen(O), etc besides metallic silver because the substance generated via biological method had a minor fraction of Carbon, Nitrogen and Oxygen, etc [ 27 ]. The same is also revealed from the elemental mapping images of nanoparticles. (S-1) The average particle size as measured from DLS measurements was found out to be 78nm 3(a). The composition of different elements present in nanoparticle by weight percentages is depicted in Fig. 3 (b). 3.4. Antibacterial properties The MIC value of biosynthesized AgNPs to inhibit bacterial growth was determined by testing zone formation with dilutions ranging from 5 to 40µg/ml. The synthesized AgNP has a MIC of 10µg/ml and 20µg/ml, for E. coli and C. aureus respectively (S-4). The antibacterial activity of AgNPs, Stachys tibetica plant extract, and Streptomycin was tested at a level of 10µg/ml, as both pathogens demonstrated sensitivity at this concentration. The freshly prepared samples were tested for antibacterial activity against two gram-positive and two gram-negative strains namely Bacillus cereus, Pseudomonas, Escherichia coli , and Staphylococcus aureus. AgNPs (10µg/ml) have shown highest zone of inhibition of about 12.1mm for Bacillus Cereus followed by Pseudomonas (11.8 mm), E. coli (9.8) and Staphylococcus aureus (7.2) (S-5). The zone of inhibition observed for plant extract was less as compared to zone of inhibition obtained for AgNPs as shown in Fig. 4 . This study developed the synergistic antibiotic activity of AgNPs, which is greater than the individual antibacterial activity of plant extract when used together[ 28 ]. AgNP behaves as a positively charged center that interacts with the negatively charged centers of bacterial cells [ 29 ]. Recent research has found that the methods of action of AgNP against microbes are dependent on the NPs' size, shape, stability, and affinity[ 30 ]. Literature has cited that the bactericidal effect of AgNPs is due to the interaction of silver with nucleosides of nucleic acids and the creation of subsequent silver compounds with bactericidal potential[ 31 ]. The organic layer-coated silver nanoparticle has shown the highest bactericidal effect towards Bacillus cereus (12.1 ± 1) which can be inferred from the relatively simple cell wall structure of G( + ) bacterial species and the presence of a thick PG layer of 30 nm[ 32 ]. The reason of bactericidal behaviour of AgNPs towards Gram negative G(-) can be inferred from the fact that G( - ) bacteria have a small PG layer of 1–5 nm between the exterior layer and the cytoplasmic film that makes these species prone to bactericidal effects[ 33 ]. 3.5. Antioxidant properties DPPH has hydrogen acceptor capability to antioxidants[ 34 ].DPPH can easily accept electron from antioxidant compounds and during this process the color of DPPH changes from violet to yellow. Thus, DPPH is a free radical that accepts a free electron from antioxidants and becomes a stable diamagnetic molecule. The DPPH radical scavenging tends to increase with increasing the concentration of AgNPs, showing a maximum at higher concentrations. At concentrations 20–200µg/mL, AgNPs showed 32.4–75.8%, Plant extract showed 8–62%, and standard ascorbic acid showed 34.5– 89.6% radical scavenging activity. the radical scavenging activity of plant extract and AgNp was lower than that of standard Ascorbic acid. The results obtained for the antioxidant activity of AgNP, plant extract, and standard ascorbic acid at different concentrations are as shown below. 3.6 Dye Degradation Dye degradation studies were performed using AgNPs as catalyst. The following formula was used to calculate the percentage of dye degradation: %Degradation = 100× ( \(\:{\varvec{C}}_{\varvec{o}}\) - C) / \(\:{\varvec{C}}_{\varvec{o}}\) where \(\:{C}_{o}\) is the initial concentration of dye solution and C is the concentration of dye solution following photocatalytic degradation.The PFO model was found to be the optimal choice in determining the rate constant and rate constant was found out to be 0.06667 min − 1 using the following rate equation: $$\:\varvec{l}\varvec{n}\frac{{\varvec{C}}_{\varvec{t}}}{{\varvec{C}}_{\varvec{o}}}=\:-\varvec{K}\varvec{t}$$ The catalytic efficiency of AgNPs was accessed from λ max peak at 550-560nm at different intervals of irradiation. In the presence of AgNPs, the primary absorption peak of dye at 550nm decreased slowly with increasing time of exposure of dye to visible light in the presence of AgNp. This indicated the photocatalytic degradation of Rhodamine-b dye in the presence of AgNPs (Fig. 5). In the Dye degradation reaction, NaBH 4 helps in the reduction of dye, the Ag nanoparticles act as an electron relay from BH 4 − (donor) to dye(acceptor). The plausible mechanism of dye degradation is believed to proceed with the decomposition of NaBH 4 into BH 4 − (nucleophilic) and subsequently produces H-Ag and Ag-BH 3 as reaction intermediates in presence of AgNP S . The intermediate Ag-H is responsible for the degradation of dye(electrophilic) into harmless degradation products[ 35 ]. Studies on photocatalytic studies demonstrated that the photocatalytic activity of metallic nanoparticles is dependent on their crystallographic structure, shape, and size[ 36 ]. J Kadam et al. employed AgNPs produced from Cauliflower waste to photodegrade methylene blue in 152 minutes [ 37 ] [ 38 ], whereas the current investigation exhibited 100% Rh-B degradation in just 1 hour As a result, this study demonstrated the efficient photodegradation capacity of AgNPs generated using S. Tibetica in visible light. 