Microwave assisted synthesis of Silver Nanoparticles using Cocculus hirsutus leaves Extract and their Anti-Urolithic activity

Microwave assisted synthesis of Silver Nanoparticles using Cocculus hirsutus leaves Extract and their Anti-Urolithic activity | 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 Microwave assisted synthesis of Silver Nanoparticles using Cocculus hirsutus leaves Extract and their Anti-Urolithic activity Muthaiah Chintha, Balaswamy Puligilla, Bala Narsimha Dhoddi This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4406241/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 3 You are reading this latest preprint version Abstract As highlighting the synergetic action of Nanoscale metal along with the biomolecules play crucial role in modern medicine. The silver nanoparticles were owed many uses in the field of medicine; one of the most important approaches of silver nitrate is Anti-urolithiasis effect. The present study focuses on the anti-urolithiasis activity of Cocculus hirsutus leaf extract capped silver nanoparticles (AgNPs). The AgNPs were synthesized by silver nitrate salt solution and Cocculus hirsutus leaf extract in microwave oven. The Microwave assisted synthesis is an efficient technique and promises more environment benign than traditional heating. The newly prepared AgNPs were characterized by various techniques. the invitro struvite crystal preparation and inhibition activity were examined by using freshly prepared silver nanoparticles. AgNPs Cocculus hirsutus Microwave in vitro urolithiasis Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 1. Introduction Nanotechnology is an emerging field of research that focuses on the creation and applications of nano particles in various domains such as catalysis, electrochemistry, biomedicine, pharmaceuticals, sensors, food technology and cosmetics etc. [ 1 – 5 ]. It is a revolutionary field that has been integrated into various domains of our daily life, many products contain nanomaterials as well as applications-based nanoparticles have emerged [ 6 ]. Nanotechnology is a key enabling technology that can offer new and innovative medical solutions for unmet medical needs so far. Nanomedicine is the term that describes the use of nanomaterials for diagnosis, monitoring, control, prevention and treatment of disease [ 7 ]. Metallic nanoparticles can be synthesized using plant extracts which is a green and safe approach [ 8 ]. The silver Nano particles produced by this method have various properties such as high chemical stability, catalytic activity and many therapeutic activities [ 9 ]. Nanomaterials have different properties and interactions depending on their size. As particle size became smaller SA\V increases which can significantly enhance the reactivity. This means that nanomaterials have surface area per unit volume which makes them more reactive, soluble and bio available than their bulk counterparts [ 10 ]. Urolithiasis commonly known as kidney stones, is indeed a multifactorial disorder characterised by the formation of urinary calculi within the renal tubules [ 11 ]. the pathogenesis of these stones is complex, involving a combination of factors such as anatomical abnormalities, metabolic disorders, and dietary habits [ 12 ]. The increased concentration of unbound ions, particularly calcium and phosphate in the urine can lead to the precipitation of these salts, farming calculi [ 13 ]. The presence or absence of endogenous inhibitors like citrate and magnesium, which prevent the stone formation, also plays a crucial role [ 14 ]. The majority of urinary calculi are composed of calcium oxalate, which accounts for approximately 75% of cases followed by struvite, uric acid and cystine stones [ 15 ]. Risk factors for urolithiasis include low fluid intake certain urinary tract malformations, urinary tract infections and dietary facts such as high sodium intake and high dietary oxalate. Management of the condition often involves increasing fluid intake, dietary modifications, and in some cases medical or surgical intervention to remove the stones. Preventative strategies are tailored to the type of stones and underlying risk factors identified in each individual case [ 16 – 17 ]. Ammonium magnesium phosphate known as struvite is produced rapidly as a result of urinary tract infection caused by the urease producing bacteria [ 18 ]. these struvites are difficult to treat because of the complications associated urolithiatic microorganisms [ 19 ]. The struvite stones account for 17% of all urinary stones and are reported to be distinctive due to their rapid growth rate persistence and high degree of recurrence (50%) [ 20 ]. The worldwide increasing in the incidences of urolithiasis and many studies has also reported that calcium oxalate stones constitute about 79% of the kidney stone disease. [ 21 ]. pathogenesis of calcium oxalate stone formation involves a series of physicochemical events which include the nucleation of the crystals, the growth and aggregation of the crystals and finally the retaining of crystal aggregates on the epithelial cell linings of the Renal tubules [ 22 ]. Cocculus hirsutus also known as broom keeper is a versatile herbal medicine used in folk medicine for various conditions such as Tuberculosis, Leprosy, Skin diseases, Dyspepsia, Pruritus [ 23 ]. The phytochemical composition and potential bioactive compounds of Cocculus hirsutus plant leaves have been the subject of study. Tannins, alkaloids, flavonoids, steroids, and saponins were found to be present in the leaves of Cocculus hirsutus. Notably, the leaves were found to contain two major copmounds:5,7-dihydroxy-3-(4’-hydroxybenzyl) chromone and kaempferide-3-O-α-rhamnosyl-7-O-α-L-rhamnoside, which belong to the homoisoflavones and flavanol triglycerides, respectively [ 24 – 25 ]. Another research endeavour focused on the leaves of Cocculus hirsutus discovered that the acetone extract exhibited antimicrobial activity against bacteria and fungi. It was further determined that O-ethyl hydroxyl amine,2-ethyl heptanoic acid and 1-nonyl cycloheptane were responsible for this observed activity [ 26 ]. Furthermore, an investigation into the potential anti-COVID-19 activity of Cocculus hirsutus identified betulin, coclaurine, and quinic acid as compounds with notable binding affinity to the main proteases of SARS-CoV-2[ 27 ]. These findings indicate that Cocculus hirsutus leaves possess a diverse range of phytochemicals with potential bioactive and therapeutic properties. The sensible and sustainable chemistry is essential for our ever-changing environment. consequently, synthesis of chemical substances started to follow green chemistry principles in recent years, Microwave assisted synthesis of Nano particles is a rapid and efficient method for producing nanostructured materials [ 28 ]. This technique offers advantages such as rapid volumetric heating, higher reaction rates, shorter reaction times, and higher product yields compared to conventional heating methods [ 29 ]. It has been applied to the synthesis of various nanoparticles including silver nanoparticles, ceria-zirconia Nano particles and iron oxide nanoparticles [ 30 – 31 ]. The microwave heating process allows for the control of particle size and morphology, resulting in nanoparticles specific properties [ 32 ]. For instance, the microwave assisted synthesis of silver nanoparticles using mint leaf extract as a reducing and stabilizing agent showed a higher peak height and smaller particle size compared to conventional heating methods [ 33 ]. Recent studies have reported the green synthesis of AgNPs using the extracts of Lantana camara, Hibiscus rosa-sinensis, Azadirachta Indica and Geranium leaf [ 34 – 36 ]. The method involves the role of plant secondary metabolites as surface active molecules which are potent reducing and capping agents involved in the formation of stable nanoparticles. The use of medicinal plants in the treatment of diseased condition has been preferred as it is considered to be safe, with minimal or less side effects. The literature attracted us to use silver nanoparticles and Cocculus hirsutus leaf extract combination for treatment of anti-struvite. 