Lemon grass Essential oil Incorporated Starch Based Active Packaging Film, Characterization and Applications

Lemon grass Essential oil Incorporated Starch Based Active Packaging Film, Characterization and Applications | 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 Lemon grass Essential oil Incorporated Starch Based Active Packaging Film, Characterization and Applications Kanak Sharma, Gargi Ghoshal This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7095535/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 These biodegradable films are typically composed of polysaccharides, proteins, lipids, or there is a growing interest in edible films for fruits and vegetables, driven by concerns over the negative impact of traditional packaging materials on health and the environment their combinations. In this study, edible film was formulated using kidney bean starch, both in its native and nano forms, with the incorporation of lemongrass essential oil. The starch, which can be sourced from various biological origins, served as the primary matrix, while the essential oil contributed its well-known antioxidant and antimicrobial properties. An acid hydrolysis technique was used to decrease the kidney bean starch's granule size and produce nanoparticles in order to improve the starch's qualities. When the physicochemical properties of native and nano starches were compared, it was found that the nano starch had better solubility, swelling power, and the ability to absorb water and oil. According to the results, kidney bean starch may be used for producing packaging films, and adding nano starch to the film formulations can enhance the functional qualities of the finished product. Starch packaging Lemongrass essential oil Nano starch Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 1. Introduction The rapid rise in global plastic production has led to significant environmental pollution. Since the 1950s, plastic manufacturing has increased from 1.5 million to 335 million tonnes [ 1 ], with 79% of plastic waste improperly disposed of in landfills or natural environments [2, 3]. The packaging industry consumes 36% of all plastics, primarily for single-use items like bags, bottles, and food packaging, generating over 150 million tonnes of waste annually. Only 9% of this waste is recycled, while 79% accumulates in the environment [2, 4]. Most plastics are derived from non-renewable petroleum, creating non-biodegradable polymers and exacerbating environmental harm. This highlights the urgent need for sustainable alternatives to reduce plastic consumption [ 4 ]. Biodegradable active packaging is emerging as an eco-friendly alternative to petroleum-based plastics due to its versatility and environmental benefits. These films are made from biopolymers like proteins, polysaccharides, lipids, or their combinations and can degrade into non-toxic molecules or harmless biomass under natural conditions. Starch-based biodegradable products particularly have gained popularity. Starch, a polysaccharide found in plant seeds, tubers, roots, and fruits [ 5 ], is commonly used due to its abundance and ease of extraction, with grains, tubers, and legumes being the primary raw materials [ 6 ]. Beans, part of the Fabaceae family, are globally significant food crops, both economically and nutritionally [ 7 ]. Kidney beans ( Phaseolus vulgaris ) contain 22–45% starch, which can be applied to both food and non-food products as a thickening agent, binder, film former, gelling agent, and more. While traditional starches from sources like maize, wheat, and potato have been well-studied, legume starches, including kidney bean starch, are less explored due to limited pulse availability in some regions. However, the rising interest in alternative starch sources is driving research into their properties. Kidney bean starch, with its high amylose content, is noted for high gelation temperatures, shear thinning resistance, rapid retrogradation, and high gel elasticity. Studying these properties is essential for expanding its industrial applications. Studies show that incorporating co-biopolymers and nanomaterials can enhance the mechanical and barrier properties of starch-based films [ 8 ]. Bio-nanocomposites, which use nanoparticles as reinforcing fillers, address the limitations of biopolymer packaging by improving rheological, physical, mechanical, and thermal properties, as well as processability and performance [ 9 , 10 ]. Native and nano starch from the same source offer better compatibility, leading to improved film properties [ 5 ]. Acid-hydrolysed starch nanoparticles are particularly useful in high-temperature food processing due to their high crystallinity and stability. However, research on nano starch derived from kidney bean starch remains limited [ 11 ]. 2. Materials and methods 2.1. Materials Kidney beans utilised in the research were sourced from Krishi Vigyan Kendra, Kukumseri, in the Lahaul & Spiti district of Himachal Pradesh, India collected through proper channel complied with local guidelines. Analytical-grade chemicals employed in the research were procured from Sisco Research Laboratories Pvt. Ltd., Loba Chemie Pvt. Ltd., and Allin Exporters. Additionally, fruits used in the study were obtained from a local market in Chandigarh, India. 2.2. Starch isolation Kidney beans' starch was extracted using a modified wet steeping technique [ 12 ] (Sandhu et al., 2005). The beans were soaked for 20 hours in 1.25 litres of distilled water containing 0.1% potassium metabisulphite. They were then ground with water to form smooth slurry. After the slurry was run through 100-mesh nylon cloth, all remaining substance was repeatedly washed with distilled water until it was clear of starch. The filtered slurry was allowed to settle for 4–5 hours, and the clear supernatant was centrifuged at 5000 rpm for 10 minutes. The top brownish protein layer was removed after each centrifugation until the supernatant was clear. A hot air oven was used to dry the final starch at 40°C, and stored in an airtight container for further use. 2.3. Preparation of Nano Starch Nano starch was produced using a modified acid hydrolysis method [ 11 ]. Fifteen grams of dry native starch were incubated in 3.16 M sulfuric acid at 40°C for seven days with magnetic stirring. After that, the mixture was centrifuged for 15 minutes at 6000 rpm. After repeatedly washing the precipitates with distilled water until they were neutralised, acetone was used to wash them. A hot air oven set to 40°C was used to dry the resultant starch. 2.4. Physicochemical properties of starch The following methods were used to measure the starch's amylose content [ 13 ], ash content [ 14 ], water and oil absorption capacity [ 13 ], swelling power, and solubility [ 14 ]. 2.5. Characterization of starch 2.5.1Surface morphology Samples of starch were examined for surface morphology using a Hitachi SU 8010 field emission scanning electron microscope. Kidney bean native and nano starches were mounted on carbon tape and gold-coated by sputtering for 60 seconds before imaging. 2.5.2 Thermal characteristics (DSC) The differential scanning calorimeter was used to evaluate the starches' thermal characteristics. A 1:3 (g/g) starch-to-water ratio was prepared using 3.1 mg of starch and distilled water in an aluminium pan, which was sealed and equilibrated for one hour. The sample was then heated at 10°C per minute, starting from 10°C for five minutes. An empty pan and indium were used for calibration. Gelatinisation onset temperature (To), peak temperature (Tp), glass transition temperature (Tg), and end set temperature (Te) were among the important thermal parameters that were measured. 2.5.3 Size distribution Using a Nano Zetasizer dynamic light scattering (DLS) device (Malvern Instruments Inc., U.K.), the size and distribution of starch particles were determined. Samples were prepared at a 0.01% (w/v) concentration in distilled water, diluted 100-fold with deionized water, and analysed at 30°C in plastic cuvettes. Measurements were performed without filtration or dust removal [ 15 ] (Sadeghi et al., 2017). 2.5.4 Rheology Rheology of starch studies the deformation and flow of starch under external forces. In 100 millilitres of distilled water, a 3% starch solution (nano/native) was heated to 90°C for 20 minutes while being constantly stirred to form a gel [ 16 ]. After cooling to room temperature, An Anton Paar MCR 102 rheometer with parallel plate geometry (PP-50; diameter: 50 mm) was used to measure the rheological properties at 25 ± 1°C. 2.5.5 X-Ray Diffraction (XRD) analysis Crystalline phases are identified via X-ray powder diffraction (XRD). An X'Pert Pro A diffractometer (Analytical, U.S.A.) operating at 45 kV and 40 mA recorded diffraction patterns using Cu Kα radiation in a 2θ configuration. 2.5.6 Fourier Transforms Infrared Spectroscopy (FTIR) Fourier Transform Infrared Spectroscopy (FTIR) was used to examine the molecular interactions of starch [ 17 ]. FTIR spectra for treated and control samples were recorded with a Shimadzu 8400 spectrometer (Japan) with a resolution of 4 cm⁻¹ and in the 500–4000 cm⁻¹ range. Samples were conditioned at 53 ± 1% relative humidity for 24 hours before analysis. Data were analyzed using the FTIR software. 2.6 Characterization of essential oil 2.6.1 Gas Chromatography–Mass Spectrometry (GCMS) Volatile compounds in essential oils are separated into individual components using Gas Chromatography (GC), which produces a linear chromatogram of these constituents. Mass Spectrometry (MS) is then used to identify and quantify each component, aiding in the detection of potential adulteration [ 18 ]. 2.7 Preparation of films Edible films were produced by casting from aqueous solutions and evaporating the liquid phase. 4g of native or nano starch were combined with 100ml of distilled water and heated to 95°C for 30 minutes in order to create kidney bean starch films. After adding 30% w/w D-sorbitol as a plasticizer, the mixture was heated for an additional 40 minutes and then homogenized with a probe sonicator for 10 minutes. Petri dishes were filled with the solution, which was then allowed to dry at room temperature. To create composite films with lemon grass essential oil, distilled water was used to make a 1% w/v Tween 80 and essential oil solution. This solution was then heated to 45°C for 40 minutes and combined with the starch solution that had already been made. The combined mixtures were stirred at 50°C for 10 minutes, transferred to petri plates and allowed to air dry at room temperature. The dried films were then removed for further characterization. 2.8 Physicochemical properties of film 2.8.1 Moisture content of films Film samples (S, S + EO, NS, NS + EO) measuring 4 × 1 cm² were heated in an oven (at 105°C) until their weight remained unchanged, indicating complete drying. The moisture content was calculated using Eq. 1 and determined using the [ 19 ] approach. All experiments were conducted in triplicate. $$\:\text{M}\text{C}=\frac{\:{\text{W}}_{0}-\:{\text{W}}_{1}}{\:{\text{W}}_{0}}\times\:100\:\:\dots\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\text{E}\text{q}\:\:1$$ Where, MC = Moisture content, W 0 = Mass of the initial sample, W 1 = Mass of the final sample. 2.8.2 Water solubility of film Film samples (S, S + EO, NS, NS + EO) measuring 4 × 1 cm² were dried for 24 hours at 105°C to achieve a constant weight (W o ). After drying, the samples were submerged for 24 hours at 25 ± 1°C in 50 ml of distilled water. The samples were reweighed (W 1 ) after being re-dried at 105°C. Eq. 2 was used to determine the film’s water solubility, with multiple tests conducted for accuracy. $$\:\text{S}\:\left(\text{%}\right)=\frac{\:{\text{W}}_{0}-\:{\text{W}}_{1}}{\:\:{\text{W}}_{0}}\times\:100\:\:\dots\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\text{E}\text{q}\:\:\:\:\:2\:\:$$ Where, S = Solubility, W 0 = Weight of the dried sample before water immersion, and W 1 = Dry weight of the insoluble sample after immersion. 2.8.3 Water Vapour Permeability (WVP) of films The film samples were put on modified beakers with calcium chloride (0% RH) after being paraffin-sealed. These beakers were then incubated at 25 ± 1°C in a desiccator filled with distilled water (100% RH). The increase in beaker weight was recorded over time [ 20 ]. These formulas were used to calculate the film's WVP. \(\:\text{W}\text{V}\text{T}\text{R}=\frac{\varDelta\:\text{W}\times\:\text{t}}{\text{A}}\) … Eq. 3 $$\:\text{W}\text{V}\text{P}=\:\frac{\text{W}\text{V}\text{T}\text{R}\times\:\text{L}}{\varDelta\:\text{P}}\:\:\dots\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:Eq\:\:\:\:\:4$$ Measurements were taken in triplicate, with the variables being water vapor transmission rate (WVTR), sample thickness (L), cross-sectional area (A), and differential pressure (ΔP) across the film. 2.8.4 Physical and optical properties Film thickness was measured with 0.01 mm accuracy using a Mitutoyo digital micrometre (Japan), taking readings at 10 points per sample and averaging the results. Each measurement was repeated three times [ 20 ]. Film color and opacity were assessed using a Hunter colorimeter (Color flex model, Hunter Lab, USA). A Texture Analyser (TA.XT Plus, Stable Microsystems) was used to assess mechanical parameters, such as tensile strength, in accordance with ASTM standard method 828 − 97 (ASTM, 2002). 