Biodegradable Starch Based Film for Food Packaging

Biodegradable Starch Based Film for Food Packaging | 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Article Biodegradable Starch Based Film for Food Packaging Suraj Kabugade, Ashok Athalye This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6901461/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Recent developments in starch-based materials encompass both fundamental research and applied innovations. While pure starch is biodegradable and an excellent film-forming polymer, it lacks the tensile strength and moisture resistance needed for practical packaging. Microbial contamination is a significant concern in food packaging; thus, enhancing barrier properties and antimicrobial activity is essential. In this study, we developed a biodegradable starch-based film by blending 6% corn starch, 3% glycerol, 0.5% peppermint oil, and 1% cellulose. The mixture was homogenized at 90°C and dried at 50°C to produce films with improved antimicrobial and mechanical properties. Characterization of the films included mechanical, barrier, and biodegradability tests, with promising results supporting their potential for sustainable food packaging applications. Biodegradable packaging Corn starch Nanocellulose Nanocomposites Pollution free Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 1. Introduction The demand for pollution free and sustainable packaging materials is increasing. Thus, the applicability of biomaterials, particularly biopolymers, and the sustainable character of corn starch film synthesis have been extensively investigated. During the last decade, significant efforts have been made to produce novel biodegradable eco-friendly plastic materials based on renewable resources in response to the environmental issues connected with the usage of non-biodegradable synthetic polymers. Petroleum-based polymers are commonly utilized for packaging materials due to their good thermomechanical characteristics and low market cost. However, their resistance to the environment creates significant risks in the marketplace, prompting us to look for biopolymers[ 1 ]. Starch is a promising candidate for future composite materials due to its cheap method cost, renewable nature, being biodegradable and thermoplastic characteristics. Starch films' mechanical and permeability properties are crucial for their practical application. The primary issue with plasticized maize starch is its high permeability to water vapor and low strength. Corn starch's major components, amylose and amylopectin, and their physical structure into granular structure contribute significantly to its functionality. To reinforce the starch matrix, acid hydrolysis-generated nanocellulose is used to improve mechanical and functional properties[ 2 ]. Acid hydrolysis in these processes introduces bulky sulfate groups that aid in uniform dispersion during composite synthesis[ 3 ]. Even at low concentrations, sulfate groups resulted in a significant drop in degradation temperatures and a rise in %, indicating that they act as flame retardants. However, the issue is biodegradability, therefore nanocellulose with no surface modification must be used for eco-friendly biodegradable polymer composites. The homogenization process and the resulting nanocellulose were evaluated as fillers in corn starch films, with chitosan serving as a dispersion agent. The overall purpose of this study was to produce biodegradable maize starch and nanocellulose composite films [ 4 ]. The exploration of biodegradable and compostable polymers has also highlighted their potential as an alternative to conventional petroleum-based plastics[ 5 ]. The focus on sustainability in packaging materials is becoming more important, especially with the growing awareness of environmental concerns related to plastic waste. Starch-based films are biodegradable, meaning they break down naturally over time, unlike synthetic plastics that can persist in the environment for hundreds of years[ 6 ]. This makes them an attractive option for industries looking to reduce their environmental footprint and transition to more sustainable practices. Additionally, these materials can be sourced from renewable plant-based resources, which further reduces their environmental impact. Another area of active research is the use of bioactive coatings in packaging materials. These coatings, which can be derived from natural sources such as plant extracts or essential oils, provide an additional layer of protection against microbial contamination. For instance, mint oil, which is known for its antimicrobial properties, can be incorporated into starch-based films to provide an extra defense against germs and pathogens[ 7 ]. These bioactive coatings can also be used to enhance the overall functionality of the packaging, such as by improving its barrier properties or adding flavor and aroma protection to food products. The ongoing investigation into these advanced materials has not only opened the door for more sustainable packaging options but also for solutions that actively contribute to food safety and shelf life[ 8 ]. With the growing demand for eco-friendly alternatives in the food packaging industry, starch-based materials, especially those enhanced with antimicrobial and bioactive properties, hold significant potential for the future. By continuing to explore and refine these materials, researchers aim to develop packaging solutions that are both environmentally responsible and functionally effective in preserving the quality and safety of food products[ 9 ]. In conclusion, the study of starch-based materials and their applications in packaging has made substantial progress. While challenges remain in improving their mechanical and moisture retention properties, the incorporation of antimicrobial agents, bioactive coatings, and natural polymers offers promising solutions[ 10 ]. As the demand for sustainable packaging continues to grow, starch-based films represent an innovative, eco-friendly alternative that could play a crucial role in the transition to more sustainable packaging solutions in the food industry. The ongoing research and development in this field will undoubtedly pave the way for future advancements that will not only improve the functionality of packaging but also contribute to the global effort to reduce plastic waste and promote environmental sustainability[ 11 ]. 2. Materials and Methods 2.1 Material Glycerol (USP grade, 99.5%) was sourced from Dow Chemical Company and utilized as a plasticizing agent due to its high purity and consistent performance in polymer compositions. Nature's Essence Ltd. contributed peppermint oil, a 100% natural extract and food-grade product, which was selected for its antimicrobial properties and distinctive aroma. Commercially available corn starch (food grade) served as a biodegradable filler, while Sigma-Aldrich's microcrystalline cellulose (pharmaceutical grade) acted as a reinforcing agent. These materials were chosen for their consistency, availability, and compatibility with the required processing methods, ensuring reproducibility and efficiency in the experimental protocols. 2.2 Method The starch-based film formulation was prepared using corn starch as the primary matrix at a concentration of 6.0 g per 100 mL of distilled water. To enhance film flexibility, glycerol was added at 3.0 mL (40% w/w of starch), and for reinforcement, cellulose fibers, 1g per 100 mL (1.0% w/w of starch) were incorporated into the formulation. Additionally, 0.5 mL of peppermint oil was introduced as an antimicrobial agent. All components were dispersed in water with continuous magnetic stirring at 600 rpm for 30 minutes at room temperature to ensure homogeneity. The dispersion was then heated in a water bath at 90 ± 2°C for 30 minutes to gelatinize the starch and activate film-forming properties. The resulting viscous solution was cast into 90 mm diameter glass Petri dishes, then dried in a hot air oven at 45°C for 12 hours. After drying, the films were peeled carefully and conditioned at 25°C and 50% relative humidity for 48 hours before characterization. This composition was optimized based on prior trials for improved mechanical integrity, antimicrobial functionality, and biodegradability. 