DEVELOPING BIOPOLYMER-BASED FUNCTIONAL NANOCOMPOSITES FROM FOREST AND AGRICULTURAL RESIDUES FOR SMART FOOD PACKAGING APPLICATIONS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1016070
• Project Start Date: Oct 1, 2018
• Project End Date: Sep 30, 2023
• Program Code: [(N/A)]- (N/A)

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

Performing Department
Agricultural & Biosystems Engineering

Non Technical Summary
The US loses or wastes roughly half the food it produces each year (~60 million tons or $160 billion) due to inadequate packaging or other losses across the field-to-fork continuum. If food wastes could be reduced by just 1% through the use of smart packaging, it would save $1.6 billion each year. Smart food packaging can maintain food freshness, decrease spoilage, and achieve longer food shelf life with better quality and safety for consumers. The smart packaging market is currently driven by increasing demand for improving health, safety, and authenticity in food & beverage, healthcare, personal care, and other industries. However, one of the biggest challenges to implementation of smart packaging is lack of functional materials that preserve their mechanical, thermal, gas barrier, and other functional properties, and are yet biodegradable and environmental friendly.The goal of this project is to develop an effective process to integrate biopolymers and nanotechnology to produce biopolymer-based functional nanocomposites (BFNs) from forest and agricultural residues for smart food packaging applications. To achieve the project goal, five supporting objectives will be accomplished: 1) Extract biopolymers (cellulose and lignin) from corn stalks and sawdust for synthesis of BFNs; 2) Determine the best performing nano-fillers that can fit into biopolymer-based matrices and form functional nanocomposites; 3) Screen the best process of BFN fabrication from three nanocomposite synthesis technologies: solvent blending, in-situ growth, thermal melting; 4) Characterize physicochemical and functional properties of the BFNs produced and evaluate their potential for use in smart food packaging applications; 5) Preliminarily estimate BFN production costs to evaluate technical and economic feasibility of the best process of BFN fabrication, and optimize the BFN fabrication process to determine the best pathway for commercialization.Successfully completing this project could provide the breakthrough needed to move smart food packaging materials closer to reality, thus meeting the increasing consumer demands for a safe and healthy food supply. The U.S. food and beverage flexible packaging industry was estimated to be $17.6 billion in annual sales in 2016, and is projected to grow to $18.5 billion by 2021. If effective BFN materials can be developed, and achieve 1% market penetration by 2021 the economic value would be $185 million in annual sales market in U.S. alone. Economic benefits would also accrue to suppliers of the raw materials used for BFN production, which could include the biofuels, pulp, and paper industries.

Animal Health Component

40%

Research Effort Categories

Basic

40%

Applied

40%

Developmental

20%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
402	5399	2020	40%
403	5010	2000	20%
501	5299	3090	40%

Knowledge Area
403 - Waste Disposal, Recycling, and Reuse; 501 - New and Improved Food Processing Technologies; 402 - Engineering Systems and Equipment;

Subject Of Investigation
5299 - Agricultural supplies, general/other; 5399 - Structures, facilities, and equipment, general/other; 5010 - Food;

Field Of Science
2020 - Engineering; 3090 - Sensory science (human senses); 2000 - Chemistry;

