MODELING IN VITRO CONTROL RELEASE AND DIFFUSION OF LOADED NANOPARTICLES (LNP) IN THE GI TRACT - IMPACT OF PROCESSING

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: EVANS-ALLEN
• Reporting Frequency: Annual
• Accession No.: 1013057
• Project Start Date: Oct 1, 2017
• Project End Date: Sep 30, 2020
• Program Code: [(N/A)]- (N/A)

Recipient Organization
ALABAMA A&M UNIVERSITY
4900 MERIDIAN STREET
NORMAL,AL 35762

Performing Department
Food & Animal Sciences

Non Technical Summary
The advancement of bioprocess engineering techniques provides unprecedented opportunities to advance new food processing techniques to enhance the nutritional content of foods. The control release information of bioactive components in foods to be generated will lead to improve design and manufacturing of novel functional food products with significant gastrointestinal tract (GIT) health. The long-term aim of the proposed project is to establish a GIT simulation and in vitro modeling program that will provide a platform to test the relationships of different food processing methods to the behavior of processed foods in the GIT and its impact on the controlled release of nutrients and their bioaccessibility. It is therefore hypothesized that Loaded Nanoparticles (LNP) subjected to different processing systems will: degrade the viability of the LNP; affect the physical properties; impact the physical and chemical properties; affect the behavior and diffusion in the GIT; and negatively impact the gut microorganisms. Hence, the main objective of the proposed project is to develop an in vitro model to study the relationship between different methods of food processing and their behavior in the GIT. In general, the in vitro digestion model will provide useful alternative platform to animal and human subject to study the behavior of LNP in the GIT. The ideal in vitro provides accurate results within relatively short period of time and can serve as a tool for screening the LNP in food systems and their impact on the GIT. The GIT comprises of the stomach, small and large intestine and from the engineering perspective, the stomach is a reservoir, blender/grinder, mixer and a prime mover (pump) that controls the human digestion system. A knowledge gap exists in the understanding of the behavior of LNP in food and its bioaccessibility and bioavailability in the GIT. Testing the efficacy of the LNP behavior in the GIT requires a digestion models that will simulates the complex physicochemical and the physiological event that occur in the GIT. The modern food industry is in pursuit of manufacturing functional foods of significant health benefits, thus, one of the primary goals. This is in line with the premise of the proposed research project, which intends to provide vital physicochemical information to advance and evolve food processing techniques by incorporating natural food ingredients with significant health benefits. Furthermore, the proposed project will provide training opportunities for underrepresented minority students with advance degrees in the area of Food and Bioprocessing Engineering concentration.

Animal Health Component

50%

Research Effort Categories

Basic

30%

Applied

50%

Developmental

20%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
501	7410	2020	80%
502	7299	1010	20%

Knowledge Area
501 - New and Improved Food Processing Technologies; 502 - New and Improved Food Products;

Subject Of Investigation
7299 - Research equipment and methods, general/other; 7410 - General technology;

Field Of Science
2020 - Engineering; 1010 - Nutrition and metabolism;

Keywords

in vitro simulation

gastrointestinal tract

food processing methods

loaded nanoparticles

in vitro digestion

bioaccessibility

control release

Goals / Objectives
The main objective of the proposed project is to develop an in vitro model to study the relationship between different methods of processing nanoparticles (NP) in food their behavior in the GIT. The following specific objectives will be the prerequisite to achieving the main objectives of the project:Design and develop custom made simulation model with an incorporated a diffusion (dialysis) stage in the GIT model;Investigate the impact of conventional thermal, microwave, Pulse Electric Field processing on the loaded nanoparticles (LNP);Study the diffusion mechanism of processed LNP in porous GI membranes (porcine) using a dual-chamber diffusion set, and five chambers mimicking the upper GI and lower GI tract, respectively;Investigate the effect of rheological, particles sizes and thermal properties on the control release of processed LNP in food emulsion;Evaluate the bioaccessibility of the processed LNP through simulated porcine GI membranes;Evaluate the antimicrobial effect of processed LNP impact and its impact on food safety;Test the effect of processed LNP on the GIT microbiome;Established a correlation between an in-vitro and in-vivo models, andProvide a baseline data to develop and seek competitive grants for the sustainable development of the project

