CELLULOSIC FRACTION-BASED FUNCTIONAL PRODUCTS FROM AGRICULTURAL BIOMASS AND AGRICULTURAL PROCESSING BY-PRODUCTS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: AFRI COMPETITIVE GRANT
• Reporting Frequency: Annual
• Accession No.: 1024185
• Grant No.: 2021-67022-33469
• Cumulative Award Amt.: $481,618.00
• Proposal No.: 2019-07089
• Project Start Date: Nov 15, 2020
• Project End Date: Nov 14, 2024
• Grant Year: 2021
• Program Code: [A1531]- Biorefining and Biomanufacturing

Recipient Organization
SOUTH DAKOTA STATE UNIVERSITY
PO BOX 2275A
BROOKINGS,SD 57007

Performing Department
Dairy and Food science

Non Technical Summary
Plastics are durable, inexpensive, lightweight, and easy to prepare, and consequently benefits are undeniable. These intrinsic traits, indeed, have catalyzed significant commercial application of plastic products compared to other materials. Their applications have grown substantially in construction, food, clothing, medicine, transportation, electronics, and household goods. Plastic production continues to grow at a rapid rate and could double by 2025 and even triple by 2050. Such an increase could mean that plastics would account for up to 20% of the world's total oil utilization by 2050.Disposable plastics, also known as "single-use" plastics, are widely used in the packaging sector. The idea was to use thus prepared products once and throw them away or recycle. Grocery bags, food packaging, bottles, straws, containers and cups, to name a few, fall in this category. Unfortunately, only 9% of plastic wastes are being recycled and 12% is incinerated, leaving 79% to fill landfills, dumps, or scatter in the environment. Inappropriate disposal of plastics, coupled with unwanted and end-of life plastics, are littering our communities, oceans, and waterways, and contribute to adverse health issues for humans and animals. In 2015, 300 million tons of plastic waste were generated globally, and plastic packaging waste accounts for 47%. Among all the nations, China is currently the world's top generator of plastic packaging waste, whereas the USA is the largest producer of plastic packaging waste on a per-capita basis. If current plastic consumption patterns and waste management practices are not been revised, improved and strictly exercised, by 2050 our landfills and the environment will be swamped with about 12 billion tons of plastic litter: a painful repercussion of plastic packaging and production.Plastics do not corrode or biodegrade, and on average 700 years are needed for a single bottle to start to decompose. Plastics also photodegrade and result in small fragments known as microplastics. These have been found in aquatic habitats of inland water, open-ocean and enclosed seas, beaches, and surface waters across the globe. Due to their small size these particles transport across large distances and accumulate in natural habitats with adverse impacts on biota and the economy. Furthermore, microparticles ingested by marine species such as fish could eventually reach humans upon consumption. The toxic monomers and oligomers in plastic bags contain get delivered to food and in-turn poison the food, leading to health complication such as adverse effects on the nervous, respiratory, and reproductive systems as well as kidneys and liver. Styrene from food containers causes the proliferation of human breast tumor cells. Similarly, styrene migration from the polystyrene containing packaging foods has been noticed in instant foods, yogurt, and milk. Fluorinated compounds are found in one-third of the U.S. fast food packaging. Fluorinated compounds are being used as dessert, bread, sandwich, and burger wrappers and in paperboard due to their water-repellant, strain-resistant, and non-stick properties. These compounds, however, are linked to kidney and testicular cancer, elevated cholesterol, decreased fertility, and thyroid problems as well as adverse developmental effects and decreased immune response in children. Their prevalence in food packaging would certainly lead to dietary exposure as well as environmental contamination during production and disposal.Plastics are versatile and cost-effective with a wide variety of functionalities but are problematic. Towards this end, cellulose from renewable agricultural residues and agriculture processing by-products stands out as a viable option. Agricultural biomass is abundant and recyclable, predominantly generated from plants (agriculture, forestry, manufacturing process, etc.), animals (animal husbandry and fishery production, food processing, rural living garbage, etc.) and microbial wastes (plant and animal wastes). On a global scale, 140 billion tons of biomass are generated annually from agriculture alone. The 2011 Billion-Ton Update (U.S. Department of Energy, 2011) estimated that in the U.S. around 205 million dry tons of primary crop residues are produced annually, with three-fourths being corn stover, followed by wheat straw, sorghum straw, barley straw, and oat straw. By the year 2030, these biomass residues are projected to reach 320 million dry tons, due to continued growth in crop yields and higher amounts of land in reduced-tillage and no-till cultivation. They constitute a valuable resource of cellulose. For example, corn stover is predominantly composed of 45% cellulose, 30% hemicellulose and 25% lignin. The isolation and marketing of cellulose would be useful to address the plastic concerns, and is practical to design and develop functional materials across a wide range of industries including packaging, food, biofuels, cosmetic, medicine, construction, and animal feed sectors.The long-term goal of this research program is to develop an economical and sustainable solution to replace plastics, and thereby improve human health. Towards this end, cellulose extract from lignocellulosic biomass is sought as a potential solution. Lignocellulose biomass is an inexpensive, environmentally friendly, and economically sustainable biomaterial that can be obtained from a range of agricultural residues and agricultural processing by-products.The overall goal of the project is to develop and establish the protocols to extract cellulose fraction from renewable agricultural residues and agricultural processing by-products to make strong and biodegradable films. The research objectives that will accomplish the project goal are:To isolate and characterize cellulose fraction from corn stover, wheat straw, soybean biomass, oat straw, switchgrass and prairie cordgrass. The amount of cellulose fraction along with the chemical composition and molecular weight variations will be established.To solubilize cellulose fraction, and to understand its interactions with salts, biodegradable polymers and biopolymers. The roles of inorganic salts in solubilizing the cellulose extract and influencing cellulose interactions and physicochemical properties with other biopolymers and biodegradable polymers will be established.To prepare cellulose fraction films and to determine tensile strength and biodegradability. These will be the starting materials intended for non-plastic packaging alternatives that could also serve as a basis to fortify bioactive compounds leading to edible films.To determine the economic and technical feasibility of the established protocols/processes at a commercial scale. The technoeconomic analysis will be carried out to estimate the commercial-scale production costs and a Life Cycle Assessment (LCA) model for the developed protocols/processes. These results will be compared to traditional petroleum-based films to ascertain and quantify the benefits.The outcome of this project offers a unique value-added proposition for the agriculture industry and farmers to use agriculture byproducts to increase the profitability of their operations. The Earth along with its current and future generations will benefit immensely with this cost-effective and environmentally sustainable solution to curb the ills associated with the use of plastics.

Animal Health Component

50%

Research Effort Categories

Basic

20%

Applied

50%

Developmental

30%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
403	1510	2000	20%
403	1549	2000	20%
403	1560	2000	20%
403	1620	2000	20%
403	1820	2000	20%

Knowledge Area
403 - Waste Disposal, Recycling, and Reuse;

Subject Of Investigation
1510 - Corn; 1549 - Wheat, general/other; 1560 - Oats; 1620 - Warm season perennial grasses; 1820 - Soybean;

Field Of Science
2000 - Chemistry;