Keywords

agricultural residues

forest residues

biopolymer

nanoparticle

nanocomposite

smart food packaging

Goals / Objectives
Smart food packaging is emerging as an effective solution to improve food quality and safety [1 - 6]. This technology integrates active and intelligent packaging processes in one system for food protection, containment, convenience, and communication. A smart food package consists of an intelligent layer and an active layer. The intelligent layer monitors and provides the user with information regarding the conditions of the food or its environment (e.g. temperature, pH, safety). The active layer protects the food from the outside environment and controls the amount of moisture, oxygen, carbon dioxide, or ethylene inside the package [4 - 7]. This packaging system can keep oxygen levels low and carbon dioxide levels high to impede cellular respiration or enzyme activity, preventing aerobic bacteria from multiplying in the foods. In addition to scavenging oxygen to reduce bacterial growth, the active layer can absorb ethylene to delay ripening of fresh fruits (e.g. apple, banana), thereby extending shelf life of the foods [5 - 7].The smart packaging market is currently driven by the increasing demand for improved health, safety, and authenticity in food & beverages, healthcare, personal care, and other industries [3 - 7]. However, one of the biggest challenges to implementation of smart packaging is lack of active and functional materials with effective mechanical, thermal, gas barrier, and other properties, yet are biodegradable and environmental friendly.The proposed project aims to develop an effective process to integrate biopolymer and nanotechnology to produce biopolymer-based functional nanocomposites (BFNs) from forest and agricultural residues for smart food packaging applications. The hypothesis is that appropriately dispersing specific nanoparticles in a biopolymer matrix could create a biodegradable functional nanocomposite. An innovative process will be developed to entrap specific nanoparticles (called nano-fillers) within a biopolymer-based matrix to form a new functional nanocomposite. Candidate nano-fillers molecules include inorganic nanoparticles (e.g. Ag TiO2, carbon fiber, or clay) and organic nanoparticles such as cellulose nanofiber (CNF) and cellulose nanocrystal (CNC). Candidate molecules for the biopolymer-based matrix are cellulose and/or lignin, which can be directly extract from forest and agricultural residues or recovered from the coproducts of biofuel, pulp, and paper industries. Wood sawdust and corn stover will be selected to extract biopolymers for BFN synthesis in this project. Three different nanocomposite synthesis processes, including solvent blending, thermal melting, or in-situ growth, will be tested for BFN fabrication. In the long-term, BFN materials can be tailored to specific applications including absorption and desorption of specific gases (e.g. O2, CO2, or C2H4), reduce oxygen and moisture permeability, increase mechanical strength and thermal stability, embedment of specific nanoparticles (e.g. Ag, TiO2) as antimicrobial agents or to signal undesirable levels of bacteria because of spoilage, (e.g. time-temperature labeling). The intent is to integrate BFNs into food packaging materials to develop new smart interaction packaging applications.The long-term goal of this project is to develop sustainable BNF materials from lignocellulosic biomass, such as woody biomass, perennial grasses, and agricultural residues, which are not only readily available, renewable, and inexpensive, but also environmental friendly, social acceptable, and economic feasible [7 - 12]. Therefore, production of BNF materials will be environmentally and economically sustainable. To achieve this long-term goal, five supporting objectives will be accomplished in this project.Extract biopolymers (cellulose and lignin) from corn stalks and sawdust for synthesis of BFNs;Determine the best performing nano-fillers that can fit into biopolymer-based matrices and form functional nanocomposites;Screen the best process of BFN fabrication from three nanocomposite synthesis technologies: solvent blending, in-situ growth, and thermal melting.Characterize physicochemical and functional properties of the BFNs produced and evaluate their potential for use in smart food packaging applications;Preliminarily estimate BFN production costs to evaluate the technical and economic feasibility of the best process of BFN fabrication. Optimize the BFN fabrication process to determine the best pathway for commercialization.Natural materials (e.g. wood, hemp, cotton, linen, etc.) have been used as food packaging materials for thousands of years and their use continues for some food products today. However, these materials lack the functionality, durability and uniformity required for modern smart food packaging. Currently, most materials used for food packaging are synthetic petroleum-based polymers (e.g. polyethylene, polyprolene), glass, and metal foils. These materials are non-degradable and can cause environmental problems. Attempts have been made to use biopolymers to create edible/inedible and biodegradable films to extend food shelf life and reduce food waste and packaging waste. Unfortunately due to poor performance characteristics (e.g., brittleness, inadequate gas and moisture barrier, low heat distortion temperature, and cost) such biopolymers have not achieved widespread commercial success [5 - 7, 10 - 12].These limitations of biopolymers can be addressed by integrating nano-fillers into the biopolymer matrix, thereby creating packaging materials with novel characteristics [5 - 7, 13 - 15]. Nano-fillers can improve the performance of the resulting nanocomposite materials because of their high surface area and ability to interact with the biopolymer-matrix. A uniform dispersion of nano-fillers added into a biopolymer can lead to a very large matrix/filler interfacial area, which changes the molecular mobility, the relaxation behavior and the consequent thermal and mechanical properties of the nanocomposites. In addition, nano-fillers can add new functional properties to the resulting nanocomposite, such as antimicrobial ability, enzyme immobilization, gas and moisture permeability reduction, oxygen scavengers, chemicals or bacterial sensing. These specific functional properties make nanocomposites possibly the best candidate used as smart food packaging materials [4 - 7, 9 - 13].Many efforts have been made to produce nanocomposites using cellulose nanocrystals or cellulose nanofibers [5 - 7, 11 - 14], but very few have reached commercial applications. There are currently no bio-based nanocomposites used in smart food packaging due to the lack of required properties and functionalities. Developing biopolymer-based and highly functional nanocomposite materials is necessary for smart food packaging. We are exploring effective processes to produce BFN materials for the needs of smart food packaging by integrating nano-fillers into biopolymer-based matrix. Our preliminary work has identified that cellulose or cellulose nanocrystal combined with metal ions (Ca2+ and Zn2+ ions) could generate a new biopolymer-based nanocomposite with significantly improved mechanical properties for applications in smart food packaging. If successfully developed, such a new biopolymer-based functional nanocomposite would represent a breakthrough in smart food packaging.