2.8.5 Swelling Power and Solubility 0.1 g of the film sample was dissolved in 10 ml of distilled water and heated for 30 minutes at 60°C, 70°C, 80°C, and 90°C for the swelling power analysis. The sample was centrifuged for 10 minutes at 2000 rpm after cooling. The precipitate was weighed, and the supernatant was dried overnight at 80°C in pre-weighed petri plates, with weight change recorded. \(\:\text{S}\text{I}=\frac{\text{W}1\:\left(\text{g}\right)}{\text{W}\text{o}\:\left(\text{g}\right)}\) … Eq. 5 \(\:\text{S}\text{o}\text{l}\text{u}\text{b}\text{i}\text{l}\text{i}\text{t}\text{y}\:\left(\text{%}\right)=\frac{\text{W}2\:\left(\text{g}\right)}{\text{W}\text{o}\:\left(\text{g}\right)}\times\:100\) … Eq. 6 Where, SI = Swelling Index, S = Solubility, W o = Starch taken (g), W 1 = Dry precipitates (g), W 2 = Weight after supernatant dried 2.9 Characterization of films The films were subjected to X-Ray Diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FTIR) using the previously described techniques. 2.9.1 SEM A scanning electron microscope (SEM) was used to analyse the films' surface morphology. The external surfaces were sputter-coated with gold prior to observation, and gold was also applied to fracture surfaces before imaging [ 21 ]. 2.9.2 Antioxidant Activity of films Essential oils are frequently incorporated to films made of starch because of their well-known antioxidant qualities. The antioxidant activity of films containing lemongrass essential oil was tested using the method described by [ 22 ] . 2.9.3 Antimicrobial activity of films Lemongrass essential oil was tested against both Gram-positive ( S. aureus ) and Gram-negative ( E. coli ) bacteria to determine the antibacterial activity of starch-based edible films. The antibacterial activity of the lemongrass essential oil was assessed using the agar well diffusion method. A 6–8 mm well in an inoculated agar plate was treated with the oil extract and then kept in a 37°C incubator for 48 hours. The oil inhibited bacterial growth by diffusing through the agar, and the clear zone diameter was measured in millimetres to determine its antibacterial activity. 2.9.4 Biodegradability Biodegradability of a film reflects its susceptibility to environmental degradation, primarily through microbial activity. To assess this, composite film specimens (1×2 cm²) were buried 2 cm below the surface in a mixture of uniformly sized soil and compost within an earthen pot. The weight loss of the films was monitored under controlled conditions (25°C) at 5-day intervals. \(\:\text{B}\left(\text{%}\right)=\:\frac{W\text{o}-W1}{W\text{o}}\times\:100\) Eq. 7 Where, B(%) = Biodegradability, Wo = Initial weight of sample, W 1 = Final weight of sample. 2.10 Applications on fruits The experiment utilized a completely randomized design with 5 samples: A fruit sample was kept unwrapped (Control), while the others were wrapped in native starch film (S), native starch film with lemongrass essential oil (S + EO), nano starch film (NS), and nano-starch film containing lemongrass essential oil (NS + EO). Fruits were kept in a refrigerator and at room temperature, and analyses were done after16 days at room temperature and 26 days in the refrigerator [ 23 ]. 2.10.1 Weight loss determination Weight loss percentages of each fruit were calculated using their initial and final weights. Grapes were weighed every four days using a digital balance. 2.10.2 Shrinkage The initial diameter of the fruits was measured at two positions near the middle using a digital micrometre (Mitutoyo, Japan), and the average diameter was recorded. During storage, diameter measurements were taken every fourth day to assess shrinkage for all samples [24] . 2.11 Statistical analysis One-way ANOVA in IBM SPSS Statistics Trial Version was used to evaluate the data. Using Duncan's multiple range test, significant variations in means were evaluated (p < 0.05). 3. Results and Discussion 3.1 Physicochemical properties of native and nano starch from kidney bean 3.1.1 Amylose Content Amylose content in starch is affected by both ambient growth circumstances and genetic variables. According to Table 1 , the amylose concentration of the native kidney bean was 12.18 ± 0.35%. On the other hand, due to climatic and ecological variations, kidney beans Table 1 Amylose content, ash content, water and oil absorption capacity of native and nano starch from kidney bean S. NO. Sample Amylose Content Ash content Water absorption capacity% Oil absorption capacity % 1 Native 12.18 ± 0.35% 0.11 ± 0.03% 2.34 ± 0.05%. 25.34 ± 0.02% 2 Nano - 0.97 ± 0.23% 53.76 ± 2.14%. 39.23 ± 0.04% had a much greater amylose content of 32.4%, according to [25, 26]. The amylose concentration of nano starch's was not ascertained. The amylose concentration may become undetected due to severe hydrolysis because the remaining linear chains are too short to create a blue iodine complex [ 27 ]. 3.1.2 Ash content Table 1 indicates that the ash level of kidney bean starch was 0.11 ± 0.03% for native starch and 0.97 ± 0.23% for nano starch. Ash content indicates starch purity and reflects the presence of non-volatile inorganic materials after high-temperature decomposition of organic compounds [ 28 ]. Ash contents are equivalent because native and nano starch have similar compositions [ 29 ]. 3.1.3 Water absorption capacity (WAC) The hydrophilic areas of starch influence its water absorption capacity (WAC), which indicates its potential to retain water through hydrogen bonding [ 28 ]. Table 1 shows that native kidney bean starch has a WAC of 2.34 ± 0.05%, within the range of 1.9 to 2.1 g/g reported by [ 28 ]. Nano starch from kidney beans exhibits a significantly higher WAC of 53.76 ± 2.14%, as also noted by Ali et al [ 30 ]. Factors affecting WAC include water binding sites, hydrogen bond availability, structural properties, and the hydrophilic-hydrophobic balance. Water binding is further influenced by the physicochemical environment, which includes temperature, vapour pressure, pH, ionic strength, and the presence of surfactants. 3.1.4 Oil absorption capacity (OAC) Native and nano kidney bean starches ability to absorb oil is shown in Table 1 . Oil absorption capacity, indicating a starch's ability to bind fat through capillary attraction, is essential for flavor preservation and improving mouthfeel [ 31 ] (Singh et al., 2006). Native kidney bean starch has an oil absorption capacity of 25.34 ± 0.02%, while nano starch shows a significantly higher capacity of 39.23 ± 0.04%. The hydrolysis of glycosidic linkages in nano starch is responsible for this improved capacity, producing smaller, more hygroscopic molecules with more free hydroxyl groups. 3.1.5 Field Emission Scanning Electron Microscopy (FE-SEM) FE-SEM was used to analyse the surface characteristics and morphology of kidney bean native and nano starches. Native starch granules were rounded and elliptical with no visible fissures. In contrast, nano starch, resulting from acid hydrolysis, appeared as agglomerated particles with irregular shapes, cracked and dented surfaces, and varying sizes (Fig. 1 ). The nanoparticles exhibited a void-filled, uneven surface and severe structural damage. The results are consistent with earlier research by [ 11 , 32 ], which observed that the morphology of starch nanoparticles is similar to the original native starch structure. Variations in granule shape arise from differences in botanical origin, amyloplast biochemistry, plant physiology, species, and age. 3.1.6 X-Ray Diffraction (XRD) For the analysis of starch granules' crystalline structure, X-ray diffraction (XRD) is essential. XRD patterns are used to classify starches into A, B, or C categories. According to Ghoshal & Kaushal [13 ], C-type starches, which are present in legumes like kidney beans, contain both A- and B-types are amorphous, whereas A- and B-type starches have different amorphous. Figure 2 shows that native kidney bean starch has peaks at 15.27º, 17.55º, 18.11º, and 23.26º 2θ, confirming its C-type structure. Nano kidney bean starch displays peaks at 15.01º, 16.56º, 18.77º, and 23.06º 2θ, which are slightly different but still indicate a C-type structure, consistent with the patterns reported by Ghoshal and Kaur [ 14 ]. Variations in XRD peaks can be influenced by factors such as starch granule size, crop type, and environmental conditions. Figure 2 indicates that both native and nano starches have similar C-type crystallinity. 3.1.7 Fourier Transform Infrared Spectroscopy (FTIR) The FTIR spectra of native and nano starch are displayed in Fig. 2 , which demonstrates similar patterns and suggests that the chemical composition of the starch has not changed significantly [ 34 ]. Peaks at 3425 cm⁻¹ and 2923 cm⁻¹ correspond to OH stretching and asymmetric C-H stretching, respectively. Peaks at 978 cm⁻¹, 2132 cm⁻¹, and 1591 cm⁻¹ are attributed to C-O, CN, and C = C stretching, while the peak at 572 cm⁻¹ represents C-H bonding. These findings align with [ 34 ]. However, FTIR only provides information on short-range order and vibration frequencies, not long-range crystallinity. The physicochemical structure of starch nanoparticles may vary slightly depending on the starch type used. 3.1.8 Swelling Power and Solubility The swelling power and solubility of native and nano starch at 60°C, 70°C, 80°C, and 90°C are displayed in Fig. 3 . Both characteristics rose as the temperature rose, peaking at 90°C. The molecular weight of amylopectin and amylose affects their swelling power and solubility. Heating causes these molecules to form hydrogen bonds with water, which weakens the crystalline structure and increases granule swelling [ 35 ]. Because to the melting of crystallites and the solvation-assisted transfer of chains from helix to coil, which raises enthalpy and breaks hydrogen bonds in the crystalline areas, nano starch showed higher solubility and swelling power than native starch. 2.5.2 Thermal characteristics (DSC) DSC measures endothermic peaks to measure gelatinization, which indicates the melting of starch [ 13 ]. The onset (To), peak (Tp), and conclusion (Tc) temperatures are influenced by the molecular makeup of amylopectin short chains (DP6–11). Because water-mediated starch crystallite melting breaks off the amorphous portions of the granule, kidney bean starches show single, narrow gelatinization endotherms. The DSC values are: To 44.57°C, Tp 66.88°C, Tc 122.93°C, and Te 167.84°C. The Differential Scanning Calorimetry (DSC) thermogram for kidney bean starch reveals key thermal properties. The Y-axis indicates heat flow (in mW/mg) and the X-axis represents temperature (in °C). The heat flow value of -12.798 J/g denotes the energy involved in an endothermic transition, likely corresponding to starch gelatinization. This transition occurs between 44.57°C and 167.84°C, with a peak at 67.861°C, reflecting the gelatinization temperature of the starch. The peak height of 1.212 mW signifies the intensity of this thermal event. Additionally, a glass transition temperature (Tg) is observed at 122.94°C, marking the transition from a glassy to a rubbery state, with a change in heat capacity (ΔCp) of 4.295 mW/°C, indicating a shift in the starch's properties at this temperature. 3.1.9 Dynamic Light Scattering (DLS) Native and nano starch sizes were determined using dynamic light scattering (DLS). Native starch had a size of 524.29 nm, while nano starch measured 282.7 nm. According to Sun [ 36 ], starch nanoparticles typically range from 10 to 300 nm and can be spherical, uneven, lamellar, or rod-like. Aggregation of starch nanocrystals is associated with hydrogen bond formation between hydroxyl groups on their surfaces [ 37 ]. 3.1.10 Rheology Figure 5 illustrates the rheological characteristics of native starch and starch nanoparticles. Nano starch suspensions exhibit higher viscosity than native starch, but viscosity decreases with increasing shear rate, indicating enhanced fluidity. This aligns with findings by Ahmad [ 16 ], which show that reduced starch size at the nano level increases viscosity. Rheological properties are influenced by granule distribution, size, shape, and intergranular contact. Flow curves (Fig. 5 b) demonstrate shear-thinning behavior, with all samples showing non-Newtonian pseudoplastic fluid characteristics. The viscosity decrease with rising shear rates is due to particle alignment in the flow direction [ 13 ]. 3.2 Characterization of essential oil 3.2.1 Gas Chromatography–Mass Spectrometry (GCMS) The volatile components of lemongrass essential oil were analyzed using GC-MS, revealing 40 compounds. Based on earlier research, eight important chemicals were identified among these [ 18 , 30 ]. Citral, neral, β-myrcene, and geraniol were the main ingredients, with small but significant levels of citronellol, 1, 3, 4-trimethyl-3-cyclohexene-1-carboxaldehyde, geranyl acetate, and D-limonene. Citral, a key quality indicator for lemongrass, was found at a concentration of 38.40%. The study noted that lemongrass essential oil contains a higher proportion of oxygenated monoterpenes compared to monoterpene hydrocarbons and sesquiterpenes, aligning with similar findings by Ali et al. and Jiang et al. [18. 30]. 