Project Methods
Objective 1: Design and develop custom made simulation model with a diffusion stage of the GIT:Task I - Design of the simulation system: Design and development of a simulation system based on the upper and lower GIT.Task II: Special diffusion module will be deigned to the incorporated into the small intestine to simulate the intestinal wall.Take III: Developing a Standard Operating Procedure (SOP)Objective 2: Investigate the impact of conventional thermal, microwave, Pulse Electric Fieldprocessing on the leaded nanoparticles (LNP);Task I - Conventional Heat Treatment: Steam retort will be used for conventional heattreatment. Loaded nanoparticles will be incorporated in a starchy material comprising of cornstarch and methyl cellulose will be used as the nanoemulsion (NAE). The NAE will be cannedin the #300 can and sealed and subsequently retort.Task II - Microwave: The microwave treatment will be done using the Microwave WorkStation-240 with FISO Commander Workstation software for sensors and result management(FISO Technologies Inc. Quebec, Canada).Task III- Pulse Electric Field (PEF): Samples (NAE) will be treated in a bench-scale, batch type, pulsed electric fields system (OSU-4J PEF, Ohio State University, Columbus, OH). The treatment module consisted of six co-fields flow, tubular chambers in series with diameters of 2.3 mm and electrode gaps of 2.92 mm. Each sample will be loaded into a sterile disposable 60 mL syringes. A bi-polar, square wave form electric pulse will be used for the experimental treatments. An oscilloscope will be used to measure the electric fields strength (30kV/cm), pulsed duration (2.9 to 485 μs), and pulse repetition rates.Objective 3: Study the diffusion mechanism of processed LNP in porous GI membranes (porcine) using a dual-chamber diffusion set, and two diffusion solutions mimicking the upper GI and lower GI tract, respectively;Task 1- Diffusion Modeling: The model will be a simplified representation of the human digestive system that mimics the segmentation motion of the GI tract. Porous GI membranes (porcine) will be used to simulate the absorption across the GI tract. Peristaltic pumps will be used to simulate the propulsive contractions of the digesta (peristalsis) movement at a velocity of 0.5 - to 2.0 cm/sec (Guyton, 1996). The simulation apparatus will be used to quantify the diffusivity of LLNP in food emulsion in the GI tract.Objective 4: Investigate the effect of rheological, particles sizes and thermal properties on the control release of processed LNP in food emulsion;Task 1 -Viscosity measurements: Viscosity measurements of the Nano-emulsions will be performed using the Discovery Hybrid Rheometer Series (Model DHR 2, TA Instruments, New Castle, DE) and to be operated via a computer software (TRIOS software V3.0, TA). A 40-mm parallel plate geometry will be used with a gap of 1000 μm.Task II - Thermophysical Property Determination: The thermal properties of the LLNP will be determined using Differential Scanning Calorimetry (DSC) (DSC 2000, TA Instruments, New Castle, DE) connected to a Refrigerated Cooling System, using a computer software (TRIOS software V3.0, TA). Twenty milligram of powdered sample will be sealed in a hermetic aluminum pan and subjected to a temperature ramp from 30 to 250°C at the heating rate of 10°C/min.Task III - Particle Size Distribution (PSD) and Poly Dispersity Index (PDI): Particle size distribution and PDI will be measured by dynamic laser scattering (DLS) technique using Malvern Zetasizer (Nano ZS90, Malvern Instruments Ltd., Enigma Business Park, UK.), based on the methods developed by Kassama et al. (2015).Task IV- Surface Charge: Zeta potential of the nanoparticle was measured from their electrophoretic mobility using a Malvern Zetasizer DLS (Nano ZS90, Malvern Instruments Ltd., Enigma Business Park, UK.), based on the methods developed by Kassama et al. (2015).Task V- Morphology: The morphology of the