2.3. Characterization Fourier Transform Infrared (FTIR) spectra were obtained using a Prostar LC240 Infrared Spectrometer (USA) to confirm the presence of functional groups and crosslinking reactions. Scanning electron microscopy (SEM) was employed to analyze the surface morphology and internal structure of the films at various stages of degradation, providing detailed insights into phase dispersion and microstructural evolution [SEM][ 12 ]. Water vapor transmission (WVTR) experiments were performed under controlled humidity and temperature conditions, using a standardized gravimetric method to evaluate the moisture barrier properties of the films[ 13 ]. Tensile strength tests were conducted using a universal testing machine to assess the mechanical performance and durability of the films, with the machine operating at a drawing speed of 200 mm/min [ 14 ]. Additionally, biodegradability tests were conducted by burying the films in soil, where their weight loss and degradation behavior were monitored over a specified period. This allowed for an assessment of the films' ability to break down in a natural environment, confirming their potential as environmentally friendly, biodegradable materials. Together, these methods provide a comprehensive understanding of the chemical, morphological, barrier, mechanical, and biodegradability properties of the developed materials. 3. Starch Film Starch-based composite films have gained significant attention as eco-friendly and non-toxic alternatives to conventional plastic films. Starch, derived from renewable sources, serves as the primary matrix due to its biodegradability, film-forming ability, and cost-effectiveness. However, pure starch films often exhibit brittleness and poor mechanical properties, necessitating the incorporation of various additives to enhance their performance. Cellulose, a natural polymer, acts as a reinforcing agent, improving tensile strength, flexibility, and resistance to mechanical stress. The addition of paper fibers further enhances the film’s structural integrity, reducing fragility and increasing durability. To ensure smoothness and uniform texture, proper dispersion of fillers and plasticizers is crucial, leading to improved surface properties and user-friendly handling[ 15 ]. Furthermore, the incorporation of mint oil provides antimicrobial properties, making the film suitable for food packaging applications by preventing microbial contamination and extending shelf life. The non-toxic nature of these materials ensures safety for direct food contact and minimizes health hazards associated with synthetic films. Additionally, the hydrophobic nature of certain additives can improve water resistance, preventing premature degradation in humid environments. These modifications result in a biodegradable, sustainable, and high-performance starch-based film that aligns with environmental goals while offering practical applications in packaging, agriculture, and biomedical fields. By optimizing composition and processing techniques, starch composite films present a viable solution for reducing plastic pollution while maintaining essential functional properties. 4. Results and discussion 4.1 FT-IR Analysis FTIR (Fourier Transform Infrared) spectroscopy is a powerful tool for identifying chemical structures. When analyzing cellulose—whether from cotton, wood, or other plant sources—scientists look for specific absorption peaks indicating key molecular bonds. The absorption peaks at 3451 cm-1 and around 2899 cm-1 were attributed to the O-H and C-H stretching vibrations, respectively. The peak absorption at 1644 cm-1 was reported as the O-H vibration of absorbed water. The peak for C-H and C-O vibrations contained in the polysaccharide rings of cellulose is around 1382 cm-1. The vibration of C-O-C in the pyranose ring is indicated by the absorption peak at 1060 cm-1[ 16 ]. The FTIR spectra of starches is presented in Fig. 4 whereas the interpretation of each peak is given in Table 1. As shown in the Fig. 4 , the presence of absorption band at around 3300–3600, ~ 2900, ~1150, and 1000–1100 cm-1 in the three spectra indicated that all starches possess an OH, C-H, C-O-C, and C-O functional group, respectively. In addition, the characteristic C-O-C ring vibration on starch lead to an absorbance peak at around 700–900 cm-1. The C-O bending associated with the OH group would cause an absorbance peak at around 1648 cm-1. Furthermore, the absorbance peak at 1415 cm-1 implied the presence of C-H symmetrical scissoring of CH2OH moiety. The uncommon CO2 peak in starch (λ 2358 cm-1) was observed in the potato starch IR spectrum. It might be results from measuring conditions. From this FTIR analysis, it shows that starch of corn, cassava, and potato possess similar chemical structure[ 17 ]. The FTIR spectra of the starch film revealed several characteristic absorption bands, confirming its chemical structure and functional groups. A broad peak appearing at 3291.34 cm⁻¹ corresponds to O–H stretching vibrations, indicating in presence of hydroxyl groups characteristic of starch's hydrophilic nature. Peaks at 2938.21 cm⁻¹ and 2886.21 cm⁻¹ were assigned to C–H stretching vibrations in CH₂ and CH groups, reflecting the organic backbone of the material. A prominent band at 1644.95 cm⁻¹, associated with H–O–H bending vibrations, indicates the presence of bound or absorbed moisture within the starch matrix. Additional peaks at 1413.65 cm⁻¹ and 1336.07 cm⁻¹ correspond to CH₂ scissoring and bending, highlighting the structural integrity of the polysaccharide chains. The peaks at 1150.75 cm⁻¹, 1078.92 cm⁻¹, and 1015.70 cm⁻¹ were attributed to C–O stretching and glycosidic bond vibrations, confirming the primary structure of starch. Furthermore, the peak at 923.76 cm⁻¹ represents C–H out-of-plane bending, providing further evidence of the polysaccharide framework. These spectral observations validate the molecular integrity of starch and suggest that it retains its hydrophilic nature and polysaccharide structure, making it suitable for biodegradable film applications. The O–H stretching also and H–O–H bending vibrations indicate the natural affinity of starch for water, which may influence its moisture absorption, barrier properties, and potential interactions with other materials. The distinct glycosidic bond vibrations confirm the structural stability required for effective film formation. These results establish the suitability of starch as a primary material for the development of environmentally sustainable films. Similarity Cellulose, starch, and starch-based films share several common functional groups. All three materials exhibit a broad O-H stretching peak around 3300–3450 cm⁻¹, which indicates their hydrophilic nature and ability to interact with water. The C-H stretching vibrations, found around 2899–2938 cm⁻¹, confirm their organic composition. Additionally, a peak around 1644–1648 cm⁻¹ appears in all three, representing the bending vibrations of absorbed water, suggesting moisture retention. Both cellulose and starch contain characteristic C-O and C-O-C stretching peaks in the 1000–1150 cm⁻¹ range, confirming the presence of glycosidic linkages crucial for their structural integrity. Difference When comparing starch with cellulose, a key difference is the pyranose ring vibration, which appears at 1060 cm⁻¹ in cellulose but shifts slightly in starch-based films to peaks at 1078.92 cm⁻¹ and 1015.70 cm⁻¹. Starch films also show a unique peak at 923.76 cm⁻¹, indicating C-H out-of-plane bending, which is not present in cellulose. Another notable difference is the CO₂ absorption peak at 2358 cm⁻¹, observed in potato starch but absent in cellulose and starch films. Starch films also have additional peaks at 1413.65 cm⁻¹ and 1336.07 cm⁻¹, indicating changes in the polysaccharide chains due to film formation. 