3.3 Characterization of films 3.3.1 Moisture Content The moisture content of native and nano starch-based films is shown in Table 2 . The moisture content of native kidney bean starch film (S) was 11.88 ± 0.07%, while the kidney bean nano starch film (NS) showed lower moisture content of 7.12 ± 2.13%. The addition of lemongrass essential oil (1% v/v) further reduced the moisture content of native starch film to 8.31 ± 0.05% and nano starch film to 5.2 ± 0.9%. The lower moisture content in films with essential oil is attributed to the oil droplets filling the gaps between starch and water chains. Similar findings were reported by [ 38 ], where the addition of essential oils like lemongrass, oregano, and black cumin reduced film moisture content. Table 2 Moisture, Water Solubility, water vapour transmission rate, Thickness & Biodegradability of kidney bean film made of native and nanostarch. S. No. Sample Moisture Content (%) Water Solubility% WVTR Thickness (mm) Biodegradability (%) 1 S 11.88 ± 0.07 21.1 ± 0.3 6.90 × 10 − 3 ± 0.5 0.37 ± 0.08 6.7605 ± 0.08 2 S + EO 8.31 ± 0.05 13.8 ± 0.23 6.89 × 10 − 3 ± 0.5 0.45 ± 0.09 3.9080 ± 0.011 3 NS 7.12 ± 2.13 62.4 ± 0.07 2.50 × 10 − 3 ± 0.5 0.25 ± 0.02 15.0470 ± 0.09 4 NS + EO 5.2 ± 0.9 56.34 ± 0.06 2.57 × 10 − 3 ± 0.5 0.35 ± 0.12 4.6116 ± 0.04 3.3.2 Water Solubility The solubility of starch films is shown in Table 2 , where nano starch films showed greater solubility compared to native starch films [ 11 ]. The native kidney bean starch film had a solubility of 21.1 ± 0.3%, but the nano starch film had a significantly higher solubility of 62.4 ± 0.07%. This increased solubility in nano starch films is attributed to higher amylopectin content, which reduces water solubility due to starch microgranule aggregation [ 39 ]. However, the addition of lemongrass essential oil reduced water solubility to 13.8 ± 0.23% in native starch films and 56.34 ± 0.06% in nano starch films. Additionally, Song et al. [ 40 ] noted a decline in water solubility with increasing essential oil content due to reduced film hydrophilicity and interactions between essential oil components and hydroxyl groups. These results are consistent with earlier research demonstrating [ 41 , 42 ] and decreases [ 40 , 43 ] in starch film solubility with essential oil addition. 3.3.3 Water vapor transmission rate (WVTR) The water vapour transmission rate (WVTR) for kidney bean starch films (native and nano) is shown in Table 2 . The native starch film exhibited a higher WVTR (6.90 × 10⁻³ ± 0.5) compared to the nano starch film (2.50 × 10⁻³ ± 0.5). The reduction in WVTR for nano starch films is attributed to the increased surface-to-volume ratio and reduced polymer chain mobility due to nanometric sizes [ 11 ]. The addition of lemongrass essential oil did not significantly affect the WVTR of the films [ 44 ]. Nanoparticles enhance film compactness, improving water vapor resistance by forming a moisture barrier, which hinders water molecule diffusion [ 45 ]. Similar effects were observed with waxy maize starch nanocrystals, reducing WVTR in films [ 45 ]. 3.3.4 Physical and optical properties Table 2 shows that the natural kidney bean starch film had a thickness of 0.37 ± 0.08 mm, while the nano starch film was thinner at 0.25 ± 0.02 mm. Adding lemongrass essential oil increased film thickness, likely due to the oil's high viscosity, as noted by [ 46 ]. The tensile strength, crucial for food packaging durability, increased from 11.7 ± 0.12 MPa in native starch films to 15.7 ± 0.23 MPa in nano starch films, due to the enhanced mechanical strength provided by nanoparticles. The addition of essential oils also affected the appearance of the films. Lemongrass essential oil increased opacity, reducing transparency by 5%, with native starch film opacity at 26.43 ± 0.83% and nano starch film at 31.36 ± 0.27%. The oil altered color properties, increasing L* and a* values while decreasing b* values, likely due to the oil droplets disrupting the film's crystalline structure and enhancing light scattering. 3.3.5 Scanning Electron Microscopy (SEM) SEM was used to examine the surface characteristics and morphology of native and nano kidney bean starch films. (Fig. 7 ). The SEM images indicated that all film components were well-blended, forming dense, pore-free, and crack-free films. However, in films containing lemongrass essential oil, small oil droplets were visible throughout the film. According to Evangelho et al. [ 42 ], the incorporation of essential oil reduced the surface homogeneity of the films, which was initially smooth without EO. Similar observations were made by Caetano et al. [ 47 ], who noted increased surface heterogeneity after adding various essential oils like borage seed, lemon, and oregano. Emulsions on nano- and micro-scales have been successfully utilized to enhance the properties of biopolymer-based food packaging by incorporating hydrophobic, bioactive, and functional compounds, as noted by Espitia et al. [ 48 ]. These emulsions help to improve surface morphology, positively impacting other film properties. 3.3.6 X – Ray Diffraction (XRD) The crystal structure of kidney bean starch is C-type, a combination of A- and B-types, in line with earlier research [ 49 ]. The intensity of diffractograms decreased after converting starch into films, indicating disorganization of the crystalline structure due to retrogradation and starch gelatinization during heating. This disorganization was influenced by sorbitol, which replaced starch hydrogen bonds with starch-plasticizer bonds, reducing crystallinity. These results align with Singh et al. [ 33 ]. X-ray diffraction analysis revealed that native kidney bean starch displayed peaks at 15.24°, 17.32°, and 23.06°. However, these peaks converged when sorbitol was added during film preparation, indicating the melting of starch crystallites. When lemongrass essential oil was added to the S + EO film, a peak at 20.9° was created, reflecting intermolecular changes in the composite matrix, as similarly reported by Ghoshal & Singh, [ 50 ]. 3.3.7 Fourier Transform Infrared Spectroscopy (FTIR) The FTIR spectra revealed that the addition of essential oil did not alter the molecular interactions of the films, as their absorbance patterns remained similar to the control due to identical elemental composition. The following were important peaks: C = O stretching at 1652 cm⁻¹, C-O stretching at 1026 cm⁻¹, C-Cl bonding at 808 cm⁻¹, C-H bending at 570 cm⁻¹, O-H stretching at 3425 cm⁻¹, and C-H stretching alkane at 2923 cm⁻¹. These findings indicate that all films were compositionally similar, as they all contained kidney bean starch [ 33 ]. 3.3.8 Antioxidant Properties of films Essential oils can decrease the reliance on artificial chemicals and prevent oxidation that impacts food quality. A DPPH radical scavenging assay evaluated the films' antioxidant properties. Lemongrass essential oil exhibited significantly higher radical scavenging activity (70.33%) compared to the starch-based film (24.55%) (Fig. 8 ). This enhanced activity is due to the ability of lemongrass essential oil to donate hydrogen atoms to reduce DPPH radicals, as reported by Kadam et al. [ 51 ]. Table 3 DPPH Inhibition% of starch film and starch + essential oil film Sample Control Sample DPPH Inhibition% S 0.782 0.590 24.55 ± 0.21 S + EO 0.782 0.232 70.33 ± 0.34 3.3.9 Antimicrobial Properties of films Both Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli were tested for their antibacterial efficacy in starch-based edible films containing lemongrass essential oil. The lemongrass essential oil-treated films demonstrated strong antibacterial properties, with inhibition zone diameters ranging from 25.3 mm to 38.9 mm against E. coli and 34.9 mm to 43.2 mm against S. aureus . In contrast, the starch films without essential oil showed no antimicrobial activity. Gram-negative bacteria's outer barrier prevents the passage of hydrophobic components, whereas Gram-positive bacteria are more sensitive to essential oils. This explains why the oil-infused films were more efficient against Gram-positive bacteria. This aligns with findings by Ahmad et al. [ 52 ], indicating that essential oils are particularly effective against food spoilage organisms and food borne pathogens caused by Gram-positive bacteria. 3.3.10 Biodegradability of Films Biodegradation involves enzymatic activity and living organisms, facilitating molecular breakdown in both aerobic and anaerobic environments and results in the partial or whole elimination of substances. After 5 days at 25 ± 1°C, films containing starch (native/nano) and essential oils showed slower degradation compared to pure starch films, with native starch and essential oil films taking the longest to degrade. Similar findings by Gonçalves et al. [ 29 ], indicated that adding starch nanoparticles accelerated film biodegradation. This enhancement is attributed to water-induced swelling in the polymer, which promotes microbial growth and accelerates degradation. 3.4 Applications on fruits Kidney bean nano starch films (NS and NS + EO) were chosen for grapefruit wrapping due to their superior mechanical, barrier, and antibacterial properties. Starch-based films were produced for comparison. After 16 days at room temperature and 26 days in refrigeration, Fig. 9 showed the appearance of wrapped and unwrapped fruits, while Fig. 10 displayed weight loss, shrinkage, and pH changes during storage. Fruits wrapped in nano starch films exhibited significantly less weight loss (p < 0.05) due to better hydrophobic and water vapor barrier properties. The wrapping did not affect the fruits' color, which is important for consumer acceptance. The superior moisture barrier of nano starch films (Table 2 ) helps to reduce water and solute migration, thereby decreasing respiration and oxidation rates during storage. 3.4.1 Weight loss determination Grapefruit weight loss is linked to respiration and water evaporation, with their thin skin making them prone to rapid water loss, shrinkage, and tissue weakening. A study evaluated the effects of wrapping films on weight loss over 28 days of refrigeration and 16 days at room temperature. All fruits lost weight during storage, but as shown in Fig. 10 , wrapped fruits exhibited less weight loss compared to unwrapped ones. 3.4.2 Shrinkage Figure 10 visually depicts the shrinkage in wrapped and unwrapped grapes, which results from moisture loss, as water constitutes about 80% of a grape's weight. The results indicate that wrapped grapes experienced less shrinkage, demonstrating the effectiveness of the wrapping films in minimizing moisture loss. 3.4.3 pH The study found that grapefruit pH increased significantly over storage, with unwrapped samples showing greater pH levels than those wrapped in NS and NS + EO films, which exhibited less pH change. Refrigerated storage further enhanced the films’ performance, mitigating pH variations and extending shelf life. These findings underscore the effectiveness of NS-based films in maintaining fruit quality under varied storage conditions. 4. CONCLUSION The study highlights the potential of using kidney bean starch, particularly in its nano form, as a viable material for active food packaging. The extraction and conversion of starch through wet steeping and acid hydrolysis yielded native and nano starches with favorable physical and chemical properties. Both types exhibited similar crystallinity, but nano starch demonstrated superior characteristics when incorporated into edible films. The addition of lemongrass essential oil further enhanced the properties of the films, improving water vapor transmission, moisture content, solubility, and biodegradability. SEM analysis revealed a homogeneous and continuous film morphology, free from defects, underscoring the suitability of nano starch-based films for practical applications. The films developed in this study showed promising performance in extending the shelf life of perishable grapes. At room temperature, grapes wrapped in the films maintained quality for up to 16 days, compared to the typical 4–5 days for unwrapped fruit. In refrigerated conditions, the shelf life extended to 26 days. These findings suggest that nano starch-based films could offer significant benefits in food packaging, enhancing preservation without compromising environmental sustainability. While kidney bean starch is not commonly explored in packaging applications, this research demonstrates its potential as a sustainable and functional material. Further studies are required to fully understand the scope of its application and optimize its use in food packaging. This work contributes to the growing field of biodegradable and edible packaging, providing a basis for future innovations in food preservation. Declarations Conflict of Interest: There is no conflict of interest from any of the author. Consent to publish declaration Ms. Kanak has done experiments and wrote the 1st draft of the paper, Dr. Gargi supervises the work and final draft was made by Dr. Gargi and both were agreed to publish in Discover Food journal. Consent to Participate declaration Sensory analysis using human trial was not done. Ethics declaration No ethical statement is needed as neither human nor animals were used for the study. Clinical trial No clinical trial was done. Funding declaration No external funding was received for the study. Author Contribution Kanak did the experiments and wrote the first draft of manuscript , Gargi supervised her work and finalized the manuscript . All author reviewed the manuscript and agreed to publish in this journal Acknowledgement I do not have anyone to acknowledge Data Availability Data availability Statement: Data will be available from corresponding author on request References Lebreton, L. Van Der Zwet, J. Damsteeg, J. W. Slat, B. Andrady, A. L. Reisser, J. River plastic emissions to the world’s oceans. Nature Communications, 8(1). (2017) https://doi.org/10.1038/ncomms15611. Geyer, R. Jambeck, J. Law, K. L. Production, use, and fate of all plastics ever made. Sci. Adv., 3(7). (2017). https://doi.org/10.1126/sciadv.1700782 Li, P. Wang, X. Su, M. Zou, X. Duan, L. Zhang, H. Characteristics of Plastic Pollution in the Environment: a review. 