4. Conclusion The green synthesis of silver nanoparticles from Stachys tibetica extract has been carried out in the present study, whose characterization revealed the face-centred cubic structure. The synthesized nanoparticles not only act as good catalyst for the degradation of rhodamine-b but also exhibited significant antibacterial and anti-oxidant properties. Therefore the versatile applicability of AgNPs could pave the way for the future applications of S.tibetica- mediated AgNPs for the prevention of microbial infection, mitigation of free radicals, and degradation of harmful chemicals like rhodamine-b that lays the pioneer steps towards control and curbing the menace of environmental pollution, especially water pollution. Declarations Funding Declaration: No funding Ethical Approval: Not applicable Informed consent: None Consent for publication: Not applicable Competing interests: The authors declare no competing interests Author Contribution Doctor syed wajaht amin shah conceptualized, drafted, and supervised the work . Mr Naseer Ahmad Dar carried out synthesis and characterization of the synthesized nanoparticles . Dr. Parvaiz wrote and thoroughly checked the manuscript. Dr Mahpara and Nighat nazir carried out biological properties and communicated the manuscript. Acknowledgement We thank Department of Chemistry, university of Kashmir, and NIT Srinagar for providing the necessary facilities to carry out biological activity and characterization during the work. Data Availability: All data and materials are present in the manuscript. References S.T. Fardood, A. Ramazani, S. 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Sarkar, Degradation of toxic textile dyes and detection of hazardous Hg2 + by low-cost bioengineered copper nanoparticles synthesized using Impatiens balsamina leaf extract, Materials Research Bulletin, 94 (2017) 257–262. Scheme 1 Scheme 1 is available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files GAagnp.tif ESI.docx Scheme1.png Scheme 1.Green synthesis of Silver nanoparticle from Aqueous extract of S. Tibetica. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5292367","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":375876540,"identity":"493f8e54-289e-451d-98aa-d334d1fa1e81","order_by":0,"name":"Naseer Ahmad Dar","email":"","orcid":"","institution":"University of Kashmir","correspondingAuthor":false,"prefix":"","firstName":"Naseer","middleName":"Ahmad","lastName":"Dar","suffix":""},{"id":375876541,"identity":"d8d48593-e836-4e65-bf38-3f40f0391b09","order_by":1,"name":"Parvaiz A Dar","email":"","orcid":"","institution":"University of Kashmir","correspondingAuthor":false,"prefix":"","firstName":"Parvaiz","middleName":"A","lastName":"Dar","suffix":""},{"id":375876542,"identity":"bc308a31-411c-44bf-a17e-306823120690","order_by":2,"name":"Mahpara Qadir","email":"","orcid":"","institution":"University of Kashmir","correspondingAuthor":false,"prefix":"","firstName":"Mahpara","middleName":"","lastName":"Qadir","suffix":""},{"id":375876543,"identity":"f94e7627-f909-4720-8657-fc5828d4994b","order_by":3,"name":"Wajaht Amin Shah","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA9ElEQVRIiWNgGAWjYDCCA1CasYEHSFYAMTNzAyEtjA1wLQfOgLQwEqmFgQGo5WAbRDdeHXzHm58/+LjnsDxz+9mDnz/Oq43mbwdq+VGxDacWyTPHDBtnPDts2NiTlyxxcNvx3BmHGRsYe87cxqnF4EYOYzPPgduMjQ05BkAtx3IbgFqYGdsIa7Fv7H9j/OPgnGO584nVktg4I8dM4mBDTe4GQlpAfpk548D/5MYZb8wszhw7kLsRqOUgPr8AQ+zBhw8H0mw39ucY36ioqcudd/7wwQc/KnBrgQPDBjB1GEweIKweCOQhVB1RikfBKBgFo2BkAQA0YWq/k8mD+gAAAABJRU5ErkJggg==","orcid":"","institution":"University of Kashmir","correspondingAuthor":true,"prefix":"","firstName":"Wajaht","middleName":"Amin","lastName":"Shah","suffix":""}],"badges":[],"createdAt":"2024-10-19 04:08:12","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5292367/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5292367/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":70028833,"identity":"17e23a5a-4d02-4478-8387-c953d6d14e40","added_by":"auto","created_at":"2024-11-27 15:57:26","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":93879,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a)\u003c/strong\u003eFTIR spectra of plant extract and silver nanoparticle \u003cstrong\u003e(b)\u003c/strong\u003e XRD pattern of AgNP\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5292367/v1/0a4b4da806540ea8a282509b.png"},{"id":70027157,"identity":"8f628540-2f06-4395-ab28-a89a28cf49c6","added_by":"auto","created_at":"2024-11-27 15:41:26","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1088627,"visible":true,"origin":"","legend":"\u003cp\u003e(\u003cstrong\u003ea \u0026amp;b\u003c/strong\u003e) SEM images of Ag nanoparticles at 100nm and 200nm. (\u003cstrong\u003eC\u003c/strong\u003e) EDX of the nanoparticle. \u003cstrong\u003e(d and e)\u003c/strong\u003e Elemental mapping images of carbon and silver respectively.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5292367/v1/31884d436c3814c0dda589b6.png"},{"id":70027617,"identity":"31ffa1e9-86ff-435c-b8ae-f9fe638c8961","added_by":"auto","created_at":"2024-11-27 15:49:26","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":43491,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a)\u003c/strong\u003e Average particle size as obtained from DLS measurements. \u003cstrong\u003e(b)\u003c/strong\u003e Percentage weight composition of different elements.