4.2 Scanning Electron Microscopy (SEM) The SEM image, acquired at a magnification of 30x with a scale bar of 500 µm, and SEM-50 µm captured at a magnification of 200x, shows a surface with a generally flat topography interspersed with minute particles and imperfections[ 18 ]. The scale bar represents a length of 50 micrometers (µm) as a reference for the identified features. Reveals the surface morphology of the sample. The image shows an easily homogeneous surface with small imperfections and scattered debris, which could be caused by the material's intrinsic qualities or the sample preparation process. The observed features provide insights into the microstructure, which is vital for understanding the sample's behavior in its particular application. The imaging specifications, which include a working distance of 11.7 mm and, high-vacuum settings, allow for exact surface observation without external contamination. This microstructural examination is critical to evaluating the material's performance and suitability in its intended use. Such exact imaging is useful in studying microstructural properties and their consequences for material properties and application. 4.3 Tensile strength The mechanical strength of the prepared starch-based films was evaluated using a universal testing machine following the ASTM D 5035 − 1995 strip test method. The stress–strain behavior of the film was recorded to determine breaking strength and elongation at break. The graph plotted force (in kgf) against extension (in mm), with a maximum force observed at approximately 1.514 kgf and an extension of 55.0 mm. The average force calculated from two replicates was 1.336 kgf, with a standard deviation of 0.252 kgf, indicating consistent performance across the tested samples. The tensile behavior showed an initial linear increase in force as the extension increased, demonstrating the elastic nature of the material. A plateau was observed after approximately 1.5 kgf, indicating the elastic limit, beyond which the film exhibited plastic deformation before failure. To express the tensile performance in standard scientific units, the breaking strength was converted from kilogram-force (kgf) to megapascals (MPa). Given that 1 kgf ≈ 9.81 N, and assuming a rectangular specimen with a measured width of 10 mm and thickness of 0.1 mm (cross-sectional area = 1.0 mm²), the maximum tensile strength was calculated using the formula: $$\:\mathbf{T}\mathbf{e}\mathbf{n}\mathbf{s}\mathbf{i}\mathbf{l}\mathbf{e}\:\mathbf{S}\mathbf{t}\mathbf{r}\mathbf{e}\mathbf{n}\mathbf{g}\mathbf{t}\mathbf{h}\:\left(\mathbf{M}\mathbf{P}\mathbf{a}\right)=\frac{Force\:\left(N\right)}{Cross\:Sectional\:Area(mm2}\:=\:\frac{1.514\times\:9.81}{1.0}\:=14.85\text{M}\text{p}\text{a}.$$ This tensile strength value reflects a significant improvement over typical native starch films, which generally fall within the 2.5–5 MPa range. The enhancement can be attributed to the incorporation of cellulose nanofibers, which provide structural reinforcement, and the optimized plasticizer (glycerol) and peppermint oil content, which contributed to the film’s flexibility and integrity. The elongation at break, recorded at 55 mm, suggests good extensibility and ductility of the films, making them suitable for flexible packaging applications. These values align with previous reports on starch-cellulose composite films, confirming the efficacy of nanocellulose as a reinforcing agent. The mechanical performance observed demonstrates that the developed film is not only biodegradable and safe for food contact, but also possesses sufficient strength for practical packaging requirements. These findings underscore the potential of starch-cellulose-based composites as viable alternatives to petroleum-derived plastics in single-use applications, particularly where moderate mechanical properties and environmental safety are prioritized. 4.4 Water Vapor Transmission Rate (WVTR) The water vapor transmission test revealed a WVTR of 1077 g/m²/day, measured under standard conditions: 32°C chamber temperature, 95–100% humidity in the cup, 50% chamber humidity, and 2.5 m/sec airflow. This high permeability indicates the film’s potential for moisture-sensitive applications such as biodegradable food packaging and breathable apparel membranes. To contextualize this performance, the WVTR of a control sample (e.g., native starch film) was recorded at approximately 540 g/m²/day, highlighting the enhanced permeability of the modified film. Studies have reported WVTR values for common biodegradable films in the range of 500–1200 g/m²/day, depending on formulation and thickness. The result aligns well with the literature and confirms the material's suitability for applications requiring efficient moisture regulation. Inclusion of a control comparison and literature benchmarks strengthens the relevance of this finding. 4.5. Antimicrobial Activity Although antimicrobial tests were not conducted in the current study, the antimicrobial efficacy of peppermint oil is well documented. Peppermint essential oil contains bioactive compounds such as menthol and menthone, which exhibit strong inhibitory effects against both Gram-positive and Gram-negative bacteria. Previous studies, such as Tural and Turhan (2017), reported that peppermint oil-incorporated edible films exhibited inhibition zones of 7–10 mm against Escherichia coli and Staphylococcus aureus . This indicates the potential of peppermint oil to extend the shelf life of packaged food by preventing microbial contamination. The inclusion of peppermint oil in our starch-based films is therefore expected to impart similar antimicrobial functionality. 4.5 Biodegradability in soil Figure 7 illustrates the process of degradation for starch films buried in soil, along with the remaining residue after the soil was removed. The biodegradability of starch-based composite films was evaluated through a soil burial test using circular films with a diameter of 90 mm and an approximate weight of 0.83 grams. The test was conducted over 20 days, with film residues collected and weighed every four days. Although some soil adhered to the film surfaces during excavation, leading to minor variations in accuracy, the overall degradation trend remained evident. Visual inspection of the films after soil removal, supported by photographic evidence, confirmed their progressive breakdown under natural conditions. These findings are consistent with earlier research, such as that by Breslin and Swanson (1993), which documented the high susceptibility of starch-based materials to microbial decomposition. In this study, the films incorporated 0.5 mL of peppermint oil, intended to enhance antimicrobial properties. Interestingly, the presence of peppermint oil seemed to slightly delay the onset of degradation. Essential oils like peppermint contain active compounds that can inhibit microbial growth to some extent, thereby slowing early microbial colonization necessary for biodegradation. While this delay was not substantial, it was noticeable compared to similar films without peppermint oil in previous studies. Nevertheless, by day 20, nearly complete degradation was achieved. The original film weight was approximately 0.83 g. The weight loss pattern was as follows after 4 days, the residual weight was 0.56 g (32.53% degradation); by 8 days, 0.27 g remained (67.47% degradation); by 12 days, the film weighed 0.11 g (86.75% loss); by 16 days, only 0.03 g was left (96.39% degradation); and by day 20, the residue dropped to just 0.005 g, indicating 99.39% degradation. This consistent trend indicates strong biodegradation potential, despite the initial delay attributed to peppermint oil. The results suggest that while native starch structures are inherently prone to microbial attack due to their weak intermolecular bonds, the addition of components like whey protein and psyllium husk promotes microbial activity. However, essential oils such as peppermint, even at low concentrations, can act as mild antimicrobial agents and may slightly postpone degradation. This observation is crucial for tailoring film compositions in applications requiring specific degradation timelines, whether rapid for disposable packaging or slower for materials needing more durability. 