2. Research methodology 2.1. Synthesis of AgNPs: Cocculus hirsutus mediated synthesis of AgNPs were synthesized by optimizing the concentration of plant material required for the formation of a stable Nanomaterial. The powdered leaf extract(0.5gr) was added to 50 ml double distilled water and kept for boiling in a house hold conventional microwave oven for 10 min to obtain the extract.it is the plant extraction that was used for the preparation of the AgNPs. The formation of AgNPs was attained by incubation of 0.01N AgNO 3 and the plant extract mixture in a ratio of 1:5 at 32 O c for 5minuts in a microwave oven. The formation of the AgNPs was confirmed from the observed colour change of solution from colourless to brown colour. 2.2 Synthesis of struvite : The hydrogel medium needed for crystal growth was attained by employing a solution containing 1M Sodium Meta Silicate (NaSiO 3 .9H 2 O) and 0.5M Ammonium dihydrogen phosphate (NH 4 H 2 PO 4 .2H 2 O) as well as a solution composed of 1M Magnesium acetate (Mg (CH 3 COO) 2 .4H 2 O) [ 37 – 38 ]. These solutions were introduced into a different test tubes and subsequently subjected to an aseptic environment at a temperature of 37 0 C until the reaction reached its culmination. Following the 24-hour duration the struvite composition of the crystals that had formed was examined. 3. Results 3.1 Characterization of the AgNPs: Initially synthesized silver nanoparticles were detected by visible colour changing from colourless to dark brown colour as well as UV-Vis Spectral data revealed the absorption peak at 320nm confirming the presence of silver nanoparticles. This was due to the green reduction of Ag + ion to Ag 0 by the phytochemicals present in the Cocculus hirsutus leaf extract. silver nanoparticles were prepared by using diverse (0.5N,1N,2N,3N &5N) concentrations of silver nitrate for optimization it was shown in the Fig. 1 & 2 . 3.2.XRD Analysis: Newly crystallized silver nanoparticle was confirmed by Bruker 5000 the Bxrd Pattern analysis given the plane indices of 111,220,311 as metallic silver XRD values are coincided. The excellent XRD spectral values promising the formation of silver nanoparticles in different concentrations of starting materials were taken shown in Fig. 3 3.3.SEM & EDX Spectroscopy We used JOEL scanning electron microscopy to analyse the shape and size of the silver nanoparticles we synthesized. The SEM images showed that the AgNPs had smooth surface and a spherical shape, with an average diameter ranging from 17 to 27nm (Fig. 6 . & Fig. 7 .). This method is widely used to study the surface characteristics of nanoparticles, such as their size, shape, morphology and distribution. Johnson and Prabhu (2015) reported similar results when they used camelina Bengalese’s to produce spherical and uniform Ag nanoparticles and examined them with SEM. This technique relies on the interaction of electrons with the sample surface, which generates secondary electrons, the signal from these electrons is then used to create an image of the sample surface morphology. The elemental composition of the AgNPs was determined by EDX analysis, which measures the energy and intensity distribution of X-ray signals generated by an electron beam on a specimen. Figure 5 . shows the EDX spectra of the AgNPs, which revealed the presence of Agathis silver nanoparticles may act as capping organic agents that bund to the surface of the silver nanoparticles. 3.4. FTIR Spectroscopy : Fourier Transform Infrared spectroscopy (FTIR) is a crucial method in the analysis of silver nanoparticles synthesized using plant extracts. The process described Cocculus hirsutus leaf extract can reduce silver nitrate, followed by centrifugation to isolate the nanoparticles. The FTIR spectra provide insight into the molecular interactions and chemical bonds present in the sample. The absorbance bands identified at various wavenumbers correspond to different correspond to different functional groups which are indicative of the biomolecules involved in the reduction and stabilization of the nanoparticles. For instance, the peak at 3304 cm-1 suggests the presence of phenols and alcohols, while the peak at 1382 cm-1 is characteristics of aldehyde or ketone. These functional groups are known to act as reducing agents, facilitating the synthesis of nanoparticles. Moreover, the carbonyl groups strong affinity for metal ions suggest that proteins may cap the nanoparticles, preventing agglomeration and stabilizing the colloidal system. This analysis underscores the complex biochemical interactions at play and highlights the role of FTIR in synthesis and stabilisation. The detailed spectral data thus serve as testament to the intricate nature of bio fabricated nanoparticles. 4. Optimization work For the optimization of the reaction, various concentrations of silver nitrate solutions like 0.5N,1N,2N,3N and 5N were prepared. thermal and microwave conditions used to evaluate the yield of products using five concentrations of starting reactant.in our observations the microwave assisted reaction procedure given excellent yields. The comparison thermal and microwave assisted products shown in Fig. 8 5. Biological Studies Anti-urolithiasis The synthesized silver nanoparticles (AgNPs) to evaluate their effects on preventing Urolithiasis invitro studies were conducted in the laboratory. Struvite was synthesized using sodium metasilicate, ammonium di hydrogen phosphate and magnesium acetate. Morphology of the crystal struvite are struvite crystals of different morphologies like X-shaped didentric, rectangular platelet, and coffin shaped were obtained from the gel medium as showed in the figure. Above formed crystals were separated and dried for break down study. In a freshly taken beaker added reaction mixture to struvite and kept for one day the struvite crystals were dissolved. After this again we went for optimization the reaction the development of stone was stopped by adding silver nanoparticles. Silver nanoparticles were added to the reaction mixture of struvite formation, AgNPs are inhibited the formation of struvite. From these two observations we confirmed that the Cocculus hirsutus mediated silver nanoparticles were excellent for antiurolithaitic activity. 