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5292367/v1/ad31ecd6c872f22791aad582.png"},{"id":70027160,"identity":"134ef2c6-a5f1-47ce-9cdc-54a779b0476c","added_by":"auto","created_at":"2024-11-27 15:41:26","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":605723,"visible":true,"origin":"","legend":"\u003cp\u003eAntibacterial bacterial activity against four bacterial strains (a) \u003cem\u003eBacillus cereus\u003c/em\u003e (b\u003cem\u003e) Pseudomonas\u003c/em\u003e (c) \u003cem\u003eE. coli\u003c/em\u003e (d) \u003cem\u003eS. aureus\u003c/em\u003e (e) Bar graph plot of zone of inhibition of four bacterial strains\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5292367/v1/c6591992dcfd5377e9cccfd8.png"},{"id":70027618,"identity":"5c250a77-7bf1-493a-a856-79a3000b3ca5","added_by":"auto","created_at":"2024-11-27 15:49:26","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":65799,"visible":true,"origin":"","legend":"\u003cp\u003eThe gradual decrease in intensity of absorption peak at 550nm with irradiation time. \u003cstrong\u003e(b)\u003c/strong\u003e The plot of -lnC/C\u003csub\u003eο\u003c/sub\u003e versus time and value of rate constant is found to be 0.06667min\u003csup\u003e-1\u003c/sup\u003e \u003cstrong\u003e(c)\u003c/strong\u003e Gradual decrease in the concentration of Rhodamine-b with time.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-5292367/v1/cf75c0a13ce48767dbe8f75d.png"},{"id":70027158,"identity":"c8810364-f354-4908-9b8e-311e27a1a12c","added_by":"auto","created_at":"2024-11-27 15:41:26","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":116830,"visible":true,"origin":"","legend":"\u003cp\u003ePercentage radical scavenging activity of ascorbic acid, plant extract and AgNP.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-5292367/v1/c3b186aa44076378658076f2.png"},{"id":70570830,"identity":"f9f8ff20-539f-4ca8-a411-be3322d4c1ce","added_by":"auto","created_at":"2024-12-04 13:40:24","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2504324,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5292367/v1/a1889a2e-f511-488f-9b9e-cb440811693e.pdf"},{"id":70027164,"identity":"2b2c436a-b6e3-43f2-808d-efe6c56bcf9f","added_by":"auto","created_at":"2024-11-27 15:41:26","extension":"tif","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":487380,"visible":true,"origin":"","legend":"","description":"","filename":"GAagnp.tif","url":"https://assets-eu.researchsquare.com/files/rs-5292367/v1/8013c76573bdf8acef0ea45f.tif"},{"id":70027166,"identity":"84325679-07b7-4df6-bd78-fe038559f350","added_by":"auto","created_at":"2024-11-27 15:41:26","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":4876199,"visible":true,"origin":"","legend":"","description":"","filename":"ESI.docx","url":"https://assets-eu.researchsquare.com/files/rs-5292367/v1/149e39892c78e6c0fd3f7c6e.docx"},{"id":70027162,"identity":"711c9e46-914d-4b50-bd0d-50c876e5978c","added_by":"auto","created_at":"2024-11-27 15:41:26","extension":"png","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":189744,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eScheme 1.\u003c/strong\u003eGreen synthesis of Silver nanoparticle from Aqueous extract of \u003cem\u003eS. Tibetica.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"Scheme1.png","url":"https://assets-eu.researchsquare.com/files/rs-5292367/v1/522ea11ea2d8fc35872278ff.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Stachys tibetica derived silver nanoparticles: A robust multifunctional material for enhanced biological activity and photocatalytic properties ","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eNanotechnology an emerging multidisciplinary field having roots in basic sciences such as physics, chemistry and biology uses science to manipulate matter at molecular level and create nanoparticles of varying sizes, shapes and chemical compositions for their potential applications to human benefit [\u003cspan additionalcitationids=\"CR2 CR3\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Physical, chemical and environmentally friendly processes are used to create nanoparticles[\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].In recent years, researchers have focused on developing efficient green chemistry procedures that use natural reducing capping and stabilizing chemicals to make metal nanoparticles with desired morphology and size. Scientists have been successful in increasing the efficacy of these materials by reducing particle size, because of their sub-microscopic size they have unique material characteristics and so synthesized nanoparticles are used in a various areas, such as engineering, medicine, environmental remediation and catalysis[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Environmental and toxicity concerns involved in some of synthetic methodologies involving the use of organic solvents, toxic reducing substances used for reduction of metals, etc have led for the demand of environmentally friendly, clean, reliable and biologically appropriate approaches. Nanoparticle synthesis from plant extracts and different types of plant biomass not only uses time conserving and environmentally friendly procedures but also these nanoparticles involve very simple and easy handling procedures[\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eStachys\u003c/em\u003e is genus of 350\u0026ndash;400 species of plants belonging to the family Lamiaceae, also called as mint family. Plants belonging to this family are found worldwide. Most members