Project Methods
Different combinations of nano-fillers and biopolymers will be investigated. The project will identify the best process for BFN fabrication by screening three different processes (solvent blending, in-situ growth, and thermal melting). The equipment and facility for extracting cellulose and lignin from biomass feedstocks have been established in the PIs' laboratory. Commercially available nano-fillers (e.g. Ag, TiO2) samples will be purchased from chemical suppliers (e.g. Sigma-Aldrich, or Fisher Scientific). Biopolymers (cellulose and lignin) will be directly extracted from corn stalk and sawdust and used as the biopolymer matrix for synthesis of BFNs to accomplish the 5 objectives.Objective 1. Extract biopolymers (cellulose and lignin) from corn stalks and sawdust for synthesis of BFNs.Extraction of cellulose and lignin from biomass are proven technologies. A green chemical method, ion liquid extraction, will be used to extract cellulose and lignin from corn stalk and sawdust to minimize environmental impacts. The yield and quality of cellulose and lignin will be determined. Characterization of the celluloses and lignin will be conducted using TEM, SEM, and XRD. The milestone of objective 1 is to achieve sufficient samples of cellulose and lignin for use in subsequent objectives.Objective 2. Determine the best performing nano-fillers that can fit into biopolymer-based matrices and form functional nanocomposites. The synthesis of BFN materials will be carried out by entrapping different ratios of nano-fillers into the biopolymer-based matrix. Commercially available nanoparticles (e.g. Ag, TiO2) will be used as nano-fillers to formulate with the biopolymers (cellulose and/or lignin) produced in objective 1 for BFN synthesis. Three processes: solvent blending, in-situ growth, and thermal melting will be individually performed to synthesize BFN samples. According to the requirements of smart food packaging, the best performing nano-filler that can fit into the biopolymer-based matrix and form functional nanocomposites with desired properties will be selected. If the produced BFN samples are not good enough to meet the requirements, new nano-fillers (e.g. clay nanoparticles, carbon nanofibers, cellulose nanocrystals or cellulose nanofibers) may also be synthesized, tested and screened. The milestone at the end of objective 2 is that appropriate BFN materials will have been generated to evaluate their potential in smart food packaging application in the objective 4.Objective 3. Screen the best process of BFN fabrication from three nanocomposite synthesis technologies: solvent blending, in-situ growth, and thermal melting. Objective 3 will determine the best process for BFN synthesis from three processes: solvent blending, in-situ growth, and thermal melting. Depending on the processing conditions and ratios between nano-fillers and biopolymer matrix, three types of nanocomposite structures may be obtained: unintercalated, intercalated, or exfoliated. These different nanostructures can result in different functionalities and physicochemical properties of the final BFN materials [17 - 25].The solvent blending process is to evenly disperse nano-fillers in a biopolymer matrix by mixing them in a solvent in which both nano-fillers and biopolymer matrix are soluble. After the solvent is removed, a multilayer sheet-like structure of nanocomposite is formed. Water, dilute NaOH (< 30%), and ethanol (< 60%) will be used as solvents. The selected nano-fillers will be individually dissolved in the solvents and then combined with the biopolymer matrixes for the BFN synthesis [19 - 26].The in-situ growth process is to evenly disperse nano-fillers in the biopolymer matrix through cationic exchange using heat, radiation, initiator diffusion, organic initiator or catalyst fixed approaches [19 - 30]. The selected nano-filler monomers then polymerize and grow in between the interlayers, forming intercalated or exfoliated nanocomposites. This in-situ growth process will be initially started at room temperature (25?C). If the nano-fillers exhibit a low polymerization rate in the biopolymer matrix, higher operating temperatures will be tested. However, the operating temperatures should be controlled within a range that doesn't damage the nano-fillers or biopolymer matrix (e.g. < 200?C) [20 - 27].The thermal melting process is to uniformly melt nano-fillers into the biopolymer matrix to produce a homogenous nanocomposites. The operating temperatures will be controlled within a range that doesn't damage the surface of the nano-fillers or biopolymer matrix (e.g. < 200?C) [31 - 40].All possible combinations of nano-fillers and biopolymers for BFN fabrications will be tested using the three processes. The produced BFN samples will be characterized in objective 4. Based on the characterization results, a comparison of the compatibility, efficacy and functionality of the resulting BFNs will be carried out to identify the best process. The selected process will be optimized in objective 5.Objective 4. Characterize physicochemical and functional properties of the BFNs produced and evaluate their potential for use in smart food packaging applications.Firstly, all produced BFN materials will individually be fabricated into thin films with the same dimensions so that they can be characterized for their physicochemical properties and functionalities. All BFN films will be characterized for their structural properties (surface area, pore volume and pore size distribution), mechanical properties (tensile, elastic modulus, linear compressibility, and shear moduli), and thermal properties (negative thermal expansion andheat capacity). The characterizations will be carried out using BET, XRD, FTIR, SEM, TEM, Tensile Analyzer, and other instruments. Based on this data the samples will be screened to select preferable BFN films that have desired physicochemical and mechanical properties for further evaluation.Secondly, functionalities including absorption and desorption of specific gases (O2, CO2, and C2H4), oxygen and moisture permeability, and water stability will be evaluated to identify the most promising BFN films for smart food packaging application. The BFN films will also be studied for their active antimicrobial activity against E. coli. Once the efficacy of the BFN films are determined, migration of the nano-fillers into foodstuffs will be studied to negate the chances of food safety concerns. Based on these preliminary results, some suggestions or recommendations for use in smart food packaging applications or further research plans may be generated.Objective 5. Preliminarily estimate BFN production costs to evaluate technical and economic feasibility of the best process of BFN fabrication. Optimize the BFN fabrication process to determine the best pathway for commercialization. The process(es) that produce the most promising BFNs will be optimized to efficiently produce BFN films at low cost. The most promising BFN films should have high adsorption and desorption capacity for specific gases (O2, CO2, and C2H4), low O2 and moisture permeability, and very good stability of mechanical strength and thermal properties, with low production cost. After optimization, the best process will be evaluated for scaling up into pilot scale production of BFN films.The energy and economic viabilities of the optimized BFN synthesis process will be analyzed to identify its potential for commercialization. Mass and energy balances, and preliminary cost estimating of the best process will be quantified in a simple scale up model, based on the laboratory results. These results will be used to evaluate the commercialization potential of the optimized BFN synthesis process. If the most optimistic commercial potential of the synthesis process is verified, the investigation of BFN material production will be conducted at pilot scale in future research.