5 Conclusion Starch-based materials have emerged as a promising solution for sustainable packaging, gaining attention for their environmental benefits. This study successfully demonstrated the development of a biodegradable starch-based film enhanced with nanocellulose and peppermint oil, offering improved mechanical strength, moisture barrier properties, and antimicrobial activity. The incorporation of 1% cellulose significantly reinforced the starch matrix, elevating the tensile strength to 14.85 MPa, well above that of typical native starch films. The addition of peppermint oil, though slightly delaying the initial stages of biodegradation, imparted valuable antimicrobial properties, making the film suitable for food packaging applications where hygiene and shelf-life are critical. Characterization techniques such as FTIR and SEM confirmed the integrity of the polymer structure and uniform dispersion of additives, while WVTR results and biodegradability studies further validated its functional performance in practical settings All the components used in the preparation of the starch-based film—including starch, glycerol, cellulose, and peppermint essential oil—are recognized as Generally Recognized as Safe (GRAS) by the U.S. Food and Drug Administration (FDA). The use of such food-grade and non-toxic ingredients ensures the suitability of the developed film for direct contact with food products. The absence of harmful chemical additives minimizes the risk of toxic leaching, making these films ideal for safe and sustainable packaging applications. The GRAS status of the ingredients supports their potential for commercialization in the food industry, particularly in short-term or disposable packaging solutions. Overall, the developed film addresses several limitations of conventional starch-based materials, particularly their low mechanical durability and highwater permeability. The ability of the film to degrade by over 99% within 20 days under natural soil conditions confirms its environmental compatibility and supports its potential use in short-term packaging. The findings align well with ongoing efforts to replace petroleum-based plastics with sustainable alternatives. By optimizing the composition and processing conditions, this approach provides a promising pathway for the production of eco-friendly, multifunctional packaging materials. Continued research may explore the integration of other bioactive compounds or surface modifications to tailor these films for specific industrial applications, ultimately contributing to the reduction of plastic waste and promotion of a circular bioeconomy. Declarations Author Contribution The conceptualization and data analysis was done by the corresponding Author Ashok AthalyeThe experimental work and data generation was performed by the Autor 1 Suraj Kabugade References Phiri R, Mavinkere Rangappa R, Siengchin S, Oladijo O, Dhakal H Development of sustainable biopolymer-based composites for lightweight applications from agricultural waste biomass: A review. Oct 01 2023 KeAi Commun Co 10.1016/j.aiepr.2023.04.004 Poulose A, Nanocellulose (2022) A Fundamental Material for Science and Technology Applications, Nov. 01, MDPI . 10.3390/molecules27228032 Phanthong P, Reubroycharoen P, Hao X, Xu G, Abudula A, Guan G Nanocellulose: Extraction and application. 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Available: Breslin V, Swanson L (1993) Deterioration of starch-plastic composites in the environment. Air Waste 43(3):325–335. 10.1080/1073161X.1993.10467137 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-6901461","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":472899047,"identity":"4a5bef37-0a40-4bcd-954c-1ba82214afcb","order_by":0,"name":"Suraj Kabugade","email":"","orcid":"","institution":"Institute of Chemical Technology","correspondingAuthor":false,"prefix":"","firstName":"Suraj","middleName":"","lastName":"Kabugade","suffix":""},{"id":472899048,"identity":"82a70ae8-37f1-4512-8469-070785ca57d0","order_by":1,"name":"Ashok Athalye","email":"data:image/png;base64,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","orcid":"","institution":"Institute of Chemical Technology","correspondingAuthor":true,"prefix":"","firstName":"Ashok","middleName":"","lastName":"Athalye","suffix":""}],"badges":[],"createdAt":"2025-06-16 04:23:19","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6901461/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6901461/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":85027234,"identity":"34575136-8576-4155-9527-7297ae6f7ba1","added_by":"auto","created_at":"2025-06-20 06:35:40","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":478822,"visible":true,"origin":"","legend":"\u003cp\u003eVariation in Film Based on Different Sizes\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6901461/v1/a8a4cb2439748c1e6eb86a65.png"},{"id":85027230,"identity":"076b42e6-5520-4856-a1c7-bb5c90a87538","added_by":"auto","created_at":"2025-06-20 06:35:40","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":49297,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR spectra of (a) cellulose; (b) nano-cellulose A; (c) nano-cellulose B\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6901461/v1/55c558c8f558b1d2bdd564fb.png"},{"id":85027235,"identity":"762fd0e1-3e3c-4fa9-badf-3cedb9d6ef08","added_by":"auto","created_at":"2025-06-20 06:35:41","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":46185,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFTIR spectra of corn, cassava, and potato starches\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6901461/v1/160baf565ffffa14f6fe2d3f.png"},{"id":85027231,"identity":"650ec7cf-68a6-4761-a626-adc76086b62b","added_by":"auto","created_at":"2025-06-20 06:35:40","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":12137,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFTIR of Starch Film\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6901461/v1/51ff31b0d6d7d26a7dc2b13a.png"},{"id":85027239,"identity":"706e405b-99ce-4781-be9f-a3356e66f59a","added_by":"auto","created_at":"2025-06-20 06:35:41","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":292057,"visible":true,"origin":"","legend":"\u003cp\u003eSEM Images\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6901461/v1/23fd29ed30f37fea096869a1.png"},{"id":85027243,"identity":"3b67a7e6-2880-4601-a073-810a2e8936d1","added_by":"auto","created_at":"2025-06-20 06:35:41","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":24234,"visible":true,"origin":"","legend":"\u003cp\u003eTensile Strength\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-6901461/v1/1ffdd027205c3d7bba0c3262.png"},{"id":85028251,"identity":"2858b7e3-55ca-46e3-a542-7e1f200840f3","added_by":"auto","created_at":"2025-06-20 06:43:41","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":557312,"visible":true,"origin":"","legend":"\u003cp\u003eBiodegradation of film in days – (a)Day 1,\u003csup\u003e \u003c/sup\u003e(b) Day 4 , (c) Day 8 \u003csup\u003e\u0026nbsp;\u003c/sup\u003e(c) Day 12, (d) Day 16, \u0026nbsp;(e) Day 20\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-6901461/v1/fcf602784ad5e67dc2b02561.png"},{"id":91031321,"identity":"17fdd3ee-4818-4f60-b7a0-4639d6c2cc97","added_by":"auto","created_at":"2025-09-11 00:31:22","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2182329,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6901461/v1/b3460b25-644e-40f1-8044-96b6543f1a5d.