of this family are either perennial or annual herbs although some species of this family are shrubs or subshrubs[\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Extensive phytochemical and pharmacological studies have been carried out on \u003cem\u003eStachys\u003c/em\u003e species, proving their ethnopharmacological benefits. Anti-inflammatory, antioxidant, analgesic, Reno protective, anxiolytic, and depressive activities are some of the prominent biological activities carried out on these species[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e] [\u003cspan additionalcitationids=\"CR12\" citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. The phytochemical content of these species is linked to a wide spectrum of medicinal benefits. As a result, the genus \u003cem\u003eStachys\u003c/em\u003e has garnered a lot of interest for the isolation and bioprospection of bioactive secondary metabolites of plant. In total, more than 200 compounds from this genus have been isolated, including terpenes (e.g., iridoids triterpenes, diterpenes,), polyphenols (e.g., lignans, flavone derivatives, phenylethanoid glycosides,), and phenolic acids etc [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan additionalcitationids=\"CR15 CR16 CR17\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eVery efficient metallic nanoparticles like Iron, nickel, Copper, and Silver have been synthesized by researchers from the plants of the lamiaceae family eg. nanoparticles synthesized from \u003cem\u003eMentha Pulegium\u003c/em\u003e, exhibited potential antibacterial, cytotoxic, antioxidant and antifungal activities[\u003cspan additionalcitationids=\"CR20\" citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Besides being used as antibacterial agents nanoparticles also play the role of a catalyst for the degradation of various environmentally hazardous chemicals. \u003cem\u003eSaliva officinalis\u003c/em\u003e mediated silver nanoparticles degrade Congo red dye very efficiently[\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. similarly, AgNPs from \u003cem\u003eendostemon viscosus\u003c/em\u003e degrade methylene blue and crystal violet dyes[\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eKeeping these findings in view, the aim of our study is the green synthesis of silver nanoparticles from \u003cem\u003eStachys tibetica\u003c/em\u003e (locally known as yaktse), which is not reported in the literature yet. we were further motivated to carry out antibacterial, anti-oxidant and dye degradation properties of these silver nanoparticles. The sample plant for this study was collected from Minjhi, Ladakh, and Silver nanoparticles were synthesized from the extract of this plant using a 20 mM AgNO\u003csub\u003e3\u003c/sub\u003e solution. The bactericidal effect of these nanoparticles was checked against four bacterial strains namely \u003cem\u003eBacillus cereus, E.Coli, S.Aeureus\u003c/em\u003e, and \u003cem\u003ePseudomonas.\u003c/em\u003e The synthesized nanoparticles were further utilized as a catalyst in the degradation of environmentally hazardous dye Rhodamine-B with the use of a UV-Vis spectrophotometer. The absorbance spectrum of the solution was examined at various wavelengths. The absorbance value at 550-560nm depicted dye concentration throughout degradation.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":" \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. \u003cem\u003eMaterials\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eThe ATR-FTIR Spectrophotometer (Agilent Technologies, USA) was used to study the structural properties of dried plant extract and silver nanoparticles at a constant temperature of 25\u0026deg;C. The wavenumber range was 500\u0026ndash;4000 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. FE-SEM was used to analyze the surface morphology of nanoparticles (FEI Quanta 200, USA) operated at 200V. XRD (Ultima-IV (Rigaku Corporation, Tokyo, Japan) uses Cu Kα radiation. The samples were scanned in a 2θ range of 10\u0026thinsp;\u0026minus;\u0026thinsp;80\u0026deg; at a scan rate of 5 min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e.\u003c/p\u003e \u003cdiv id=\"Sec4\" class=\"Section3\"\u003e \u003ch2\u003e2.1.1. \u003cem\u003eCollection of plant material\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eThe aerial parts of \u003cem\u003eStachys tibetica\u003c/em\u003e were collected from Minjhi region of Ladakh, india. The proper identification of material was done by curator Akhter Ahmed Malik of the Department of Taxonomy, Kashmir University. The voucher no [9062-(KASH)] has been deposited at the herbarium, Department of Botany, Kashmir University, Srinagar.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section3\"\u003e \u003ch2\u003e2.1.2. \u003cem\u003eExtract preparation\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eThe collected material was shade-dried for ten days. 30 grams of ground plant material was soaked in distilled water and kept on standing for 5 days. The final solution was filtered through Whatman filter paper No.1 to obtain a uniform yellowish aqua extract of \u003cem\u003eStachys tibetica\u003c/em\u003e for further experimentation.