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bean\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7095535/v1/6f7b41097b3250a707d891e9.png"},{"id":88215120,"identity":"5772083d-da24-4d7a-a26e-08499af1c775","added_by":"auto","created_at":"2025-08-04 06:39:14","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":140931,"visible":true,"origin":"","legend":"\u003cp\u003e(a) XRD (b) FTIR for kidney bean native (S) and nano starch (NS)\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7095535/v1/06a0fd699cfd3f07cb1a6a93.png"},{"id":88215162,"identity":"cc1c4411-84df-4138-8cec-d966e3977d20","added_by":"auto","created_at":"2025-08-04 06:39:21","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":83550,"visible":true,"origin":"","legend":"\u003cp\u003e(a)Swelling Power and (b) Solubility of native (S) and nano (NS) starch from kidney bean\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7095535/v1/2bd1ebef55b97bb812a0bab0.png"},{"id":88215155,"identity":"39837815-7cc9-45f3-92e4-50d13a24aff4","added_by":"auto","created_at":"2025-08-04 06:39:20","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":90205,"visible":true,"origin":"","legend":"\u003cp\u003eDSC of native starch\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-7095535/v1/6d2dcfc5d68ccc11dfabea31.png"},{"id":88215152,"identity":"4b44021d-909b-4d35-b520-cde2b55cc515","added_by":"auto","created_at":"2025-08-04 06:39:19","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":126443,"visible":true,"origin":"","legend":"\u003cp\u003e(a) Viscosity (b) Shear stress of native (S) and nano (NS) starch particles against shear rate\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-7095535/v1/ba882d2086eaaa10ead4cbf3.png"},{"id":88215119,"identity":"f1920815-4c09-40ce-b261-a972fcd362b2","added_by":"auto","created_at":"2025-08-04 06:39:14","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":102878,"visible":true,"origin":"","legend":"\u003cp\u003eGCMS of Lemon Grass Essential Oil\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-7095535/v1/b7e12bbf25bb02c62e5b9f03.png"},{"id":88215146,"identity":"7995d8d4-6a76-45bc-a68c-dae7078752d8","added_by":"auto","created_at":"2025-08-04 06:39:18","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":449021,"visible":true,"origin":"","legend":"\u003cp\u003eSEM images of Starch Films (a) Native Starch (b) Native Starch + Essential Oil (c) Nano Starch (d) Nano Starch + Essential Oil.\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-7095535/v1/bb6a5db620041289c15561c4.png"},{"id":88215141,"identity":"b77acd7d-8110-4e78-aa75-f4f2335d8ac5","added_by":"auto","created_at":"2025-08-04 06:39:17","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":69979,"visible":true,"origin":"","legend":"\u003cp\u003eAntioxidant values (%)\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-7095535/v1/83bb73d61ab253ed330f2c79.png"},{"id":88215164,"identity":"bf349c0a-658e-4f0d-8516-2b3f5cb89d72","added_by":"auto","created_at":"2025-08-04 06:39:22","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":1865598,"visible":true,"origin":"","legend":"\u003cp\u003eEffect on grapes (a) at room temperature (b) in refrigerator\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-7095535/v1/369f0ae28ab76339d5f0fdbf.png"},{"id":88215427,"identity":"2ec5f1fd-81bc-4754-a2d8-53919367c8cf","added_by":"auto","created_at":"2025-08-04 06:47:20","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":223121,"visible":true,"origin":"","legend":"\u003cp\u003e(a) Shrinkage (b) Weight loss (c) pH of grape fruit at room temperature\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-7095535/v1/2c3c62ed45de01a9633eb6f2.png"},{"id":88215163,"identity":"407d1b06-7ec9-471e-9d27-a66896ed4369","added_by":"auto","created_at":"2025-08-04 06:39:21","extension":"png","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":203019,"visible":true,"origin":"","legend":"\u003cp\u003e(a) Shrinkage (b) Weight loss (c) pH of grape fruit in refrigerator\u003c/p\u003e","description":"","filename":"11.png","url":"https://assets-eu.researchsquare.com/files/rs-7095535/v1/812a756917f63b13e7605cf2.png"},{"id":100888220,"identity":"6c527b99-9dfd-43aa-8568-286237ef5044","added_by":"auto","created_at":"2026-01-22 12:42:03","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":5257869,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7095535/v1/ed9b7c9a-2391-4080-99f7-bec4a5ba65bc.pdf"},{"id":88215134,"identity":"00fe04ff-3369-40e5-8718-edbcd1627b28","added_by":"auto","created_at":"2025-08-04 06:39:17","extension":"doc","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":897024,"visible":true,"origin":"","legend":"","description":"","filename":"GraphicalAbstract.doc","url":"https://assets-eu.researchsquare.com/files/rs-7095535/v1/0b92b5cdf5124ce33bdef3ec.doc"}],"financialInterests":"No competing interests reported.","formattedTitle":"Lemon grass Essential oil Incorporated Starch Based Active Packaging Film, Characterization and Applications","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eThe rapid rise in global plastic production has led to significant environmental pollution. Since the 1950s, plastic manufacturing has increased from 1.5\u0026nbsp;million to 335\u0026nbsp;million tonnes [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e], with 79% of plastic waste improperly disposed of in landfills or natural environments [2, 3]. The packaging industry consumes 36% of all plastics, primarily for single-use items like bags, bottles, and food packaging, generating over 150\u0026nbsp;million tonnes of waste annually. Only 9% of this waste is recycled, while 79% accumulates in the environment [2, 4]. Most plastics are derived from non-renewable petroleum, creating non-biodegradable polymers and exacerbating environmental harm. This highlights the urgent need for sustainable alternatives to reduce plastic consumption [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eBiodegradable active packaging is emerging as an eco-friendly alternative to petroleum-based plastics due to its versatility and environmental benefits. These films are made from biopolymers like proteins, polysaccharides, lipids, or their combinations and can degrade into non-toxic molecules or harmless biomass under natural conditions. Starch-based biodegradable products particularly have gained popularity. Starch, a polysaccharide found in plant seeds, tubers, roots, and fruits [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e5\u003c/span\u003e], is commonly used due to its abundance and ease of extraction, with grains, tubers, and legumes being the primary raw materials [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eBeans, part of the Fabaceae family, are globally significant food crops, both economically and nutritionally [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Kidney beans (\u003cem\u003ePhaseolus vulgaris\u003c/em\u003e) contain 22\u0026ndash;45% starch, which can be applied to both food and non-food products as a thickening agent, binder, film former, gelling agent, and more. While traditional starches from sources like maize, wheat, and potato have been well-studied, legume starches, including kidney bean starch, are less explored due to limited pulse availability in some regions. However, the rising interest in alternative starch sources is driving research into their properties. Kidney bean starch, with its high amylose content, is noted for high gelation temperatures, shear thinning resistance, rapid retrogradation, and high gel elasticity. Studying these properties is essential for expanding its industrial applications.\u003c/p\u003e\u003cp\u003eStudies show that incorporating co-biopolymers and nanomaterials can enhance the mechanical and barrier properties of starch-based films [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Bio-nanocomposites, which use nanoparticles as reinforcing fillers, address the limitations of biopolymer packaging by improving rheological, physical, mechanical, and thermal properties, as well as processability and performance [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Native and nano starch from the same source offer better compatibility, leading to improved film properties [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Acid-hydrolysed starch nanoparticles are particularly useful in high-temperature food processing due to their high crystallinity and stability. However, research on nano starch derived from kidney bean starch remains limited [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e"},{"header":"2. Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1. Materials\u003c/h2\u003e\u003cp\u003eKidney beans utilised in the research were sourced from Krishi Vigyan Kendra, Kukumseri, in the Lahaul \u0026amp; Spiti district of Himachal Pradesh, India collected through proper channel complied with local guidelines. Analytical-grade chemicals employed in the research were procured from Sisco Research Laboratories Pvt. Ltd., Loba Chemie Pvt. Ltd., and Allin Exporters. Additionally, fruits used in the study were obtained from a local market in Chandigarh, India.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2. Starch isolation\u003c/h2\u003e\u003cp\u003eKidney beans' starch was extracted using a modified wet steeping technique [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e12\u003c/span\u003e] (Sandhu et al., 2005). The beans were soaked for 20 hours in 1.25 litres of distilled water containing 0.1% potassium metabisulphite. They were then ground with water to form smooth slurry. After the slurry was run through 100-mesh nylon cloth, all remaining substance was repeatedly washed with distilled water until it was clear of starch. The filtered slurry was allowed to settle for 4\u0026ndash;5 hours, and the clear supernatant was centrifuged at 5000 rpm for 10 minutes. The top brownish protein layer was removed after each centrifugation until the supernatant was clear. A hot air oven was used to dry the final starch at 40\u0026deg;C, and stored in an airtight container for further use.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e2.3. Preparation of Nano Starch\u003c/h2\u003e\u003cp\u003eNano starch was produced using a modified acid hydrolysis method [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Fifteen grams of dry native starch were incubated in 3.16 M sulfuric acid at 40\u0026deg;C for seven days with magnetic stirring. After that, the mixture was centrifuged for 15 minutes at 6000 rpm. After repeatedly washing the precipitates with distilled water until they were neutralised, acetone was used to wash them. A hot air oven set to 40\u0026deg;C was used to dry the resultant starch.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003e2.4. Physicochemical properties of starch\u003c/h2\u003e\u003cp\u003eThe following methods were used to measure the starch's amylose content [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e13\u003c/span\u003e], ash content [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e14\u003c/span\u003e], water and oil absorption capacity [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e13\u003c/span\u003e], swelling power, and solubility [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003e2.5. Characterization of starch\u003c/h2\u003e\u003cdiv id=\"Sec8\" class=\"Section3\"\u003e\u003ch2\u003e2.5.1Surface morphology\u003c/h2\u003e\u003cp\u003eSamples of starch were examined for surface morphology using a Hitachi SU 8010 field emission scanning electron microscope. Kidney bean native and nano starches were mounted on carbon tape and gold-coated by sputtering for 60 seconds before imaging.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec9\" class=\"Section3\"\u003e\u003ch2\u003e2.5.2 Thermal characteristics (DSC)\u003c/h2\u003e\u003cp\u003eThe differential scanning calorimeter was used to evaluate the starches' thermal characteristics. A 1:3 (g/g) starch-to-water ratio was prepared using 3.1 mg of starch and distilled water in an aluminium pan, which was sealed and equilibrated for one hour. The sample was then heated at 10\u0026deg;C per minute, starting from 10\u0026deg;C for five minutes. An empty pan and indium were used for calibration. Gelatinisation onset temperature (To), peak temperature (Tp), glass transition temperature (Tg), and end set temperature (Te) were among the important thermal parameters that were measured.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec10\" class=\"Section3\"\u003e\u003ch2\u003e2.5.3 Size distribution\u003c/h2\u003e\u003cp\u003eUsing a Nano Zetasizer dynamic light scattering (DLS) device (Malvern Instruments Inc., U.K.), the size and distribution of starch particles were determined. Samples were prepared at a 0.01% (w/v) concentration in distilled water, diluted 100-fold with deionized water, and analysed at 30\u0026deg;C in plastic cuvettes. Measurements were performed without filtration or dust removal [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e15\u003c/span\u003e] (Sadeghi et al., 2017).