6. Conclusions The Cocculus hirsutus mediated silver nanoparticles (AgNPs) are synthesized by using microwave oven. The Cocculus hirsutus leaf extract capped AgNPs were prevented the struvite crystal nucleation and also dissolved the struvite crystal formed in the gel medium. The urolithiasis inhibitory effect might have been duo to rich content of bioactive phenols, flavonoid and terpenoid content of the Cocculus hirsutus the above studies conclude that the both Cocculus hirsutus and AgNPs can be employed as an alternative medicine for the prophylactic treatment of urolithiasis condition. Declarations Ethical approval: This work not evaluated on animals and ethical committee’s approval not applicable. Funding: Not applicable, any organisation not providing fund. Author Contribution Chintha Muthaiah did all the work and writing manuscriptDr.P.Balaswamy supervised the whole work and prepared tables in the manuscript.Dr.Bala Narsimha Dhoddi provided supporting information explained the supplementary data Acknowledgement: We, the authors, are thankful to Mahatma Gandhi University, Nalgonda, Telangana, India. for providing facilities and generous support. References Nasrollahzadeh, M., Sajadi, S. M., Sajjadi, M., & Issaabadi, Z. (2019). An introduction to nanotechnology. Interface science and technology (Vol. 28, pp. 1–27). Elsevier. Pathakoti, K., Goodla, L., Manubolu, M., & Hwang, H. M. (2019). Nanoparticles and their potential applications in agriculture, biological therapies, food, biomedical, and pharmaceutical industry: a review. Nanotechnology and Nanomaterial Applications in Food Health and Biomedical Sciences , 121–162. Mandal, A., & Ray Banerjee, E. (2020). Introduction to nanoscience, nanotechnology and nanoparticles. Nanomaterials and Biomedicine: Therapeutic and Diagnostic Approach , 1–39. Sun-Waterhouse, D., & Waterhouse, G. I. (2016). Recent advances in the application of nanomaterials and nanotechnology in food research. Novel approaches of nanotechnology in food , 21–66. Khot, L. R., Sankaran, S., Maja, J. M., Ehsani, R., & Schuster, E. W. (2012). Applications of nanomaterials in agricultural production and crop protection: a review. Crop protection , 35 , 64–70. Sriram, P., & Suttee, A. (2020). Nanotechnology advances, benefits, and applications in daily life. Nanotechnology (pp. 23–44). CRC. Moghimi, S. M., Hunter, A. C., & Murray, J. C. (2005). Nanomedicine: current status and future prospects. The FASEB journal , 19 (3), 311–330. Mittal, A. K., Chisti, Y., & Banerjee, U. C. (2013). Synthesis of metallic nanoparticles using plant extracts. Biotechnology advances , 31 (2), 346–356. Kästner, C., & Thünemann, A. F. (2016). Catalytic reduction of 4-nitrophenol using silver nanoparticles with adjustable activity. Langmuir , 32 (29), 7383–7391. Lowry, G. V., Gregory, K. B., Apte, S. C., & Lead, J. R. (2012). Transformations of nanomaterials in the environment. Coe, F. L., Parks, J. H., & Asplin, J. R. (1992). The pathogenesis and treatment of kidney stones. New England Journal of Medicine , 327 (16), 1141–1152. Coe, F. L., Evan, A. P., Worcester, E. M., & Lingeman, J. E. (2010). Three pathways for human kidney stone formation. Urological research , 38 , 147–160. Evan, A. P., Worcester, E. M., Coe, F. L., Williams, J., & Lingeman, J. E. (2015). Mechanisms of human kidney stone formation. Urolithiasis , 43 , 19–32. Basavaraj, D. R., Biyani, C. S., Browning, A. J., & Cartledge, J. J. (2007). The role of urinary kidney stone inhibitors and promoters in the pathogenesis of calcium containing renal stones. EAU-EBU update series , 5 (3), 126–136. Moe, O. W. (2006). Kidney stones: pathophysiology and medical management. The lancet , 367 (9507), 333–344. Coe, F. L., Parks, J. H., & Asplin, J. R. (1992). The pathogenesis and treatment of kidney stones. New England Journal of Medicine , 327 (16), 1141–1152. Morgan, M. S., & Pearle, M. S. (2016). Medical management of renal stones. Bmj , 352 . Stroup, S. P., & Auge, B. K. (2010). Urinary infection and struvite stones. Urinary tract stone disease (pp. 217–224). Springer London. Wang, L. P., Wong, H. Y., & Griffith, D. P. (1997). Treatment options in struvite stones. Urologic Clinics of North America , 24 (1), 149–162. Daudon, M., Jungers, P., Bazin, D., & Williams, J. C. (2018). Recurrence rates of urinary calculi according to stone composition and morphology. Urolithiasis , 46 , 459–470. Li, S., Huang, X., Liu, J., Yue, S., Hou, X., Hu, L., & Wu, J. (2022). Trends in the incidence and DALYs of urolithiasis from 1990 to 2019: results from the global burden of disease study 2019. Frontiers in Public Health , 10 , 825541. O’Kell, A. L., Grant, D. C., & Khan, S. R. (2017). Pathogenesis of calcium oxalate urinary stone disease: species comparison of humans, dogs, and cats. Urolithiasis , 45 (4), 329–336. Logesh, R., Das, N., Adhikari-Devkota, A., & Devkota, H. P. (2020). Cocculus hirsutus (L.) W. Theob. (Menispermaceae): a review on traditional uses, phytochemistry and pharmacological activities. Medicines , 7 (11), 69. Madhavan, V., Ullah, M. S., Gurudeva, M. R., & Yoganarasimhan, S. N. (2010). Pharmacognostical studies on the leaves of Cocculus hirsutus . Diels–Chilahinta . Linn.. an Ayurvedic drug. Prasad, R. Phytochemical screening for various secondary metabolites of Cocculus hirsutus (Stem, Leaf and Fruit Ethanolic Extracts). Jeyachandran, R., Mahesh, A., Cindrella, L., & Baskaran, X. (2008). Screening Antibacterial Activity of Root Extract of Cocculus hirsutus (L). Journal of Plant Sciences , 3 (3), 194–198. Rajan, M., Prabhakaran, S., Prusty, J. S., Chauhan, N., Gupta, P., & Kumar, A. (2023). Phytochemicals of Cocculus hirsutus deciphered SARS-CoV-2 inhibition by targeting main proteases in molecular docking, simulation, and pharmacological analyses. Journal of Biomolecular Structure and Dynamics , 41 (15), 7406–7420. Nadagouda, M. N., Speth, T. F., & Varma, R. S. (2011). Microwave-assisted green synthesis of silver nanostructures. Accounts of Chemical Research , 44 (7), 469–478. Kaur, N., Singh, A., & Ahmad, W. (2023). Microwave assisted green synthesis of silver nanoparticles and its application: a review. Journal of Inorganic and Organometallic Polymers and Materials , 33 (3), 663–672. Devaiah, D., Reddy, L. H., Park, S. E., & Reddy, B. M. (2018). Ceria–zirconia mixed oxides: Synthetic methods and applications. Catalysis Reviews , 60 (2), 177–277. Devi, N., Sahoo, S., Kumar, R., & Singh, R. K. (2021). A review of the microwave-assisted synthesis of carbon nanomaterials, metal oxides/hydroxides and their composites for energy storage applications. Nanoscale , 13 (27), 11679–11711. Chin, C. D. W., Treadwell, L. J., & Wiley, J. B. (2021). Microwave synthetic routes for shape-controlled catalyst nanoparticles and nanocomposites. Molecules , 26 (12), 3647. Gabriela, Á. M., Gabriela, M. D. O. V., Luis, A. M., Reinaldo, P. R., Michael, H. M., Rodolfo, G. P., & Roberto, V. B. J. (2017). Biosynthesis of silver nanoparticles using mint leaf extract (Mentha piperita) and their antibacterial activity. Advanced Science Engineering and Medicine , 9 (11), 914–923. Girish, K. (2017). Antimicrobial activities of Lantana camara Linn. Asian Journal of Pharmaceutical and Clinical Research , 10 (3), 57–67. Vanlalveni, C., Lallianrawna, S., Biswas, A., Selvaraj, M., Changmai, B., & Rokhum, S. L. (2021). Green synthesis of silver nanoparticles using plant extracts and their antimicrobial activities: A review of recent literature. RSC advances , 11 (5), 2804–2837. Nadaf, S. J., Jadhav, N. R., Naikwadi, H. S., Savekar, P. L., Sapkal, I. D., Kambli, M. M., & Desai, I. A. (2022). Green synthesis of gold and silver nanoparticles: Updates on research, patents, and future prospects. OpenNano , 8 , 100076. Das, P., Kumar, K., Nambiraj, A., Awasthi, R., Dua, K., & Malipeddi, H. (2018). Antibacterial and in vitro growth inhibition study of struvite urinary stones using Oxalis corniculata Linn. Leaf extract and its biofabricated silver nanoparticles. Recent Patents on Drug Delivery & Formulation , 12 (3), 170–178. McLean, R. J. C., Downey, J., Clapham, L., & Nickel, J. C. (1990). A simple technique for studying struvite crystal growth in vitro. Urological research , 18 , 39–43. Additional Declarations No competing interests reported. Supplementary Files Graphicalabstract.png Graphical abstract Cite Share Download PDF Status: Under Review Version 1 posted Editor assigned by journal 22 May, 2024 Submission checks completed at journal 