Novel Pla / Chitosan Blends With Bio-released Ag-nps Nano Composites for Eco-friendly Food Packaging.

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Malarvizhi Nithya, Rajeswari Balaji, Tharani Sundarajan, Vennila Sivalingam This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4788416/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 12 Jun, 2025 Read the published version in Environmental Science and Pollution Research → Version 1 posted 5 You are reading this latest preprint version Abstract Polylactic acid / Chitosan blends with bio-released Ag nanoparticles nano composite films were prepared for eco-friendly food packaging materials. These food packaging materials were prepared to enhance the biodegradability of packaging materials as well as to increase the shelf life of food products against microbial action. The preparation was made by dissolving PLA and chitosan using suitable solvents with PEG 400. To this blend silver nanoparticles obtained from Tulsi leaves extract were added and homogenized. PLA/Chitosan blend with Ag-NPs were prepared and characterized their spectral, morphological and antimicrobial properties. The spectral and morphological studies showed the blending of PLA/Chitosan as well as Ag nanoparticles in the nanocomposites. The antibacterial activity of the PLA/Chitosan/AgNPs nanocomposites (PCAg) showed greater inhibitory effect against Staphylococcus aureus and also against Escherichia coli. Polylactic acid Chitosan Silver nanoparticles food packaging Leaf extract Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Bio-polymer based food packaging materials have fascinated many researchers for the sustainable growth of pollution free environment. Packaging the food materials in a proper manner ensure the quality as well as its information about its contents. Packaging has become an essential part in food industries. The main objective of food packaging is to preserve its nutritional value against attack from ultraviolet light, oxygen as well as microbial action (Fathima, et.al 2018 ). Most of the food industries use petroleum based plastic materials like polyethylene terephthalate (PET), polypropylene (PP), low- and high-density polyethylene and polyvinyl chloride (PVC) for packaging owing to their strength, flexibility and resistivity (Tajeddin et al.,2020 and Mangaraj et al., 2019 ) The use these polymers for the long term creates accretion of plastic waste in the environment due to disposal and nonbiodegradability. In recent years more than 30% of the solid plastic waste was land-filled (Gan & Chow 2018 and Singh et al., 2020 ) In order to minimise the accumulation of plastic waste materials on the landscape, recycling of plastic has become an inspiring option. Nevertheless, these recycled plastics have chemical contamination than pure plastics due to the interaction of food components with the plastic material (Ingrao et al., 2017 ). The retrieval of plastic from food packaging waste is more tough than glass or metals. The rising environmental issues related to plastics and depletion of fossil fuel driven many researchers to go into an alternative food packaging material (Ahmed et al., 2018 ). For this purpose, compostable and biodegradable polymers have attracted many scientists owing to its renewable, nontoxic, biodegradable and biocompatible properties (Abdulla et al., 2024 and Salarbashi et al., 2019 ). Bio-degradable polymers can be regarded as polysaccharides such as starch, chitosan, cellulose and protein-based polymers are collagen, zein, soy protein. There are some aliphatic polyesters like polybutylene adipate terephthalate (PBAT), polylactic acid (PLA) and so on (Araújo, et al.,2014 and Balaji et al.,2018) Among these biodegradable polymers PLA has attracted much attention by the food packaging industries due to its biodegradability, biocompatibility, thermoplasticity and easy processability. Since it pertains to the class of aliphatic polyesters, it can be produced by chemical polymerisation of lactic acid (LA) monomer (Choksi et al., 2017) as well as by carbohydrate fermentation process (Ahmad et al., 2024 ). The choice of anti-microbial agents for food packaging materials along with bio-polymers must follow the regulations, mainly with reference to toxicological effects (Khaneghah et al., 2018 ) Chitosan, a biodegradable polymer has been researched and used extensively in the field of food packaging films due to its non-toxic and environmentally friendly properties (Singh et al.,2021 and Flórez et al., 2022 ). Chitosan shows an exceptional antioxidant and antimicrobial activity against a wide range of pathogenic and spoilage microorganisms (Chandran et al.,2024 and Friedman et al.,2010). The antimicrobial activity of chitosan has chosen as a probable natural food preservative in food packaging industry. Research on interactions of chitosan with cellular DNA of microorganisms hindered DNA transcription, RNA translation as well as protein synthesis and resulting in weakening of bacterial activities (Raafat et al., 2009). Food safety and quality are the two most important factors with regard to food industries and also for consumers worldwide. Fresh and hygienically prepared food with extended shelf-life and without the use of chemical preservatives are the major concerns in food industries nowadays (Kumar et al., 2021 ). To meet these requirements, antimicrobial packaging methods have been used by industries to keep the freshness as well as the nutritional value of the packed food. In this research work we aim to synthesis such kind of antimicrobial packaging material which will serve as a promising material for the food stuff. AgNPs incorporated antimicrobial packaging along with biodegradable polymers is an able form of active food packaging material would encompass the shelf-life of foods and also decrease the risk of pathogens. We were able to obtain the plant-mediated AgNPs synthesis and their incorporation with biodegradable polymers like