Keywords

cellulose-extract

biodegradable

biomass

inorganic salts

plastic replacing

Goals / Objectives
Plastics are versatile and cost-effective materials that can be manufactured with a variety of functionalities. Their advantages are particularly apparent in medicine and public health with several industrial applications. The global production of plastics has increased considerably over the last decades from 5 million tons per year during the 1960s to 280 million tons per year in 2011. Without a doubt, plastics bring many benefits to society and offer widespread technological advances. However, concerns about plastic usage and disposal are immense. Accumulation of plastic waste in landfills and in natural habitats, physical problems for wildlife resulting from ingestion or entanglement in plastic debris, and leaching of chemicals from plastic products and transfer to humans and wildlife are but a few. Plastics can also disrupt marine habitats and have a negative impact on the biodiversity.Plastics are strong, flexible, and low-cost and thus are desirable for packaging. It has been established, however, that these manmade materials are harmful and have potential to migrate their organic and inorganic constituents. The use of plastics, especially for food packaging, could be catastrophic if toxic monomers and oligomers from the plastic migrate into the food. For example, polystyrene is routinely used to package foods, and styrene migration is detected in instant foods, yogurt, and milk. More recent findings on contamination of water in plastic bottles further accentuate these concerns. Plastic debris such as polypropylene and nylon are found in bottled water. These compounds, not perceivable by the naked eyes, end up in the human body upon consumption, and have the ability to induce a wide range of harmful and carcinogenic effects.In the United States, packaging accounts for around 25% of plastics and 13.7% of the weight of waste in landfills. The lack of biodegradability and associated detrimental impacts of plastics on the environment are concerns of great magnitude. The possible endocrine-disrupting properties and the long-term pollution complications undeniably warrant developing alternatives to plastics. Towards this end, cellulose from renewable agricultural biomass such as corn stover, wheat straw, sorghum biomass, switchgrass, and prairie cordgrass, to name a few, stands out as a viable option to create biodegradable plastic packaging materials.Cellulose is a low-density biomaterial with strong and stiff structure, and certainly meets the desirable qualities of plastics. Cellulose is abundant and manifests good biocompatibility, biodegradability, and low-toxicity. Cellulose is a suitable and desirable alternative for plastics.The long-term goal of the PD's research program is to develop an economical and sustainable solution to alleviate plastic contamination and to improve human health. Towards this end, cellulose extracted from lignocellulosic biomass is sought as a potential solution. Lignocellulose biomass is inexpensive, environmentally friendly and economically sustainable biomaterial that can be obtained from a range of agricultural residues and agricultural processing by-products.The overall goal of the project is to develop and establish the protocols to extract the cellulose fraction from renewable agricultural residues and agricultural processing by-products to make strong and biodegradable films.The research objectives that will accomplish the project goal are:To isolate and characterize the cellulose fraction from corn stover, wheat straw, soybean biomass, oat straw, switchgrass, and prairie cordgrass. The amount of cellulose fraction, along with the chemical composition and molecular weight variations, will be established.To solubilize the cellulose fraction, and to understand its interactions with salts, biodegradable polymers, and biopolymers. The roles of inorganic salts in solubilizing the cellulose extract and influencing cellulose interactions and physicochemical properties with other biopolymers and biodegradable polymers will be established.To prepare cellulose fraction films and to determine tensile strength and biodegradability of the films. These will be the starting materials intended for non-plastic packaging alternatives that could also serve as a basis to fortify bioactive compounds leading to edible films.To determine the economic and technical feasibility of the established protocols/processes at a commercial scale. The technoeconomic analysis will be carried out to estimate the commercial-scale production costs and a Life Cycle Assessment (LCA) model for the developed protocols/processes. These results will be compared to traditional petroleum-based films to ascertain and quantify the benefits.

Project Methods
Objective 1: To isolate and characterize cellulose extract from corn stover, wheat straw, soybean straw, oat straw, switchgrass, and prairie cordgrass.Biomass feedstocks (corn stover, wheat straw, soybean straw, oat straw, switchgrass, and prairie cordgrass) will be obtained from research farms operated by the South Dakota Agricultural Experiment Station. Samples will be dried and ground to 40-mesh particles and stored at room temperature until use. In order to isolate cellulose extract, biomass material will be mechanically stirred in hot water (~85°C) and the pH will be adjusted to 6.8 by adding NaOH solution. The pH will be raised to 11.5 and then 3% H2O2 will be added to remove lignin from the cellulose fraction. Subsequently, the hot slurry will be sheared at 10,000 rpm for 1 h, cooled, centrifuged, and the solid residue will be separated. The supernatant that contains hemicellulose fraction will be saved to isolate hemicellulose. The washing of the solid residue with hot water and shearing cycles will be repeated until clear supernatant is obtained. The final solid residue will be collected, suspended in water, and dried by drum or spray drying to obtain the final cellulose extract. Hemicellulose from the supernatant will be separated by adjusting pH to 4 and centrifuging. The supernatant will then be treated with 3 volumes of 100% ethanol to precipitate the hemicellulose, which will be collected and dried. The chemical composition, glycosyl-linkage, and molecular size distribution of the cellulose extracts will be determined. The crystalline structure of extracted cellulose will be determined using X-ray powder diffraction protocols. The type of biomass and its influence on chemical composition and molecular weight distribution will be revealed.Objective 2: To solubilize cellulose fraction, and to understand its interactions with salts, other biodegradable polymers and biopolymers.The role of inorganic salts in solubilizing the cellulose extracts and influencing cellulose interactions and physicochemical properties with other biopolymers and biodegradable polymers will be established. As the molecular weight of cellulose extracts from different feedstocks will be different, a range of ZnCl2 solutions at different temperatures will be tested and viscosity measured to establish suitable salt concentration. To further compare the results, cellulose extract will be solubilized in ionic solvents (e.g. 1-butyl-3-methyl-imidazolium chloride) and film properties will be compared. Effects of cations on the viscosity and rheology of Zn-cellulose fraction solutions will be established. Later, interactions with biopolymers such as chitosan, alginate, iota-carrageenan, gelatinized corn starch, and biodegradable polymer polylactic acid will be assessed using Fourier transformed infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) to understand the interaction mechanisms.Objective 3: To prepare cellulose fraction films and to determine tensile strength and biodegradability. The highest viscosity solutions from Objective 2 will be selected in order to reduce the number films to be prepared. Bubbles, if any, will first be vacuumed out of the solutions. Films will be hand cast on a glass plate using a thin-layer chromatographic plate applicator. The clearance between the applicator and glass plate will be set to 1 mm. The glass plate along with the film will then be immersed in 500 mL ethanol for 30 min to coagulate the film. This will be followed by immersion in a fresh ethanol bath (500 mL) for another 30 min. The plate/film will then be removed and air dried at room temperature. It will then be placed in a water bath for 30 min to remove any excess salt, and subsequently soaked in glycerol (5% v/v) for another 30 min, and then air dried. The films will be characterized using X-ray powder diffraction, FTIR, DSC, and Scanning Electron Microscopy (SEM). Film properties such as color, water vapor permeability, water uptake, water solubility, tensile strength, and biodegradation will be established.FilmsTensile strength: Films will be cut into rectangular strips 8 cm long and 1 cm wide, and the tensile strengths was measured using the MTS EM Tensile with mechanical grip at room temperature. Average values from triplicate measurements was used to calculate the film strength.Biodegradation: Natural soil will be used as the biodegradation environment at ambient temperature under moisture-controlled conditions. Six to eight films (from each set) of the size 20 x 20 cm, weight measured, will be enclosed in a nylon mesh netting (2 x 2 mm mesh size) and will be buried 15 cm beneath the surface of the soil. Every day, soil will be moistened with distilled water. From 2 to 60 days after burying the degraded films and fragments will be collected, rinsed in water several times to stop degradation, dried at room temperature, and weighed. Tensile strength will be measured as described in the previous section. SEM images will be recorded to understand the surface changes. The half-life t1/2 and degradation rate constant k will be calculated from the double logarithmic plots of weight loss against the burring period (t) in soil.Objective 4: To determine the economic and technical feasibility of the established protocols/processes at a commercial scale. Technoeconomic analysis (TEA): Mass and energy balances will be focal points of this of the study to initially estimate processing parameters, capital and operating costs. Energy and cost analyses will be performed on the developed methods. The operational parameters, costs, and energy balances for each method will be determined and will be used to assess commercial viability. Value engineering performed concurrently with Objective 1-3 research will provide commercialization and cost minimization feedback.Estimated commercial-scale cost of production: The TEA results from the developed protocols/processes will be analyzed and transformed from lab-scale to production-scale in order to estimate the production cost. Deliverables will include: a process model, estimated capital/operating costs, energy requirements, and the primary inputs for determining the environmental impacts to use in the LCA model. These results will be compared to traditional petroleum-based films to ascertain and quantify the benefits of the developed protocols/processes.Life Cycle Assessment (LCA) model: The inputs and processing requirements determined in this objective will be used to develop the LCA model. These results will be compared to traditional petroleum-based films to ascertain and quantify the benefits of the developed protocols/processes.