22 May, 2024 First submitted to journal 11 May, 2024 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-4406241","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":305610905,"identity":"e2b43740-262c-4daa-bd6e-77c3ee474548","order_by":0,"name":"Muthaiah Chintha","email":"","orcid":"","institution":"Mahatma Gandhi University","correspondingAuthor":false,"prefix":"","firstName":"Muthaiah","middleName":"","lastName":"Chintha","suffix":""},{"id":305610906,"identity":"2c0759dc-6b04-4251-b590-ebe412673544","order_by":1,"name":"Balaswamy Puligilla","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABB0lEQVRIiWNgGAWjYFAC5oYDDAYWMgwSDIwHGP/ZAEUYGw/g18II0iLBA9TCcICBLQ0qQkALkIBrOQwWw6vFXCKx8eCXAgke/tnNDw7z8Jy3W9t+GGhLjU00Li2WMxIbDssAHSZx55jBYR6J28nbziQCtRxLy23AocXgBlCLBMgvNxKAWgxuJ5sdAGphbDhMWIv8jfQPh3kSziWbnX9IWMvBD0AtBjdygLYcOGBndoOALZY9DxsOgwLZ8M6ZgoNzG5ITzG4AbUnA4xdz9uTDH3/8sZGTu92+8cHbBjt7s/PpDx98qLHB7TAgZuZBEkgEq0zAoRymhfEHkoA9HsWjYBSMglEwQgEAVPdp33b83g8AAAAASUVORK5CYII=","orcid":"","institution":"Mahatma Gandhi University","correspondingAuthor":true,"prefix":"","firstName":"Balaswamy","middleName":"","lastName":"Puligilla","suffix":""},{"id":305610907,"identity":"3563e56a-fcaf-4725-99e6-327cb3a09627","order_by":2,"name":"Bala Narsimha Dhoddi","email":"","orcid":"","institution":"government degree college( mahata","correspondingAuthor":false,"prefix":"","firstName":"Bala","middleName":"Narsimha","lastName":"Dhoddi","suffix":""}],"badges":[],"createdAt":"2024-05-11 17:08:27","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4406241/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4406241/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":57707844,"identity":"4026bb62-0b66-4f48-b925-c200959f9383","added_by":"auto","created_at":"2024-06-04 15:13:15","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":137767,"visible":true,"origin":"","legend":"\u003cp\u003eChange in colour of reaction mixture\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4406241/v1/71ac632cb86cbca820357a5f.png"},{"id":57707843,"identity":"8bc0e044-15e4-466a-a5f0-5bb7f1cd6026","added_by":"auto","created_at":"2024-06-04 15:13:15","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":10799,"visible":true,"origin":"","legend":"\u003cp\u003eUV-VIS spectrum of AgNPS\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4406241/v1/d7a8a795a7084fd3027c011a.png"},{"id":57707837,"identity":"73b40690-ca21-4b19-a91f-96c3e53108f2","added_by":"auto","created_at":"2024-06-04 15:13:15","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":20295,"visible":true,"origin":"","legend":"\u003cp\u003eXRD spectrum of AgNPs\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4406241/v1/0d2f3e070a34abf820588e14.png"},{"id":57708644,"identity":"48a792bf-57b7-467b-a68f-5def4ef14a2c","added_by":"auto","created_at":"2024-06-04 15:21:15","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":32848,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 5\u003c/strong\u003e.EDX spectrum of AgNPs\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-4406241/v1/1a12eb16f7849c4f2155fbcb.png"},{"id":57709162,"identity":"89131666-9f92-4f75-b208-429150f04f29","added_by":"auto","created_at":"2024-06-04 15:29:15","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":129204,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 6\u003c/strong\u003e. SEM image of AgNPs at 10µm\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-4406241/v1/6ee6dbcbdf201baac5f0d4fa.png"},{"id":57707839,"identity":"b87c2721-01b9-4f9e-a539-80e37e210aae","added_by":"auto","created_at":"2024-06-04 15:13:15","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":94985,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 7\u003c/strong\u003e.SEM image of AgNPs (100nm)\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-4406241/v1/b502b9167fac982f57cef5f9.png"},{"id":57708647,"identity":"d7070c78-1934-45b0-9e63-957f7d426474","added_by":"auto","created_at":"2024-06-04 15:21:15","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":8649,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 8.\u003c/strong\u003eFT-IR Spectrum of AgNPs\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-4406241/v1/936307809955c12a5d48cb5f.png"},{"id":57707840,"identity":"bc586a5a-bd82-4cef-889e-ef49003da2d8","added_by":"auto","created_at":"2024-06-04 15:13:15","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":59019,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 9\u003c/strong\u003e. comparison between thermal and microwave reaction\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-4406241/v1/9c98ffe22c6b0d3e4f69a7b1.png"},{"id":57707846,"identity":"c6c0ae24-faf5-483d-8c24-59671a143d83","added_by":"auto","created_at":"2024-06-04 15:13:15","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":283862,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 10\u003c/strong\u003e. Urolithiatic activity\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-4406241/v1/9c05045a0b5bb74ad73ae206.png"},{"id":57709217,"identity":"cd4b982b-4fb8-4e0a-b773-f830bc02ebf4","added_by":"auto","created_at":"2024-06-04 15:29:20","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1129905,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4406241/v1/2afb11e7-0424-4af0-844b-57a1f9a4d08c.pdf"},{"id":57708645,"identity":"cee122f1-c9f2-496b-ad97-c162f663d32d","added_by":"auto","created_at":"2024-06-04 15:21:15","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":89873,"visible":true,"origin":"","legend":"\u003cp\u003eGraphical abstract\u003c/p\u003e","description":"","filename":"Graphicalabstract.png","url":"https://assets-eu.researchsquare.com/files/rs-4406241/v1/7181065565818fda15153736.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Microwave assisted synthesis of Silver Nanoparticles using Cocculus hirsutus leaves Extract and their Anti-Urolithic activity","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eNanotechnology is an emerging field of research that focuses on the creation and applications of nano particles in various domains such as catalysis, electrochemistry, biomedicine, pharmaceuticals, sensors, food technology and cosmetics etc. [\u003cspan additionalcitationids=\"CR2 CR3 CR4\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. It is a revolutionary field that has been integrated into various domains of our daily life, many products contain nanomaterials as well as applications-based nanoparticles have emerged [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Nanotechnology is a key enabling technology that can offer new and innovative medical solutions for unmet medical needs so far. Nanomedicine is the term that describes the use of nanomaterials for diagnosis, monitoring, control, prevention and treatment of disease [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Metallic nanoparticles can be synthesized using plant extracts which is a green and safe approach [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. The silver Nano particles produced by this method have various properties such as high chemical stability, catalytic activity and many therapeutic activities [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Nanomaterials have different properties and interactions depending on their size. As particle size became smaller SA\\V increases which can significantly enhance the reactivity. This means that nanomaterials have surface area per unit volume which makes them more reactive, soluble and bio available than their bulk counterparts [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eUrolithiasis commonly known as kidney stones, is indeed a multifactorial disorder