Polylactic acid and chitosan. Here we have chosen ( Ocimum sanctum) Tulsi leaves extract for the AgNPs synthesis owing to its remarkable properties such as optical, catalytic, electric and most important medicinal properties (Goyal et al., 2020) AgNPs disturb the cell permeability and cell respiration of the microorganisms by attaching itself with the cell walls and leads to cell death (Ramkumar et al.,2017). Furthermore, the free silver ions can hinder the protein synthesis by denaturing ribosomes in the cytoplasm (Durán et al., 2016 ). Owing to these properties AgNPs have been extensively used in the field of medicinal and packaging industries. Thus, in this research work, we aim to achieve a bio-composite of PLA/Chitosan/Ag-NPs derived from bio-polymers as well as bio-synthesised Ag nanoparticles to optimize the shelf life and its properties. PLA/Chitosan blend with Ag-NPs can be prepared and characterized its spectral, morphological and antimicrobial activities as a natural food packaging material to serve as a potential component for the sustainable development and pollution free environment. Materials and methodology Materials Poly-Lactide (PLA) (Resomer1 L210S) was received from SRL Pvt Ltd and the low molecular weight (MW-50,000-190,000) 75–85% deacetylated chitosan and polyethylene glycol 400 MW were purchased from Sigma-Aldrich (Bangalore, India). Glacial acetic acid and dichloromethane solvents with HPLC grade were procured from Spectrochem Pvt. Ltd., (Mumbai, India). Tulsi leaves were collected from our nearby place. Methodology. Preparation of AgNPs from Tulsi leaves: 20 g of fresh Tulsi leaves were washed thoroughly with double-distilled water and cut into small pieces. The finely chopped leaves were then mixed with 100 mL double distilled water and the mixture was boiled for 30 minutes. After boiling, the mixture was filtered with Whatman Filter paper no.1. 10 mL of aqueous extract of Tulsi leaves was added to 90 mL of silver nitrate solution so that its final concentration to 10 -3 M. The solution is allowed to react at room temperature. After adding with silver nitrate solution, the formation of AgNPs was monitored for every 5 minutes. The reduction of silver nanoparticles by Tulsi leaves was performed as per the report suggested by Saifuddin et al. (Ramteke et al.,2013) Preparation of PLA/Chitosan / AgNPs nanocomposite; Fig 1 shows the representation of preparation of PLA/Chitosan/AgNPs blends. To prepare PLA/Chitosan / AgNPs nanocomposite film, solution of PLA and chitosan were dissolved in different solutions and then added together for blending. 3% (w/v) of PLA is dissolved in dichloromethane solvent and 3% (w/v) chitosan is dissolved in 3% (v/v) acetic acid under room temperature and stirred under magnetic stirrer for 2 hours. Then both solutions are blended homogeneously by continuous stirring in a magnetic stirrer. PEG 400 (w/w) is then added and stirred for 1 hour. 10 mL of silver nanoparticles obtained from Tulsi extract was added and blended homogeneously by continuous stirring in a magnetic stirrer for 2 hours. The prepared blend of PLA/Chitosan/AgNPs is allowed to be cast on a glass petridsh and evaporated at 30 o C for 24 hrs. Table 1 describes the sample code and composition of the prepared PLA/CS/AgNPs nanocomposites. Table-1. The sample code and composition of the prepared PLA/CS/AgNPs nanocomposites. Sample code PLA (wt. %) Chitosan (wt. %) PEG (wt. %) Silver nanoparticle solution 10 -3 M PLA neat 3 ----- ----- ----- PC 3 3 10 ----- PCAg 3 3 10 10 mL PC- Polylactic acid and Chitosan: PCAg- Polylactic acid/Chitosan/AgNPs Characterization Methods. The UV-Visible spectrophotometer (Cary 60, Agilent) was used to measure the absorbance of the synthesised silver nanoparticles in the range of 200–800 nm. Attenuated Total Reflection (ATR-FTIR) spectra of PLA, Chitosan and PLA/Chitosan /AgNPs nanocomposites were recorded using ABB MB3000 FTIR spectrophotometer (Zurich, Switzerland) in the spectral range from 600 to 4000 cm -1 . A background run was done before recording each spectrum. Antibacterial activity of the samples was determined by disc diffusion method on Muller Hinton agar (MHA) medium. The surface morphology of the prepared samples was analysed using Field Emission Scanning Electron Microscope (FESEM) by ZEISS sigma 300 model instrument operating at 20 kV with different magnifications. The X-ray diffraction pattern of PCAg nanocomposites were characterized by Malvern PANanalytical – empyrean series 3 diffractometer operated at 40 kV. The patterns were recorded in the region of 2θ = 5–80 ◦ . Results and Discussion ATR-FTIR spectroscopy. FT-IR spectroscopic technique reveals the structure, functional groups and their interactions in the PLA/chitosan/AgNPs nanocomposites. Fig 2 shows the spectral data arises due to the interactions of PLA/chitosan/AgNPs nanocomposites. The neat PLA displays a sharp absorption band at 1745cm -1 assigned to C=O stretching vibration and the peak at 1452cm -1 is regarded to C-H asymmetric stretching in CH 3 . The absorption peak at 1082 cm -1 is assigned for C-O-C stretching of PLA. The absorption peaks in the region 3400 cm -1 and 2860 cm -1 are due to -OH and -NH stretching vibrations of PEG incorporated PLA/CS matrix. The peak at 1082 cm-1in neat PLA and 1092cm -1 in PLA/PEG/CS are attributed to C-O-C antisymmetric stretching linkage of polysaccharide groups. The peak intensity is increased with the incorporation of chitosan. The absorption peak at 1647 cm -1 for amide in PLA/PEG/CS is close to the spectra of PLA/PEG/CS/AgNPs at 1653 cm -1 specifies that the interactions of green synthesised AgNPs with bio-polymers was not affected the secondary structure after binding with AgNPs (Das et al.,2017). UV-Visible spectral analysis The initial characterization of the AgNPs was conducted using UV-Visible spectroscopy. Silver ions get reduced in to silver nanoparticles was continuously monitored by taking solutions at different concentrations