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec11\" class=\"Section3\"\u003e\u003ch2\u003e2.5.4 Rheology\u003c/h2\u003e\u003cp\u003eRheology of starch studies the deformation and flow of starch under external forces. In 100 millilitres of distilled water, a 3% starch solution (nano/native) was heated to 90\u0026deg;C for 20 minutes while being constantly stirred to form a gel [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. After cooling to room temperature, An Anton Paar MCR 102 rheometer with parallel plate geometry (PP-50; diameter: 50 mm) was used to measure the rheological properties at 25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section3\"\u003e\u003ch2\u003e2.5.5 X-Ray Diffraction (XRD) analysis\u003c/h2\u003e\u003cp\u003eCrystalline phases are identified via X-ray powder diffraction (XRD). An X'Pert Pro A diffractometer (Analytical, U.S.A.) operating at 45 kV and 40 mA recorded diffraction patterns using Cu Kα radiation in a 2θ configuration.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section3\"\u003e\u003ch2\u003e2.5.6 Fourier Transforms Infrared Spectroscopy (FTIR)\u003c/h2\u003e\u003cp\u003eFourier Transform Infrared Spectroscopy (FTIR) was used to examine the molecular interactions of starch [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. FTIR spectra for treated and control samples were recorded with a Shimadzu 8400 spectrometer (Japan) with a resolution of 4 cm⁻\u0026sup1; and in the 500\u0026ndash;4000 cm⁻\u0026sup1; range. Samples were conditioned at 53\u0026thinsp;\u0026plusmn;\u0026thinsp;1% relative humidity for 24 hours before analysis. Data were analyzed using the FTIR software.\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003ch2\u003e2.6 Characterization of essential oil\u003c/h2\u003e\u003cdiv id=\"Sec15\" class=\"Section3\"\u003e\u003ch2\u003e2.6.1 Gas Chromatography\u0026ndash;Mass Spectrometry (GCMS)\u003c/h2\u003e\u003cp\u003eVolatile compounds in essential oils are separated into individual components using Gas Chromatography (GC), which produces a linear chromatogram of these constituents. Mass Spectrometry (MS) is then used to identify and quantify each component, aiding in the detection of potential adulteration [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\u003ch2\u003e2.7 Preparation of films\u003c/h2\u003e\u003cp\u003eEdible films were produced by casting from aqueous solutions and evaporating the liquid phase. 4g of native or nano starch were combined with 100ml of distilled water and heated to 95\u0026deg;C for 30 minutes in order to create kidney bean starch films. After adding 30% w/w D-sorbitol as a plasticizer, the mixture was heated for an additional 40 minutes and then homogenized with a probe sonicator for 10 minutes. Petri dishes were filled with the solution, which was then allowed to dry at room temperature.\u003c/p\u003e\u003cp\u003eTo create composite films with lemon grass essential oil, distilled water was used to make a 1% w/v Tween 80 and essential oil solution. This solution was then heated to 45\u0026deg;C for 40 minutes and combined with the starch solution that had already been made. The combined mixtures were stirred at 50\u0026deg;C for 10 minutes, transferred to petri plates and allowed to air dry at room temperature. The dried films were then removed for further characterization.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\u003ch2\u003e2.8 Physicochemical properties of film\u003c/h2\u003e\u003cdiv id=\"Sec18\" class=\"Section3\"\u003e\u003ch2\u003e2.8.1 Moisture content of films\u003c/h2\u003e\u003cp\u003eFilm samples (S, S\u0026thinsp;+\u0026thinsp;EO, NS, NS\u0026thinsp;+\u0026thinsp;EO) measuring 4 \u0026times; 1 cm\u0026sup2; were heated in an oven (at 105\u0026deg;C) until their weight remained unchanged, indicating complete drying. The moisture content was calculated using Eq.\u0026nbsp;1 and determined using the [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e19\u003c/span\u003e] approach. All experiments were conducted in triplicate.\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\:\\text{M}\\text{C}=\\frac{\\:{\\text{W}}_{0}-\\:{\\text{W}}_{1}}{\\:{\\text{W}}_{0}}\\times\\:100\\:\\:\\dots\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\text{E}\\text{q}\\:\\:1$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eWhere, MC\u0026thinsp;=\u0026thinsp;Moisture content, W\u003csub\u003e0\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Mass of the initial sample, W\u003csub\u003e1\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Mass of the final sample.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec19\" class=\"Section3\"\u003e\u003ch2\u003e2.8.2 Water solubility of film\u003c/h2\u003e\u003cp\u003eFilm samples (S, S\u0026thinsp;+\u0026thinsp;EO, NS, NS\u0026thinsp;+\u0026thinsp;EO) measuring 4 \u0026times; 1 cm\u0026sup2; were dried for 24 hours at 105\u0026deg;C to achieve a constant weight (W\u003csub\u003eo\u003c/sub\u003e). After drying, the samples were submerged for 24 hours at 25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C in 50 ml of distilled water. The samples were reweighed (W\u003csub\u003e1\u003c/sub\u003e) after being re-dried at 105\u0026deg;C. Eq.\u0026nbsp;2 was used to determine the film\u0026rsquo;s water solubility, with multiple tests conducted for accuracy.\u003cdiv id=\"Equb\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equb\" name=\"EquationSource\"\u003e\n$$\\:\\text{S}\\:\\left(\\text{%}\\right)=\\frac{\\:{\\text{W}}_{0}-\\:{\\text{W}}_{1}}{\\:\\:{\\text{W}}_{0}}\\times\\:100\\:\\:\\dots\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\text{E}\\text{q}\\:\\:\\:\\:\\:2\\:\\:$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eWhere, S\u0026thinsp;=\u0026thinsp;Solubility, W\u003csub\u003e0\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Weight of the dried sample before water immersion, and\u003c/p\u003e\u003cp\u003eW\u003csub\u003e1\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Dry weight of the insoluble sample after immersion.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec20\" class=\"Section3\"\u003e\u003ch2\u003e2.8.3 Water Vapour Permeability (WVP) of films\u003c/h2\u003e\u003cp\u003eThe film samples were put on modified beakers with calcium chloride (0% RH) after being paraffin-sealed. These beakers were then incubated at 25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C in a desiccator filled with distilled water (100% RH). The increase in beaker weight was recorded over time [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. These formulas were used to calculate the film's WVP.\u003c/p\u003e\u003cp\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\text{W}\\text{V}\\text{T}\\text{R}=\\frac{\\varDelta\\:\\text{W}\\times\\:\\text{t}}{\\text{A}}\\)\u003c/span\u003e\u003c/span\u003e \u0026hellip; Eq.\u0026nbsp;3\u003cdiv id=\"Equc\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equc\" name=\"EquationSource\"\u003e\n$$\\:\\text{W}\\text{V}\\text{P}=\\:\\frac{\\text{W}\\text{V}\\text{T}\\text{R}\\times\\:\\text{L}}{\\varDelta\\:\\text{P}}\\:\\:\\dots\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:Eq\\:\\:\\:\\:\\:4$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eMeasurements were taken in triplicate, with the variables being water vapor transmission rate (WVTR), sample thickness (L), cross-sectional area (A), and differential pressure (ΔP) across the film.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec21\" class=\"Section3\"\u003e\u003ch2\u003e2.8.4 Physical and optical properties\u003c/h2\u003e\u003cp\u003eFilm thickness was measured with 0.01 mm accuracy using a Mitutoyo digital micrometre (Japan), taking readings at 10 points per sample and averaging the results. Each measurement was repeated three times [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Film color and opacity were assessed using a Hunter colorimeter (Color flex model, Hunter Lab, USA). A Texture Analyser (TA.XT Plus, Stable Microsystems) was used to assess mechanical parameters, such as tensile strength, in accordance with ASTM standard method 828\u0026thinsp;\u0026minus;\u0026thinsp;97 (ASTM, 2002).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec22\" class=\"Section3\"\u003e\u003ch2\u003e2.8.5 Swelling Power and Solubility\u003c/h2\u003e\u003cp\u003e0.1 g of the film sample was dissolved in 10 ml of distilled water and heated for 30 minutes at 60\u0026deg;C, 70\u0026deg;C, 80\u0026deg;C, and 90\u0026deg;C for the swelling power analysis. The sample was centrifuged for 10 minutes at 2000 rpm after cooling. The precipitate was weighed, and the supernatant was dried overnight at 80\u0026deg;C in pre-weighed petri plates, with weight change recorded.\u003c/p\u003e\u003cp\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\text{S}\\text{I}=\\frac{\\text{W}1\\:\\left(\\text{g}\\right)}{\\text{W}\\text{o}\\:\\left(\\text{g}\\right)}\\)\u003c/span\u003e\u003c/span\u003e\u0026hellip; Eq.\u0026nbsp;5\u003c/p\u003e\u003cp\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\text{S}\\text{o}\\text{l}\\text{u}\\text{b}\\text{i}\\text{l}\\text{i}\\text{t}\\text{y}\\:\\left(\\text{%}\\right)=\\frac{\\text{W}2\\:\\left(\\text{g}\\right)}{\\text{W}\\text{o}\\:\\left(\\text{g}\\right)}\\times\\:100\\)\u003c/span\u003e\u003c/span\u003e \u0026hellip; Eq.\u0026nbsp;6\u003c/p\u003e\u003cp\u003eWhere, SI\u0026thinsp;=\u0026thinsp;Swelling Index, S\u0026thinsp;=\u0026thinsp;Solubility, W\u003csub\u003eo\u003c/sub\u003e= Starch taken (g), W\u003csub\u003e1\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Dry precipitates (g), W\u003csub\u003e2\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Weight after supernatant dried\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec23\" class=\"Section2\"\u003e\u003ch2\u003e2.9 Characterization of films\u003c/h2\u003e\u003cp\u003eThe films were subjected to X-Ray Diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FTIR) using the previously described techniques.\u003c/p\u003e\u003cdiv id=\"Sec24\" class=\"Section3\"\u003e\u003ch2\u003e2.9.1 SEM\u003c/h2\u003e\u003cp\u003eA scanning electron microscope (SEM) was used to analyse the films' surface morphology. The external surfaces were sputter-coated with gold prior to observation, and gold was also applied to fracture surfaces before imaging [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e21\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec25\" class=\"Section3\"\u003e\u003ch2\u003e2.9.2 Antioxidant Activity of films\u003c/h2\u003e\u003cp\u003eEssential oils are frequently incorporated to films made of starch because of their well-known antioxidant qualities. The antioxidant activity of films containing lemongrass essential oil was tested using the method described by [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e22\u003c/span\u003e] .\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec26\" class=\"Section3\"\u003e\u003ch2\u003e2.9.3 Antimicrobial activity of films\u003c/h2\u003e\u003cp\u003eLemongrass essential oil was tested against both Gram-positive (\u003cem\u003eS. aureus\u003c/em\u003e) and Gram-negative (\u003cem\u003eE. coli\u003c/em\u003e) bacteria to determine the antibacterial activity of starch-based edible films. The antibacterial activity of the lemongrass essential oil was assessed using the agar well diffusion method. A 6\u0026ndash;8 mm well in an inoculated agar plate was treated with the oil extract and then kept in a 37\u0026deg;C incubator for 48 hours. The oil inhibited bacterial growth by diffusing through the agar, and the clear zone diameter was measured in millimetres to determine its antibacterial activity.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec27\" class=\"Section3\"\u003e\u003ch2\u003e2.9.4 Biodegradability\u003c/h2\u003e\u003cp\u003eBiodegradability of a film reflects its susceptibility to environmental degradation, primarily through microbial activity. To assess this, composite film specimens (1\u0026times;2 cm\u0026sup2;) were buried 2 cm below the surface in a mixture of uniformly sized soil and compost within an earthen pot. The weight loss of the films was monitored under controlled conditions (25\u0026deg;C) at 5-day intervals.\u003c/p\u003e\u003cp\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\text{B}\\left(\\text{%}\\right)=\\:\\frac{W\\text{o}-W1}{W\\text{o}}\\times\\:100\\)\u003c/span\u003e\u003c/span\u003e Eq.\u0026nbsp;7\u003c/p\u003e\u003cp\u003eWhere, B(%)\u0026thinsp;=\u0026thinsp;Biodegradability, Wo\u0026thinsp;=\u0026thinsp;Initial weight of sample, W\u003csub\u003e1\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;Final weight of sample.\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec28\" class=\"Section2\"\u003e\u003ch2\u003e2.10 Applications on fruits\u003c/h2\u003e\u003cp\u003eThe experiment utilized a completely randomized design with 5 samples: A fruit sample was kept unwrapped (Control), while the others were wrapped in native starch film (S), native starch film with lemongrass essential oil (S\u0026thinsp;+\u0026thinsp;EO), nano starch film (NS), and nano-starch film containing lemongrass essential oil (NS\u0026thinsp;+\u0026thinsp;EO). Fruits were kept in a refrigerator and at room temperature, and analyses were done after16 days at room temperature and 26 days in the refrigerator [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e\u003cdiv id=\"Sec29\" class=\"Section3\"\u003e\u003ch2\u003e2.10.1 Weight loss determination\u003c/h2\u003e\u003cp\u003eWeight loss percentages of each fruit were calculated using their initial and final weights. Grapes were weighed every four days using a digital balance.