Progress 11/15/20 to 11/14/24

Outputs
Target Audience:The target audience for this research includes scientists and industry representatives working in areas where biomass constituents, particularly cellulose, could serve as plastic alternatives. Students interested in this field of study are also part of the target audience. The public could benefit from this work through the development of efficient, safe, and biodegradable films that could eventually replace plastic bags and other products. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Four graduate students (Shafaet Ahmed, PhD degree, 20 h/week; Sandeep Paudel, PhD degree, 20 h/week; Sumi Regmi, PhD degree, 20 h/week; Sharad Bhattarai, MS degree, 20 h/week) and two undergraduate students (Kylie Rosenau, 10 h/week; Cornelius Chong Zhao Yu, 10 h/week) have engaged in the research. They have been introduced to new research areas involving experimental protocols. They are presenting results at national and international conferences. These experiences will prepare them for challenging scientific roles and responsibilities in the near future. Graduate students Shafaet Ahmed: Functional biobased materials from lignocellulosic biomass Sandeep Paudel: Biodegradable packaging films from corn biomass and corncob Sumi Regmi: Biodegradable packaging films from soybean biomass and soy hulls Sharad Bhattarai: Biodegradable packaging films from wheat and rice biomass How have the results been disseminated to communities of interest?The results have been presented at 17 state and international meetings, and published in 11 international peer-reviewed journals and in one book chapter. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Objective 1: To isolate and characterize the cellulose fraction from corn stover, wheat straw, soybean biomass, oat straw, switchgrass, and prairie cordgrass. 100% Accomplished We collected corn stover, wheat straw, soybean biomass, oat straw, switchgrass, and prairie cordgrass from farms on the SDSU campus and the nearby Brookings area. The materials were dried, ground using a Glen Mills Inc. hammer mill, sieved to 100 microns, and stored at room temperature for future use. A sequential treatment with KOH and NaClO2 was employed to obtain white cellulose. In this process, the biomass was treated with 5% KOH at a 1:20 biomass-to-base ratio at room temperature for 14 hours. The insoluble residue was then separated and washed with water. The slurry was treated with 1% NaClO2 at a biomass-to-chemical ratio of 1:20 at pH 5, which was adjusted with 10% acetic acid. The mixture was maintained at 70 degrees Celsius for 6 hours. The residue was separated and treated with 1% NaClO2 for effective discoloration. The white residue was washed with distilled water, vacuum-dried, and stored at room temperature for further use and characterization. The yield ranged from 25% to 39%. The extracts of wheat, soybean, switchgrass, and prairie cordgrass were analyzed for neutral sugar composition and glycosyl linkage. The composition analysis results suggest that the cellulose extract is predominantly made up of 58% xylose, 30.6% cellulose, 8.3% arabinose, 1.4% galactose, 0.7% rhamnose, 0.6% mannose, 0.2% fructose, and 0.2% ribose. Among the four samples, the relative monosaccharide percentages varied from 5 to 20. In all the samples, the largest linkage corresponded to the 4-linked glucopyranosyl residue, which mainly originates from cellulose. The next largest non-cellulosic linkage involved the 2-linked and 4-linked xylopyranosyl residues. Minor amounts of 4,6-linked glucopyranosyl and 3,4-linked xylopyranosyl are also observed. The molecular weight of the cellulosic fractions could not be determined because they could not be solubilized in any ionic solvents. Objective 2: To solubilize the cellulose fraction, and to understand its interactions with salts, biodegradable polymers, and biopolymers. 100% Accomplished During the fourth year, we focused on extracting white cellulose from alfalfa and soyhull biomass. The yield of cellulose residue was found to be 38.3%. We solubilized the cellulose using a 68% ZnCl2 solution. Initially, 16.24 g of ZnCl2 was dissolved in 6.0 mL of distilled water. The cellulose extract was mixed with 1.6 mL of distilled water in a separate container. Both vials were kept at 83 °C for 30 minutes. Then, the ZnCl2 solution was added to the cellulose solution, vortexed, and maintained at 83 °C for 10 minutes with constant stirring. Subsequently, the required amounts of CaCl2 and sorbitol were added, and films were cast. Box-Behnken Design (BBD) was used to optimize the amounts of cellulose, CaCl2, and glycerol. Initially, various amounts of cellulose, CaCl2, and the plasticizer sorbitol were tested to establish ranges for the BBD analysis, which were 0.3-0.5 g, 200-500 mM, and 0.5-1.5%, respectively. For alfalfa, the optimum amounts were determined to be 0.5 g of cellulose, 461.3 mM CaCl2, and 1.05% plasticizer. Conversely, in the case of soyhulls, these amounts were found to be 0.4 g of cellulose, 500 mM CaCl2, and 1.5% plasticizer. The optimized films were characterized by their water-solubility, moisture absorption, moisture content, transparency, water vapor permeability, tensile strength, and biodegradability. A plastic bag from a local grocery store served as the control. The optimized films were then employed to evaluate the shelf life of strawberries. The alfalfa and soyhull films were white and transparent. Their water solubility ranged from 9.2% to 69.6%, the moisture content was 10.0% and 8.4%, and the water-vapor permeability (WVP) was 0.9±0.3×10^-10 gm^-1s^-1Pa^-1 and 0.47±0.11×10^-10 gm^-1s^-1Pa^-1 respectively. The shelf life of strawberries was extended by 2 days and 3 days with alfalfa and soyhull films, respectively. Objective 3: To prepare cellulose fraction films and to determine tensile strength and biodegradability. 100% Accomplished The tensile strengths of the alfalfa and soyhull cellulose films were 16.9 MPa and 6.3 MPa, respectively. In comparison, the tensile strength of the commercial plastic film was lower at 2.56 MPa. Films were buried in soil for 60 days to assess their biodegradability, and their weight was monitored on alternate dates. The films were cut into 8 x 8 cm strips, and the initial weight was measured. Three strips from each film type were tested. The soil was sourced from SDSU farms, with a moisture content of approximately 24%. Biodegradability experiments were conducted in the lab by adding the soil to several glass jars. The films were buried at least 4 cm beneath the soil surface. Soil moisture was monitored regularly, and to maintain the required 24% moisture, water was added as needed. The alfalfa cellulose-based films lost 87% of their total weight due to decomposition over 35 days. Further observations revealed that the films fully decomposed within 40 days. For the soyhull films, these values were a 90% loss within 25 days and total decomposition within 30 days. Objective 4: To determine the economic and technical feasibility of the established protocols/processes at a commercial scale. 100% Accomplished We conducted a technoeconomic analysis and developed a Life Cycle Assessment model for the process. Our analysis reveals that the final cost to produce 0.055 metric tons of biomass films is $2.35 billion. In comparison, synthetic film production costs between $110 million and $ 330 million for the same amount. Therefore, the production cost of biodegradable films is over seven times higher than that of commercial films.