and also at different time intervals by UV-Visible spectrophotometer. The baseline was adjusted using Millipore water as a blank. Fig. 3 shows the colour of the Tulsi leaves extract before and after adding with silver nitrate solution. The colour changes in the leaf extract are shown in Fig 3. The colour changes from light green to brown in the reaction solution indicates the formation of AgNPs during the reduction process. The free electrons in metal nanoparticles mutually vibrate in resonance with light waves, which results a surface plasmon resonance (SPR) absorption band (Mulvaney et al.,1996). This research work, the biosynthesized silver nanoparticles were periodically analysed for UV–vis spectroscopic studies (Cary 60, Agilent) at room temperature operated at a resolution of 1 nm between 250 and 800 nm ranges. The surface plasmon resonance (SPR) of silver nanoparticles produced a peak centered near 430 nm. UV–vis absorbance of reaction mixture was taken from 5 to 20 min. It was observed that absorbance peak was centered near 430 nm indicating reduction of 0.01M AgNO 3 into silver nanoparticles using Tulsi extract. It was also observed that bio-reduction of silver ions into nanoparticles started at 0 min and bio-reduction was completed at almost 20 min indicating rapid biosynthesis of silver nanoparticles. An absorption band at 280 nm and 320 nm is evidenced for Tulsi extract. Antimicrobial activity The current research work examines the antibacterial properties of prepared PLA/CS/AgNPs nanocomposites against Escherichia coli and staphylococcus aureus bacteria. Fig 4 illustrates the incubation zone surrounding the PLA/CS/AgNPs bacterial culture. Table -II Microorganisms/Sample Zone of Inhibition in mm 1 2 Escherichia coli PC 10 20 PC Ag 15 20 Staphylococcus aureus PC 12 22 PC Ag 16 22 Polylactic acid /Chitosan composites shown significant antibacterial properties against Escherichia coli and Staphylococcus aureus. It has the ability to impede the growth of bacteria. The negatively charged cell walls of E. coli would bind to the positively charged -NH 3 + , breaking down their enzymes in the process. The bacteria are subsequently unable to proliferate and eventually become destroyed as the enzymes are unable to generate energy (Rabea et al.,2003 and Liu et al., 2005). From table II, it shows the addition of green synthesised AgNPs to the PLA/CS composite increased the inhibition zone to a higher level in both E. coli and Staphylococcus aureus. AgNPs has exceptional bactericidal activity which can be explained by their interaction with the bacterial strain's peptidoglycan cell wall and plasma membrane (Sondi et al.,2004) Additionally, it has been claimed that AgNPs' interaction with cell walls enhances membrane permeability by producing pits or pores that kill bacteria (Morones et al., 2005). The incorporation of silver nanoparticles with biopolymers for food packaging applications, extended a substantial increase in antibacterial action against common foodborne pathogens like E. Coli and S. Aureus, lead to an increased food shelf life. FESEM analysis The morphology of PLA/Chitosan / AgNPs nanocomposite film has been analysed by FESEM at different magnifications as shown in the Fig 5. The PLA /Chitosan /AgNPs nanocomposite film shows the complete blending of PLA and Chitosan as shown in Figure 5a and 5b. With higher magnification shows some pores and voids in the nanocomposite film. The previous studies also reported that the chitosan incorporated metal nanocomposite blends will lead to porous structure which depends on concentration of chitosan (Ikeda et al.,2014). Fig 5c shows the existence of silver nanoparticles, which is white, spherical in shape blended in the PLA-Chitosan film. The agglomeration of silver nanoparticles is shown in the FESEM images. The incorporation of silver nanoparticles in PLA-Chitosan matrix may introduce amorphous nature in the nanocomposite film resulting in the increased porosity evidenced by the increased antimicrobial activity of nanocomposite film. XRD Analysis. The XRD pattern of the neat PLA, PLA/chitosan and PLA/Chitosan/Ag biosynthesised nanocomposites were studied and shown in Fig 6. The neat biopolymer film as well as PLA/Chitosan blends shown the characteristics 2θ reflections at 21.4 ◦ and 21.6 ◦ owing to the crystalline structures of PLA and chitosan as reported in the literatures (Rubilar et al 2013). After adding biosynthesised silver ions to the nanocomposite, a small increase in the intensity of 16.6 ◦ peak was observed due to the presence of silver ions with fcc crystalline structure confirmed from Braggs diffraction pattern (Vishwajeet Singh et al 2015). Conclusion PLA/Chitosan blends with bio-synthesised AgNPs nanocomposites for potential food packaging material were successfully synthesised and characterized. Ag nanoparticles was effectively synthesised from Tulsi (Ocimum sanctum) extract using 0.01 M AgNO 3 and it was confirmed by UV-visible spectrophotometer, morphology study as well as XRD. The spectral studies show better interactions between Polylactic acid, chitosan and biosynthesised Ag nanoparticles. Antibacterial study showed an increased zone of inhibition value for bio-synthesised Ag nanoparticle incorporated PCAg films against Staphylococcus aureus and E. coli. FESEM study confirmed the presence of silver nanoparticles along with the polymer composition. The prepared polylactic acid/Chitosan /bio-synthesised nanocomposites showed excellent properties specifically the antibacterial property which makes the matrix a suitable for food packaging material which could reduce the food wastage and food spoilage. These nanocomposites could serve as a prominent alternative for the sustainable and pollution free environment. Declarations Ethical Approval - Not applicable Consent to Participate – Not applicable Consent to Publish - Not applicable Data Availability Statement - The datasets used and analysed during this study are available from the corresponding author on reasonable request. Authors Contributions - Study conception and design: Dr. N. Malarvizhi: data collection – Dr Malarvizhi Nithya and Dr. Tharani sundarajan: Analysis and interpretation of results- Dr. Malarvizhi Nithya and Rajeswari Balaji: Draft manuscript preparation – Dr. Malarvizhi Nithya, Rajeswari Balaji, Dr. Tharani sundarajan and Ms. Vennila sivalingam. All authors reviewed the results and approved the final version of the manuscript . Funding – No funds, grants or other support was received. Competing Interests - The authors declare that no competing interests. References Abdulla, S. F., Shams, R., & Dash, K. K. (2024). 