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:Undergraduate and graduate students: Undergraduate students have been targeted through formal and/or informal classroom instruction, research workshops/symposia, and other outreach activities. Many undergraduate students will choose agricultural production, food engineering, and biomass processing as an occupation. The activities in this project will help the students prepare for graduate school or careers in sustainable agriculture, food engineering, and biomass refinery industries. Graduate students have also been targeted to receive professional training to prepare them for their future academic or industrial careers by directly participating in all aspects of research activities, including experimental design, running tests, collecting data, and publishing research results. Research community: Agricultural engineering and food processing scientists and professionals were targeted through publishing the research results and new discoveries in peer-reviewed professional journals and/or presentations at scientific conferences or symposia. An understanding of our research results will help them to develop hypotheses and effective processes that will advance their own research programs. Farmers and food producers:This audience was targeted because biomass feedstocks for the production of biopolymer-based nanocomposites will be supplied by these farmers, while the products of produced nanocomposites will be used in food processing by the producers. They were targeted through formal and informal instruction or training in technical workshops, research symposia, and other extension/outreach activities.? Public andcustomers: This audience was targeted because the smart packaging and food products developed from this research will ultimately be consumed or used by the general public. The customers' needs, likes, and acceptances should be considered in the research.The audience was targeted through publications of research results, websites, conferences, workshops, and extension/outreach activities. Changes/Problems:Activities in goal 1 were impacted most significantly by the disruptions caused by Covid-19. What opportunities for training and professional development has the project provided?The project provided great opportunities for professional education and training of the next generation workforce for local communities and industries by involving 4 graduate and 2 undergraduate students through research and education activities. The research results and discoveries were published in 2 peer-reviewed journal papers, two manuscripts that are under review, and 3 conference papers/posters/presentations during project year 2. The website established at: https://lw9898.wixsite.com/linweiwebsite in project year 1 was updated to disseminate the latest research results and newest knowledge obtained in this project. These outreach activities are not only promoting public awareness of sustainable technologies, but also disseminating the new discoveries and knowledge to improve agricultural production, food safety, and smart packaging materials. The details of the 4 graduate, 2 undergraduate students, and 1 postdoc who were involved or trained in this project are listed below: Abdus Sobhan is a current Ph.D. student of Agriculture and Biosystems Engineering (ABE). His focus is to develop activated carbon based functional nanocomposite films. He also aided the PI in guiding other graudate and undergraduate students in the research projects. Zhisheng Cen is a current M.S. student of ABE. His worked sought to fabricate PLA-based nanocomposite coatings for applications in control-release fertilizer development. Emmanual Arkoh-Mensah is a current M.S. student of ABE. He started to participate in the project on January 10, 2020 and worked to fabricate and characterize biochar-based control release fertilizers. Nadee Kaluwahandi is a current M.S. student of ABE. He started to participate in the project on January 10, 2020 and worked to investigate the effect of cold plasma treatment on food packaging and food safety. Jace Jerome is a current undergraduate student at Univesity of South Dakota. He started to participate in the project on July 10, 2020 and worked to develop a production cost analysis model for new products developed in the project. Stoen Mollman is a current undergraduate student at Univesity of South Dakota. He started to participate in the project on May 10, 2020 and worked on marketing analysis of new products developed in the project. Shouyun Cheng was a Ph.D. student of ABE at SDSU and graduated on December 10, 2018. After graduation he worked as a postdoc and helped the PI develop the nanocellulose extraction process during this evaluation period. He also helped the PI to guide graduate and undergraduate students in the research project. How have the results been disseminated to communities of interest?Six publications (including peer-reviewed journal papers, conference paper, and presentations) were achieved in the project year 2. Research results were also disseminated to local communities and the public through workshops, conferences, and other outreach activities. What do you plan to do during the next reporting period to accomplish the goals?We will continue to develop biopolymer-based nanocomposites for applications in biosensors, precision fertilization, sustainable agriculture, and food health and safety. Our long-term goal is to establish nationally and internationally recognized research programs at SDSU. We will further extend the network of research collaborations across not only SDSU, but also to national /international institutes, agencies, and industrial partners. We will explore all possible internal and external sources to secure research funding to improve the research facility and analytic capability, and continue to move the research to a higher level with more positive impacts. To achieve the goal of this project, we plan the following activities in the next evaluating period. Goal 1: Extract biopolymers (cellulose and lignin) from corn stalks and sawdust for synthesis of BFNs. According to the performance of nanocomposite films produced in the project, we will optimize the 2H process for production of more effective cellulose and nanocelluloses at low cost in project year 3. Goal 2: Determine the best performing nano-fillers that can fit into biopolymer-based matrices and form functional nanocomposites. Additional biomass feedstocks such as woody sawdust, switchgrass, soybean straw will be examined for their suitability for use as nano-fillers in production of biopolymer-based nanocomposites. New applications of nanocomposites for food processing and sustainable agriculture will be explored in the coming year. Goal 3: The nanocomposite film fabrication process, combined with the solvent blending method, will be upgraded and optimized to improve the properties of nanocomposite films at low cost. In-situ growth and thermal melting will also be tested in next project year. Goal 4: We will continue to characterize the physicochemical and functional properties of biopolymer-based nanocomposites produced in the projects. We will evaluate the suitability of these produced films for applications in food processing and sustainable agriculture. Goal 5: Preliminarily estimate BFN production costs to evaluate the technical and economic feasibility of the best process of BFN fabrication. Optimize the BFN fabrication process to determine the best pathway for commercialization. We will estimate production costs of the produced nanocomposites and other products made by the nanocomposites, and then evaluate the technical and economic feasibility of nanocomposites for uses in not only smart food packaging, but also other relate areas such as precision agriculture.