Publications

• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Regmi, S., Janaswamy, S. 2024. Soyhulls lignocellulose: An industrial by-product to develop biodegradable packaging films and to preserve fruits. USDA S1075 Multi-state meeting, July 25.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Paudel, S., Janaswamy, S. 2024. Alfalfa: A forage to sustainable biodegradable food packaging. Institute of Food Technologies (IFT) Annual Meeting & Food Expo, Chicago, IL, July 14-17.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Regmi, S., Janaswamy, S. 2024. Optimization of biodegradable and UV-blocking packaging films from soybean hulls for the preservation of raspberries. Institute of Food Technologies (IFT) Annual Meeting & Food Expo, Chicago, IL, July 14-17.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Ahmed, S., Janaswamy, S. 2024. A sustainable and green technique for biodegradable food packaging films. Institute of Food Technologies (IFT) Annual Meeting & Food Expo, Chicago, IL, July 14-17.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Regmi, S., Janaswamy, S. 2024. Valorization of soybean hulls: A sustainable approach to biodegradable packaging films. South Dakota State University Graduate Research, Scholarship, and Creative Activity Day, April 23.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Regmi, S., Janaswamy, S. 2024. Cellulosic residue of soybean hulls for developing eco-friendly packaging films and raspberries preservation. Recent advances and future prospects in formatting a healthier food system. 7th AMIFOST-2024, March 19-20.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Paudel, S., Janaswamy, S. 2024. Valorization of corncob by preparing cellulosic residue-based biodegradable packaging films. Recent advances and future prospects in formatting a healthier food system. 7th AMIFOST-2024, March 19-20.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Regmi, S., Janaswamy, S. 2024. Fabrication of Biodegradable Films from Soyhulls Lignocellulose to Address Plastic Perils. 2024 Hydrochar and Biochar Conference, South Dakota School of Mines, Rapid City, July 26-28.
• Type: Book Chapters Status: Accepted Year Published: 2025 Citation: Regmi, S., Paudel, S., Bhattarai, S., Janaswamy, S. 2025. Circular Food Packaging: Opportunities, Challenges, and Consumer Behavior. In Sustainable Food Systems and Circular Economy: Current State and Future Prospects; Kumar, S., Boojh, R., Dutta, J. Eds. Scrivener Publishing LLC. In Press.
• Type: Peer Reviewed Journal Articles Status: Published Year Published: 2025 Citation: Ahmed, S., Janaswamy, S. 2025. Green fabrication of biodegradable films: Harnessing the cellulosic residue of oat straw. International Journal of Biological Macromolecules, 140656. https://doi.org/10.1016/j.ijbiomac.2025.140656
• Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Regmi, S., Janaswamy, S. 2025. Biodegradable packaging films from the alkali-extracted lignocellulosic residue of soyhulls extend the shelf life of strawberries. Food Bioscience, 106016. https://doi.org/10.1016/j.fbio.2025.106016
• Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Paudel, S., Janaswamy, S. 2025. Use of alfalfa cellulose for formulation of strong, biodegradable film to extend the shelf life of strawberries. International Journal of Biological Macromolecules. 290: 139004.
• Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Regmi, S., Paudel, S., Janaswamy, S. 2024. Development of eco-friendly packaging films from soyhulls lignocellulose: Towards valorizing agro-industrial byproducts. Foods. 13: 4000. DOI: 10.3390/foods13244000
• Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Regmi, S., Janaswamy, S. 2024. Biodegradable films from soyhull cellulosic residue with UV protection and antioxidant properties improve the shelf-life of post-harvested raspberries. Food Chemistry, 460: 140672.DOI: 10.1016/j.foodchem.2024.140672
• Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Paudel, S., Janaswamy, S. 2024. Corncob-derived biodegradable packaging films: A sustainable solution for raspberry post-harvest preservation. Food Chemistry, 454: 139749. https://doi.org/10.1016/j.foodchem.2024.139749
• Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Bhattarai, S., Janaswamy, S. 2024. Biodegradable, UV-blocking, and antioxidant films from alkali-digested lignocellulosic residue fibers of switchgrass. Chemosphere, 359: 142393. https://doi.org/10.1016/j.chemosphere.2024.142393
• Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Ahmed, S., Janaswamy, S., Yadav, M.P. 2024. Biodegradable films form the lignocellulosic fibers of wheat straw and the effect of calcium ions. International Journal of Biological Macromolecules, 264, 130601. https://doi.org/10.1016/j.ijbiomac.2024.130601
• Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Mominul, H., Janaswamy, S. 2024. Biodegradable packaging films from banana peel fiber. Sustainable Chemistry and Pharmacy. 37: 101400. https://doi.org/10.1016/j.scp.2023.101400
• Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Bhattarai, S., Janaswamy, S. 2024. Biodegradable films from the lignocellulosic residue of switchgrass. Resources, Conservation & Recycling. 201: 107322. https://doi.org/10.1016/j.resconrec.2023.107322
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Bhattarai, S., Janaswamy, S. 2024. Developing high tensile strength films from rice biomass derived lignocellulosic residue. Minnesota Section of the IFT Great Plains Subsection 2024 Poster Competition, SDSU, Brookings, SD, October 25
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Paudel, S., Janaswamy, S. 2024. UV-blocking, biodegradable, and strong films made from alfalfa cellulosic residue extend the shelf-life of strawberries. Minnesota Section of the IFT Great Plains Subsection 2024 Poster Competition, SDSU, Brookings, SD, October 25.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Regmi, S., Janaswamy, S. 2024. Eco-friendly films from alkali-digested lignocellulosic fibers of soyhulls: Towards valorization of agro-industrial by-products. Minnesota Section of the IFT Great Plains Subsection 2024 Poster Competition, SDSU, Brookings, SD, October 25.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Paudel, S., Janaswamy, S. 2024. Corncob: A sustainable cellulosic resource for biodegradable films. ACS Fall Denver, CO, August 18-22.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Regmi, S., Janaswamy, S. 2024. Soybean hulls cellulosic extract: A novel material for biodegradable packaging films and raspberries preservation. ACS Fall Denver, CO, August 18-22.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Paudel, S., Janaswamy, S. 2024. Biodegradable packaging films from alfalfa cellulosic residue: A facile route to address plastic concerns and extend the shelf-life of strawberries. SD EPSCoR, Research Symposium. Sioux Falls, SD, July 31-August 1.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Regmi, S., Janaswamy, S. 2024. A sustainable approach to develop biodegradable films: Towards valorization of soyhulls. SD EPSCoR, Research Symposium. Sioux Falls, SD, July 31-August 1.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Paudel, S., Janaswamy, S. 2024. Strong and Biodegradable Films from Alfalfa Cellulosic Residue. 2024 Hydrochar and Biochar Conference, South Dakota School of Mines, Rapid City, July 26-28.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2024 Citation: Paudel, S., Janaswamy, S. 2024. Queen forage Alfalfa: A sustainable source for biodegradable packaging film. USDA S1075 Multi-state meeting, July 25.

Progress 11/15/22 to 11/14/23

Outputs
Target Audience:The target audience for this research includes scientists and industry representatives working in areas in which biomass constituents, especially cellulose, could be used as plastic alternatives. Students interested in this field of study are also included in the target audience. The public could benefit from this work by the development of efficient, safe, and biodegradable films that eventually replace plastic bags and other products. Changes/Problems:The graduate students are still mastering the techniques and experimental protocols and gaining research data and will be published in peer-reviewed journals. What opportunities for training and professional development has the project provided?Three graduate students (Shafaet Ahmed, PhD degree, 20 h/week; Sandeep Paudel, PhD degree, 20 h/week; Sumi Regmi, PhD degree, 20 h/week) have been involved in the research. They have been exposed to new research areas involving a series of experimental protocols. They are presenting results national and international conferences. These experiences will help them to take up challenging scientific roles and responsibilities in the near future. Graduate students Shafaet Ahmed: Functional biobased materials from lignocellulosic biomass Sandeep Paudel: Biodegradable packaging films from corn biomass and corncob Sumi Regmi: Biodegradable packaging films from soybean biomass and soy hulls How have the results been disseminated to communities of interest?The results have been presented at three state and international meetings, and published in three international peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals?Objective 1: To isolate and characterize cellulose fraction from corn stover, wheat straw, soybean biomass, oat straw, switchgrass and prairie cordgrass. We have completed this objective. Objective 2: To solubilize the cellulose fraction, and to understand its interactions with salts, biodegradable polymers and biopolymers. We will complete role of other crosslinking cations such as Fe2+, Mn2+, Cd2+, Ag2+, Sr2+, and Al3+ and biopolymers starch, alginate and PLA on film properties, especially water-vapor-permeability (WVP), tensile strength, and biodegradability. Objective 3: To prepare cellulose fraction films and to determine tensile strength and biodegradability. We will complete the tensile strength, elongation and biodegradability of all the films prepared. Objective 4: To determine the economic and technical feasibility of the established protocols/processes at a commercial scale. We will complete the techno-economic and LCA analyses.