characterised by the formation of urinary calculi within the renal tubules [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. the pathogenesis of these stones is complex, involving a combination of factors such as anatomical abnormalities, metabolic disorders, and dietary habits [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. The increased concentration of unbound ions, particularly calcium and phosphate in the urine can lead to the precipitation of these salts, farming calculi [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. The presence or absence of endogenous inhibitors like citrate and magnesium, which prevent the stone formation, also plays a crucial role [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. The majority of urinary calculi are composed of calcium oxalate, which accounts for approximately 75% of cases followed by struvite, uric acid and cystine stones [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Risk factors for urolithiasis include low fluid intake certain urinary tract malformations, urinary tract infections and dietary facts such as high sodium intake and high dietary oxalate. Management of the condition often involves increasing fluid intake, dietary modifications, and in some cases medical or surgical intervention to remove the stones. Preventative strategies are tailored to the type of stones and underlying risk factors identified in each individual case [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAmmonium magnesium phosphate known as struvite is produced rapidly as a result of urinary tract infection caused by the \u003cem\u003eurease\u003c/em\u003e producing bacteria [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. these struvites are difficult to treat because of the complications associated urolithiatic microorganisms [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. The struvite stones account for 17% of all urinary stones and are reported to be distinctive due to their rapid growth rate persistence and high degree of recurrence (50%) [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. The worldwide increasing in the incidences of urolithiasis and many studies has also reported that calcium oxalate stones constitute about 79% of the kidney stone disease. [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. pathogenesis of calcium oxalate stone formation involves a series of physicochemical events which include the nucleation of the crystals, the growth and aggregation of the crystals and finally the retaining of crystal aggregates on the epithelial cell linings of the Renal tubules [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003eCocculus hirsutus\u003c/em\u003e also known as broom keeper is a versatile herbal medicine used in folk medicine for various conditions such as Tuberculosis, Leprosy, Skin diseases, Dyspepsia, Pruritus [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. The phytochemical composition and potential bioactive compounds of \u003cem\u003eCocculus hirsutus\u003c/em\u003e plant leaves have been the subject of study. Tannins, alkaloids, flavonoids, steroids, and saponins were found to be present in the leaves of \u003cem\u003eCocculus hirsutus.\u003c/em\u003e Notably, the leaves were found to contain two major copmounds:5,7-dihydroxy-3-(4\u0026rsquo;-hydroxybenzyl) chromone and kaempferide-3-O-α-rhamnosyl-7-O-α-L-rhamnoside, which belong to the homoisoflavones and flavanol triglycerides, respectively [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Another research endeavour focused on the leaves of \u003cem\u003eCocculus hirsutus\u003c/em\u003e discovered that the acetone extract exhibited antimicrobial activity against bacteria and fungi. It was further determined that O-ethyl hydroxyl amine,2-ethyl heptanoic acid and 1-nonyl cycloheptane were responsible for this observed activity [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Furthermore, an investigation into the potential anti-COVID-19 activity of \u003cem\u003eCocculus hirsutus\u003c/em\u003e identified betulin, coclaurine, and quinic acid as compounds with notable binding affinity to the main proteases of SARS-CoV-2[\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. These findings indicate that \u003cem\u003eCocculus hirsutus\u003c/em\u003e leaves possess a diverse range of phytochemicals with potential bioactive and therapeutic properties.\u003c/p\u003e \u003cp\u003eThe sensible and sustainable chemistry is essential for our ever-changing environment. consequently, synthesis of chemical substances started to follow green chemistry principles in recent years, Microwave assisted synthesis of Nano particles is a rapid and efficient method for producing nanostructured materials [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. This technique offers advantages such as rapid volumetric heating, higher reaction rates, shorter reaction times, and higher product yields compared to conventional heating methods [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. It has been applied to the synthesis of various nanoparticles including silver nanoparticles, ceria-zirconia Nano particles and iron oxide nanoparticles [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. The microwave heating process allows for the control of particle size and morphology, resulting in nanoparticles specific properties [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. For instance, the microwave assisted synthesis of silver nanoparticles using mint leaf extract as a reducing and stabilizing agent showed a higher peak height and smaller particle size compared to conventional heating methods [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eRecent studies have reported the green synthesis of AgNPs using the extracts of \u003cem\u003eLantana camara, Hibiscus rosa-sinensis, Azadirachta Indica and Geranium leaf\u003c/em\u003e [\u003cspan additionalcitationids=\"CR35\" citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. The method involves the role of plant secondary metabolites as surface active molecules which are potent reducing and capping agents involved in the formation of stable nanoparticles. The use of medicinal plants in the treatment of diseased condition has been preferred as it is considered to be safe, with minimal or less side effects. The literature attracted us to use silver nanoparticles and \u003cem\u003eCocculus hirsutus\u003c/em\u003e leaf extract combination for treatment of anti-struvite.\u003c/p\u003e"},{"header":"2. Research methodology","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Synthesis of AgNPs:\u003c/h2\u003e \u003cp\u003e \u003cem\u003eCocculus hirsutus\u003c/em\u003e mediated synthesis of AgNPs were synthesized by optimizing the concentration of plant material required for the formation of a stable Nanomaterial. The powdered leaf extract(0.5gr) was added to 50 ml double distilled water and kept for boiling in a house hold conventional microwave oven for 10 min to obtain the extract.it is the plant extraction that was used for the preparation of the AgNPs. The formation of AgNPs was attained by incubation of 0.01N AgNO\u003csub\u003e3\u003c/sub\u003e and the plant extract mixture in a ratio of 1:5 at 32\u003csup\u003eO\u003c/sup\u003ec for 5minuts in a microwave oven. The formation of the AgNPs was confirmed from the observed colour change of solution from colourless to brown colour.