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Methods\u003c/h2\u003e \u003cdiv id=\"Sec7\" class=\"Section3\"\u003e \u003ch2\u003e2.2.1. \u003cem\u003ePreparation of silver Nanoparticle\u003c/em\u003e\u003c/h2\u003e \u003cp\u003e \u003cem\u003eStachtys tibetica\u003c/em\u003e extract-based nanoparticles were synthesized as per the green synthetic methodology reported for the synthesis of nanoparticles[\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. The aqueous extract of \u003cem\u003eStachys tibetica\u003c/em\u003e was dried on a water bath till the solution became deeply yellow. 20 mmol solution of AgNO\u003csub\u003e3\u003c/sub\u003e was filled in the burette and added to water-dissolved aqueous extract of \u003cem\u003eStachys tibetica\u003c/em\u003e dropwise the solution was continuously stirred on hotplate set at a temperature of 50 \u003csup\u003e\u0026ordm;\u003c/sup\u003eC. Silver nitrate solution was continuously added till the colour of solution changes from yellowish to deep brown which marked the end point of reaction. The final solution was centrifuged at 8000 rpm for 15 minutes. The final nanoparticle was obtained by discarding the supernant. The silver nanoparticle obtained was washed with distilled water 3\u0026ndash;4 times to obtain a pure mass of nano scale particles which was further dried in oven at 50\u003csup\u003e\u0026ordm;\u003c/sup\u003eC to obtain a powdery mass of nanoparticle.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e2.2.2. \u003cem\u003eAntibacterial Activity\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eA panel of four bacterial strains namely \u003cem\u003eStaphylococcus aureus, Bacillus cereus, Pseudomonas\u003c/em\u003e and \u003cem\u003eE.coli\u003c/em\u003e, obtained from Microbial type culture collection (MTCC), Institute of Microbial Technology, Chandigarh, India. These bacterial strains were cultivated on nutrient agar plates and kept on agar slants at 37\u0026ordm;C. Microorganism cell suspension in 0.9% NaCl was adjusted to 0.5 Mc Farland to obtain approximately 106 cfu/ml.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003e2.2.3. \u003cem\u003eAntioxidant properties\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eThe free radical scavenging potential of AgNP, plant extract, and standard ascorbic acid was checked against DPPH(2,2-Diphenyl-1-Picrylhydrazyl) using a modified approach proposed by W brand Williams et al[\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. In different test tubes solutions with concentrations (20,40,60,80,100,120,140,160,180,200 \u0026micro;g/ml) of AgNP, plant extract, and ascorbic acid were prepared). To each solution, DPPH(1mM) prepared in methanol was added and mixed thoroughly. The solution obtained after the addition of DPPH was incubated in the dark for 30 minutes at room temperature before being measured for UV absorbance at 517nm with a UV-Vis spectrophotometer (perkin-Elmer Lambda 950:UK). The DPPH solution obtained without the addition of a sample was used as a control to measure radical scavenging activity. The Free radical scavenging activity of all samples was calculated using the formula;\u003c/p\u003e \u003cp\u003eRadical scavenging activity = \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{{AB}_{C-{AB}_{S}}}{{AB}_{C}}\\times\\:100\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003cp\u003eWhere AB\u003csub\u003eC\u003c/sub\u003e is the absorbance of methanol\u0026thinsp;+\u0026thinsp;DPPH, and AB\u003csub\u003eS\u003c/sub\u003e is the absorbance of sample solution\u0026thinsp;+\u0026thinsp;DPPH.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003e2.2.4. \u003cem\u003eDegradation of rhodamine-B dye\u003c/em\u003e\u003c/h2\u003e \u003cp\u003e10mg of Rhodamine-b dye were typically added to 1000 mL of double distilled water as a stock solution. A control of dye and NaBH\u003csub\u003e4\u003c/sub\u003e(0.06M) without the inclusion of silver nanoparticles was also kept. 5ml freshly prepared NaBH\u003csub\u003e4\u003c/sub\u003e solution was added to 20ml of rhodamine B solution and subsequently 5ml of nano catalyst of a definite concentration was added to it. Prior to irradiation, the reaction suspension was thoroughly mixed to ensure that the working solution was clearly balanced. Following that, the dispersion was exposed to visible light and continuously monitored. Aliquots of 2\u0026ndash;3 mL solution were filtered and utilized to analyze the photocatalytic degradation of dye at particular time intervals. The absorbance spectrum of the solution was examined with a UV-Vis spectrophotometer at various wavelengths. The absorbance value at 550-560nm depicted dye concentration throughout degradation.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"3. Results and Discussion","content":"\u003cp\u003eNanoparticles serve an effective scientific and research tools having an excellent catalytic efficiency due to their large surface area. Silver nanoparticles synthesized through green approach act as functional materials that have been explored for curbing of various modern biological, industrial and environmental challenges. These nanoparticles synthesized from the aqueous extract of \u003cem\u003eStachys tibetica\u003c/em\u003e were characterised using various techniques. Initial screening was carried out through FT-IR analysis, that revealed the presence of various functional groups. P-XRD revealed the crystallinity, and the morphology was screened using SEM analysis. The synthesised nanoparticles were explored in various directions ranging from bioactivity, antioxidant mitigation to water pollution remediation. The extensive discussions are addressed in various sections\u003c/p\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n\u003ch2\u003e3.1. FTIR