Impacts
What was accomplished under these goals? Objective 1: To isolate and characterize the cellulose fraction from corn stover, wheat straw, soybean biomass, oat straw, switchgrass, and prairie cordgrass. 100% Accomplished We collected corn stover, wheat straw, soybean biomass, oat straw, switchgrass, and prairie cordgrass from farms on the SDSU campus and nearby Brooking's area. They were dried, ground (using Glen Mills Inc. hammer mill), sieved (100 microns), and stored at room temperature for future use. A sequential treatment with KOH and NaClO2 was used to obtain white cellulose. Herein, the biomass was treated with 5% KOH at a 1:20 biomass-to-base ratio at room temperature for 14 h. The insoluble residue was then separated and washed with water. The slurry was treated with 1% NaClO2 at biomass to the chemical ratio 1:20 at pH 5. The pH was adjusted with 10% acetic acid. The mixture was kept at 70 deg C for 6 h. The residue was separated and treated with 1% NaClO2 for effective discoloration. The white residue was washed with distilled water, vacuum dried, and stored at room temperature for further use and characterization. The yield was 25-39%. The wheat, soybean, switchgrass, and prairie cordgrass extract were subjected to neutral sugar composition and glycosyl linkage analysis. The composition analysis results suggest that the cellulose extract is predominantly composed of 58% xylose, 30.6% cellulose, 8.3% arabinose, 1.4% galactose, 0.7% rhamnose, 0.6% mannose, 0.2% fructose, and 0.2% ribose. Among the four samples, the relative monosaccharide relative percentage varied from 5 to 20. In all the samples, the largest linkage in all the samples corresponded to the 4-linked glucopyranosyl residue, which originated mainly from cellulose. The next largest non-cellulosic linkage was of the 2-linked and 4-linked xylopyranosyl residues. Minor amounts of 4,6-linked glucopyranosyl, and 3,4-linked xylopyranosyl are also seen. The molecular weight of the cellulosic fractions could not be determined as they could not be solubilized in any ionic solvents. Objective 2: To solubilize the cellulose fraction, and to understand its interactions with salts, biodegradable polymers, and biopolymers. 70% Accomplished During the 3rd year, we focused on the white cellulose extract from oat biomass. The cellulose residue yield was found to be 38.3%. We solubilized the cellulose using a 68% ZnCl2 solution. Initially, 16.24 g of ZnCl2 was dissolved in 6.0 mL of distilled water. In a separate container, 0.4 g of the cellulose extract was mixed with 1.6 mL of distilled water. Both these vials were kept at 65 deg C for 30 min. Then the ZnCl2 solution was added to the cellulose solution, vortexed, and held at 65 deg C for 30 min with constant stirring. The solubilized cellulose chains were crosslinked using 200, 400, 600, and 800 mM CaCl2 salt. After adding the calcium salt, the solution vial was left at 65 deg C for 30 min with constant stirring. These solutions were then hand-cast into films by pouring the hot solution into the applicator on a glass plate in a tray. Later, 500 mL of ethanol was added to the tray, and the glass plate was shaken gently. After 5 mins of coagulation, the tray was emptied, and a fresh 500 mL of ethanol was added and further coagulated for 5 min. Later, the film was placed onto a wooden frame and secured with tight clips. The frame containing the film was immersed in another tray containing 500 mL distilled water with gentle shaking for 5 min. The water was changed and continued shaking for 5 more min. Subsequently, the film and the frame were dipped in 5% glycerol in another tray for 10 min. Later, the film was separated from the frame and dried at room temperature. The films were characterized by their water-solubility, moisture absorption, moisture content, transparency, water-vapor-permeability, tensile strength, and biodegradability. A plastic film bag from a local grocery store was used as the control. The films were white in color and transparent. The films' water solubility was from 43.9 to 68.5%. Their moisture absorption ranged from 42.2-58.9%, and the moisture content ranged from 12.8-18.6%. These parameters showed an inverse relationship with added calcium ion concentration. The water-vapor-permeability (WVP) of films was 6.54±0.3×10-11 gm-1s-1Pa-1 at the 200 mM CaCl2 level and decreased to 4.96±0.14×10-11 gm-1s-1Pa-1 at the 800 mM CaCl2 level. The differences are due to increased chain immobility resulting from higher levels of calcium crosslinking in the cellulose-extract films. Objective 3: To prepare cellulose fraction films and to determine tensile strength and biodegradability. 75% Accomplished The tensile strength of the oat biomass cellulose films in the presence of 200 mM CaCl2 was 8.20 MPa. Increasing the CaCl2 level to 800 mM increases tensile strength to 17.24 MPa. Comparatively the commercial plastic film was lower at 2.56 MPa. Films were buried in the soil for 30 days to determine biodegradability, and their weight was monitored every week. The films were cut into 8 x 8 cm strips, and the initial weight was measured. Three strips from each film type were tested. The soil was procured from SDSU farms, and the moisture content was around 20%. The biodegradability experiments were carried out in the lab by adding the soil to several glass jars. The films were buried at least 4 cm beneath the soil surface. The soil moisture was monitored regularly, and to maintain the required 20% moisture, an adequate amount of water was added as necessary. The oat cellulose-based films lost 82% of their total weight due to decomposition over 30 days. Further continuation revealed that films decomposed fully within 60 days. We also observed that films generated with higher levels of crosslinking calcium ions decomposed slower. Objective 4: To determine the economic and technical feasibility of the established protocols/processes at a commercial scale. 50% Accomplished We initiated the technoeconomic analysis and Life Cycle Assessment model for the process. To this point, we have established a price range for most of the experimental steps.

Publications

• Type: Journal Articles Status: Published Year Published: 2023 Citation: Paudel, S., Regmi, S., Janaswamy, S. 2023. Effect of glycerol and sorbitol on cellulose-based biodegradable films. Food Packaging and Shelf Life 37: 101090.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Shafaet, A., Janaswamy, S. 2023. Strong and biodegradable films from avocado peel fiber. Industrial Crops and Products. 201: 116926
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Bhattarai, S., Janaswamy, S. 2023. Biodegradable, UV-blocking, and antioxidant films from lignocellulosic fibers of spent coffee grounds. International Journal of Biological Macromolecules. 253: 126798.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Janaswamy, S. Sustainable solutions for addressing plastic perils. Institute of Food Technologies (IFT) Annual Meeting & Food Expo, Chicago, IL, July 16-19, 2023
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Paudel, S., Janaswamy, S. 2023. Corncob cellulose: A sustainable source for biodegradable packaging films. Institute of Food Technologies (IFT) Annual Meeting & Food Expo, Chicago, IL, July 16-19.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Regmi, S., Janaswamy, S. 2023. Biodegradable packaging films from soybean hulls. Institute of Food Technologies (IFT) Annual Meeting & Food Expo, Chicago, IL, July 16-19.