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 \u003cb\u003eSynthesis of struvite\u003c/b\u003e:\u003c/h2\u003e \u003cp\u003eThe hydrogel medium needed for crystal growth was attained by employing a solution containing 1M Sodium Meta Silicate (NaSiO\u003csub\u003e3\u003c/sub\u003e.9H\u003csub\u003e2\u003c/sub\u003eO) and 0.5M Ammonium dihydrogen phosphate (NH\u003csub\u003e4\u003c/sub\u003eH\u003csub\u003e2\u003c/sub\u003ePO\u003csub\u003e4\u003c/sub\u003e.2H\u003csub\u003e2\u003c/sub\u003eO) as well as a solution composed of 1M Magnesium acetate (Mg (CH\u003csub\u003e3\u003c/sub\u003eCOO)\u003csub\u003e2\u003c/sub\u003e.4H\u003csub\u003e2\u003c/sub\u003eO) [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. These solutions were introduced into a different test tubes and subsequently subjected to an aseptic environment at a temperature of 37\u003csup\u003e0\u003c/sup\u003eC until the reaction reached its culmination. Following the 24-hour duration the struvite composition of the crystals that had formed was examined.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e3.1 Characterization of the AgNPs:\u003c/h2\u003e \u003cp\u003eInitially synthesized silver nanoparticles were detected by visible colour changing from colourless to dark brown colour as well as UV-Vis Spectral data revealed the absorption peak at 320nm confirming the presence of silver nanoparticles. This was due to the green reduction of Ag\u003csup\u003e\u003cb\u003e+\u003c/b\u003e\u003c/sup\u003e ion to Ag\u003csup\u003e0\u003c/sup\u003e by the phytochemicals present in the \u003cem\u003eCocculus hirsutus\u003c/em\u003e leaf extract. silver nanoparticles were prepared by using diverse (0.5N,1N,2N,3N \u0026amp;5N) concentrations of silver nitrate for optimization it was shown in the Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e\u0026amp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e3.2.XRD Analysis:\u003c/h2\u003e \u003cp\u003eNewly crystallized silver nanoparticle was confirmed by Bruker 5000 the Bxrd Pattern analysis given the plane indices of 111,220,311 as metallic silver XRD values are coincided. The excellent XRD spectral values promising the formation of silver nanoparticles in different concentrations of starting materials were taken shown in Fig.\u0026nbsp;3\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e3.3.SEM \u0026amp; EDX Spectroscopy\u003c/h2\u003e \u003cp\u003eWe used JOEL scanning electron microscopy to analyse the shape and size of the silver nanoparticles we synthesized. The SEM images showed that the AgNPs had smooth surface and a spherical shape, with an average diameter ranging from 17 to 27nm (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e6\u003c/span\u003e. \u0026amp; Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e7\u003c/span\u003e.). This method is widely used to study the surface characteristics of nanoparticles, such as their size, shape, morphology and distribution. Johnson and Prabhu (2015) reported similar results when they used camelina Bengalese\u0026rsquo;s to produce spherical and uniform Ag nanoparticles and examined them with SEM. This technique relies on the interaction of electrons with the sample surface, which generates secondary electrons, the signal from these electrons is then used to create an image of the sample surface morphology.\u003c/p\u003e \u003cp\u003eThe elemental composition of the AgNPs was determined by EDX analysis, which measures the energy and intensity distribution of X-ray signals generated by an electron beam on a specimen. Figure\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e5\u003c/span\u003e. shows the EDX spectra of the AgNPs, which revealed the presence of Agathis silver nanoparticles may act as capping organic agents that bund to the surface of the silver nanoparticles.\u003c/p\u003e\u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e\u003cb\u003e3.4. FTIR Spectroscopy\u003c/b\u003e:\u003c/h2\u003e \u003cp\u003eFourier Transform Infrared spectroscopy (FTIR) is a crucial method in the analysis of silver nanoparticles synthesized using plant extracts. The process described \u003cem\u003eCocculus hirsutus\u003c/em\u003e leaf extract can reduce silver nitrate, followed by centrifugation to isolate the nanoparticles. The FTIR spectra provide insight into the molecular interactions and chemical bonds present in the sample. The absorbance bands identified at various wavenumbers correspond to different correspond to different functional groups which are indicative of the biomolecules involved in the reduction and stabilization of the nanoparticles. For instance, the peak at 3304 cm-1 suggests the presence of phenols and alcohols, while the peak at 1382 cm-1 is characteristics of aldehyde or ketone. These functional groups are known to act as reducing agents, facilitating the synthesis of nanoparticles. Moreover, the carbonyl groups strong affinity for metal ions suggest that proteins may cap the nanoparticles, preventing agglomeration and stabilizing the colloidal system. This analysis underscores the complex biochemical interactions at play and highlights the role of FTIR in synthesis and stabilisation. The detailed spectral data thus serve as testament to the intricate nature of bio fabricated nanoparticles.\u003c/p\u003e\u003c/div\u003e"},{"header":"4. Optimization work","content":"\u003cp\u003eFor the optimization of the reaction, various concentrations of silver nitrate solutions like 0.5N,1N,2N,3N and 5N were prepared. thermal and microwave conditions used to evaluate the yield of products using five concentrations of starting reactant.in our observations the microwave assisted reaction procedure given excellent yields. The comparison thermal and microwave assisted products shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e8\u003c/span\u003e\u003c/p\u003e"},{"header":"5. Biological Studies","content":"\u003cp\u003e\u003cstrong\u003eAnti-urolithiasis\u003c/strong\u003e\u003c/p\u003e \u003cp\u003eThe synthesized silver nanoparticles (AgNPs) to evaluate their effects on preventing Urolithiasis invitro studies were conducted in the laboratory. Struvite was synthesized using sodium metasilicate, ammonium di hydrogen phosphate and magnesium acetate. Morphology of the crystal struvite are struvite crystals of different morphologies like X-shaped didentric, rectangular platelet, and coffin shaped were obtained from the gel medium as showed in the figure. Above formed crystals were separated and dried for break down study. In a freshly taken beaker added reaction mixture to struvite and kept for one day the struvite crystals were dissolved. After this again we went for optimization the reaction the development of stone was stopped by adding silver nanoparticles. Silver nanoparticles were added to the reaction mixture of struvite formation, AgNPs are inhibited the formation of struvite. From these two observations we confirmed that the \u003cem\u003eCocculus hirsutus\u003c/em\u003e mediated silver nanoparticles were excellent for antiurolithaitic activity.\u003c/p\u003e "},{"header":"6. Conclusions","content":"\u003cp\u003eThe \u003cem\u003eCocculus hirsutus\u003c/em\u003e mediated silver nanoparticles (AgNPs) are synthesized by using microwave oven. The \u003cem\u003eCocculus hirsutus\u003c/em\u003e leaf extract capped AgNPs were prevented the struvite crystal nucleation and also dissolved the struvite crystal formed in the gel medium. The urolithiasis inhibitory effect might have been duo to rich content of bioactive phenols, flavonoid and terpenoid content of the \u003cem\u003eCocculus hirsutus\u003c/em\u003e the above studies conclude that the both \u003cem\u003eCocculus hirsutus\u003c/em\u003e and AgNPs can be employed as an alternative medicine for the prophylactic treatment of urolithiasis condition.