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African journal of biotechnology Vol. 14(33), pp. 2554-2567, Cite Share Download PDF Status: Published Journal Publication published 12 Jun, 2025 Read the published version in Environmental Science and Pollution Research → Version 1 posted Editorial decision: Major Revision 12 Dec, 2024 Reviewers agreed at journal 31 Aug, 2024 Reviewers invited by journal 29 Aug, 2024 Editor assigned by journal 06 Aug, 2024 First submitted to journal 04 Aug, 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-4788416","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":346796265,"identity":"01ea8f76-3f77-4d7a-b352-5181bd0e9faa","order_by":0,"name":"Malarvizhi 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College","correspondingAuthor":false,"prefix":"","firstName":"Rajeswari","middleName":"","lastName":"Balaji","suffix":""},{"id":346796267,"identity":"41dce849-0933-40bb-8ca3-32738d1daec4","order_by":2,"name":"Tharani Sundarajan","email":"","orcid":"","institution":"Bharathi Women's College","correspondingAuthor":false,"prefix":"","firstName":"Tharani","middleName":"","lastName":"Sundarajan","suffix":""},{"id":346796268,"identity":"e1374933-f381-4ed6-8560-86948a0c291a","order_by":3,"name":"Vennila Sivalingam","email":"","orcid":"","institution":"Bharathi Women's College","correspondingAuthor":false,"prefix":"","firstName":"Vennila","middleName":"","lastName":"Sivalingam","suffix":""}],"badges":[],"createdAt":"2024-07-23 11:35:15","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4788416/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4788416/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11356-025-36610-1","type":"published","date":"2025-06-12T15:57:36+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":65442578,"identity":"2a4c1fcf-0e84-434b-bf90-be330fa44a29","added_by":"auto","created_at":"2024-09-27 13:22:01","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":144352,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSchematic representation of preparation PLA/Chitosan/AgNPs blends\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4788416/v1/6317377a56e11e7d46526083.png"},{"id":65443534,"identity":"3b7df8a0-b9c3-4cd3-af8d-02d5e79f21e2","added_by":"auto","created_at":"2024-09-27 13:30:01","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":96018,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFTIR spectra of (a) neat PLA (b) PLA/CS (c) PLA/CS/AgNPs nanocomposites\u003c/strong\u003e.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4788416/v1/f38bcd804062d536256b7862.png"},{"id":65443535,"identity":"2fc7110b-b32a-4a91-bdb9-31cff8d0b59c","added_by":"auto","created_at":"2024-09-27 13:30:01","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":211620,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eUV-visible absorption spectra of Tulsi extract and Ag nano particle.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4788416/v1/f14da71803c7f3eaae080ea4.png"},{"id":65442584,"identity":"6d65f7cf-0bbf-4f4e-b357-cc5137307a41","added_by":"auto","created_at":"2024-09-27 13:22:01","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":966166,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAntibacterial activity of PC and PCAg NPs against Escherichia coli and Staphylococcus aureus\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4788416/v1/73d4fa8a18070eef95bed97d.png"},{"id":65442583,"identity":"dc28077e-e95f-48d0-bc09-a8154f1e3c9c","added_by":"auto","created_at":"2024-09-27 13:22:01","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1104715,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFESEM images of PCAg with different magnifications\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-4788416/v1/49b7f1f23ca87e90e54bd298.png"},{"id":65442580,"identity":"af342bdb-1b0f-4f88-8e1d-528b4977d4ff","added_by":"auto","created_at":"2024-09-27 13:22:01","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":34026,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eXRD patterns of PLA neat, PC and PCAg nanocomposite\u003c/strong\u003e.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-4788416/v1/f05640fc1d187839e0b8529b.png"},{"id":84726838,"identity":"12fe37b1-1055-4996-ba06-fee82ee4fd80","added_by":"auto","created_at":"2025-06-16 16:08:31","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3863489,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4788416/v1/41c9419f-f69f-4d06-839c-10631d84118e.pdf"}],"financialInterests":"","formattedTitle":"\u003cp\u003eNovel Pla / Chitosan Blends With Bio-released Ag-nps Nano Composites for Eco-friendly Food Packaging.