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec30\" class=\"Section3\"\u003e\u003ch2\u003e2.10.2 Shrinkage\u003c/h2\u003e\u003cp\u003eThe initial diameter of the fruits was measured at two positions near the middle using a digital micrometre (Mitutoyo, Japan), and the average diameter was recorded. During storage, diameter measurements were taken every fourth day to assess shrinkage for all samples [24] .\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec31\" class=\"Section2\"\u003e\u003ch2\u003e2.11 Statistical analysis\u003c/h2\u003e\u003cp\u003eOne-way ANOVA in IBM SPSS Statistics Trial Version was used to evaluate the data. Using Duncan's multiple range test, significant variations in means were evaluated (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\u003c/div\u003e"},{"header":"3. Results and Discussion","content":"\u003cdiv id=\"Sec33\" class=\"Section2\"\u003e\u003ch2\u003e3.1 Physicochemical properties of native and nano starch from kidney bean\u003c/h2\u003e\u003cdiv id=\"Sec34\" class=\"Section3\"\u003e\u003ch2\u003e3.1.1 Amylose Content\u003c/h2\u003e\u003cp\u003eAmylose content in starch is affected by both ambient growth circumstances and genetic variables. According to Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, the amylose concentration of the native kidney bean was 12.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35%. On the other hand, due to climatic and ecological variations, kidney beans\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eAmylose content, ash content, water and oil absorption capacity of native and nano starch from kidney bean\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"6\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eS. NO.\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSample\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAmylose Content\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eAsh content\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eWater absorption capacity%\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eOil absorption capacity %\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNative\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e0.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e2.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05%.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e25.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNano\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e-\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e0.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23%\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e53.76\u0026thinsp;\u0026plusmn;\u0026thinsp;2.14%.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e39.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003ehad a much greater amylose content of 32.4%, according to [25, 26]. The amylose concentration of nano starch's was not ascertained. The amylose concentration may become undetected due to severe hydrolysis because the remaining linear chains are too short to create a blue iodine complex [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e27\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec35\" class=\"Section3\"\u003e\u003ch2\u003e3.1.2 Ash content\u003c/h2\u003e\u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e indicates that the ash level of kidney bean starch was 0.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03% for native starch and 0.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23% for nano starch. Ash content indicates starch purity and reflects the presence of non-volatile inorganic materials after high-temperature decomposition of organic compounds [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Ash contents are equivalent because native and nano starch have similar compositions [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e29\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec36\" class=\"Section3\"\u003e\u003ch2\u003e3.1.3 Water absorption capacity (WAC)\u003c/h2\u003e\u003cp\u003eThe hydrophilic areas of starch influence its water absorption capacity (WAC), which indicates its potential to retain water through hydrogen bonding [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e shows that native kidney bean starch has a WAC of 2.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05%, within the range of 1.9 to 2.1 g/g reported by [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Nano starch from kidney beans exhibits a significantly higher WAC of 53.76\u0026thinsp;\u0026plusmn;\u0026thinsp;2.14%, as also noted by Ali et al [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Factors affecting WAC include water binding sites, hydrogen bond availability, structural properties, and the hydrophilic-hydrophobic balance. Water binding is further influenced by the physicochemical environment, which includes temperature, vapour pressure, pH, ionic strength, and the presence of surfactants.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec37\" class=\"Section3\"\u003e\u003ch2\u003e3.1.4 Oil absorption capacity (OAC)\u003c/h2\u003e\u003cp\u003eNative and nano kidney bean starches ability to absorb oil is shown in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. Oil absorption capacity, indicating a starch's ability to bind fat through capillary attraction, is essential for flavor preservation and improving mouthfeel [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e31\u003c/span\u003e] (Singh et al., 2006). Native kidney bean starch has an oil absorption capacity of 25.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02%, while nano starch shows a significantly higher capacity of 39.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04%. The hydrolysis of glycosidic linkages in nano starch is responsible for this improved capacity, producing smaller, more hygroscopic molecules with more free hydroxyl groups.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec38\" class=\"Section3\"\u003e\u003ch2\u003e3.1.5 Field Emission Scanning Electron Microscopy (FE-SEM)\u003c/h2\u003e\u003cp\u003eFE-SEM was used to analyse the surface characteristics and morphology of kidney bean native and nano starches. Native starch granules were rounded and elliptical with no visible fissures. In contrast, nano starch, resulting from acid hydrolysis, appeared as agglomerated particles with irregular shapes, cracked and dented surfaces, and varying sizes (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The nanoparticles exhibited a void-filled, uneven surface and severe structural damage. The results are consistent with earlier research by [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e32\u003c/span\u003e], which observed that the morphology of starch nanoparticles is similar to the original native starch structure. Variations in granule shape arise from differences in botanical origin, amyloplast biochemistry, plant physiology, species, and age.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec39\" class=\"Section3\"\u003e\u003ch2\u003e3.1.6 X-Ray Diffraction (XRD)\u003c/h2\u003e\u003cp\u003eFor the analysis of starch granules' crystalline structure, X-ray diffraction (XRD) is essential. XRD patterns are used to classify starches into A, B, or C categories. According to Ghoshal \u0026amp; Kaushal [13 ], C-type starches, which are present in legumes like kidney beans, contain both A- and B-types are amorphous, whereas A- and B-type starches have different amorphous. Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e shows that native kidney bean starch has peaks at 15.27\u0026ordm;, 17.55\u0026ordm;, 18.11\u0026ordm;, and 23.26\u0026ordm; 2θ, confirming its C-type structure. Nano kidney bean starch displays peaks at 15.01\u0026ordm;, 16.56\u0026ordm;, 18.77\u0026ordm;, and 23.06\u0026ordm; 2θ, which are slightly different but still indicate a C-type structure, consistent with the patterns reported by Ghoshal and Kaur [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Variations in XRD peaks can be influenced by factors such as starch granule size, crop type, and environmental conditions. Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e indicates that both native and nano starches have similar C-type crystallinity.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec40\" class=\"Section3\"\u003e\u003ch2\u003e3.1.7 Fourier Transform Infrared Spectroscopy (FTIR)\u003c/h2\u003e\u003cp\u003eThe FTIR spectra of native and nano starch are displayed in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, which demonstrates similar patterns and suggests that the chemical composition of the starch has not changed significantly [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Peaks at 3425 cm⁻\u0026sup1; and 2923 cm⁻\u0026sup1; correspond to OH stretching and asymmetric C-H stretching, respectively. Peaks at 978 cm⁻\u0026sup1;, 2132 cm⁻\u0026sup1;, and 1591 cm⁻\u0026sup1; are attributed to C-O, CN, and C\u0026thinsp;=\u0026thinsp;C stretching, while the peak at 572 cm⁻\u0026sup1; represents C-H bonding. These findings align with [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. However, FTIR only provides information on short-range order and vibration frequencies, not long-range crystallinity. The physicochemical structure of starch nanoparticles may vary slightly depending on the starch type used.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec41\" class=\"Section3\"\u003e\u003ch2\u003e3.1.8 Swelling Power and Solubility\u003c/h2\u003e\u003cp\u003eThe swelling power and solubility of native and nano starch at 60\u0026deg;C, 70\u0026deg;C, 80\u0026deg;C, and 90\u0026deg;C are displayed in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. Both characteristics rose as the temperature rose, peaking at 90\u0026deg;C. The molecular weight of amylopectin and amylose affects their swelling power and solubility. Heating causes these molecules to form hydrogen bonds with water, which weakens the crystalline structure and increases granule swelling [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Because to the melting of crystallites and the solvation-assisted transfer of chains from helix to coil, which raises enthalpy and breaks hydrogen bonds in the crystalline areas, nano starch showed higher solubility and swelling power than native starch.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec42\" class=\"Section3\"\u003e\u003ch2\u003e2.5.2 Thermal characteristics (DSC)\u003c/h2\u003e\u003cp\u003eDSC measures endothermic peaks to measure gelatinization, which indicates the melting of starch [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. The onset (To), peak (Tp), and conclusion (Tc) temperatures are influenced by the molecular makeup of amylopectin short chains (DP6\u0026ndash;11). Because water-mediated starch crystallite melting breaks off the amorphous portions of the granule, kidney bean starches show single, narrow gelatinization endotherms. The DSC values are: To 44.57\u0026deg;C, Tp 66.88\u0026deg;C, Tc 122.93\u0026deg;C, and Te 167.84\u0026deg;C.\u003c/p\u003e\u003cp\u003eThe Differential Scanning Calorimetry (DSC) thermogram for kidney bean starch reveals key thermal properties. The Y-axis indicates heat flow (in mW/mg) and the X-axis represents temperature (in \u0026deg;C). The heat flow value of -12.798 J/g denotes the energy involved in an endothermic transition, likely corresponding to starch gelatinization. This transition occurs between 44.57\u0026deg;C and 167.84\u0026deg;C, with a peak at 67.861\u0026deg;C, reflecting the gelatinization temperature of the starch. The peak height of 1.212 mW signifies the intensity of this thermal event. Additionally, a glass transition temperature (Tg) is observed at 122.94\u0026deg;C, marking the transition from a glassy to a rubbery state, with a change in heat capacity (ΔCp) of 4.295 mW/\u0026deg;C, indicating a shift in the starch's properties at this temperature.