Analysis of Synthesized Nanoparticle\u003c/h2\u003e\n\u003cp\u003eThe FTIR (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003ea) spectra of produced AgNPs were examined to determine the existence of potential functional groups inside biomolecules that bind to the surface of silver for bioreduction as shown in the Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e(a). IR spectrum of impregnated silver nanoparticle shows absorption bands at 2975cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e corresponding to N-H Stretching, 2150cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e corresponding to C-N double bond stretching, 1609cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e corresponding to N-H bending, 1380cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e corresponding to N-O stretching whereas plant extract absorbs at 2945cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e,2041cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e,1609cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e,1392cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n\u003ch2\u003e3.2. XRD Analysis\u003c/h2\u003e\n\u003cp\u003eThe XRD pattern (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eb) of silver nanoparticles shows strong peaks at 2Ɵ = 33.21\u0026ordm;, 38.47\u0026ordm;, 47.37\u0026ordm; and 77\u0026ordm; which correspond to the (1 1 1), (2 0 0), (2 2 0), (3 1 1), Bragg\u0026rsquo;s reflections of the face-centered cubic structure of silver. In addition, unassigned peaks that were weaker than those of silver could be related to the bioorganic chemicals forming on the surface of the AgNPs[\u003cspan class=\"CitationRef\"\u003e26\u003c/span\u003e].\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n\u003ch2\u003e3.3. SEM And EDS Analysis of synthesized nanoparticle\u003c/h2\u003e\n\u003cp\u003eField-Emission Scanning electron Microscope (FE-SEM) was used to analyze the surface topography and morphology of \u003cem\u003eS.Tibetica\u003c/em\u003e based AgNPs. It is apparent from the SEM monograps that the material has developed into regular nanospheres with a rugged surface which increases the surface area of the catalyst Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e(a and b). The existence of silver metal in the produced nanoparticles was confirmed using energy dispersive X-ray spectroscopy (EDS). As seen in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e(C). A sharp peak at 3 KeV established the existence of elemental silver in the nanoparticles. The EDS examination of produced AgNPs revealed indications of elements like Carbon(C), Nitrogen(N), and Oxygen(O), etc besides metallic silver because the substance generated via biological method had a minor fraction of Carbon, Nitrogen and Oxygen, etc [\u003cspan class=\"CitationRef\"\u003e27\u003c/span\u003e]. The same is also revealed from the elemental mapping images of nanoparticles. (S-1)\u003c/p\u003e\n\u003cp\u003eThe average particle size as measured from DLS measurements was found out to be 78nm 3(a). The composition of different elements present in nanoparticle by weight percentages is depicted in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e(b).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n\u003ch2\u003e3.4. Antibacterial properties\u003c/h2\u003e\n\u003cp\u003eThe MIC value of biosynthesized AgNPs to inhibit bacterial growth was determined by testing zone formation with dilutions ranging from 5 to 40\u0026micro;g/ml. The synthesized AgNP has a MIC of 10\u0026micro;g/ml and 20\u0026micro;g/ml, for \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eC. aureus\u003c/em\u003e respectively (S-4). The antibacterial activity of AgNPs, \u003cem\u003eStachys tibetica\u003c/em\u003e plant extract, and Streptomycin was tested at a level of 10\u0026micro;g/ml, as both pathogens demonstrated sensitivity at this concentration. The freshly prepared samples were tested for antibacterial activity against two gram-positive and two gram-negative strains namely \u003cem\u003eBacillus cereus, Pseudomonas, Escherichia coli\u003c/em\u003e, and \u003cem\u003eStaphylococcus aureus.\u003c/em\u003e AgNPs (10\u0026micro;g/ml) have shown highest zone of inhibition of about 12.1mm for \u003cem\u003eBacillus Cereus\u003c/em\u003e followed by \u003cem\u003ePseudomonas\u003c/em\u003e (11.8 mm), \u003cem\u003eE. coli\u003c/em\u003e (9.8) and \u003cem\u003eStaphylococcus aureus\u003c/em\u003e (7.2) (S-5). The zone of inhibition observed for plant extract was less as compared to zone of inhibition obtained for AgNPs as shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e.\u003c/p\u003e\n\u003cp\u003eThis study developed the synergistic antibiotic activity of AgNPs, which is greater than the individual antibacterial activity of plant extract when used together[\u003cspan class=\"CitationRef\"\u003e28\u003c/span\u003e]. AgNP behaves as a positively charged center that interacts with the negatively charged centers of bacterial cells [\u003cspan class=\"CitationRef\"\u003e29\u003c/span\u003e]. Recent research has found that the methods of action of AgNP against microbes are dependent on the NPs' size, shape, stability, and affinity[\u003cspan class=\"CitationRef\"\u003e30\u003c/span\u003e]. Literature has cited that the bactericidal effect of AgNPs is due to the interaction of silver with nucleosides of nucleic acids and the creation of subsequent silver compounds with bactericidal potential[\u003cspan class=\"CitationRef\"\u003e31\u003c/span\u003e]. The organic layer-coated silver nanoparticle has shown the highest bactericidal effect towards \u003cem\u003eBacillus cereus\u003c/em\u003e (12.