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003cstrong\u003eEthical approval:\u003c/strong\u003e \u003cp\u003eThis work not evaluated on animals and ethical committee\u0026rsquo;s approval not applicable.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding:\u003c/h2\u003e \u003cp\u003eNot applicable, any organisation not providing fund.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eChintha Muthaiah did all the work and writing manuscriptDr.P.Balaswamy supervised the whole work and prepared tables in the manuscript.Dr.Bala Narsimha Dhoddi provided supporting information explained the supplementary data\u003c/p\u003e\u003ch2\u003eAcknowledgement:\u003c/h2\u003e \u003cp\u003eWe, the authors, are thankful to Mahatma Gandhi University, Nalgonda, Telangana, India. for providing facilities and generous support.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eNasrollahzadeh, M., Sajadi, S. M., Sajjadi, M., \u0026amp; Issaabadi, Z. (2019). An introduction to nanotechnology. \u003cem\u003eInterface science and technology\u003c/em\u003e (Vol. 28, pp. 1\u0026ndash;27). Elsevier.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePathakoti, K., Goodla, L., Manubolu, M., \u0026amp; Hwang, H. M. (2019). Nanoparticles and their potential applications in agriculture, biological therapies, food, biomedical, and pharmaceutical industry: a review. \u003cem\u003eNanotechnology and Nanomaterial Applications in Food Health and Biomedical Sciences\u003c/em\u003e, 121\u0026ndash;162.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMandal, A., \u0026amp; Ray Banerjee, E. (2020). Introduction to nanoscience, nanotechnology and nanoparticles. \u003cem\u003eNanomaterials and Biomedicine: Therapeutic and Diagnostic Approach\u003c/em\u003e, 1\u0026ndash;39.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSun-Waterhouse, D., \u0026amp; Waterhouse, G. I. (2016). Recent advances in the application of nanomaterials and nanotechnology in food research. \u003cem\u003eNovel approaches of nanotechnology in food\u003c/em\u003e, 21\u0026ndash;66.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKhot, L. R., Sankaran, S., Maja, J. M., Ehsani, R., \u0026amp; Schuster, E. W. (2012). Applications of nanomaterials in agricultural production and crop protection: a review. \u003cem\u003eCrop protection\u003c/em\u003e, \u003cem\u003e35\u003c/em\u003e, 64\u0026ndash;70.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSriram, P., \u0026amp; Suttee, A. (2020). Nanotechnology advances, benefits, and applications in daily life. \u003cem\u003eNanotechnology\u003c/em\u003e (pp. 23\u0026ndash;44). CRC.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMoghimi, S. M., Hunter, A. C., \u0026amp; Murray, J. C. (2005). Nanomedicine: current status and future prospects. \u003cem\u003eThe FASEB journal\u003c/em\u003e, \u003cem\u003e19\u003c/em\u003e(3), 311\u0026ndash;330.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMittal, A. K., Chisti, Y., \u0026amp; Banerjee, U. C. (2013). Synthesis of metallic nanoparticles using plant extracts. \u003cem\u003eBiotechnology advances\u003c/em\u003e, \u003cem\u003e31\u003c/em\u003e(2), 346\u0026ndash;356.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eK\u0026auml;stner, C., \u0026amp; Th\u0026uuml;nemann, A. F. (2016). Catalytic reduction of 4-nitrophenol using silver nanoparticles with adjustable activity. \u003cem\u003eLangmuir\u003c/em\u003e, \u003cem\u003e32\u003c/em\u003e(29), 7383\u0026ndash;7391.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLowry, G. V., Gregory, K. B., Apte, S. C., \u0026amp; Lead, J. R. (2012). Transformations of nanomaterials in the environment.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCoe, F. L., Parks, J. H., \u0026amp; Asplin, J. R. (1992). The pathogenesis and treatment of kidney stones. \u003cem\u003eNew England Journal of Medicine\u003c/em\u003e, \u003cem\u003e327\u003c/em\u003e(16), 1141\u0026ndash;1152.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCoe, F. L., Evan, A. P., Worcester, E. M., \u0026amp; Lingeman, J. E. (2010). Three pathways for human kidney stone formation. \u003cem\u003eUrological research\u003c/em\u003e, \u003cem\u003e38\u003c/em\u003e, 147\u0026ndash;160.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEvan, A. P., Worcester, E. M., Coe, F. L., Williams, J., \u0026amp; Lingeman, J. E. (2015). Mechanisms of human kidney stone formation. \u003cem\u003eUrolithiasis\u003c/em\u003e, \u003cem\u003e43\u003c/em\u003e, 19\u0026ndash;32.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBasavaraj, D. R., Biyani, C. S., Browning, A. J., \u0026amp; Cartledge, J. J. (2007). The role of urinary kidney stone inhibitors and promoters in the pathogenesis of calcium containing renal stones. \u003cem\u003eEAU-EBU update series\u003c/em\u003e, \u003cem\u003e5\u003c/em\u003e(3), 126\u0026ndash;136.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMoe, O. W. (2006). Kidney stones: pathophysiology and medical management. \u003cem\u003eThe lancet\u003c/em\u003e, \u003cem\u003e367\u003c/em\u003e(9507), 333\u0026ndash;344.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCoe, F. L., Parks, J. H., \u0026amp; Asplin, J. R. (1992). The pathogenesis and treatment of kidney stones. \u003cem\u003eNew England Journal of Medicine\u003c/em\u003e, \u003cem\u003e327\u003c/em\u003e(16), 1141\u0026ndash;1152.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMorgan, M. S., \u0026amp; Pearle, M. S. (2016). Medical management of renal stones. \u003cem\u003eBmj\u003c/em\u003e, \u003cem\u003e352\u003c/em\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eStroup, S. P., \u0026amp; Auge, B. K. (2010). Urinary infection and struvite stones. \u003cem\u003eUrinary tract stone disease\u003c/em\u003e (pp. 217\u0026ndash;224). Springer London.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, L. P., Wong, H. Y., \u0026amp; Griffith, D. P. (1997). Treatment options in struvite stones. \u003cem\u003eUrologic Clinics of North America\u003c/em\u003e, \u003cem\u003e24\u003c/em\u003e(1), 149\u0026ndash;162.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDaudon, M., Jungers, P., Bazin, D., \u0026amp; Williams, J. C. (2018). Recurrence rates of urinary calculi according to stone composition and morphology. \u003cem\u003eUrolithiasis\u003c/em\u003e, \u003cem\u003e46\u003c/em\u003e, 459\u0026ndash;470.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi, S., Huang, X., Liu, J., Yue, S., Hou, X., Hu, L., \u0026amp; Wu, J. (2022). Trends in the incidence and DALYs of urolithiasis from 1990 to 2019: results from the global burden of disease study 2019. \u003cem\u003eFrontiers in Public Health\u003c/em\u003e, \u003cem\u003e10\u003c/em\u003e, 825541.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eO\u0026rsquo;Kell, A. L., Grant, D. C., \u0026amp; Khan, S. R. (2017). Pathogenesis of calcium oxalate urinary stone disease: species comparison of humans, dogs, and cats. \u003cem\u003eUrolithiasis\u003c/em\u003e, \u003cem\u003e45\u003c/em\u003e(4), 329\u0026ndash;336.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLogesh, R., Das, N., Adhikari-Devkota, A., \u0026amp; Devkota, H. P. (2020). \u003cem\u003eCocculus hirsutus\u003c/em\u003e (L.) W. Theob. (Menispermaceae): a review on traditional uses, phytochemistry and pharmacological activities. \u003cem\u003eMedicines\u003c/em\u003e, \u003cem\u003e7\u003c/em\u003e(11), 69.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMadhavan, V., Ullah, M. S., Gurudeva, M. R., \u0026amp; Yoganarasimhan, S. N. (2010). Pharmacognostical studies on the leaves of \u003cem\u003eCocculus hirsutus\u003c/em\u003e. \u003cem\u003eDiels\u0026ndash;Chilahinta\u003c/em\u003e. Linn.. an Ayurvedic drug.