\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eBio-polymer based food packaging materials have fascinated many researchers for the sustainable growth of pollution free environment. Packaging the food materials in a proper manner ensure the quality as well as its information about its contents. Packaging has become an essential part in food industries. The main objective of food packaging is to preserve its nutritional value against attack from ultraviolet light, oxygen as well as microbial action (Fathima, et.al \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Most of the food industries use petroleum based plastic materials like polyethylene terephthalate (PET), polypropylene (PP), low- and high-density polyethylene and polyvinyl chloride (PVC) for packaging owing to their strength, flexibility and resistivity (Tajeddin et al.,2020 and Mangaraj et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) The use these polymers for the long term creates accretion of plastic waste in the environment due to disposal and nonbiodegradability. In recent years more than 30% of the solid plastic waste was land-filled (Gan \u0026amp; Chow \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2018\u003c/span\u003e and Singh et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2020\u003c/span\u003e)\u003c/p\u003e \u003cp\u003eIn order to minimise the accumulation of plastic waste materials on the landscape, recycling of plastic has become an inspiring option. Nevertheless, these recycled plastics have chemical contamination than pure plastics due to the interaction of food components with the plastic material (Ingrao et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The retrieval of plastic from food packaging waste is more tough than glass or metals. The rising environmental issues related to plastics and depletion of fossil fuel driven many researchers to go into an alternative food packaging material (Ahmed et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). For this purpose, compostable and biodegradable polymers have attracted many scientists owing to its renewable, nontoxic, biodegradable and biocompatible properties (Abdulla et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2024\u003c/span\u003e and Salarbashi et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Bio-degradable polymers can be regarded as polysaccharides such as starch, chitosan, cellulose and protein-based polymers are collagen, zein, soy protein. There are some aliphatic polyesters like polybutylene adipate terephthalate (PBAT), polylactic acid (PLA) and so on (Ara\u0026uacute;jo, et al.,2014 and Balaji et al.,2018) Among these biodegradable polymers PLA has attracted much attention by the food packaging industries due to its biodegradability, biocompatibility, thermoplasticity and easy processability. Since it pertains to the class of aliphatic polyesters, it can be produced by chemical polymerisation of lactic acid (LA) monomer (Choksi et al., 2017) as well as by carbohydrate fermentation process (Ahmad et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe choice of anti-microbial agents for food packaging materials along with bio-polymers must follow the regulations, mainly with reference to toxicological effects (Khaneghah et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) Chitosan, a biodegradable polymer has been researched and used extensively in the field of food packaging films due to its non-toxic and environmentally friendly properties (Singh et al.,2021 and Fl\u0026oacute;rez et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Chitosan shows an exceptional antioxidant and antimicrobial activity against a wide range of pathogenic and spoilage microorganisms (Chandran et al.,2024 and Friedman et al.,2010). The antimicrobial activity of chitosan has chosen as a probable natural food preservative in food packaging industry. Research on interactions of chitosan with cellular DNA of microorganisms hindered DNA transcription, RNA translation as well as protein synthesis and resulting in weakening of bacterial activities (Raafat et al., 2009). Food safety and quality are the two most important factors with regard to food industries and also for consumers worldwide. Fresh and hygienically prepared food with extended shelf-life and without the use of chemical preservatives are the major concerns in food industries nowadays (Kumar et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). To meet these requirements, antimicrobial packaging methods have been used by industries to keep the freshness as well as the nutritional value of the packed food.\u003c/p\u003e \u003cp\u003eIn this research work we aim to synthesis such kind of antimicrobial packaging material which will serve as a promising material for the food stuff. AgNPs incorporated antimicrobial packaging along with biodegradable polymers is an able form of active food packaging material would encompass the shelf-life of foods and also decrease the risk of pathogens. We were able to obtain the plant-mediated AgNPs synthesis and their incorporation with biodegradable polymers like Polylactic acid and chitosan.\u003c/p\u003e \u003cp\u003eHere we have chosen (\u003cem\u003eOcimum sanctum)\u003c/em\u003e Tulsi leaves extract for the AgNPs synthesis owing to its remarkable properties such as optical, catalytic, electric and most important medicinal properties (Goyal et al., 2020) AgNPs disturb the cell permeability and cell respiration of the microorganisms by attaching itself with the cell walls and leads to cell death (Ramkumar et al.,2017). Furthermore, the free silver ions can hinder the protein synthesis by denaturing ribosomes in the cytoplasm (Dur\u0026aacute;n et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Owing to these properties AgNPs have been extensively used in the field of medicinal and packaging industries. Thus, in this research work, we aim to achieve a bio-composite of PLA/Chitosan/Ag-NPs derived from bio-polymers as well as bio-synthesised Ag nanoparticles to optimize the shelf life and its properties. PLA/Chitosan blend with Ag-NPs can be prepared and characterized its spectral, morphological and antimicrobial activities as a natural food packaging material to serve as a potential component for the sustainable development and pollution free environment.\u003c/p\u003e "},{"header":"Materials and methodology","content":"\u003cp\u003e\u003cstrong\u003eMaterials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePoly-Lactide (PLA) (Resomer1 L210S) was received from SRL Pvt Ltd and the low molecular weight (MW-50,000-190,000) 75\u0026ndash;85% deacetylated chitosan and polyethylene glycol 400 MW were purchased from Sigma-Aldrich (Bangalore, India). Glacial acetic acid and dichloromethane solvents with HPLC grade were procured from Spectrochem Pvt. Ltd., (Mumbai, India). Tulsi leaves were collected from our nearby place.