NP will be determined by transmission electron microscope (TEM) at Auburn University at an accelerating voltage of 60 kV. A sample size of 10 µL of aqueous suspension of particles will be placed on a 300 mesh copper grids and stained with a 2 % (w/v) phosphotungstic acid in dH2O to provide contrast under magnification. The suspension will be allowed to dry before viewing at 40,000 to 100,000 times magnification.Task VI - Entrapment efficiency: The entrapment efficiency was determined as a function of the DPPH concentration decrease over time. The NP will be dispersed in 95 % acetonitrile solution for a period of 72 h with periodic mixing to allow time for all bioactive compounds to dissolve in the solution based on the modified method developed by Misir and Kassama (2017).Task VII -Time Release Kinetics: The release kinetics will be determined by our own developed method which is different from the standard method commonly used (HPLC, UV-Vis & Flouresce spectroscopy). 1 mg/mL concentration of the LNP will be dropped in a 3 mL of 0.1 mM DPPH solution based on the modified method developed by Misir and Kassama (2017. Task IIX-Antioxidant activity of the LNP: According to the method proposed by Braca et al. (2001) with some modifications DPPH free radical scavenging activity will be determined. The mixture will be shake for 45 min at room temperature. The decrease of solution absorbance due to proton donating activity of components NP will be determined at 517 nm. Lower absorbance of the reaction mixture indicates higher free radical scavenging activity. Vitamin C (L-ascorbic acid) will be used as the positive control. Objective 5: Evaluate the bioavailability of processed LNP through simulated porcine GI membranes;Task 1 - Bioavailability: The digestibility of samples will be determined by the extent of diffusion of LNP through the dialysis membrane. The dialysis tubing will be purchased from Sigma Aldrich Inc. (St Louis, MO, USA) with a molecular weight cutoff of 12- 14 kDa. The dialysis tubing will be sized to 10 cm and soaked in distilled water at 4°C before use. The digesta in the stomach will be transported through the dialysis membrane based on the method described in Tharakan et al. (2010).Objective 6: Evaluate the antimicrobial effect of processed LNP impact and its impact on food safety;Task I-Efficacy of Antimicrobial Activity of LNP: Agar Diffusion method will be used to test the potency of the antimicrobial potential of the bioactive extract. This method will apply nanoemulsion to evaluate an inhibition zone as described by Lavalle´et al. (2010); Hanson and Martin (1978).Objective 7: Test the effect of processed LNP on the GIT microbiome;Task I - Test the effect of NAE on the GIT Microbiome: The following strains will be used for all experiments: Freeze Dried ampoule of 19829, Type strain; Ruminococcus gauvreauii CCRI-16110 (R. gauvreauii), Freeze Dried ampoule of 19114, Type strain; Enterococcus caccae SS-1777 (E.caccae), Freeze Dried ampoule of 16992, Type strain; Bifidobacterium catenulatum B669 (B.catenulatum). Frozen aliquots of bacteria will be recovered through inoculation in strain specific broth and grown overnight in the anaerobic chamber at 37°C. Each bacterial strain will be cultured and grown overnight sequentially at least twice prior to use to recovery from freezing before inoculating the bioreactors simulating the colon. Post inoculation microbial growth will be determined by measuring the McFarland Units (MU) using a densitometer, hence the MU will be correlated to microbial population.Objective 8: Established a correlation between an in-vitro and in-vivo models, andTask I- Correlation of in Vitro to in vivo: Six weeks old Sprague Dawley rats (Harlan, IN) will be maintained in a climate controlled room with temperature and relative humidity at 21°C and 50%, respectively and a 12 h light and dark cycle based on the modified method developed by Boateng et al. (2013).