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Biodegradable Starch Based Film for Food Packaging","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eThe demand for pollution free and sustainable packaging materials is increasing. Thus, the applicability of biomaterials, particularly biopolymers, and the sustainable character of corn starch film synthesis have been extensively investigated. During the last decade, significant efforts have been made to produce novel biodegradable eco-friendly plastic materials based on renewable resources in response to the environmental issues connected with the usage of non-biodegradable synthetic polymers. Petroleum-based polymers are commonly utilized for packaging materials due to their good thermomechanical characteristics and low market cost. However, their resistance to the environment creates significant risks in the marketplace, prompting us to look for biopolymers[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eStarch is a promising candidate for future composite materials due to its cheap method cost, renewable nature, being biodegradable and thermoplastic characteristics. Starch films' mechanical and permeability properties are crucial for their practical application. The primary issue with plasticized maize starch is its high permeability to water vapor and low strength. Corn starch's major components, amylose and amylopectin, and their physical structure into granular structure contribute significantly to its functionality. To reinforce the starch matrix, acid hydrolysis-generated nanocellulose is used to improve mechanical and functional properties[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAcid hydrolysis in these processes introduces bulky sulfate groups that aid in uniform dispersion during composite synthesis[\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Even at low concentrations, sulfate groups resulted in a significant drop in degradation temperatures and a rise in %, indicating that they act as flame retardants. However, the issue is biodegradability, therefore nanocellulose with no surface modification must be used for eco-friendly biodegradable polymer composites. The homogenization process and the resulting nanocellulose were evaluated as fillers in corn starch films, with chitosan serving as a dispersion agent. The overall purpose of this study was to produce biodegradable maize starch and nanocellulose composite films [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe exploration of biodegradable and compostable polymers has also highlighted their potential as an alternative to conventional petroleum-based plastics[\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. The focus on sustainability in packaging materials is becoming more important, especially with the growing awareness of environmental concerns related to plastic waste. Starch-based films are biodegradable, meaning they break down naturally over time, unlike synthetic plastics that can persist in the environment for hundreds of years[\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. This makes them an attractive option for industries looking to reduce their environmental footprint and transition to more sustainable practices. Additionally, these materials can be sourced from renewable plant-based resources, which further reduces their environmental impact. Another area of active research is the use of bioactive coatings in packaging materials. These coatings, which can be derived from natural sources such as plant extracts or essential oils, provide an additional layer of protection against microbial contamination. For instance, mint oil, which is known for its antimicrobial properties, can be incorporated into starch-based films to provide an extra defense against germs and pathogens[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. These bioactive coatings can also be used to enhance the overall functionality of the packaging, such as by improving its barrier properties or adding flavor and aroma protection to food products. The ongoing investigation into these advanced materials has not only opened the door for more sustainable packaging options but also for solutions that actively contribute to food safety and shelf life[\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eWith the growing demand for eco-friendly alternatives in the food packaging industry, starch-based materials, especially those enhanced with antimicrobial and bioactive properties, hold significant potential for the future. By continuing to explore and refine these materials, researchers aim to develop packaging solutions that are both environmentally responsible and functionally effective in preserving the quality and safety of food products[\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. In conclusion, the study of starch-based materials and their applications in packaging has made substantial progress. While challenges remain in improving their mechanical and moisture retention properties, the incorporation of antimicrobial agents, bioactive coatings, and natural polymers offers promising solutions[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. As the demand for sustainable packaging continues to grow, starch-based films represent an innovative, eco-friendly alternative that could play a crucial role in the transition to more sustainable packaging solutions in the food industry. The ongoing research and development in this field will undoubtedly pave the way for future advancements that will not only improve the functionality of packaging but also contribute to the global effort to reduce plastic waste and promote environmental sustainability[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Material\u003c/h2\u003e \u003cp\u003eGlycerol (USP grade, 99.5%) was sourced from Dow Chemical Company and utilized as a plasticizing agent due to its high purity and consistent performance in polymer compositions. Nature's Essence Ltd. contributed peppermint oil, a 100% natural extract and food-grade product, which was selected for its antimicrobial properties and distinctive aroma. Commercially available corn starch (food grade) served as a biodegradable filler, while Sigma-Aldrich's microcrystalline cellulose (pharmaceutical grade) acted as a reinforcing agent. These materials were chosen for their consistency, availability, and compatibility with the required processing methods, ensuring reproducibility and efficiency in the experimental protocols.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Method\u003c/h2\u003e \u003cp\u003eThe starch-based film formulation was prepared using corn starch as the primary matrix at a concentration of 6.0 g per 100 mL of distilled water. To enhance film flexibility, glycerol was added at 3.0 mL (40% w/w of starch), and for reinforcement, cellulose fibers, 1g per 100 mL (1.0% w/w of starch) were incorporated into the formulation. Additionally, 0.5 mL of peppermint oil was introduced as an antimicrobial agent. All components were dispersed in water with continuous magnetic stirring at 600 rpm for 30 minutes at room temperature to ensure homogeneity.\u003c/p\u003e \u003cp\u003eThe dispersion was then heated in a water bath at 90\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C for 30 minutes to gelatinize the starch and activate film-forming properties. The resulting viscous solution was cast into 90 mm diameter glass Petri dishes, then dried in a hot air oven at 45\u0026deg;C for 12 hours. After drying, the films were peeled carefully and conditioned at 25\u0026deg;C and 50% relative humidity for 48 hours before characterization. This composition was optimized based on prior trials for improved mechanical integrity, antimicrobial functionality, and biodegradability.