Progress 11/15/21 to 11/14/22

Outputs
Target Audience:The target audience for this research includes scientists and industry representatives working in areas in which biomass constituents, especially cellulose, could be used as plastic alternatives. Students interested in this field of study are also included in the target audience. The public could benefit from this work by the development of efficient, safe, and biodegradable films that eventually replace plastic bags and other products. Changes/Problems:The graduate students are still mastering the techniques and experimental protocols and gaining research data to be suitable for publishing in peer-reviewed journals. What opportunities for training and professional development has the project provided?Five graduate students (Sajal Bhattarai, MS degree, 20 h/week; Mominul Hoque, MS degree, 20 h/week; Shafaet Ahmed, PhD degree, 20 h/week; Sandeep Paudel, MS degree, 20 h/week; Sumi Regmi, MS degree, 20 h/week) and two undergraduate students (Darratu Salesa; 10 h/week; Kate Eastlund, 10 h/week) have been involved in the research. They have been exposed to new research areas involving a series of experimental protocols. The obtained results were presented at fourteen state and international conferences. These experiences will help them to take up challenging scientific roles and responsibilities in the near future. Graduate students Sajal Bhattarai: Biodegradable materials from biowaste Mominul Hoque: Biodegradable films from agricultural byproducts Shafaet Ahmed: Biodegradable materials from agricultural biomass Sandeep Paudel: Biodegradable packaging films from corn biomass and corncob Sumi Regmi: Biodegradable packaging films from soybean biomass and soy hulls Undergraduate students Darratu Salesa: Biodegradable films from spent coffee grounds Kate Estlund: Value-added functional products from agricultural byproducts How have the results been disseminated to communities of interest?The results so far have been presented at four state and international meetings. What do you plan to do during the next reporting period to accomplish the goals?Objective 1: To isolate and characterize cellulose fraction from corn stover, wheat straw, soybean biomass, oat straw, switchgrass and prairie cordgrass. We will continue to extract white cellulose from corn stover, wheat straw, soybean biomass, oat straw, switchgrass, and prairie cordgrass to generate the materials needed for the other objectives. We will determine the molecular weight variations of the resultant cellulose fractions. Objective 2: To solubilize the cellulose fraction, and to understand its interactions with salts, biodegradable polymers and biopolymers. We will continue to work to solubilize the extracts, prepare films, and characterize them. We will investigate the role of other crosslinking cations such as Fe2+, Mn2+, Cd2+, Ag2+, Sr2+, and Al3+ and biopolymers starch, alginate and PLA on film properties, especially water-vapor-permeability (WVP), tensile strength, and biodegradability. Objective 3: To prepare cellulose fraction films and to determine tensile strength and biodegradability. The cellulose-based films generated above will be tested for tensile strength and biodegradability. Objective 4: To determine the economic and technical feasibility of the established protocols/processes at a commercial scale. We will complete the techno-economic and LCA analyses.

Impacts
What was accomplished under these goals? Objective 1: To isolate and characterize the cellulose fraction from corn stover, wheat straw, soybean biomass, oat straw, switchgrass, and prairie cordgrass. 70% Accomplished We collected corn stover, wheat straw, soybean biomass, oat straw, switchgrass, and prairie cordgrass from farms on the SDSU campus and Brookings area. They were dried, ground (using Glen Mills Inc. hammer mill), sieved (100 microns), and stored at room temperature for future use. A sequential treatment with KOH and NaClO2 was used to obtain white cellulose. Herein, the biomass was treated with 5% KOH at a 1:20 biomass-to-base ratio at room temperature for 14 h. The insoluble residue was then separated and washed with water. The slurry was treated with 1% NaClO2 at biomass to the chemical ratio 1:20 at pH 5. The pH was adjusted with 10% acetic acid. The mixture was kept at 70 deg C for 6 h. The residue was separated and treated with 1% NaClO2 for effective discoloration. The white residue was washed with distilled water, vacuum dried, and stored at room temperature for further use and characterization. The yield was 25-30%. We also extracted cellulose using pure NaOH solutions (20 or 50%). The biomass was added to 20 or 50% NaOH solutions at a 1:20 biomass-to-base ratio, and the reaction was carried out for 3 h. Later, the solution was neutralized with 10% acetic acid, and the residue was washed with distilled water several times and vacuum dried and stored at room temperature for further use and characterization. This process retains a significant amount of hemicellulose and lignin in the extracted sample, thus giving it a light golden brown. The extracted amount corresponded to roughly 30-40%. The wheat, soybean, switchgrass, and prairie cordgrass extract were subjected to neutral sugar composition and glycosyl linkage analysis. The composition analysis results suggest that the cellulose extract is predominantly composed of 58% xylose, 30.6% cellulose, 8.3% arabinose, 1.4% galactose, 0.7% rhamnose, 0.6% mannose, 0.2% fructose, and 0.2% ribose. Among the four samples, the relative monosaccharide relative percentage varied from 5 to 20. In all the samples, the largest linkage in all the samples corresponded to the 4-linked glucopyranosyl residue, which originated mainly from cellulose. The next largest non-cellulosic linkage was of the 2-linked and 4-linked xylopyranosyl residues. Minor amounts of 4,6-linked glucopyranosyl, and 3,4-linked xylopyranosyl are also seen. The molecular weight of the cellulosic fractions will be determined during the 3rd year. Objective 2: To solubilize the cellulose fraction, and to understand its interactions with salts, biodegradable polymers, and biopolymers. 50% Accomplished During the 2nd year, we focused on the white cellulose extract from prairie cordgrass. We solubilized it using a 68% ZnCl2 solution. Initially, 16.24 g of ZnCl2 was dissolved in 6.0 mL of distilled water. In a separate container, 0.4 g of the cellulose extract was mixed with 1.6 mL of distilled water. Both these vials were kept at 65 deg C for 30 min. Then the ZnCl2 solution was added to the cellulose solution, vortexed, and held at 65 deg C for 30 min with constant stirring. The solubilized cellulose chains were crosslinked using 200, 300, 400, and 500 mM CaCl2 salt. After adding the calcium salt, the solution vial was left at 65 deg C for 30 min with constant stirring. These solutions were then hand-cast into films by pouring the hot solution into the applicator on a glass plate in a tray. Later, 400 mL of ethanol was added to the tray, and the glass plate was shaken gently. After 5 mins of coagulation, the tray was emptied, and a fresh 400 mL of ethanol was added and further coagulated for 5 min. Later, the film was placed onto a wooden frame and secured with tight clips. The frame containing the film was immersed in another tray containing 500 mL distilled water with gentle shaking for 5 min. The water was changed and continued shaking for 5 more min. Subsequently, the film and the frame were dipped in 5% glycerol in another tray for 10 min. Later, the film was separated from the frame and dried at room temperature. The films were characterized by their color, thickness, water-solubility, moisture absorption, moisture content, transparency, water-vapor-permeability, tensile strength, and biodegradability. A plastic film bag from a local grocery store was used as the control. The films were white in color. Their L values (Hunter lab color scale) ranged from 82.9 to 85.8. Comparatively, the commercial plastic films were whiter with L>99. The a and b values, the total color difference, whiteness index, and yellowness index of the films did not show dependence on calcium chloride concentration. The thickness of the prepared films was in the range of 0.08 to 0.12 mm, compared to 0.08 mm of commercial plastic films. The films' water solubility was 41.2-56.3%, showing an inverse relationship with calcium ion concentration. The moisture absorption of the films ranged from 210-330% compared to 19% for commercial plastic films. The moisture content ranged from 16.1-27.7%, compared to 5% for commercial plastic films. These films were transparent, and transparency was inversely related to calcium concentration. The water-vapor-permeability (WVP) of films was 4.7 x 10-8 gm-1s-1Pa-1 at the 200 mM CaCl2 level and decreased to 4.3 x 10-8 gm-1s-1Pa-1 at the 500 mM CaCl2 level. Comparatively, commercial plastic films had a WVP of 0.8 x 10-10 gm-1s-1Pa-1. The differences are due to increased chain immobility resulting from higher levels of calcium crosslinking in the cellulose-extract films. Objective 3: To prepare cellulose fraction films and to determine tensile strength and biodegradability. 50% Accomplished The tensile strength of the films in the presence of 200 mM CaCl2 was 9.64 MPa. Increasing the CaCl2 level to 500 mM increases tensile strength to 23.24 MPa. We are currently measuring the tensile strength of the commercial plastic film was 2.56 MPa. comparison. Films were buried in the soil for 30 days to determine biodegradability, and their weight was monitored every week. The films were cut into 8 x 8 cm strips, and the initial weight was measured. Three strips from each film type were tested. The soil was procured from SDSU farms, and the moisture content was around 20%. The biodegradability experiments were carried out in the lab by adding the soil to several glass jars. The films were buried at least 10 cm beneath the soil surface. The soil moisture was monitored regularly, and to maintain the required 24% moisture, an adequate amount of water was added as necessary. The cellulose-based films lost 90% of their total weight due to decomposition over 30 days. Further continuation revealed that films decomposed fully within 60 d. We also observed that films generated with higher levels of crosslinking calcium ions decomposed slower. On the other hand, the commercial plastic films were intact without substantial weight change. Objective 4: To determine the economic and technical feasibility of the established protocols/processes at a commercial scale. 25% Accomplished We initiated the technoeconomic analysis and Life Cycle Assessment model for the process. To this point, we have established a price range for most of the experimental steps.