Impacts
What was accomplished under these goals? Goal 1: Extract biopolymers (cellulose and lignin) from corn stalks and sawdust for synthesis of BFNs. (30% Accomplished) Due to the impact of Covid-19, many supplies and equipment/tools became unavailable because of the business shutdown. This made it impossible to optimize the new 2H process in year 2, even though the process to combine hydrolysis and homogenization of biomass to extract cellulose and nanocellulose had undergone preliminary testing in year 1. Thus we modified our work plans, and instead of producing celluloses and nanocelluloses ourselves, we used commercial nanocelluloses to fabricate functional nanocomposites and develop new applications of nanocomposites in the Goal 2. We will perform optimization of the 2H process in year 3 after the Covid-19 issue is resolved. Goal 2: Determine the best performing nano-fillers that can fit into biopolymer-based matrices and form functional nanocomposites. (50% Accomplished) Research focused on developing nanocomposite films based on activated carbon (AC) and cellulose nanofibers (CNF), and then integrating silver nanoparticles (AgNPs). These films were then characterized for functional properties related to smart food packaging. A new nanocomposite film was produced using a casting method. An ultrasonic suspension of CNF was impregnated with AC and AgNPs, creating what we call activated carbon-based silver-cellulose film (AgNPs/AC-CNF). The antimicrobial properties of this film were tested against food pathogens, Staphylococcus aureus and Escherichia coli. We found that AgNPs concentrations in the range of 140-450 ppm inhibited the growth of S. aureus and E. coli on the agar plates. Moreover, when AgNPs increased from 70 to 450 ppm, the conductivity of AgNPs/AC-CNF film was siginificantly improved. The peak current increased from 0.2 to 1.16 μA, while the electrical resistivity decreased from 6.8 to 1.9 kΩ.cm. This AgNPs/AC-CNF film showed good promise to serve as an effective antimicrobial conductive film for applications in smart food packaging. The new discoveries have been published in a peer-reviewed journal: Materials Today Communications, in April 2020. Goal 3: Screen the best process of BFN fabrication from three nanocomposite synthesis technologies: solvent blending, in-situ growth, and thermal melting. (45% Accomplished) To develop an effective process to fabricate biopolymer-based functional nanocomposites (BFNs), a novel water solution casting method was created to produce the AgNPs/AC-CNF film used in Goal 2. First, AgNPs suspensions were prepared by mixing different concentrations of AgNPs (e.g. 70, 140, 250, 350 and 450 ppm) in deionized water. Second, 2.5 g of CNF gels (3% of solid) were then added individually into each of the prepared AgNPs solutions. These were then gently agitated at 300 rpm and 50°C for 20 minutes to thoroughly mix CNF with AgNPs. After that, 1.4 g of AC was individually added to each solution to obtain the AgNPs/AC-CNF mixture, which consisted of 70% AC and 30% CNF on a dry basis. Solutions were then homogenized for 30 minutes using a handheld homogenizer at 4,000 rpm. The homogenized mixtures were then poured into Petri dishes (diameter 13 cm) and dried at 60°C for 16 h to create the AgNPs/AC-CNF films containing different AgNPs concentrations. The dried AgNPs/ACCNF films were peeled from the Petri dishes and conditioned in a desiccator with 50-56% relative humdity for 2 days prior to the analysis or characterization. This process produced antimicrobial conductive films that have been verified as very promising for application in smart food packaging in goal 1. Goal 4: Characterize physicochemical and functional properties of the BFNs produced and evaluate their potential for use in smart food packaging applications. (60% Accomplished) Characterization of physicochemical and functional properties of the AgNPs/AC-CNF films was conducted. The micro-structure of the films was analyzed using a transmission electron microscope, and the images of surface morphology showed CNF elements bound with AC particles, while the AgNPs were aggregated together and anchored with AC and CNF. The agglomerated AgNPs particles were exposed on the surface of the film, enabling the potential for antibacterial effects. To identify the possible functional groups of AgNPs/AC-CNF films and interactions between CNF and AC, fourier transform infrared spectroscopy (FTIR) analysis was performed. The FTIR results indicated that there is no significant band difference between the 15%, 30% and 50% levels of CNF in the AgNP/AC-CNF films, but determined that aromatic double carbon bonds (C=C), amine groups (C-N), carbondioxide bonds (O=C=O), and alcohol groups (O-H) were formed inside the films during casting. The mechanical properties of AgNPs/AC-CNF films were strengthened by the strong bonding and show promise for smart packaging applications. The crystalline phase and surface composition of AgNPs/AC-CNF films were analyzed using a X-ray diffractometer (XRD). The results showed that there were no obvious differences in the peaks between different concentrations of AgNPs/AC-CNF films. XRD confirmed that AgNPs has a face-centered cubic (FCC) structure. A higher degree of crystallinity of AgNPs with CNF and AC was formed. Both ionic and physical crosslinking of polymeric mixtures influenced the stabilization of AgNPs in the polymeric nanocomposites. The thermal properties of AgNPs/AC-CNF films were analyzed using a thermogravimetric analyzer (TGA) (Q5000 SA, TA Instruments). The results showed that the AgNPs served as the heat barrier components with activated carbon and nanocellulose. When the concentration of AgNPs inside the films increased from 70 to 450 ppm, the films' thermal stability was also increased. Compared to the other films, the film containing 450 ppm of AgNPs had 60% higher thermal stability. Electrical properties of AgNPs/AC-CNF films were characterized by determining their cyclic voltammetry (CV), differential plus voltammetry (DPV), and linear sweep voltammetry (LSV) using a potentiostat. The CV of AgNPs/AC-CNF films increased from 0.2 to 1.1 μA when AgNPs content increased from 70 to 450 ppm, meanwhile, the resistivity values fluctuated from 6.8-1.9 kΩ.cm. The results confirmed that the conductivity of the films was improved as AgNPs content increased from 70 to 450 ppm. The antimicrobial activities of the AgNPs/AC-CNF films were demonstrated against food pathogens, Staphylococcus aureus and Escherichia coli, as described earlier. The results obtained to this point show that the nanocomposite films have promising potential for use in smart food packaging. We also see exciting possibilities for precision agriculture. For example, in preliminary trials we have used the mechanism of bio-polymer-based nanocomposite to create "smart" biochar-based control release fertilizers for precision agriculture. We were awarded $83,500 in research funding to extend the applications of nanocomposites to develop biochar-based control release nitrogen fertilizers. Goal 5: Preliminarily estimate BFN production costs to evaluate the technical and economic feasibility of the best process of BFN fabrication. Optimize the BFN fabrication process to determine the best pathway for commercialization. (30% Accomplished) The executive director of South Dakota Enterprise Institute, Mr. Tim Weelborg is serving as a business mentor for commercialization of new technologies developed in our research. Two undergraduate students (Jace Jerome and Stoen Mollman) from Univesity of South Dakota established a cost analysis model for commercialization of the new technologies and products during the second year of project. They have established a preliminary production cost analysis model and provided very useful information to develop key business concepts, skills, and resources for successfully commercialization of the technologies and products being developed in this project.