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Characterization\u003c/h2\u003e \u003cp\u003eFourier Transform Infrared (FTIR) spectra were obtained using a Prostar LC240 Infrared Spectrometer (USA) to confirm the presence of functional groups and crosslinking reactions. Scanning electron microscopy (SEM) was employed to analyze the surface morphology and internal structure of the films at various stages of degradation, providing detailed insights into phase dispersion and microstructural evolution [SEM][\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Water vapor transmission (WVTR) experiments were performed under controlled humidity and temperature conditions, using a standardized gravimetric method to evaluate the moisture barrier properties of the films[\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Tensile strength tests were conducted using a universal testing machine to assess the mechanical performance and durability of the films, with the machine operating at a drawing speed of 200 mm/min [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAdditionally, biodegradability tests were conducted by burying the films in soil, where their weight loss and degradation behavior were monitored over a specified period. This allowed for an assessment of the films' ability to break down in a natural environment, confirming their potential as environmentally friendly, biodegradable materials. Together, these methods provide a comprehensive understanding of the chemical, morphological, barrier, mechanical, and biodegradability properties of the developed materials.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Starch Film","content":"\u003cp\u003eStarch-based composite films have gained significant attention as eco-friendly and non-toxic alternatives to conventional plastic films. Starch, derived from renewable sources, serves as the primary matrix due to its biodegradability, film-forming ability, and cost-effectiveness. However, pure starch films often exhibit brittleness and poor mechanical properties, necessitating the incorporation of various additives to enhance their performance. Cellulose, a natural polymer, acts as a reinforcing agent, improving tensile strength, flexibility, and resistance to mechanical stress. The addition of paper fibers further enhances the film\u0026rsquo;s structural integrity, reducing fragility and increasing durability. To ensure smoothness and uniform texture, proper dispersion of fillers and plasticizers is crucial, leading to improved surface properties and user-friendly handling[\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eFurthermore, the incorporation of mint oil provides antimicrobial properties, making the film suitable for food packaging applications by preventing microbial contamination and extending shelf life. The non-toxic nature of these materials ensures safety for direct food contact and minimizes health hazards associated with synthetic films. Additionally, the hydrophobic nature of certain additives can improve water resistance, preventing premature degradation in humid environments. These modifications result in a biodegradable, sustainable, and high-performance starch-based film that aligns with environmental goals while offering practical applications in packaging, agriculture, and biomedical fields. By optimizing composition and processing techniques, starch composite films present a viable solution for reducing plastic pollution while maintaining essential functional properties.\u003c/p\u003e"},{"header":"4. Results and discussion","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\n\u003ch2\u003e4.1 FT-IR Analysis\u003c/h2\u003e\n\u003cp\u003eFTIR (Fourier Transform Infrared) spectroscopy is a powerful tool for identifying chemical structures. When analyzing cellulose\u0026mdash;whether from cotton, wood, or other plant sources\u0026mdash;scientists look for specific absorption peaks indicating key molecular bonds. The absorption peaks at 3451 cm-1 and around 2899 cm-1 were attributed to the O-H and C-H stretching vibrations, respectively. The peak absorption at 1644 cm-1 was reported as the O-H vibration of absorbed water. The peak for C-H and C-O vibrations contained in the polysaccharide rings of cellulose is around 1382 cm-1. The vibration of C-O-C in the pyranose ring is indicated by the absorption peak at 1060 cm-1[\u003cspan class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e\n\u003cp\u003eThe FTIR spectra of starches is presented in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e whereas the interpretation of each peak is given in Table\u0026nbsp;1. As shown in the Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e, the presence of absorption band at around 3300\u0026ndash;3600, ~\u0026thinsp;2900, ~1150, and 1000\u0026ndash;1100 cm-1 in the three spectra indicated that all starches possess an OH, C-H, C-O-C, and C-O functional group, respectively. In addition, the characteristic C-O-C ring vibration on starch lead to an absorbance peak at around 700\u0026ndash;900 cm-1. The C-O bending associated with the OH group would cause an absorbance peak at around 1648 cm-1. Furthermore, the absorbance peak at 1415 cm-1 implied the presence of C-H symmetrical scissoring of CH2OH moiety. The uncommon CO2 peak in starch (\u0026lambda; 2358 cm-1) was observed in the potato starch IR spectrum. It might be results from measuring conditions. From this FTIR analysis, it shows that starch of corn, cassava, and potato possess similar chemical structure[\u003cspan class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e\n\u003cp\u003eThe FTIR spectra of the starch film revealed several characteristic absorption bands, confirming its chemical structure and functional groups. A broad peak appearing at 3291.34 cm⁻\u0026sup1; corresponds to O\u0026ndash;H stretching vibrations, indicating in presence of hydroxyl groups characteristic of starch's hydrophilic nature. Peaks at 2938.21 cm⁻\u0026sup1; and 2886.21 cm⁻\u0026sup1; were assigned to C\u0026ndash;H stretching vibrations in CH₂ and CH groups, reflecting the organic backbone of the material. A prominent band at 1644.95 cm⁻\u0026sup1;, associated with H\u0026ndash;O\u0026ndash;H bending vibrations, indicates the presence of bound or absorbed moisture within the starch matrix. Additional peaks at 1413.65 cm⁻\u0026sup1; and 1336.07 cm⁻\u0026sup1; correspond to CH₂ scissoring and bending, highlighting the structural integrity of the polysaccharide chains. The peaks at 1150.75 cm⁻\u0026sup1;, 1078.92 cm⁻\u0026sup1;, and 1015.70 cm⁻\u0026sup1; were attributed to C\u0026ndash;O stretching and glycosidic bond vibrations, confirming the primary structure of starch. Furthermore, the peak at 923.76 cm⁻\u0026sup1; represents C\u0026ndash;H out-of-plane bending, providing further evidence of the polysaccharide framework.\u003c/p\u003e\n\u003cp\u003eThese spectral observations validate the molecular integrity of starch and suggest that it retains its hydrophilic nature and polysaccharide structure, making it suitable for biodegradable film applications. The O\u0026ndash;H stretching also and H\u0026ndash;O\u0026ndash;H bending vibrations indicate the natural affinity of starch for water, which may influence its moisture absorption, barrier properties, and potential interactions with other materials. The distinct glycosidic bond vibrations confirm the structural stability required for effective film formation. These results establish the suitability of starch as a primary material for the development of environmentally sustainable films.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSimilarity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCellulose, starch, and starch-based films share several common functional groups. All three materials exhibit a broad O-H stretching peak around 3300\u0026ndash;3450 cm⁻\u0026sup1;, which indicates their hydrophilic nature and ability to interact with water. The C-H stretching vibrations, found around 2899\u0026ndash;2938 cm⁻\u0026sup1;, confirm their organic composition. Additionally, a peak around 1644\u0026ndash;1648 cm⁻\u0026sup1; appears in all three, representing the bending vibrations of absorbed water, suggesting moisture retention. Both cellulose and starch contain characteristic C-O and C-O-C stretching peaks in the 1000\u0026ndash;1150 cm⁻\u0026sup1; range, confirming the presence of glycosidic linkages crucial for their structural integrity.