Progress 10/01/17 to 09/30/20

Outputs
Target Audience:1.Graduate minority students: will be trained to develop expertise in food processing with emphasis on in vitro stimulation of the GIT; 2. Undergraduate minority students: experiential learning program to develop skillsets for research; 3. The information will be vital for the food industry, particularly the functional food manufacturers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One doctoral student was being trained on the design and development of the simulation system and he had successfully submitted and defended his doctoral dissertation proposal based on the project objectives. He concluded with the research objectives of the project.Two (2) undergraduate students were also recruited and were provided with experiential learning opportunities on developing research skills. How have the results been disseminated to communities of interest?The research outcomes were presented at international conferences i.e., IFT, ASABE annual virtual meetings. Local conferences, i.e., AAMU-STEM Day and graduate seminars. The undergraduate student also submitted an abstract at the AAMU-STEM-Day. However, the STEM-DAY was canceled due to the COVID-19 effect on early campus closure. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Much was accomplished on the project goals, however, the advent of the COVID-19 significantly diminishes the progress of the project. Unfortunately, none of the project goals were fully completed. I have applied in November 2020 for a project extension to complete the goals. However, there was inconsistency in the decisions of the no-cost extension as to why the application was neglected upon the submission of a no-cost extension in November 2020.

Publications

• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Mohammad Anwar Ul Alam and L.S. Kassama. 2020. Characterization and Comparative Analysis of Polylactic Acid vs Polylactic Co-glycolic Acid for Nanoencapsulation. Abstract Submission ID# 26166 virtually presented at the SHIFT20 IFT Annual, Virtual, July 12-15.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Misir, J., L. S. Kassama. 2020. Paper title: Thermal stability and In vitro Controlled Release Kinetics of Lycopene Nanoparticles in the GIT. Submission ASABE: ID # 2001266, presented at the AIM-2020 Virtual Agricultural and Biological Engineers Annual International Meetings, Virtual, July 12 to 15.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Ogechukwu Tasie, L. S. Kassama. 2020. Paper title: Bioavailability of Phytochemical and Anti-Oxidant derived from Enzymatic Hydrolyzed Canned Red Kidney Beans (Phaseolus vulgaris L.) in the GIT. Submission ASABE: ID # 2001286 presented at the AIM-2020 Virtual Agricultural and Biological Engineers Annual International Meetings, Virtual, July 12 to 15.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Mohammad Anwar Ul Alam and L.S. Kassama. 2020. Characterization of Polylactic Acid vs Polylactic Co-glycolic Acid for Controlled Release Kinetics of Encapsulated Bioactive Compounds. Abstract submitted for lecture presentation at the 2020-AAMU-STEM-DAY, Health, and Wellness Center. Normal, Alabama, April 16, 2020.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Cydney Jackson, Mohammad Anwar Ul Alam, and L.S. Kassama. 2020. Synthesis and Characterization of Poly Caprolactone (PCL) Nanoparticles (NP) for the Delivery of Lycopene. Abstract submitted for poster presentation at the 2020-AAMU-STEM-DAY, Health, and Wellness Center. Normal, Alabama, April 16, 2020.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Nina Biggs, Mohammad Anwar Ul Alam, Edwin Ochieng, and L.S. Kassama. 2020. Properties of Encapsulated Lycopene Nano-Particle (NP) Extracted from Raw Tomatoes by Column Chromatography Methods. Abstract submitted for poster presentation at the 2020-AAMU-STEM-DAY, Health, and Wellness Center. Normal, Alabama, April 16, 2020.