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec43\" class=\"Section3\"\u003e\u003ch2\u003e3.1.9 Dynamic Light Scattering (DLS)\u003c/h2\u003e\u003cp\u003eNative and nano starch sizes were determined using dynamic light scattering (DLS). Native starch had a size of 524.29 nm, while nano starch measured 282.7 nm. According to Sun [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e36\u003c/span\u003e], starch nanoparticles typically range from 10 to 300 nm and can be spherical, uneven, lamellar, or rod-like. Aggregation of starch nanocrystals is associated with hydrogen bond formation between hydroxyl groups on their surfaces [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e37\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec44\" class=\"Section3\"\u003e\u003ch2\u003e3.1.10 Rheology\u003c/h2\u003e\u003cp\u003eFigure \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e illustrates the rheological characteristics of native starch and starch nanoparticles. Nano starch suspensions exhibit higher viscosity than native starch, but viscosity decreases with increasing shear rate, indicating enhanced fluidity. This aligns with findings by Ahmad [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e16\u003c/span\u003e], which show that reduced starch size at the nano level increases viscosity. Rheological properties are influenced by granule distribution, size, shape, and intergranular contact. Flow curves (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eb) demonstrate shear-thinning behavior, with all samples showing non-Newtonian pseudoplastic fluid characteristics. The viscosity decrease with rising shear rates is due to particle alignment in the flow direction [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec45\" class=\"Section2\"\u003e\u003ch2\u003e3.2 Characterization of essential oil\u003c/h2\u003e\u003cdiv id=\"Sec46\" class=\"Section3\"\u003e\u003ch2\u003e3.2.1 Gas Chromatography\u0026ndash;Mass Spectrometry (GCMS)\u003c/h2\u003e\u003cp\u003eThe volatile components of lemongrass essential oil were analyzed using GC-MS, revealing 40 compounds. Based on earlier research, eight important chemicals were identified among these [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Citral, neral, β-myrcene, and geraniol were the main ingredients, with small but significant levels of citronellol, 1, 3, 4-trimethyl-3-cyclohexene-1-carboxaldehyde, geranyl acetate, and D-limonene. Citral, a key quality indicator for lemongrass, was found at a concentration of 38.40%. The study noted that lemongrass essential oil contains a higher proportion of oxygenated monoterpenes compared to monoterpene hydrocarbons and sesquiterpenes, aligning with similar findings by Ali et al. and Jiang et al. [18. 30].\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec47\" class=\"Section2\"\u003e\u003ch2\u003e3.3 Characterization of films\u003c/h2\u003e\u003cdiv id=\"Sec48\" class=\"Section3\"\u003e\u003ch2\u003e3.3.1 Moisture Content\u003c/h2\u003e\u003cp\u003eThe moisture content of native and nano starch-based films is shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The moisture content of native kidney bean starch film (S) was 11.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07%, while the kidney bean nano starch film (NS) showed lower moisture content of 7.12\u0026thinsp;\u0026plusmn;\u0026thinsp;2.13%. The addition of lemongrass essential oil (1% v/v) further reduced the moisture content of native starch film to 8.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05% and nano starch film to 5.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9%. The lower moisture content in films with essential oil is attributed to the oil droplets filling the gaps between starch and water chains. Similar findings were reported by [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e38\u003c/span\u003e], where the addition of essential oils like lemongrass, oregano, and black cumin reduced film moisture content.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eMoisture, Water Solubility, water vapour transmission rate, Thickness \u0026amp; Biodegradability of kidney bean film made of native and nanostarch.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026times;\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eS. No.\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSample\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMoisture Content (%)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eWater Solubility%\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eWVTR\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eThickness (mm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eBiodegradability (%)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e11.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e21.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026times;\" colname=\"c5\"\u003e\u003cp\u003e6.90 \u0026times; 10\u003csup\u003e\u0026minus;\u0026thinsp;3\u003c/sup\u003e\u0026plusmn; 0.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e6.7605\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eS\u0026thinsp;+\u0026thinsp;EO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e8.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e13.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026times;\" colname=\"c5\"\u003e\u003cp\u003e6.89 \u0026times; 10\u003csup\u003e\u0026minus;\u0026thinsp;3\u003c/sup\u003e\u0026plusmn; 0.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e3.9080\u0026thinsp;\u0026plusmn;\u0026thinsp;0.011\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e7.12\u0026thinsp;\u0026plusmn;\u0026thinsp;2.13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e62.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026times;\" colname=\"c5\"\u003e\u003cp\u003e2.50 \u0026times; 10\u003csup\u003e\u0026minus;\u0026thinsp;3\u003c/sup\u003e\u0026plusmn; 0.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e15.0470\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNS\u0026thinsp;+\u0026thinsp;EO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e5.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e56.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026times;\" colname=\"c5\"\u003e\u003cp\u003e2.57 \u0026times; 10\u003csup\u003e\u0026minus;\u0026thinsp;3\u003c/sup\u003e\u0026plusmn; 0.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e4.6116\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec49\" class=\"Section3\"\u003e\u003ch2\u003e3.3.2 Water Solubility\u003c/h2\u003e\u003cp\u003eThe solubility of starch films is shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, where nano starch films showed greater solubility compared to native starch films [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. The native kidney bean starch film had a solubility of 21.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3%, but the nano starch film had a significantly higher solubility of 62.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07%. This increased solubility in nano starch films is attributed to higher amylopectin content, which reduces water solubility due to starch microgranule aggregation [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. However, the addition of lemongrass essential oil reduced water solubility to 13.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23% in native starch films and 56.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06% in nano starch films. Additionally, Song et al. [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e40\u003c/span\u003e] noted a decline in water solubility with increasing essential oil content due to reduced film hydrophilicity and interactions between essential oil components and hydroxyl groups. These results are consistent with earlier research demonstrating [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e41\u003c/span\u003e, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e42\u003c/span\u003e] and decreases [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e40\u003c/span\u003e, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e43\u003c/span\u003e] in starch film solubility with essential oil addition.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec50\" class=\"Section3\"\u003e\u003ch2\u003e3.3.3 Water vapor transmission rate (WVTR)\u003c/h2\u003e\u003cp\u003eThe water vapour transmission rate (WVTR) for kidney bean starch films (native and nano) is shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The native starch film exhibited a higher WVTR (6.90 \u0026times; 10⁻\u0026sup3; \u0026plusmn; 0.5) compared to the nano starch film (2.50 \u0026times; 10⁻\u0026sup3; \u0026plusmn; 0.5). The reduction in WVTR for nano starch films is attributed to the increased surface-to-volume ratio and reduced polymer chain mobility due to nanometric sizes [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. The addition of lemongrass essential oil did not significantly affect the WVTR of the films [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. Nanoparticles enhance film compactness, improving water vapor resistance by forming a moisture barrier, which hinders water molecule diffusion [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. Similar effects were observed with waxy maize starch nanocrystals, reducing WVTR in films [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e45\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec51\" class=\"Section3\"\u003e\u003ch2\u003e3.3.4 Physical and optical properties\u003c/h2\u003e\u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e shows that the natural kidney bean starch film had a thickness of 0.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 mm, while the nano starch film was thinner at 0.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02 mm. Adding lemongrass essential oil increased film thickness, likely due to the oil's high viscosity, as noted by [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. The tensile strength, crucial for food packaging durability, increased from 11.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12 MPa in native starch films to 15.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23 MPa in nano starch films, due to the enhanced mechanical strength provided by nanoparticles.\u003c/p\u003e\u003cp\u003eThe addition of essential oils also affected the appearance of the films. Lemongrass essential oil increased opacity, reducing transparency by 5%, with native starch film opacity at 26.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.83% and nano starch film at 31.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27%. The oil altered color properties, increasing L* and a* values while decreasing b* values, likely due to the oil droplets disrupting the film's crystalline structure and enhancing light scattering.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec52\" class=\"Section3\"\u003e\u003ch2\u003e3.3.5 Scanning Electron Microscopy (SEM)\u003c/h2\u003e\u003cp\u003eSEM was used to examine the surface characteristics and morphology of native and nano kidney bean starch films. (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e). The SEM images indicated that all film components were well-blended, forming dense, pore-free, and crack-free films. However, in films containing lemongrass essential oil, small oil droplets were visible throughout the film. According to Evangelho et al. [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e42\u003c/span\u003e], the incorporation of essential oil reduced the surface homogeneity of the films, which was initially smooth without EO. Similar observations were made by Caetano et al. [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e47\u003c/span\u003e], who noted increased surface heterogeneity after adding various essential oils like borage seed, lemon, and oregano. Emulsions on nano- and micro-scales have been successfully utilized to enhance the properties of biopolymer-based food packaging by incorporating hydrophobic, bioactive, and functional compounds, as noted by Espitia et al. [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. These emulsions help to improve surface morphology, positively impacting other film properties.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec53\" class=\"Section3\"\u003e\u003ch2\u003e3.3.6 X \u0026ndash; Ray Diffraction (XRD)\u003c/h2\u003e\u003cp\u003eThe crystal structure of kidney bean starch is C-type, a combination of A- and B-types, in line with earlier research [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e49\u003c/span\u003e]. The intensity of diffractograms decreased after converting starch into films, indicating disorganization of the crystalline structure due to retrogradation and starch gelatinization during heating. This disorganization was influenced by sorbitol, which replaced starch hydrogen bonds with starch-plasticizer bonds, reducing crystallinity. These results align with Singh et al. [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. X-ray diffraction analysis revealed that native kidney bean starch displayed peaks at 15.24\u0026deg;, 17.32\u0026deg;, and 23.06\u0026deg;. However, these peaks converged when sorbitol was added during film preparation, indicating the melting of starch crystallites. When lemongrass essential oil was added to the S\u0026thinsp;+\u0026thinsp;EO film, a peak at 20.9\u0026deg; was created, reflecting intermolecular changes in the composite matrix, as similarly reported by Ghoshal \u0026amp; Singh, [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e50\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec54\" class=\"Section3\"\u003e\u003ch2\u003e3.3.7 Fourier Transform Infrared Spectroscopy (FTIR)\u003c/h2\u003e\u003cp\u003eThe FTIR spectra revealed that the addition of essential oil did not alter the molecular interactions of the films, as their absorbance patterns remained similar to the control due to identical elemental composition. The following were important peaks: C\u0026thinsp;=\u0026thinsp;O stretching at 1652 cm⁻\u0026sup1;, C-O stretching at 1026 cm⁻\u0026sup1;, C-Cl bonding at 808 cm⁻\u0026sup1;, C-H bending at 570 cm⁻\u0026sup1;, O-H stretching at 3425 cm⁻\u0026sup1;, and C-H stretching alkane at 2923 cm⁻\u0026sup1;. These findings indicate that all films were compositionally similar, as they all contained kidney bean starch [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e33\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec55\" class=\"Section3\"\u003e\u003ch2\u003e3.3.8 Antioxidant Properties of films\u003c/h2\u003e\u003cp\u003eEssential oils can decrease the reliance on artificial chemicals and prevent oxidation that impacts food quality. A DPPH radical scavenging assay evaluated the films' antioxidant properties. Lemongrass essential oil exhibited significantly higher radical scavenging activity (70.33%) compared to the starch-based film (24.55%) (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e). This enhanced activity is due to the ability of lemongrass essential oil to donate hydrogen atoms to reduce DPPH radicals, as reported by Kadam et al. [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e51\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eDPPH Inhibition% of starch film and starch\u0026thinsp;+\u0026thinsp;essential oil film\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSample\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eControl\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSample\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eDPPH Inhibition%\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.782\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.590\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e24.