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1) which can be inferred from the relatively simple cell wall structure of G(\u003cem\u003e+\u003c/em\u003e) bacterial species and the presence of a thick PG layer of 30 nm[\u003cspan class=\"CitationRef\"\u003e32\u003c/span\u003e]. The reason of bactericidal behaviour of AgNPs towards Gram negative G(-) can be inferred from the fact that G(\u003cem\u003e-\u003c/em\u003e) bacteria have a small PG layer of 1\u0026ndash;5 nm between the exterior layer and the cytoplasmic film that makes these species prone to bactericidal effects[\u003cspan class=\"CitationRef\"\u003e33\u003c/span\u003e].\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n\u003ch2\u003e3.5. Antioxidant properties\u003c/h2\u003e\n\u003cp\u003eDPPH has hydrogen acceptor capability to antioxidants[\u003cspan class=\"CitationRef\"\u003e34\u003c/span\u003e].DPPH can easily accept electron from antioxidant compounds and during this process the color of DPPH changes from violet to yellow. Thus, DPPH is a free radical that accepts a free electron from antioxidants and becomes a stable diamagnetic molecule. The DPPH radical scavenging tends to increase with increasing the concentration of AgNPs, showing a maximum at higher concentrations. At concentrations 20\u0026ndash;200\u0026micro;g/mL, AgNPs showed 32.4\u0026ndash;75.8%, Plant extract showed 8\u0026ndash;62%, and standard ascorbic acid showed 34.5\u0026ndash; 89.6% radical scavenging activity. the radical scavenging activity of plant extract and AgNp was lower than that of standard Ascorbic acid. The results obtained for the antioxidant activity of AgNP, plant extract, and standard ascorbic acid at different concentrations are as shown below.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n\u003ch2\u003e3.6 Dye Degradation\u003c/h2\u003e\n\u003cp\u003eDye degradation studies were performed using AgNPs as catalyst. The following formula was used to calculate the percentage of dye degradation:\u003c/p\u003e\n\u003cp\u003e%Degradation\u0026thinsp;=\u0026thinsp;\u003cstrong\u003e100\u0026times; (\u003c/strong\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\varvec{C}}_{\\varvec{o}}\\)\u003c/span\u003e\u003c/span\u003e\u003cstrong\u003e- C) /\u003c/strong\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\varvec{C}}_{\\varvec{o}}\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003ewhere \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{C}_{o}\\)\u003c/span\u003e\u003c/span\u003eis the initial concentration of dye solution and C is the concentration of dye solution following photocatalytic degradation.The PFO model was found to be the optimal choice in determining the rate constant and rate constant was found out to be 0.06667 min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e using the following rate equation:\u003c/p\u003e\n\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\n\u003cdiv id=\"FileID_Equa\" class=\"mathdisplay\"\u003e$$\\:\\varvec{l}\\varvec{n}\\frac{{\\varvec{C}}_{\\varvec{t}}}{{\\varvec{C}}_{\\varvec{o}}}=\\:-\\varvec{K}\\varvec{t}$$\u003c/div\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe catalytic efficiency of AgNPs was accessed from \u0026lambda;\u003csub\u003emax\u003c/sub\u003e peak at 550-560nm at different intervals of irradiation. In the presence of AgNPs, the primary absorption peak of dye at 550nm decreased slowly with increasing time of exposure of dye to visible light in the presence of AgNp. This indicated the photocatalytic degradation of Rhodamine-b dye in the presence of AgNPs (Fig.\u0026nbsp;5). In the Dye degradation reaction, \u003cstrong\u003eNaBH\u003c/strong\u003e\u003csub\u003e\u003cstrong\u003e4\u003c/strong\u003e\u003c/sub\u003e helps in the reduction of dye, the Ag nanoparticles act as an electron relay from \u003cstrong\u003eBH\u003c/strong\u003e\u003csub\u003e\u003cstrong\u003e4\u003c/strong\u003e\u003c/sub\u003e\u003csup\u003e\u003cstrong\u003e\u0026minus;\u003c/strong\u003e\u003c/sup\u003e(donor) to dye(acceptor). The plausible mechanism of dye degradation is believed to proceed with the decomposition of \u003cstrong\u003eNaBH\u003c/strong\u003e\u003csub\u003e\u003cstrong\u003e4\u003c/strong\u003e\u003c/sub\u003e into \u003cstrong\u003eBH\u003c/strong\u003e\u003csub\u003e\u003cstrong\u003e4\u003c/strong\u003e\u003c/sub\u003e\u003csup\u003e\u003cstrong\u003e\u0026minus;\u003c/strong\u003e\u003c/sup\u003e (nucleophilic) and subsequently produces \u003cstrong\u003eH-Ag\u003c/strong\u003e and \u003cstrong\u003eAg-BH\u003c/strong\u003e\u003csub\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/sub\u003e as reaction intermediates in presence of AgNP\u003csub\u003eS\u003c/sub\u003e. The intermediate \u003cstrong\u003eAg-H\u003c/strong\u003e is responsible for the degradation of dye(electrophilic) into harmless degradation products[\u003cspan class=\"CitationRef\"\u003e35\u003c/span\u003e].