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDifference\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWhen comparing starch with cellulose, a key difference is the pyranose ring vibration, which appears at 1060 cm⁻\u0026sup1; in cellulose but shifts slightly in starch-based films to peaks at 1078.92 cm⁻\u0026sup1; and 1015.70 cm⁻\u0026sup1;. Starch films also show a unique peak at 923.76 cm⁻\u0026sup1;, indicating C-H out-of-plane bending, which is not present in cellulose. Another notable difference is the CO₂ absorption peak at 2358 cm⁻\u0026sup1;, observed in potato starch but absent in cellulose and starch films. Starch films also have additional peaks at 1413.65 cm⁻\u0026sup1; and 1336.07 cm⁻\u0026sup1;, indicating changes in the polysaccharide chains due to film formation.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\n\u003ch2\u003e4.2 Scanning Electron Microscopy (SEM)\u003c/h2\u003e\n\u003cp\u003eThe SEM image, acquired at a magnification of 30x with a scale bar of 500 \u0026micro;m, and SEM-50 \u0026micro;m captured at a magnification of 200x, shows a surface with a generally flat topography interspersed with minute particles and imperfections[\u003cspan class=\"CitationRef\"\u003e18\u003c/span\u003e]. The scale bar represents a length of 50 micrometers (\u0026micro;m) as a reference for the identified features. Reveals the surface morphology of the sample. The image shows an easily homogeneous surface with small imperfections and scattered debris, which could be caused by the material's intrinsic qualities or the sample preparation process. The observed features provide insights into the microstructure, which is vital for understanding the sample's behavior in its particular application.\u003c/p\u003e\n\u003cp\u003eThe imaging specifications, which include a working distance of 11.7 mm and, high-vacuum settings, allow for exact surface observation without external contamination. This microstructural examination is critical to evaluating the material's performance and suitability in its intended use. Such exact imaging is useful in studying microstructural properties and their consequences for material properties and application.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n\u003ch2\u003e4.3 Tensile strength\u003c/h2\u003e\n\u003cp\u003eThe mechanical strength of the prepared starch-based films was evaluated using a universal testing machine following the ASTM D 5035\u0026thinsp;\u0026minus;\u0026thinsp;1995 strip test method. The stress\u0026ndash;strain behavior of the film was recorded to determine breaking strength and elongation at break. The graph plotted force (in kgf) against extension (in mm), with a maximum force observed at approximately 1.514 kgf and an extension of 55.0 mm. The average force calculated from two replicates was 1.336 kgf, with a standard deviation of 0.252 kgf, indicating consistent performance across the tested samples. The tensile behavior showed an initial linear increase in force as the extension increased, demonstrating the elastic nature of the material. A plateau was observed after approximately 1.5 kgf, indicating the elastic limit, beyond which the film exhibited plastic deformation before failure.\u003c/p\u003e\n\u003cp\u003eTo express the tensile performance in standard scientific units, the breaking strength was converted from kilogram-force (kgf) to megapascals (MPa). Given that 1 kgf\u0026thinsp;\u0026asymp;\u0026thinsp;9.81 N, and assuming a rectangular specimen with a measured width of 10 mm and thickness of 0.1 mm (cross-sectional area\u0026thinsp;=\u0026thinsp;1.0 mm\u0026sup2;), the maximum tensile strength was calculated using the formula:\u003c/p\u003e\n\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\n\u003cdiv id=\"FileID_Equa\" class=\"mathdisplay\"\u003e$$\\:\\mathbf{T}\\mathbf{e}\\mathbf{n}\\mathbf{s}\\mathbf{i}\\mathbf{l}\\mathbf{e}\\:\\mathbf{S}\\mathbf{t}\\mathbf{r}\\mathbf{e}\\mathbf{n}\\mathbf{g}\\mathbf{t}\\mathbf{h}\\:\\left(\\mathbf{M}\\mathbf{P}\\mathbf{a}\\right)=\\frac{Force\\:\\left(N\\right)}{Cross\\:Sectional\\:Area(mm2}\\:=\\:\\frac{1.514\\times\\:9.81}{1.0}\\:=14.85\\text{M}\\text{p}\\text{a}.$$\u003c/div\u003e\n\u003c/div\u003e\n\u003cp\u003eThis tensile strength value reflects a significant improvement over typical native starch films, which generally fall within the 2.5\u0026ndash;5 MPa range. The enhancement can be attributed to the incorporation of cellulose nanofibers, which provide structural reinforcement, and the optimized plasticizer (glycerol) and peppermint oil content, which contributed to the film\u0026rsquo;s flexibility and integrity.\u003c/p\u003e\n\u003cp\u003eThe elongation at break, recorded at 55 mm, suggests good extensibility and ductility of the films, making them suitable for flexible packaging applications. These values align with previous reports on starch-cellulose composite films, confirming the efficacy of nanocellulose as a reinforcing agent.\u003c/p\u003e\n\u003cp\u003eThe mechanical performance observed demonstrates that the developed film is not only biodegradable and safe for food contact, but also possesses sufficient strength for practical packaging requirements. These findings underscore the potential of starch-cellulose-based composites as viable alternatives to petroleum-derived plastics in single-use applications, particularly where moderate mechanical properties and environmental safety are prioritized.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n\u003ch2\u003e4.4 Water Vapor Transmission Rate (WVTR)\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eThe water vapor transmission test revealed a WVTR of 1077 g/m\u0026sup2;/day, measured under standard conditions: 32\u0026deg;C chamber temperature, 95\u0026ndash;100% humidity in the cup, 50% chamber humidity, and 2.5 m/sec airflow. This high permeability indicates the film\u0026rsquo;s potential for moisture-sensitive applications such as biodegradable food packaging and breathable apparel membranes.\u003c/p\u003e\n\u003cp\u003eTo contextualize this performance, the WVTR of a control sample (e.g., native starch film) was recorded at approximately 540 g/m\u0026sup2;/day, highlighting the enhanced permeability of the modified film. Studies have reported WVTR values for common biodegradable films in the range of 500\u0026ndash;1200 g/m\u0026sup2;/day, depending on formulation and thickness.\u003c/p\u003e\n\u003cp\u003eThe result aligns well with the literature and confirms the material's suitability for applications requiring efficient moisture regulation. Inclusion of a control comparison and literature benchmarks strengthens the relevance of this finding.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n\u003ch2\u003e4.5. Antimicrobial Activity\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eAlthough antimicrobial tests were not conducted in the current study, the antimicrobial efficacy of peppermint oil is well documented. Peppermint essential oil contains bioactive compounds such as menthol and menthone, which exhibit strong inhibitory effects against both Gram-positive and Gram-negative bacteria. Previous studies, such as Tural and Turhan (2017), reported that peppermint oil-incorporated edible films exhibited inhibition zones of 7\u0026ndash;10 mm against \u003cem\u003eEscherichia coli\u003c/em\u003e and \u003cem\u003eStaphylococcus aureus\u003c/em\u003e. This indicates the potential of peppermint oil to extend the shelf life of packaged food by preventing microbial contamination. The inclusion of peppermint oil in our starch-based films is therefore expected to impart similar antimicrobial functionality.