Publications

• Type: Journal Articles Status: Published Year Published: 2020 Citation: Cheng, S., L. Wei, K. Muthukumarappan, S.I. Mart�nez-Monteagudo. 2020. Kinetic analysis of non-isothermal oxidation of bioactive milk Lipids. J Food Process Eng. e13519. https://doi.org/10.1111/jfpe.13519
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Kaluwahandi, N., L. Wei, K. Muthukumarappan. 2020. Opportunities and challenges of cold plasma in food processing. Paper #: 2000969. ASABE Annual International Meeting. Virtual. July 13-15.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Sobhan, A., K. Muthukumarappan, L. Wei. 2020. Development of a novel PLA coated bio-nanocomposite film indicator for monitoring meat freshness. Paper #: 2000568. ASABE Annual International Meeting. Virtual. July 13-15.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Sobhan, A., K. Muthukumarappan, L. Wei, T. Van Den Top, R. Zhou. 2020. Development of an activated carbon-based nanocomposite film with antibacterial property for smart food packaging. Materials Today Communications DOI: https://doi.org/10.1016/j.mtcomm.2020.101124.
• Type: Journal Articles Status: Under Review Year Published: 2020 Citation: Sobhan, A., K. Muthukumarappan, L. Wei. 2020. Biosensors and biopolymer based nanocomposites for smart food packaging: Challenges and opportunities. J. Food Control. (submitted).
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Sobhan, A., K. Muthukumarappan, L. Wei. 2020. Development of bio-nanocomposite films by combination of PLA and biochar for smart food packaging. Paper #: 2000566. ASABE Annual International Meeting. Virtual. July 13-15.

Progress 10/01/18 to 09/30/19

Outputs
Target Audience:Undergraduate and graduate students: Undergraduate students have been targeted through formal and/or informal classroom instruction, research workshops/symposia, and other outreach activities. Many undergraduate students will choose agricultural production, food engineering, and biomass processing as an occupation. The activities in this project will help the students prepare for graduate school or careers in sustainable agriculture, food engineering, and biomass refinery industries. Graduate students have also been targeted to receive professional training to prepare for their future academic or industrial careers. Research community: Agricultural engineering and food processing scientists and professionals were targeted through publishing the research results and new discoveries in peer-reviewed professional journals and/or presentations at scientific conferences or symposia. An understanding of our research results will help them to develop hypotheses and effective processes that will advance their own research programs. Farmers and biomass producers:This audience was targeted because biomass feedstocks for the production of biopolymer-based nanocomposites will be supplied by these farmers and producers.They were targeted through formal and informal instruction or training in technical workshop, research symposium, and other extension/outreach activities.? Public andcustomers: This audience was targeted because the smart packaging and food products developed from this research will ultimately be consumed or used by the general public. The customers' needs, likes, and acceptances should be considered in the research.The audience was targeted through publications of research results, websites, conferences, workshops, and extension/outreach activities. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project provided great opportunities for professional education and training of the next generation workforce for local communities and industries by involving four graduate students through the research and education activities. One PhD and one M.S. student have grained their degrees. The research results and discoveries were published in 7 conference papers/posters/presentation and 2 peer-reviewed journal papers. A website was established to disseminate the research results and newest knowledge of biomass conversion and bioproduct development at: https://lw9898.wixsite.com/linweiwebsite. These outreach activities are not only promoting public awareness of sustainable technologies, but also disseminating the new discoveries and knowledge to improve agricultural production, food safety, and smart packaging materials. The details of the 4 graduate students who were involved or trained in this project are listed as following: Abdus Sobhan is a current Ph.D. student of ABME. He helped the PI fabricate activated carbon based functional nanocomposite films. He aided the PI to guide undergraduate students in the research project. Zhisheng Cen is a current M.S. student of ABE. He worked on the study of fabricating PLA-based nanocomposite coatings for applications in control-release fertilizer development. Shouyun Cheng was a Ph.D. student of ABME, He helped the PI to develop the nanocellulose extraction process. He also aided the PI to guide undergraduate students in the research project. This student graduated on December 10, 2018. Zeyad Albahr was a M.S. student of ABE. He worked on fabrication and characterization of PLA-based nanocellulose composite films for food packaging applications. This student graduated on December 10, 2018. How have the results been disseminated to communities of interest?Nine publications (including peer-reviewed journal papers and presentations) were achieved in the first year of project. Research results were also disseminated to the local communities and public through workshops, conferences, or other outreach activities. What do you plan to do during the next reporting period to accomplish the goals?We will continue to develop biopolymer-based nanocomposites for applications in biosensors, precision fertilization, sustainable agriculture, and food health and safety. Our long-term goal is to establish nationally and internationally recognized research programs at SDSU. We will further extend the network of research collaborations across not only SDSU, but also to national /international institutes, agencies, and industrial partners. We will explore all possible internal and external sources to secure research funding to improve the research facility and analytic capability, and continue to move the research to a higher level with more positive impacts. To achieve the goal of this project, we plan the following activities in the next evaluating period. Goal 1: Extract biopolymers (cellulose and lignin) from corn stalks and sawdust for synthesis of BFNs. According to the performance of CNF and CNC used as nanofiller in the produced nanocomposite films, we will improve and optimize the effectiveness of 2H process for production of cellulose and nanocelluloses at low cost. Goal 2: Determine the best performing nano-fillers that can fit into biopolymer-based matrices and form functional nanocomposites. Additional biomass feedstocks such as woody sawdust, switchgrass, soybean straw will be examined for their suitability for use as nano-fillers in production of biopolymer-based nanocomposites; Goal 3: The nanocomposite film fabrication process combined with the solvent blending method will be upgraded and optimized to improve the properties of nanocomposite films at low cost. In-situ growth and thermal melting will also be tested in next period. Goal 4: We will continue to characterize the biopolymer-based nanocomposite films for physicochemical and functional properties. We will evaluate the suitability of the produced films for use in smart food packaging applications. Goal 5: Preliminarily estimate BFN production costs to evaluate the technical and economic feasibility of the best process of BFN fabrication. Optimize the BFN fabrication process to determine the best pathway for commercialization. We will estimate production costs of the produced nanocomposite films, and then evaluate the technical and economic feasibility of nanocomposite films for use in smart food packaging.