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

Outputs
Target Audience:1. Graduate minority students: will be trained to develop expertise in food processing with emphasis on in vitro simulation of the GIT; 2. Undergraduate minority students: experiential learning program to develop skillsets for research; 3. The information will be vital for the food industry, particularly the functional food manufacturers. Changes/Problems:The Specific Objective 2 is to investigate the impact of the conventional thermal, microwave, Pulsed Electric Field (PEF) processing on the loaded nanoparticles (LNP). Due to the inoperable conditions of the PEF system, the use of the PEF will be dropped. Hence, only the effect of the conventional thermal, and microwave pasteurization systems will be applied. What opportunities for training and professional development has the project provided?Currently, one doctoral student is being trained on the design and development of the simulation system and he has successfully submitted and defended his doctoral dissertation proposal based on the project objectives. He is currently implementing the research objectives of the project. Two (2) undergraduate students were also recruited and were provided with experiential learning opportunities on developing research skills. How have the results been disseminated to communities of interest?The research outcomes were presented at international conferences i.e., IFT, ASABE annual virtual meetings. Local conferences, i.e., AAMU-STEM Day and graduate seminars. The undergraduate student also submitted an abstract at the AAMU-STEM-Day. However, the STEM-DAY was canceled due to the COVID-19 effect on early campus closure. What do you plan to do during the next reporting period to accomplish the goals?Due to the delay in obtaining a graduate student in the execution of the stipulated project goals and objectives and coupled with the delay of campus lockdown from March to August 2020 due to the COVID-19 effect. During this period, no laboratory study was conducted, hence, there will be a need for a no-cost extension to be able to complete the planned task to accomplish the specific objectives 7 & 8.

Impacts
What was accomplished under these goals? The project provides a simple method of replacing the human gastrointestinal tract (GIT) with bioreactors that mimic the physiological functions of the human GIT system. Based on this model, we will be able to provide training to students on how to test various processed foods on human physiology, without the concern of using human/animal subjects in research. Specific Objective 1. Design and develop a custom-made simulation model with an incorporated diffusion (dialysis) stage in the GIT model: Outcome: The simulation model is complete. Hence, ready for experimentation. Specific Objective 2. Investigate the impact of the conventional thermal, microwave, Pulsed Electric Field (PEF) processing on the loaded nanoparticles (LNP): Outcome: The method of synthesizing loaded nanoparticles is complete. Hence, the impact of conventional thermal and microwave processing on loaded nanoparticles is currently ongoing. The use of PEF processing method was dropped because of the inoperable conditions of the PEF system. Specific Objective 3. Study the diffusion mechanism of processed LNP in porous GI membranes (porcine) using a dual-chamber diffusion set, and five chambers mimicking the upper GI and lower GI tract, respectively: Outcome: Currently conducting the preliminary test on the use of different naturally occurring porcine collagen membranes to adopt for this objective. Specific Objective 4. Investigate the effect of rheological, particles sizes, and thermal properties on the controlled release of processed LNP in food emulsion: Outcome: A new beverage product was developed with emulsified nanoparticles and the effect of rheological properties is currently ongoing. Specific Objective 5. Evaluate the bioaccessibility of the processed LNP through simulated porcine GI membranes; Outcome: This objective is currently ongoing. Specific Objective 6. Evaluate the antimicrobial effect of processed LNP impact and its impact on food safety: Outcome: This objective is contingent on the completion of specific objective 5. Specific Objective 7. Test the effect of processed LNP on the GIT microbiome: Outcome: This objective is contingent on the completion of specific objective 5. Specific Objective 8. Established a correlation between in vitro and in vivo models, and Outcome: This objective is contingent on the completion of specific objective 5. and Specific Objective 9. Provide baseline data to develop and seek competitive grants for the sustainable development of the project: Outcome: After the completion of the project.