\u003c/p\u003e\n\u003cp\u003eStudies on photocatalytic studies demonstrated that the photocatalytic activity of metallic nanoparticles is dependent on their crystallographic structure, shape, and size[\u003cspan class=\"CitationRef\"\u003e36\u003c/span\u003e]. J Kadam et al. employed AgNPs produced from Cauliflower waste to photodegrade methylene blue in 152 minutes [\u003cspan class=\"CitationRef\"\u003e37\u003c/span\u003e] [\u003cspan class=\"CitationRef\"\u003e38\u003c/span\u003e], whereas the current investigation exhibited 100% Rh-B degradation in just 1 hour As a result, this study demonstrated the efficient photodegradation capacity of AgNPs generated using \u003cem\u003eS. Tibetica\u003c/em\u003e in visible light.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4. Conclusion","content":"\u003cp\u003eThe green synthesis of silver nanoparticles from \u003cem\u003eStachys tibetica\u003c/em\u003e extract has been carried out in the present study, whose characterization revealed the face-centred cubic structure. The synthesized nanoparticles not only act as good catalyst for the degradation of rhodamine-b but also exhibited significant antibacterial and anti-oxidant properties. Therefore the versatile applicability of AgNPs could pave the way for the future applications of \u003cem\u003eS.tibetica-\u003c/em\u003emediated AgNPs for the prevention of microbial infection, mitigation of free radicals, and degradation of harmful chemicals like rhodamine-b that lays the pioneer steps towards control and curbing the menace of environmental pollution, especially water pollution.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding Declaration:\u003c/strong\u003e No funding\u003c/p\u003e\n\u003cp\u003eEthical Approval: Not applicable\u003c/p\u003e\n\u003cp\u003eInformed consent: None\u003c/p\u003e\n\u003cp\u003eConsent for publication: Not applicable\u003c/p\u003e\n\u003cp\u003eCompeting interests: The authors declare no competing interests\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eDoctor syed wajaht amin shah conceptualized, drafted, and supervised the work . Mr Naseer Ahmad Dar carried out synthesis and characterization of the synthesized nanoparticles . Dr. Parvaiz wrote and thoroughly checked the manuscript. Dr Mahpara and Nighat nazir carried out biological properties and communicated the manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eWe thank Department of Chemistry, university of Kashmir, and NIT Srinagar for providing the necessary facilities to carry out biological activity and characterization during the work.\u003c/p\u003e\u003ch2\u003eData Availability:\u003c/h2\u003e \u003cp\u003eAll data and materials are present in the manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e \u003cli\u003e\u003cspan\u003eS.T. Fardood, A. Ramazani, S. 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Sarkar, Degradation of toxic textile dyes and detection of hazardous Hg2\u0026thinsp;+\u0026thinsp;by low-cost bioengineered copper nanoparticles synthesized using Impatiens balsamina leaf extract, Materials Research Bulletin, 94 (2017) 257\u0026ndash;262.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Scheme 1","content":"\u003cp\u003eScheme 1 is available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"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":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Stachys tibetica, silver nanoparticles, FE-SEM, XRD, Rhodamine-b","lastPublishedDoi":"10.21203/rs.3.rs-5292367/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5292367/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e \u003cem\u003eStachys tibetica\u003c/em\u003e, a significant medicinal plant was collected from Ladakh (India) and subjected to extraction. The extract was titrated with AgNO\u003csub\u003e3\u003c/sub\u003e at 50\u0026ordm;C to obtain silver nanoparticles with plant extract acts as a reducing and capping agent. The eco-friendly synthetic procedure utilizes the phytochemicals in \u003cem\u003eStachys tibetica\u003c/em\u003e, ensuring a sustainable and non-toxic approach to nanoparticle production. The characterization of the silver nanoparticles was conducted using various techniques, including Fourier-transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), and dynamic light scattering (DLS). Field emission scanning electron microscopy (FE-SEM) was utilized to examine the surface morphology, revealing the irregular and rugged spherical shape of the synthesized nanoparticles. \u003cem\u003eStachys tibetica\u003c/em\u003e-derived silver nanoparticles (AgNPs) act as a versatile and robust multifunctional material with proficient bioactivity and catalytic properties. The synthesized silver nanoparticles displayed effective antioxidant and antimicrobial properties against \u003cem\u003eBacillus Cereus, Pseudomonas, Escherichia coli, and S.Aeureus\u003c/em\u003e. Additionally, the green nanoparticles degraded Rhodamine-B under sunlight irradiation within 1 hour at a rate constant of 0.066 min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e.\u003c/p\u003e","manuscriptTitle":"Stachys tibetica derived silver nanoparticles: A robust multifunctional material for enhanced biological activity and photocatalytic properties ","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-27 15:41:21","doi":"10.21203/rs.3.rs-5292367/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"ff42a2e7-59ae-4b17-930a-7191f89f50eb","owner":[],"postedDate":"November 27th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-12-04T13:40:01+00:00","versionOfRecord":[],"versionCreatedAt":"2024-11-27 15:41:21","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5292367","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5292367","identity":"rs-5292367","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}