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePrasad, R. Phytochemical screening for various secondary metabolites of \u003cem\u003eCocculus hirsutus\u003c/em\u003e (Stem, Leaf and Fruit Ethanolic Extracts).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJeyachandran, R., Mahesh, A., Cindrella, L., \u0026amp; Baskaran, X. (2008). Screening Antibacterial Activity of Root Extract of \u003cem\u003eCocculus hirsutus\u003c/em\u003e (L). \u003cem\u003eJournal of Plant Sciences\u003c/em\u003e, \u003cem\u003e3\u003c/em\u003e(3), 194\u0026ndash;198.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRajan, M., Prabhakaran, S., Prusty, J. S., Chauhan, N., Gupta, P., \u0026amp; Kumar, A. (2023). Phytochemicals of \u003cem\u003eCocculus hirsutus\u003c/em\u003e deciphered SARS-CoV-2 inhibition by targeting main proteases in molecular docking, simulation, and pharmacological analyses. \u003cem\u003eJournal of Biomolecular Structure and Dynamics\u003c/em\u003e, \u003cem\u003e41\u003c/em\u003e(15), 7406\u0026ndash;7420.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNadagouda, M. N., Speth, T. F., \u0026amp; Varma, R. S. (2011). Microwave-assisted green synthesis of silver nanostructures. \u003cem\u003eAccounts of Chemical Research\u003c/em\u003e, \u003cem\u003e44\u003c/em\u003e(7), 469\u0026ndash;478.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKaur, N., Singh, A., \u0026amp; Ahmad, W. (2023). Microwave assisted green synthesis of silver nanoparticles and its application: a review. \u003cem\u003eJournal of Inorganic and Organometallic Polymers and Materials\u003c/em\u003e, \u003cem\u003e33\u003c/em\u003e(3), 663\u0026ndash;672.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDevaiah, D., Reddy, L. H., Park, S. E., \u0026amp; Reddy, B. M. (2018). Ceria\u0026ndash;zirconia mixed oxides: Synthetic methods and applications. \u003cem\u003eCatalysis Reviews\u003c/em\u003e, \u003cem\u003e60\u003c/em\u003e(2), 177\u0026ndash;277.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDevi, N., Sahoo, S., Kumar, R., \u0026amp; Singh, R. K. (2021). A review of the microwave-assisted synthesis of carbon nanomaterials, metal oxides/hydroxides and their composites for energy storage applications. \u003cem\u003eNanoscale\u003c/em\u003e, \u003cem\u003e13\u003c/em\u003e(27), 11679\u0026ndash;11711.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChin, C. D. W., Treadwell, L. J., \u0026amp; Wiley, J. B. (2021). Microwave synthetic routes for shape-controlled catalyst nanoparticles and nanocomposites. \u003cem\u003eMolecules\u003c/em\u003e, \u003cem\u003e26\u003c/em\u003e(12), 3647.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGabriela, \u0026Aacute;. M., Gabriela, M. D. O. V., Luis, A. M., Reinaldo, P. R., Michael, H. M., Rodolfo, G. P., \u0026amp; Roberto, V. B. J. (2017). Biosynthesis of silver nanoparticles using mint leaf extract (Mentha piperita) and their antibacterial activity. \u003cem\u003eAdvanced Science Engineering and Medicine\u003c/em\u003e, \u003cem\u003e9\u003c/em\u003e(11), 914\u0026ndash;923.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGirish, K. (2017). Antimicrobial activities of Lantana camara Linn. \u003cem\u003eAsian Journal of Pharmaceutical and Clinical Research\u003c/em\u003e, \u003cem\u003e10\u003c/em\u003e(3), 57\u0026ndash;67.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVanlalveni, C., Lallianrawna, S., Biswas, A., Selvaraj, M., Changmai, B., \u0026amp; Rokhum, S. L. (2021). Green synthesis of silver nanoparticles using plant extracts and their antimicrobial activities: A review of recent literature. \u003cem\u003eRSC advances\u003c/em\u003e, \u003cem\u003e11\u003c/em\u003e(5), 2804\u0026ndash;2837.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNadaf, S. J., Jadhav, N. R., Naikwadi, H. S., Savekar, P. L., Sapkal, I. D., Kambli, M. M., \u0026amp; Desai, I. A. (2022). Green synthesis of gold and silver nanoparticles: Updates on research, patents, and future prospects. \u003cem\u003eOpenNano\u003c/em\u003e, \u003cem\u003e8\u003c/em\u003e, 100076.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDas, P., Kumar, K., Nambiraj, A., Awasthi, R., Dua, K., \u0026amp; Malipeddi, H. (2018). Antibacterial and in vitro growth inhibition study of struvite urinary stones using Oxalis corniculata Linn. Leaf extract and its biofabricated silver nanoparticles. \u003cem\u003eRecent Patents on Drug Delivery \u0026amp; Formulation\u003c/em\u003e, \u003cem\u003e12\u003c/em\u003e(3), 170\u0026ndash;178.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMcLean, R. J. C., Downey, J., Clapham, L., \u0026amp; Nickel, J. C. (1990). A simple technique for studying struvite crystal growth in vitro. \u003cem\u003eUrological research\u003c/em\u003e, \u003cem\u003e18\u003c/em\u003e, 39\u0026ndash;43.\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":"bionanoscience","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bnsc","sideBox":"Learn more about [BioNanoScience](http://link.springer.com/journal/12668)","snPcode":"12668","submissionUrl":"https://submission.nature.com/new-submission/12668/3","title":"BioNanoScience","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"AgNPs, Cocculus hirsutus, Microwave, in vitro urolithiasis","lastPublishedDoi":"10.21203/rs.3.rs-4406241/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4406241/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eAs highlighting the synergetic action of Nanoscale metal along with the biomolecules play crucial role in modern medicine. The silver nanoparticles were owed many uses in the field of medicine; one of the most important approaches of silver nitrate is Anti-urolithiasis effect. The present study focuses on the anti-urolithiasis activity of \u003cem\u003eCocculus hirsutus \u003c/em\u003eleaf extract capped silver nanoparticles (AgNPs). The AgNPs were synthesized by silver nitrate salt solution and \u003cem\u003eCocculus hirsutus \u003c/em\u003eleaf extract in microwave oven. The Microwave assisted synthesis is an efficient technique and promises more environment benign than traditional heating. The newly prepared AgNPs were characterized by various techniques. the invitro struvite crystal preparation and inhibition activity were examined by using freshly prepared silver nanoparticles.\u003c/p\u003e","manuscriptTitle":"Microwave assisted synthesis of Silver Nanoparticles using Cocculus hirsutus leaves Extract and their Anti-Urolithic activity","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-06-04 15:13:10","doi":"10.21203/rs.3.rs-4406241/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"editorAssigned","content":"","date":"2024-05-22T17:06:02+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-05-22T16:39:13+00:00","index":"","fulltext":""},{"type":"submitted","content":"BioNanoScience","date":"2024-05-11T17:06:51+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bionanoscience","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bnsc","sideBox":"Learn more about [BioNanoScience](http://link.springer.com/journal/12668)","snPcode":"12668","submissionUrl":"https://submission.nature.com/new-submission/12668/3","title":"BioNanoScience","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"a8cadd29-8a2e-4513-802a-78142b807ada","owner":[],"postedDate":"June 4th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2024-06-04T15:13:10+00:00","versionOfRecord":[],"versionCreatedAt":"2024-06-04 15:13:10","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4406241","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4406241","identity":"rs-4406241","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}