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eS\u0026thinsp;+\u0026thinsp;EO\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e0.782\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e0.232\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e70.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec56\" class=\"Section3\"\u003e\u003ch2\u003e3.3.9 Antimicrobial Properties of films\u003c/h2\u003e\u003cp\u003eBoth Gram-positive \u003cem\u003eStaphylococcus aureus\u003c/em\u003e and Gram-negative \u003cem\u003eEscherichia coli\u003c/em\u003e were tested for their antibacterial efficacy in starch-based edible films containing lemongrass essential oil. The lemongrass essential oil-treated films demonstrated strong antibacterial properties, with inhibition zone diameters ranging from 25.3 mm to 38.9 mm against \u003cem\u003eE. coli\u003c/em\u003e and 34.9 mm to 43.2 mm against \u003cem\u003eS. aureus\u003c/em\u003e. In contrast, the starch films without essential oil showed no antimicrobial activity. Gram-negative bacteria's outer barrier prevents the passage of hydrophobic components, whereas Gram-positive bacteria are more sensitive to essential oils. This explains why the oil-infused films were more efficient against Gram-positive bacteria. This aligns with findings by Ahmad et al. [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e52\u003c/span\u003e], indicating that essential oils are particularly effective against food spoilage organisms and food borne pathogens caused by Gram-positive bacteria.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec57\" class=\"Section3\"\u003e\u003ch2\u003e3.3.10 Biodegradability of Films\u003c/h2\u003e\u003cp\u003eBiodegradation involves enzymatic activity and living organisms, facilitating molecular breakdown in both aerobic and anaerobic environments and results in the partial or whole elimination of substances. After 5 days at 25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C, films containing starch (native/nano) and essential oils showed slower degradation compared to pure starch films, with native starch and essential oil films taking the longest to degrade. Similar findings by Gon\u0026ccedil;alves et al. [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e29\u003c/span\u003e], indicated that adding starch nanoparticles accelerated film biodegradation. This enhancement is attributed to water-induced swelling in the polymer, which promotes microbial growth and accelerates degradation.\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Sec58\" class=\"Section2\"\u003e\u003ch2\u003e3.4 Applications on fruits\u003c/h2\u003e\u003cp\u003eKidney bean nano starch films (NS and NS\u0026thinsp;+\u0026thinsp;EO) were chosen for grapefruit wrapping due to their superior mechanical, barrier, and antibacterial properties. Starch-based films were produced for comparison. After 16 days at room temperature and 26 days in refrigeration, Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e9\u003c/span\u003e showed the appearance of wrapped and unwrapped fruits, while Fig.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e10\u003c/span\u003e displayed weight loss, shrinkage, and pH changes during storage. Fruits wrapped in nano starch films exhibited significantly less weight loss (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) due to better hydrophobic and water vapor barrier properties. The wrapping did not affect the fruits' color, which is important for consumer acceptance. The superior moisture barrier of nano starch films (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) helps to reduce water and solute migration, thereby decreasing respiration and oxidation rates during storage.\u003c/p\u003e\u003cdiv id=\"Sec59\" class=\"Section3\"\u003e\u003ch2\u003e3.4.1 Weight loss determination\u003c/h2\u003e\u003cp\u003eGrapefruit weight loss is linked to respiration and water evaporation, with their thin skin making them prone to rapid water loss, shrinkage, and tissue weakening. A study evaluated the effects of wrapping films on weight loss over 28 days of refrigeration and 16 days at room temperature. All fruits lost weight during storage, but as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e10\u003c/span\u003e, wrapped fruits exhibited less weight loss compared to unwrapped ones.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec60\" class=\"Section3\"\u003e\u003ch2\u003e3.4.2 Shrinkage\u003c/h2\u003e\u003cp\u003eFigure \u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e10\u003c/span\u003e visually depicts the shrinkage in wrapped and unwrapped grapes, which results from moisture loss, as water constitutes about 80% of a grape's weight. The results indicate that wrapped grapes experienced less shrinkage, demonstrating the effectiveness of the wrapping films in minimizing moisture loss.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec61\" class=\"Section3\"\u003e\u003ch2\u003e3.4.3 pH\u003c/h2\u003e\u003cp\u003eThe study found that grapefruit pH increased significantly over storage, with unwrapped samples showing greater pH levels than those wrapped in NS and NS\u0026thinsp;+\u0026thinsp;EO films, which exhibited less pH change. Refrigerated storage further enhanced the films\u0026rsquo; performance, mitigating pH variations and extending shelf life. These findings underscore the effectiveness of NS-based films in maintaining fruit quality under varied storage conditions.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e"},{"header":"4. CONCLUSION","content":"\u003cp\u003eThe study highlights the potential of using kidney bean starch, particularly in its nano form, as a viable material for active food packaging. The extraction and conversion of starch through wet steeping and acid hydrolysis yielded native and nano starches with favorable physical and chemical properties. Both types exhibited similar crystallinity, but nano starch demonstrated superior characteristics when incorporated into edible films. The addition of lemongrass essential oil further enhanced the properties of the films, improving water vapor transmission, moisture content, solubility, and biodegradability. SEM analysis revealed a homogeneous and continuous film morphology, free from defects, underscoring the suitability of nano starch-based films for practical applications.\u003c/p\u003e\u003cp\u003eThe films developed in this study showed promising performance in extending the shelf life of perishable grapes. At room temperature, grapes wrapped in the films maintained quality for up to 16 days, compared to the typical 4\u0026ndash;5 days for unwrapped fruit. In refrigerated conditions, the shelf life extended to 26 days. These findings suggest that nano starch-based films could offer significant benefits in food packaging, enhancing preservation without compromising environmental sustainability.\u003c/p\u003e\u003cp\u003eWhile kidney bean starch is not commonly explored in packaging applications, this research demonstrates its potential as a sustainable and functional material. Further studies are required to fully understand the scope of its application and optimize its use in food packaging. This work contributes to the growing field of biodegradable and edible packaging, providing a basis for future innovations in food preservation.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003ch2\u003eConflict of Interest:\u003c/h2\u003e\u003cp\u003eThere is no conflict of interest from any of the author.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConsent to publish declaration\u003c/strong\u003e\u003cp\u003eMs. Kanak has done experiments and wrote the 1st draft of the paper, Dr. Gargi supervises the work and final draft was made by Dr. Gargi and both were agreed to publish in Discover Food journal.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003ch2\u003eConsent to Participate declaration\u003c/h2\u003e\u003cp\u003eSensory analysis using human trial was not done.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eEthics declaration\u003c/strong\u003e\u003cp\u003eNo ethical statement is needed as neither human nor animals were used for the study.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eClinical trial\u003c/strong\u003e\u003cp\u003eNo clinical trial was done.\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eFunding declaration\u003c/h2\u003e\u003cp\u003eNo external funding was received for the study.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eKanak did the experiments and wrote the first draft of manuscript , Gargi supervised her work and finalized the manuscript . All author reviewed the manuscript and agreed to publish in this journal\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eI do not have anyone to acknowledge\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eData availability Statement: Data will be available from corresponding author on request\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eLebreton, L. Van Der Zwet, J. Damsteeg, J. W. Slat, B. Andrady, A. L. Reisser, J. River plastic emissions to the world\u0026rsquo;s oceans. Nature Communications, 8(1). (2017) https://doi.org/10.1038/ncomms15611.\u003c/li\u003e\n\u003cli\u003eGeyer, R. Jambeck, J. Law, K. L. Production, use, and fate of all plastics ever made. Sci. Adv., 3(7). (2017). https://doi.org/10.1126/sciadv.1700782 \u003c/li\u003e\n\u003cli\u003eLi, P. Wang, X. Su, M. Zou, X. Duan, L. Zhang, H. Characteristics of Plastic Pollution in the Environment: a review. 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Food Control, 124, 107877. https://doi.org/10.1016/j.foodcont.2021.107877.\u003c/li\u003e\n\u003cli\u003eAhmad, M. Benjakul, S. Prodpran, T. Agustini, T. W. Physico-mechanical and antimicrobial properties of gelatin film from the skin of unicorn leatherjacket incorporated with essential oils, Food Hydrocol. 28(1) (2012) 189\u0026ndash;199, https://doi.org/10.1016/j.foodhyd.2011.12.003.\u003c/li\u003e\n\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":"Starch, packaging, Lemongrass essential oil, Nano starch","lastPublishedDoi":"10.21203/rs.3.rs-7095535/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7095535/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThese biodegradable films are typically composed of polysaccharides, proteins, lipids, or there is a growing interest in edible films for fruits and vegetables, driven by concerns over the negative impact of traditional packaging materials on health and the environment their combinations. In this study, edible film was formulated using kidney bean starch, both in its native and nano forms, with the incorporation of lemongrass essential oil. The starch, which can be sourced from various biological origins, served as the primary matrix, while the essential oil contributed its well-known antioxidant and antimicrobial properties. An acid hydrolysis technique was used to decrease the kidney bean starch's granule size and produce nanoparticles in order to improve the starch's qualities. When the physicochemical properties of native and nano starches were compared, it was found that the nano starch had better solubility, swelling power, and the ability to absorb water and oil. According to the results, kidney bean starch may be used for producing packaging films, and adding nano starch to the film formulations can enhance the functional qualities of the finished product.\u003c/p\u003e","manuscriptTitle":"Lemon grass Essential oil Incorporated Starch Based Active Packaging Film, Characterization and Applications","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-04 06:38:19","doi":"10.21203/rs.3.rs-7095535/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":"f5125431-02d9-49c9-b8f8-d01ad04c130a","owner":[],"postedDate":"August 4th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-01-22T12:41:01+00:00","versionOfRecord":[],"versionCreatedAt":"2025-08-04 06:38:19","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7095535","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7095535","identity":"rs-7095535","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}