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethodology.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of AgNPs from Tulsi leaves:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e20 g of fresh Tulsi leaves were washed thoroughly with double-distilled water and cut into small pieces. The finely chopped leaves were then mixed with 100 mL double distilled water and the mixture was boiled for 30 minutes. After boiling, the mixture was filtered with Whatman Filter paper no.1. 10 mL of aqueous extract of Tulsi leaves was added to 90 mL of silver nitrate solution so that its final concentration to 10\u003csup\u003e-3\u003c/sup\u003e M. The solution is allowed to react at room temperature. After adding with silver nitrate solution, the formation of AgNPs was monitored for every 5 minutes. The reduction of silver nanoparticles by Tulsi leaves was performed as per the report suggested by Saifuddin et al. \u0026nbsp;(Ramteke et al.,2013)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of PLA/Chitosan / AgNPs nanocomposite;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFig 1 shows the representation of preparation of PLA/Chitosan/AgNPs blends. To prepare PLA/Chitosan \u003cstrong\u003e/\u003c/strong\u003eAgNPs nanocomposite film, solution of PLA and chitosan were dissolved in different solutions and then added together for blending. \u0026nbsp;3% (w/v) of PLA is dissolved in dichloromethane solvent and 3% (w/v) chitosan is dissolved in 3% (v/v) acetic acid under room temperature and stirred under magnetic stirrer for 2 hours. Then both solutions are blended homogeneously by continuous stirring in a magnetic stirrer. PEG 400 (w/w) is then added and stirred for 1 hour. 10 mL of silver nanoparticles obtained from Tulsi extract was added and blended homogeneously by continuous stirring in a magnetic stirrer for 2 hours. The prepared blend of PLA/Chitosan/AgNPs is allowed to be cast on a glass petridsh and evaporated at 30\u003csup\u003eo\u003c/sup\u003eC for 24 hrs. Table 1 describes the sample code and composition of the prepared PLA/CS/AgNPs nanocomposites.\u003c/p\u003e\n\u003cp\u003eTable-1. The sample code and composition of the prepared PLA/CS/AgNPs nanocomposites.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"614\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSample code\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePLA\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(wt. %)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eChitosan\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(wt. %)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePEG\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(wt. %)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6966%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSilver nanoparticle solution\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;10\u003csup\u003e-3\u003c/sup\u003eM\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003ePLA neat\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003e-----\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003e-----\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6966%;\"\u003e\n \u003cp\u003e-----\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003ePC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6966%;\"\u003e\n \u003cp\u003e-----\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003ePCAg\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.5759%;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21.6966%;\"\u003e\n \u003cp\u003e10 mL\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003ePC- Polylactic acid and Chitosan: \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; PCAg- Polylactic acid/Chitosan/AgNPs\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCharacterization Methods.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe UV-Visible spectrophotometer (Cary 60, Agilent) was used to measure the absorbance of the synthesised silver nanoparticles in the range of 200\u0026ndash;800 nm. Attenuated Total Reflection (ATR-FTIR) spectra of PLA, Chitosan and PLA/Chitosan /AgNPs nanocomposites were recorded using ABB MB3000 FTIR spectrophotometer (Zurich, Switzerland) in the spectral range from 600 to 4000 cm\u003csup\u003e-1\u003c/sup\u003e. A background run was done before recording each spectrum. Antibacterial activity of the samples was determined by disc diffusion method on Muller Hinton agar (MHA) medium. The surface morphology of the prepared samples was analysed using\u0026nbsp;Field Emission Scanning Electron Microscope (FESEM) by ZEISS sigma 300 model instrument operating at 20 kV with different magnifications.\u0026nbsp;The X-ray diffraction pattern of PCAg nanocomposites were characterized by Malvern PANanalytical \u0026ndash; empyrean series 3 diffractometer operated at 40 kV. The patterns were recorded in the region of 2\u0026theta; = 5\u0026ndash;80\u003csup\u003e◦\u003c/sup\u003e.\u003c/p\u003e"},{"header":"Results and Discussion","content":"\u003cp\u003e\u003cstrong\u003eATR-FTIR spectroscopy.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFT-IR spectroscopic technique reveals the structure, functional groups and their interactions in the PLA/chitosan/AgNPs nanocomposites. Fig 2 shows the spectral data arises due to the interactions of PLA/chitosan/AgNPs nanocomposites.