Impacts
What was accomplished under these goals? Goal 1: Extract biopolymers (cellulose and lignin) from corn stalks and sawdust for synthesis of BFNs. (25% Accomplished) A new process combining hydrolysis and homogenization (2H process) for cellulose and nanocellulose extraction was designed and tested. Biomass feedstocks (e.g. corn stover and prairie cordgrass) were first ground into powder and then hydrolyzed by acid solutions. The slurry was then subject to high speed (>8000 RPM) homogenization to extract cellulose nanofiber (CNF) and cellulose nanocrystals (CNC). The intermolecular and intra-molecular bonds of cellulose chains were destructed by the shearing forces of the high-speed homogenizer when the homogenization time was long enough. The smaller particle size of celluloses ranged from 0.03 to 0.23 μm (30 - 230 nm). The mean of nanocellulose dimension was 0.10μm (100nm), which meets the criteria for commercial nanocellulose products. After optimization, this 2H process will be consided for U.S. patent application. Goal 2: Determine the best performing nano-fillers that can fit into biopolymer-based matrices and form functional nanocomposites. (30% Accomplished) To screen the best nanofillers (CNF or CNC) for fabrication of biopolymer-based nanocomposites, the CNF and CNC produced by 2H process were combined with different substrates. First, CNF was individually combined with rice starch flour, potato starch powder, biochar, and O2Si powder to produce CNF-based nanocomposite films through a water solution casting method. Second, different concentrations (1%, 3%, and 5%) of CNC were intermixed with polylactic acid (PLA) to produce PLA-based nanocomposite (PBN) films using a solvent blending and casting approach. Third, carbon-based nanocomposite (CBN) films were generated by intermingling different concentrations (15%, 30%, 50%) of CNF into biochar (BC), activated carbon (AC), or biochar-derived-activated-carbon (BAC). To enphance anti-bacteria properties, different concentrations (140, 220, 300, 380, and 450 ppm) of silver nanoparticles (AgNPs) were impregnated into the CBN films to produce a new antimicrobial conductive and thermal stable films (named AgNPs/CBN film) . As a control, neat CNF and PLA films were also generated under the same conditions. Performance of all resulting films is described in Goal 4. Goal 3: Screen the best process of BFN fabrication from three nanocomposite synthesis technologies: solvent blending, in-situ growth, and thermal melting. (30% Accomplished) Two different processes were designed and tested to produce the nanocomposite films described in Goal 2. One is the water solution casting method. CNF was individually mixed with water solutions of rice starch flour, potato starch powder, biochar, or O2Si powder. The ratio of CNF to the substrate was 1:10. The prepared solutions were homogenized at 10,000 RPM for 15 min. The mixtures were then put into a 3-in. petri dish, uniformed to the same thickness, and placed into a drying oven at 50 oC for 24 h to form CNF-based nanocomposite films. The second process was the solvent blending and casting method, which was used to generate PBN films. PLA was first dissolved in methane chloride for 3 hours and then different concentrations (0, 1, 3, or 5%) of CNC along with 20 ml distilled water were added. These slurries were blended using a homogenizer at 8,000 RPM for 5 min. The mixtures were then put into 3-in. petri dishes, uniformed to the same thickness, and placed into a drying oven at 35oC for 20 h to form PBN films. The water solution casting method was also used to fabricate CBN films, but higher percentages (15, 30, 50%) of CNF were individually formulated with BC, AC, and BAC. The new AgNPs/CBN films were made by impregnating AgNPs into the CBN films and drying at 60 oC for 30 h. Performance of all resulting films is described in Goal 4. Optimizations of these two processes will be carried out after evaluating film performance. Goal 4: Characterize physicochemical and functional properties of the BFNs produced and evaluate their potential for use in smart food packaging applications. (30% Accomplished) The CNF-based nanocomposite films produced by combining CNF with rice starch flour, potato starch powder, biochar, or O2Si powder through water solution casting method were characterized for physicochemical properties, including water absorption rate (WAR), water uptake rate (WUR), and water vapor permeability rate (WVPR). These new nanocomposite films were compared to the neat PLA film. The CNF + BC film had the highest WAR (255%), following by the films of CNF+PLA, neat CNF, and CNF + Rice flour. The film of CNF + Potato starch had the lowest WAR. The WUR of CNF + AC films were significantly higher than that of CNF + BC films, but there was no significant difference among CNF + AC films or CNF + BC films when the CNF contents increased from 5% to 15%. The methylene blue (MB) absorption