Publications

• Type: Journal Articles Status: Published Year Published: 2022 Citation: Janaswamy, S., Yadav, M. P., Ahmed, S., Hoque, M., Bhattarai, S. 2022. Cellulosic fraction from agricultural biomass as a viable alternative for plastics and plastic products. Industrial Crops and Products, 179, 114692.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Janaswamy, S. 2022. From field to film  Developing biodegradable plastics to clean up the plastic mess. MN-IFT and Phi Tau Sigma Joint Kick-off meeting, University of Minnesota, MN. September 22.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Janaswamy, S. 2022. Ag biomass: A versatile and sustainable tool to develop biodegradable plastics and clean up the plastic mess. Whistler Center for Carbohydrate Research, Purdue University, West Lafayette, IN. October 6.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Janaswamy, S. 2022. Ag biomass: A versatile and sustainable tool to develop biodegradable packaging films and clean up the plastic mess. Sustainable Biofuels and Co-Products Research Unit, Eastern Regional Research Center, ARS, USDA, Wyndmoor, PA. November 10.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Eastlund, K. A., Ahmed, S., Yadav, M. P., Janaswamy, S. 2022. Plastic replacing cellulosic films from wheat biomass. Minnesota Section of the IFT Great Plains Subsection 2022 Poster Competition, SDSU, Brookings, SD, March 24.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Hoque, M., Janaswamy, S. 2022. Biodegradable films from banana peel cellulose. Minnesota Section of the IFT Great Plains Subsection 2022 Poster Competition, SDSU, Brookings, SD, March 24.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Hoque, M., Janaswamy, S. 2022. Banana peel cellulose: A valuable resource to develop biodegradable plastics. Food sustainability: Challenges and opportunities for the future, 5th AMIFOST-2022, March 29-31.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Bhattarai, S., Janaswamy, S. 2022. Spent coffee grounds as a viable source of plastic replacing biodegradable films. Food sustainability: Challenges and opportunities for the future, 5th AMIFOST-2022, March 29-31.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Eastlund, K. A., Ahmed, S., Yadav, M. P., Janaswamy, S. 2022. Plastic replacing and biodegradable films from wheat biomass. Undergraduate Research, Scholarship and Creative Activity Day (URSCAD), SDSU, Brookings, SD, April 12.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Bhattarai, S., Janaswamy, S. 2022. Transparent, strong and UV blocking and biodegradable films from the spent coffee grounds extract. Gamma Sigma Delta Honor Society of Agriculture Poster context, South Dakota State University, Brookings, SD, April 20.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Eastlund, K. A., Ahmed, S., Yadav, M. P., Janaswamy, S. 2022. Cellulose extract from hard wheat biomass: A viable biomaterial to create biodegradable plastics. Gamma Sigma Delta Honor Society of Agriculture Poster context, South Dakota State University, Brookings, SD, April 20.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Hoque, M., Yadav, M. P., Janaswamy, S. 2022. Biodegradable packaging films based on cellulose extract from prairie cordgrass. Institute of Food Technologies (IFT) Annual Meeting & Food Expo, Chicago, IL, July 10-13.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Salesa, D., Bhattarai, S., Janaswamy, S. 2022. UV Blocking, High tensile strength and Biodegradable films composed of avicel and spent coffee grounds. SD EPSCoR Undergraduate Research Symposium, July 28.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Bhattarai, S., Janaswamy, S. 2022. Biodegradable films from switchgrass cellulose-extract. Virtual International Conference on Agri-Food Innovations in the quest for Food and Environmental Security, November 7.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: Regmi, S., Paudel, S., Janaswamy, S. 2022. Effect of plasticizers glycerol and sorbitol on cellulose films. Virtual International Conference on Agri-Food Innovations in the quest for Food and Environmental Security, November 7.

Progress 11/15/20 to 11/14/21

Outputs
Target Audience:The target audience for this research includes scientists and industry representatives working in areas in which biomass constituents, especially cellulose, could be used as plastic alternatives. Students interested in this field of study are also included in the target audience. The general public could benefit from this work by the development of efficient, safe, and biodegradable films that eventually replace plastic bags and other products. Changes/Problems:The graduate students are still mastering the techniques and experimental protocols and gaining research data to be suitable to publish in peer-reviewed journals. What opportunities for training and professional development has the project provided?Three graduate students (Sajal Bhattarai, MS degree, 20 h/week; Mominul Hoque, MS degree, 20 h/week; Shafaet Ahmed, PhD degree, 20 h/week) and two undergraduate students (Jake Larsen; 10 h/week; Ethan Berg, 10 h/week) have been involved in the research. They have been exposed to new areas of research involving a series of experimental protocols. The obtained results were presented at four state and international conferences. These experiences will help them to take up challenging scientific roles and responsibilities in the near future. Graduate students Sajal Bhattarai: Biodegradable materials from biowaste Mominul Hoque: Biodegradable films from agricultural byproducts Shafaet Ahmed: Biodegradable materials from agricultural biomass Undergraduate students Jake Larsen: Biodegradable films from agricultural biomass Ethan Berg: Life cycle analysis on agricultural byproducts based functional materials How have the results been disseminated to communities of interest?The results so far have been presented at four state and international meetings. What do you plan to do during the next reporting period to accomplish the goals?Objective 1: To isolate and characterize cellulose fraction from corn stover, wheat straw, soybean biomass, oat straw, switchgrass and prairie cordgrass. We will continue to extract white cellulose from corn stover, wheat straw, soybean biomass, oat straw, switchgrass, and prairie cordgrass to generate the quantities of materials needed for the other objectives. We will establish the chemical composition and molecular weight variations of the resultant cellulose fractions. Objective 2: To solubilize the cellulose fraction, and to understand its interactions with salts, biodegradable polymers and biopolymers. We will continue work to solubilize the extracts, prepare films, and characterize them. We will investigate the role of other crosslinking cations such as Fe2+, Mn2+, Cd2+, Ag2+, Sr2+, Al3+ and Fe3+ and biopolymers starch, alginate and PLA on film properties, especially water-vapor-permeability (WVP), tensile strength, and biodegradability. Objective 3: To prepare cellulose fraction films and to determine tensile strength and biodegradability. The cellulose-based films generated above will be subjected to tensile strength and biodegradability testing. Objective 4: To determine the economic and technical feasibility of the established protocols/processes at a commercial scale. We will continue to work on the technoeconomic and LCA analyses.