\u003c/p\u003e\n\u003cp\u003eThe neat PLA displays a sharp absorption band at 1745cm\u003csup\u003e-1\u003c/sup\u003e assigned to C=O stretching vibration and the peak at 1452cm\u003csup\u003e-1\u003c/sup\u003e is regarded to C-H asymmetric stretching in CH\u003csub\u003e3\u003c/sub\u003e. The absorption peak at 1082 cm\u003csup\u003e-1\u003c/sup\u003e is assigned for C-O-C stretching of PLA. The absorption peaks in the region 3400 cm\u003csup\u003e-1\u0026nbsp;\u003c/sup\u003eand 2860 cm\u003csup\u003e-1\u003c/sup\u003e are due \u0026nbsp; \u0026nbsp; to -OH and -NH stretching vibrations of PEG incorporated PLA/CS matrix. The peak at 1082 cm-1in neat PLA and 1092cm\u003csup\u003e-1\u003c/sup\u003e in PLA/PEG/CS are attributed to C-O-C antisymmetric stretching linkage of polysaccharide groups. The peak intensity is increased with the incorporation of chitosan. The absorption peak at 1647 cm\u003csup\u003e-1\u003c/sup\u003e for amide in PLA/PEG/CS is close to the spectra of PLA/PEG/CS/AgNPs at 1653 cm\u003csup\u003e-1\u003c/sup\u003e specifies that the interactions of green synthesised AgNPs with bio-polymers was not affected the secondary structure after binding with AgNPs (Das et al.,2017).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eUV-Visible spectral analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe initial characterization of the AgNPs was conducted using UV-Visible spectroscopy. Silver ions get reduced in to silver nanoparticles was continuously monitored by taking solutions at different concentrations and also at different time intervals by UV-Visible spectrophotometer. The baseline was adjusted using Millipore water as a blank.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFig. 3 shows the colour of the Tulsi leaves extract before and after adding with silver nitrate solution. The colour changes in the leaf extract are shown in Fig 3. The colour changes from light green to brown in the reaction solution indicates the formation of AgNPs during the reduction process.\u0026nbsp;The free electrons in metal nanoparticles mutually vibrate in resonance with light waves, which results a surface plasmon resonance (SPR) absorption band (Mulvaney et al.,1996). This research work, the biosynthesized silver nanoparticles were periodically analysed for UV\u0026ndash;vis spectroscopic studies (Cary 60, Agilent) at room temperature operated at a resolution of 1 nm between 250 and 800 nm ranges. The surface plasmon resonance (SPR) of silver nanoparticles produced a peak centered near 430 nm. UV\u0026ndash;vis absorbance of reaction mixture was taken from 5 to 20 min. It was observed that absorbance peak was centered near 430 nm indicating reduction of 0.01M AgNO\u003csub\u003e3\u003c/sub\u003e into silver nanoparticles using Tulsi extract. It was also observed that bio-reduction of silver ions into nanoparticles started at 0 min and bio-reduction was completed at almost 20 min indicating rapid biosynthesis of silver nanoparticles. An absorption band at 280 nm and 320 nm is evidenced for Tulsi extract.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntimicrobial activity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe current research work examines the antibacterial properties of prepared PLA/CS/AgNPs nanocomposites against Escherichia coli\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eand staphylococcus aureus bacteria. Fig 4 illustrates the incubation zone surrounding the PLA/CS/AgNPs bacterial culture.\u003c/p\u003e\n\u003cp\u003eTable -II \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 210px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMicroorganisms/Sample\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 391px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eZone of Inhibition in mm\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" style=\"width: 601px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eEscherichia coli \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 210px;\"\u003e\n \u003cp\u003ePC\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 210px;\"\u003e\n \u003cp\u003ePC Ag\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" style=\"width: 601px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eStaphylococcus aureus \u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 210px;\"\u003e\n \u003cp\u003ePC\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 210px;\"\u003e\n \u003cp\u003ePC Ag\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 196px;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003ePolylactic acid /Chitosan composites shown significant antibacterial properties against Escherichia coli and\u0026nbsp;Staphylococcus aureus. It has\u0026nbsp;the ability to impede the growth of bacteria. The negatively charged cell walls of E. coli would bind to the positively charged -NH\u003csub\u003e3\u003c/sub\u003e \u003cstrong\u003e\u003csup\u003e+\u003c/sup\u003e\u003c/strong\u003e, breaking down their enzymes in the process. The bacteria are subsequently unable to proliferate and eventually become destroyed as the enzymes are unable to generate energy (Rabea et al.,2003 and Liu et al., 2005). From table II, it shows the addition of green synthesised AgNPs to the PLA/CS composite increased the inhibition zone to a higher level in both E. coli and\u0026nbsp;Staphylococcus aureus.\u0026nbsp;AgNPs has exceptional bactericidal activity which can be explained by their interaction with the bacterial strain\u0026apos;s peptidoglycan cell wall and plasma membrane (Sondi et al.,2004)\u0026nbsp;Additionally, it has been claimed that AgNPs\u0026apos; interaction with cell walls enhances membrane permeability by producing pits or pores that kill bacteria (Morones et al., 2005).\u0026nbsp;The incorporation of silver nanoparticles with biopolymers for food packaging applications, extended a substantial increase in antibacterial action against\u0026nbsp;common foodborne pathogens like E. Coli and S. Aureus, lead to an increased food shelf life.