Publications

• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Mohammad Anwar Ul Alam and L.S. Kassama. 2020. Characterization and Comparative Analysis of Polylactic Acid vs Polylactic Co-glycolic Acid for Nanoencapsulation. Abstract Submission ID# 26166 virtually presented at the SHIFT20 IFT Annual, Virtual, July 12-15.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Misir, J., L. S. Kassama. 2020. Paper title: Thermal stability and In vitro Controlled Release Kinetics of Lycopene Nanoparticles in the GIT. Submission ASABE: ID # 2001266, presented at the AIM-2020 Virtual Agricultural and Biological Engineers Annual International Meetings, Virtual, July 12 to 15.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Ogechukwu Tasie, L. S. Kassama. 2020. Paper title: Bioavailability of Phytochemical and Anti-Oxidant derived from Enzymatic Hydrolyzed Canned Red Kidney Beans (Phaseolus vulgaris L.) in the GIT. Submission ASABE: ID # 2001286 presented at the AIM-2020 Virtual Agricultural and Biological Engineers Annual International Meetings, Virtual, July 12 to 15.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Mohammad Anwar Ul Alam and L.S. Kassama. 2020. Characterization of Polylactic Acid vs Polylactic Co-glycolic Acid for Controlled Release Kinetics of Encapsulated Bioactive Compounds. Abstract submitted for lecture presentation at the 2020-AAMU-STEM-DAY, Health, and Wellness Center. Normal, Alabama, April 16, 2020.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Cydney Jackson, Mohammad Anwar Ul Alam, and L.S. Kassama. 2020. Synthesis and Characterization of Poly Caprolactone (PCL) Nanoparticles (NP) for the Delivery of Lycopene. Abstract submitted for poster presentation at the 2020-AAMU-STEM-DAY, Health, and Wellness Center. Normal, Alabama, April 16, 2020.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: Nina Biggs, Mohammad Anwar Ul Alam, Edwin Ochieng, and L.S. Kassama. 2020. Properties of Encapsulated Lycopene Nano-Particle (NP) Extracted from Raw Tomatoes by Column Chromatography Methods. Abstract submitted for poster presentation at the 2020-AAMU-STEM-DAY, Health, and Wellness Center. Normal, Alabama, April 16, 2020

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

Outputs
Target Audience:1. Graduate minority students: will be trained to develop expertise in food processing with emphasis on in vitro simulation of the GIT; 2. Undergraduate minority students: experiential learning program to develop skillsets for research; 3. The information will be vital for the food industry, particularly the functional food manufacturers. Changes/Problems:Due to the delay in obtaining a graduate student in the execution of the stipulated project goals and objectives, there is a need for a no-cost extension to be able to complete the planned task to accomplish the objectives. What opportunities for training and professional development has the project provided?Currently, one graduate student was recruited and being trained on the design and development of the simulation system and also currently developing a doctoral dissertation proposal based on the project objectives. How have the results been disseminated to communities of interest?The research outcomes were presented at international conferences i.e., IFT, ASABE annual meetings. Local conferences, i.e., AAMU-STEM Day and Dean's Conferences and graduate seminars. What do you plan to do during the next reporting period to accomplish the goals?The next reporting period coincides with the last year of the project. Due to the delay in obtaining a graduate student in the execution ofthe stipulated project goals and objectives, there might be the need for a no-cost extension to be able to complete the planned task to accomplish the objectives.