rate of CNF + BAC films increased when the BC activation temperature increased from 700 oC to 900 oC. The MB absorption rate of CNF + BAC films was very closed to the CNF + AC films if the BC was activated at 900 oC. The PBN films showed significant improvements in mechanical tensile strength and physical properties. The tensile strength of neat PLA films was 1.96 kg/cm2, but was 49% higher in the PBNF film containing 3% CNC. The tensile strengths of films containing 1% and 5% CNC were similar to the neat PLA film. The WUR of PLA+CNC nanocomposite films were lower than that of neat PLA film when the CNF content was 1% or 3%, but higher when CNC content was 5%. The neat PLA film had the highest WVPR, and as the percentage of CNC increased, the WVPR of PBNF films decreased. The PBNF looks very promising as a renewable and biodegradable material for smart packaging. The CBN films were characterized for their physicochemical and functional properties. The electrical conductivity of CBN film decreased when the CNF content increased, but the tensile strength, strain, and Young's modulus of CBN films increased significantly from 0.03 to 4.78 MPa, 0.13 to 1.94% and 97.64 to 247.3 MPa, respectively. The thermal stability of CBN films was also improved significantly. The linear sweep voltammetry (LSV), cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical spectroscopy (EIS) of CBN films were examined. The Rs values significantly increased from 1.5 k? to 12.5 k? when CNF contents in the CBN films increased from 15% to 50%. The CBN film with less than 50% of CNF content was suitable for developing biosensor. The antimicrobial properties of AgNPs/CBN film at different AgNPs concentrations (140-450 ppm) were evaluated against the Gram positive bacterium S. aureus and the Gram-negative bacterium E. coli. The higher AgNPs concentrations were more effective in inhibiting microbial growth. Moreover, the resistivity change of AgNPs/CBN films occurred significantly at higher AgNP concentrations. This AgNPs/CBN film shows not only significant potential for use in electronic devices and biosensors, as well as smart food packaging. Goal 5: Preliminarily estimate BFN production costs to evaluate the technical and economic feasibility of the best process of BFN fabrication. Optimize the BFN fabrication process to determine the best pathway for commercialization. (0% Accomplished) No activities were conducted in this goal 5 during the first year of project.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Abdus Sobhan, Kasiviswanathan Muthukumarappan, Zhisheng Cen, Lin Wei. 2019. Characterization of nanocellulose and activated carbon nanocomposite films biosensing properties for smart packaging. Carbohydrate Polymers. Volume 225: page115189.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Abdus Sobhan, Kasiviswanathan Muthukumarappan, Lin Wei. 2019. Characterization of bionanocomposite films based on nanocellulose and activated carbon. ASABE Annual International Meeting. July 7  10. Boston, MA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Zhisheng Cen, Lin Wei, Yajun Wu. 2019. Developing a control-release nitrogenous fertilizer by combination of biochar and sodium alginate. ASABE Annual International Meeting. July 7  10. Boston, MA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Shouyun Cheng, Lin Wei, Kasiviswanathan Muthukumarappan, and Sergio I. Martinez- Monteagudo. 2019. Oxidation kinetics of bioactive milk lipids using differential scanning calorimetry. American Dairy Science Association (ADSA) Annual Meeting. June 23  26. Cincinnati, Ohio.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Lin Wei, Abdus Sobhan, Kasiviswanathan Muthukumarappan. 2019. Development of nanocellulose based activated carbon film for smart food packaging applications. Paper ID: P04  092, IFT annual Meeting. June 2  5. New Orleans, LA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Zhisheng Cen, Lin Wei, 2019. Develop biochar-based control release nitrogenous fertilizers by coating PLA. SDSU Research Forum. May 21. South Dakota State University, Brookings, SD.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Zhisheng Cen, Lin Wei, 2018. Develop an effective biochar-based nanocomposite for removal of nitrogen and phosphorus from agricultural wastewater. SD Eastern Water Conference. October 17. South Dakota State University, Brookings, SD.
• Type: Journal Articles Status: Submitted Year Published: 2019 Citation: Abdus Sobhan, Kasiviswanathan Muthukumarappan, Lin Wei, Trevor Van Den Top, Ruanbao Zhou. 2019. Characterization of activated carbon based nanocomposite films for smart food packaging. Composite Science and Technology.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Lin Wei, Zeyad Ali Albahr. 2019. Develop biopolymer-based nanocomposite films by combination of cellulose nanocrystal (CNC) and polylactic acid (PLA). ASABE Annual International Meeting. July 7  10. Boston, MA.