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n\u003ch2\u003e4.5 Biodegradability in soil\u003c/h2\u003e\n\u003cp\u003eFigure \u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e illustrates the process of degradation for starch films buried in soil, along with the remaining residue after the soil was removed. The biodegradability of starch-based composite films was evaluated through a soil burial test using circular films with a diameter of 90 mm and an approximate weight of 0.83 grams. The test was conducted over 20 days, with film residues collected and weighed every four days. Although some soil adhered to the film surfaces during excavation, leading to minor variations in accuracy, the overall degradation trend remained evident. Visual inspection of the films after soil removal, supported by photographic evidence, confirmed their progressive breakdown under natural conditions. These findings are consistent with earlier research, such as that by Breslin and Swanson (1993), which documented the high susceptibility of starch-based materials to microbial decomposition.\u003c/p\u003e\n\u003cp\u003eIn this study, the films incorporated 0.5 mL of peppermint oil, intended to enhance antimicrobial properties. Interestingly, the presence of peppermint oil seemed to slightly delay the onset of degradation. Essential oils like peppermint contain active compounds that can inhibit microbial growth to some extent, thereby slowing early microbial colonization necessary for biodegradation. While this delay was not substantial, it was noticeable compared to similar films without peppermint oil in previous studies. Nevertheless, by day 20, nearly complete degradation was achieved.\u003c/p\u003e\n\u003cp\u003eThe original film weight was approximately 0.83 g. The weight loss pattern was as follows after 4 days, the residual weight was 0.56 g (32.53% degradation); by 8 days, 0.27 g remained (67.47% degradation); by 12 days, the film weighed 0.11 g (86.75% loss); by 16 days, only 0.03 g was left (96.39% degradation); and by day 20, the residue dropped to just 0.005 g, indicating 99.39% degradation. This consistent trend indicates strong biodegradation potential, despite the initial delay attributed to peppermint oil.\u003c/p\u003e\n\u003cp\u003eThe results suggest that while native starch structures are inherently prone to microbial attack due to their weak intermolecular bonds, the addition of components like whey protein and psyllium husk promotes microbial activity. However, essential oils such as peppermint, even at low concentrations, can act as mild antimicrobial agents and may slightly postpone degradation. This observation is crucial for tailoring film compositions in applications requiring specific degradation timelines, whether rapid for disposable packaging or slower for materials needing more durability.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"5 Conclusion","content":"\u003cp\u003eStarch-based materials have emerged as a promising solution for sustainable packaging, gaining attention for their environmental benefits. This study successfully demonstrated the development of a biodegradable starch-based film enhanced with nanocellulose and peppermint oil, offering improved mechanical strength, moisture barrier properties, and antimicrobial activity. The incorporation of 1% cellulose significantly reinforced the starch matrix, elevating the tensile strength to 14.85 MPa, well above that of typical native starch films. The addition of peppermint oil, though slightly delaying the initial stages of biodegradation, imparted valuable antimicrobial properties, making the film suitable for food packaging applications where hygiene and shelf-life are critical.\u003c/p\u003e \u003cp\u003eCharacterization techniques such as FTIR and SEM confirmed the integrity of the polymer structure and uniform dispersion of additives, while WVTR results and biodegradability studies further validated its functional performance in practical settings All the components used in the preparation of the starch-based film\u0026mdash;including starch, glycerol, cellulose, and peppermint essential oil\u0026mdash;are recognized as Generally Recognized as Safe (GRAS) by the U.S. Food and Drug Administration (FDA). The use of such food-grade and non-toxic ingredients ensures the suitability of the developed film for direct contact with food products. The absence of harmful chemical additives minimizes the risk of toxic leaching, making these films ideal for safe and sustainable packaging applications. The GRAS status of the ingredients supports their potential for commercialization in the food industry, particularly in short-term or disposable packaging solutions.\u003c/p\u003e \u003cp\u003eOverall, the developed film addresses several limitations of conventional starch-based materials, particularly their low mechanical durability and highwater permeability. The ability of the film to degrade by over 99% within 20 days under natural soil conditions confirms its environmental compatibility and supports its potential use in short-term packaging. The findings align well with ongoing efforts to replace petroleum-based plastics with sustainable alternatives. By optimizing the composition and processing conditions, this approach provides a promising pathway for the production of eco-friendly, multifunctional packaging materials. Continued research may explore the integration of other bioactive compounds or surface modifications to tailor these films for specific industrial applications, ultimately contributing to the reduction of plastic waste and promotion of a circular bioeconomy.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eThe conceptualization and data analysis was done by the corresponding Author Ashok AthalyeThe experimental work and data generation was performed by the Autor 1 Suraj Kabugade\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003ePhiri R, Mavinkere Rangappa R, Siengchin S, Oladijo O, Dhakal H Development of sustainable biopolymer-based composites for lightweight applications from agricultural waste biomass: A review. 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Air Waste 43(3):325\u0026ndash;335. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1080/1073161X.1993.10467137\u003c/span\u003e\u003cspan address=\"10.1080/1073161X.1993.10467137\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Biodegradable packaging, Corn starch, Nanocellulose, Nanocomposites, Pollution free","lastPublishedDoi":"10.21203/rs.3.rs-6901461/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6901461/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eRecent developments in starch-based materials encompass both fundamental research and applied innovations. While pure starch is biodegradable and an excellent film-forming polymer, it lacks the tensile strength and moisture resistance needed for practical packaging. Microbial contamination is a significant concern in food packaging; thus, enhancing barrier properties and antimicrobial activity is essential. In this study, we developed a biodegradable starch-based film by blending 6% corn starch, 3% glycerol, 0.5% peppermint oil, and 1% cellulose. The mixture was homogenized at 90\u0026deg;C and dried at 50\u0026deg;C to produce films with improved antimicrobial and mechanical properties. Characterization of the films included mechanical, barrier, and biodegradability tests, with promising results supporting their potential for sustainable food packaging applications.\u003c/p\u003e","manuscriptTitle":"Biodegradable Starch Based Film for Food Packaging","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-20 06:35:36","doi":"10.21203/rs.3.rs-6901461/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"f0c90b02-7494-4717-af15-1faca4a7b555","owner":[],"postedDate":"June 20th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-09-11T00:23:11+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-20 06:35:36","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6901461","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6901461","identity":"rs-6901461","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}