Impacts
What was accomplished under these goals? Objective 1: To isolate and characterize the cellulose fraction from corn stover, wheat straw, soybean biomass, oat straw, switchgrass and prairie cordgrass. 25% Accomplished We collected corn stover, wheat straw, soybean biomass, oat straw, switchgrass, and prairie cordgrass from farms on the SDSU campus and Brookings area. They were dried, ground (using Glen Mills Inc. hammer mill), sieved (100 microns), and stored at room temperature for future use. Initially, we extracted white cellulose using the acid hydrolysis protocol (30% H2O2 + 70% glacial acetic acid). The biomass to acid hydrolysis solution was maintained at 1:10 ratio. The reaction was carried out for 24 h at 95 deg C. Later, the solution was neutralized using 20% NaOH (at a ratio of 1:12 biomass to NaOH) and left at room temperature for 1 h. The product was then washed several times until neutralization. The white cellulose was filtered and vacuum dried and was stored at room temperature for further use and characterization. Each of the biomass types resulted in 25-30% of the biomass being recovered as cellulose extract. The acid hydrolysis protocol releases strong gases, and so we also explored other procedures to recover white cellulose. A sequential treatment with KOH and NaClO2 was also used to obtain white cellulose. Herein, the biomass was treated with 5% KOH at 1:20 biomass to base ratio at room temperature for 14 h. The insoluble residue was then separated and washed with water. The slurry was treated with 1% NaClO2 at biomass to chemical ratio of 1:20 at pH 5. The pH was adjusted with 10% acetic acid. The mixture was kept at 70 deg C for 6 h. The residue was separated and further treated with 1% NaClO2 for effective discoloration. The white residue was washed with distilled water and vacuum dried and stored at room temperature for further use and characterization. This protocol also yielded 25-30% of the biomass as white cellulose. We also extracted cellulose using pure NaOH solutions (20 or 50%). The biomass was added to 20 or 50% NaOH solutions at 1:20 biomass to base ratio and the reaction was carried out for 3 h. Later, the solution was neutralized with 10% acetic acid, and the residue was washed with distilled water several times and vacuum dried and stored at room temperature for further use and characterization. This process retains a significant amount of hemicellulose and lignin in the extracted sample, thus giving it a light golden brown color. The extracted amount corresponded to roughly 30-40%. The chemical compositions and molecular weights of these samples will be completed when the lab of our collaborator (Dr. Madhav Yadav, Eastern Regional Research Center, ARS) reopens following Covid. Objective 2: To solubilize the cellulose fraction, and to understand its interactions with salts, biodegradable polymers and biopolymers. 25% Accomplished Our initial focus was on the cellulose extract from corn stalks using the NaOH protocol. We solubilized it using a 68% ZnCl2 solution. Initially, 16.24 g of ZnCl2 was dissolved in 6.0 mL distilled water. In a separate container, 0.4 g of the cellulose extract was mixed with 1.6 mL of distilled water. Both these vials were kept at 65 deg C for 30 min. Then the ZnCl2 solution was added to the cellulose solution, vortexed, and held at 65 deg C for 30 min with constant stirring. The solubilized cellulose chains were then crosslinked using 20, 30, or 40 mM of CaCl2 salt. After adding the calcium salt, the solution vial was left at 65 deg C for 30 min with constant stirring. These solutions were then hand-cast into films by pouring the hot solution into the applicator on a glass plate in a tray. Later, 400 mL of ethanol was added to the tray and the glass plate was shaken gently. After 5 mins of coagulation, the tray was emptied and a fresh 400 mL of ethanol was added and further coagulated for 5 min. Later, the film was placed onto a wooden frame and secured with tight clips. The frame containing the film was immersed in another tray containing 500 mL distilled water with gentle shaking for 5 min. The water was changed and continued shaking for 5 more min. Subsequently, the film along with the frame was dipped in 5% glycerol, in another tray, for 10 min. Later, the film was separated from the frame and dried at room temperature. The films were characterized for their color, thickness, water-solubility, moisture absorption, moisture content, transparency, water-vapor-permeability, tensile strength, and biodegradability. A plastic film bag from a local grocery store was used as the control. The NaOH extracted cellulose retained a significant amount of hemicellulose and lignin, and thus the prepared films were brownish in color. Their L values (Hunter lab color scale) were in the range 47.4 to 59.0. Comparatively, the commercial plastic films were whiter with L>99. Interestingly, the a and b values, total color difference (?E), whiteness index (WI), and yellowness index (YI) of the films increased with calcium chloride concentration increase. The thickness of the prepared films was in the range 0.11 to 0.15 mm, compared to 0.08 mm of commercial plastic films. The water solubility of the NaOH-extracted cellulose films was in the range 45.8-58.7%, showing an inverse relationship with calcium ion concentration. The moisture absorption of the films ranged from 79-83% compared to 19% for the commercial plastic films. The moisture content ranged from 54.5-59.9%, compared to 5% for the commercial plastic films. These films were transparent, and transparency was inversely related to calcium concentration. The water-vapor-permeability (WVP) of films was 2.5 x 10-10 gm-1s-1Pa-1 at the 20 mM CaCl2 level and rose to 4.1 x 10-10 gm-1s-1Pa-1 at the 40 mM CaCl2 level. Comparatively, commercial plastic films had a WVP of 0.8 x 10-10 gm-1s-1Pa-1. The differences are due to increased chain immobility resulting from higher levels of calcium crosslinking in the cellulose-extract films. Objective 3: To prepare cellulose fraction films and to determine tensile strength and biodegradability. 25% Accomplished Using the NaOH extracted cellulose from corn stalks, subsequently mixed with CaCl2 and cast to produce films, we determined that the 20 mM CaCl2 level created films with a tensile strength of 1.16 MPa. Increasing the CaCl2 level to 40 mM, boosts the tensile strength to 2.61 MPa. We are currently measuring the tensile strength of the commercial plastic films for comparison. To determine biodegradability, films were buried in the soil for 30 days and their weight was monitored on the weekly basis. The films were cut into 8 x 8 cm strips and the initial weight was measured. Three strips from each film type were tested. The soil was procured from SDSU farms and the moisture content was around 20%. The biodegradability experiments were carried out in the lab by adding the soil to several individual glass jars. The films were buried at least 10 cm beneath the soil surface. The soil moisture was monitored regularly and to maintain the required 20% moisture, an adequate amount of water was added as necessary. The cellulose-based films lost 90% of their total weight due to decomposition over the 30-day period. Further continuation revealed that films decomposed fully within 60 d. We also observed that films generated with higher levels of crosslinking calcium ions decomposed at a slower rate. On the other hand, the commercial plastic films were intact without substantial weight change. Objective 4: To determine the economic and technical feasibility of the established protocols/processes at a commercial scale. 5% Accomplished We initiated the technoeconomic analysis and Life Cycle Assessment model for the process. To this point, we have established a price range for the majority of the experimental steps.

Publications

• Type: Journal Articles Status: Published Year Published: 2021 Citation: Nie, G., Zang, Y., Yue, W., Wang, M., Baride, A., Sigdel, A., Janaswamy, S. 2021. Cellulose-based hydrogel beads: Preparation and characterization. Carbohydrate Polymer Technologies and Applications. 2: 100074.
• Type: Journal Articles Status: Accepted Year Published: 2022 Citation: Janaswamy, S., Yadav, M. P., Ahmed, S., Hoque, M., Bhattarai, S. 2022. Cellulosic fraction from agricultural biomass as a viable alternative for plastics and plastic products. Industrial Crops and Products. (Accepted for publication)
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Janaswamy, S. 2021. Polysaccharides: Molecular structure to functional products. Sri Padmavati Mahila Visvavidyalam. Tirupati, India. July 31.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Janaswamy, S. 2021. Cellulose-based functional materials from agriculture biomass. Innovations and Sustainability in Food Processing. Amit Institute of Food Technology. Uttar Pradesh, India. June 23.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Larsen, J., Hoque, M., Janaswamy, S. 2021. Extracting corn stover cellulose to create functional and biodegradable films. USDA-Research and Extension Experience for Undergraduates (REEU). Brookings, SD. August 5.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Larsen, J., Hoque, M., Janaswamy, S. 2021. Cellulose extract from corn stover: Its potential to reduce plastic pollution. 8th Annual Research Symposium featuring Undergraduates in South Dakota. Brookings, SD. SD EPSCoR. July 29.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Ahmed, S., Mominul, H., Sajal, B., Janaswamy, S. 2021. Physical, mechanical and biodegradable properties of films from avocado peel cellulose. 2nd International E-meeting on Biopolymers and Bioplastics. Virtual. October 28-29.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Mominul, H., Ahmed, S., Janaswamy, S. 2021. Cellulose-based biodegradable films from lawn grass as an alternative to plastics. 2nd International E-meeting on Biopolymers and Bioplastics. Virtual. October 28-29.