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFESEM analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe morphology of PLA/Chitosan \u003cstrong\u003e/\u003c/strong\u003eAgNPs nanocomposite film has been analysed by FESEM at different magnifications as shown in the Fig 5. The PLA /Chitosan /AgNPs nanocomposite film shows the complete blending of PLA and Chitosan as shown in Figure 5a and 5b. \u0026nbsp;With higher magnification shows some pores and voids in the nanocomposite film. \u0026nbsp; \u0026nbsp;The previous studies also reported that the chitosan incorporated metal nanocomposite blends will lead to porous structure which depends on concentration of chitosan (Ikeda et al.,2014). \u0026nbsp;Fig 5c shows the existence of silver nanoparticles, which is white, spherical in shape blended in the PLA-Chitosan film. \u0026nbsp;The agglomeration of silver nanoparticles is shown in the FESEM images. The incorporation of silver nanoparticles in PLA-Chitosan matrix may introduce amorphous nature in the nanocomposite film resulting in the increased porosity evidenced by the increased antimicrobial activity of nanocomposite film.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eXRD Analysis.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe XRD pattern of the neat PLA, PLA/chitosan and PLA/Chitosan/Ag biosynthesised nanocomposites were studied and shown in Fig 6. \u0026nbsp;The neat biopolymer film as well as PLA/Chitosan blends shown the characteristics 2\u0026theta; reflections at 21.4\u003csup\u003e◦\u003c/sup\u003e and 21.6\u003csup\u003e◦\u003c/sup\u003e owing to the crystalline structures of PLA and chitosan as reported in the literatures (Rubilar et al 2013). After adding biosynthesised silver ions to the nanocomposite, a small increase in the intensity of 16.6\u003csup\u003e◦\u003c/sup\u003e peak was observed due to the presence of silver ions with fcc crystalline structure confirmed from Braggs diffraction pattern (Vishwajeet Singh et al 2015).\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003ePLA/Chitosan blends with bio-synthesised AgNPs nanocomposites for potential food packaging material were successfully synthesised and characterized. Ag nanoparticles was effectively synthesised from Tulsi (Ocimum sanctum) extract using 0.01 M AgNO\u003csub\u003e3\u003c/sub\u003e and it was confirmed by UV-visible spectrophotometer, morphology study as well as XRD. The spectral studies show better interactions between Polylactic acid, chitosan and biosynthesised Ag nanoparticles. Antibacterial study showed an increased zone of inhibition value for bio-synthesised Ag nanoparticle incorporated PCAg films against Staphylococcus aureus and E. coli. FESEM study confirmed the presence of silver nanoparticles along with the polymer composition. The prepared polylactic acid/Chitosan /bio-synthesised nanocomposites showed excellent properties specifically the antibacterial property which makes the matrix a suitable for food packaging material which could reduce the food wastage and food spoilage. These nanocomposites could serve as a prominent alternative for the sustainable and pollution free environment.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthical Approval \u003c/strong\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp;- \u0026nbsp; \u0026nbsp;Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Participate \u003c/strong\u003e\u0026ndash; Not applicable\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Publish \u0026nbsp;\u003c/strong\u003e - \u0026nbsp; \u0026nbsp; Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement\u003c/strong\u003e - The datasets used and analysed during this study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors Contributions \u003c/strong\u003e- Study conception and design: Dr. N. Malarvizhi: \u0026nbsp;data collection \u0026ndash; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Dr Malarvizhi Nithya and Dr. Tharani sundarajan: Analysis and interpretation of results- \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Dr. Malarvizhi Nithya and Rajeswari Balaji: Draft manuscript preparation \u0026ndash; Dr. Malarvizhi Nithya, Rajeswari Balaji, Dr. Tharani sundarajan and Ms. Vennila sivalingam. \u0026nbsp;All authors reviewed the results and approved the final version of the manuscript\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding \u003c/strong\u003e\u0026ndash; No funds, grants or other support was received.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e - The authors declare that no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbdulla, S. 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Biosynthesis of silver nanoparticles by plants crude extracts and their characterization using UV, XRD, TEM and EDX. \u003cem\u003eAfrican journal of biotechnology\u003c/em\u003e Vol. 14(33), pp. 2554-2567, \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"environmental-science-and-pollution-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"espr","sideBox":"Learn more about [Environmental Science and Pollution Research](https://www.springer.com/journal/11356)","snPcode":"11356","submissionUrl":"https://submission.nature.com/new-submission/11356/3","title":"Environmental Science and Pollution Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Polylactic acid, Chitosan, Silver nanoparticles, food packaging, Leaf extract","lastPublishedDoi":"10.21203/rs.3.rs-4788416/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4788416/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003ePolylactic acid / Chitosan blends with bio-released Ag nanoparticles nano composite films were prepared for eco-friendly food packaging materials. These food packaging materials were prepared to enhance the biodegradability of packaging materials as well as to increase the shelf life of food products against microbial action. The preparation was made by dissolving PLA and chitosan using suitable solvents with PEG 400. To this blend silver nanoparticles obtained from Tulsi leaves extract were added and homogenized. PLA/Chitosan blend with Ag-NPs were prepared and characterized their spectral, morphological and antimicrobial properties. The spectral and morphological studies showed the blending of PLA/Chitosan as well as Ag nanoparticles in the nanocomposites. 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