Impacts
What was accomplished under these goals? The project provides a simple method of replacing the human gastrointestinal tract (GIT) with bioreactors that mimic the physiological functions of the human GIT system. Based on this model, we will be able to provide training to students on how to test various processed foods on human physiology, without the concern of using human/animal subjects in research. Specific Objective 1. Design and develop a custom-made simulation model with an incorporated diffusion (dialysis) stage in the GIT model: Outcome: The simulation model is in the final stage of completion. Pumps, valves, motors and process control components are being purchased to semi-automate the system. Specific Objective 2. Investigate the impact of conventional thermal, microwave, Pulsed Electric Field processing on the loaded nanoparticles (LNP): Outcome: The method of synthesizing loaded nanoparticles is complete, hence this objective is contingent on the completion of the specific objective 1. Specific Objective 3. Study the diffusion mechanism of processed LNP in porous GI membranes (porcine) using a dual-chamber diffusion set, and five chambers mimicking the upper GI and lower GI tract, respectively: Outcome: Currently reviewing different natural occurring porcine collagen membranes to adopt for this objective. Specific Objective 4. Investigate the effect of rheological, particles sizes and thermal properties on the control release of processed LNP in food emulsion: Outcome: This objective is contingent on the completion of specific objectives 1 and 2. Specific Objective 5. Evaluate the bioaccessibility of the processed LNP through simulated porcine GI membranes; Outcome: This objective is contingent on the completion of specific objectives 1, 2, and 3. Specific Objective 6. Evaluate the antimicrobial effect of processed LNP impact and its impact on food safety: Outcome: This objective is contingent on the completion of specific objectives 1, 2, and 3. Specific Objective 7. Test the effect of processed LNP on the GIT microbiome: Outcome: This objective is contingent on the completion of specific objectives 1, 2, and 3. Specific Objective 8. Established a correlation between an in vitro and in vivo models, and Outcome: This objective is contingent on the completion of specific objectives 1, 2, 3, 4, and 5. Specific Objective 9. Provide baseline data to develop and seek competitive grants for the sustainable development of the project: Outcome: After the completion of the project.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Ogechukwu Tasie, Lamin S. Kassama, Regine Mankolo, Judith Boateng . 2019. Instrumental Inductively Coupled Plasma profiling of mineral in enzymatic hydrolyzed red kidney beans (Phaseolus vulgaris L.). EC Nutrition, 14(10): 893-901.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Mohammad Anwar Ul Alam, L. S. Kassama. 2019 Comparative Study of Lycopene Encapsulation Efficiency in Polycapprolactone Vs Poly Lactic Coglycolic Acid. (Paper #: 1901707). Precedings the American Society of Agricultural and Biological Engineers Annual International Meetings, Boston Marriott Copley Place, Boston, Massachusetts, July 07 to 10. Ogechukwu Tasie, L. S. Kassama. 2019. Paper title: The Antioxidants potentials of Enzymatically Hydrolyzed Canned Red Kidney Beans (Phaseolus vulgaris L.) in the in vitro Simulated Gastrointestinal Tract. (Paper # 1901721). Precedings the American Society of Agricultural and Biological Engineers Annual International Meetings, Boston Marriott Copley Place, Boston, Massachusetts, July 07 to 10.
• Type: Conference Papers and Presentations Status: Accepted Year Published: 2019 Citation: Al-Alam, M.A. and L.S. Kassama. 2019. Comparative study of Lycopene encapsulation efficiency in Polycapprolactone Vs Poly Lactic Co-glycolic Acid. Paper ID # 1901707. Presented at the ASABE Annual International Meeting, Marriott Copley Place Boston, Boston, Massachusetts, July 07-10. Tasie, O.B., J. Boateng and L.S. Kassama. 2019. The antioxidant potentials of enzymatically hydrolyzed canned red kidney beans (Phaseolus vulgaris L.) in the in vitro simulated gastrointestinal tract. Paper ID # 1901721. Presented at the ASABE Annual International Meeting, Marriott Copley Place Boston, Boston, Massachusetts, July 07-10. Tasie, O.B., J. Boateng, L.S. Kassama. 2019. ICP Profiling of Mineral Contents in Hydrolyzed Red Kidney Beans (Phaseolus vulgaris L.). Presented (Abstract ID #: 23728) at IFT19 in New Orleans, LA, June 2 - 5, 2019. Tasie, O.B., J. Boateng, L.S. Kassama. 2019. Bioavailability of Nutrients During in vitro Fermentation of Hydrolyzed Canned Red Kidney Beans (Phaseolus vulgaris L.) in a Simulated Gastrointestinal Tract System. Presented (Abstract ID #: 23761) at IFT19 in New Orleans, LA, June 2 - 5, 2019. Muhammad Anwar Ul Alam & Kassama, L.S. 2019. Comparative study of Lycopene encapsulation efficiency in Polycapprolactone Vs Poly Lactic Co-glycolic Acid. Abstract #17 and Poster presented at the AAMU-STEM-DAY, Health and wellness Center. Normal, Alabama, April 12, 2019. Kassama, L.S. 2019. The impact of processing on food quality. College of Agriculture and Natural Sciences Dean and Research Directors Seminar for Principal Investigators. Dawson Building, AAMU, Normal, Alabama, November 12. Mohammad Anwar Ul Alam and Kassama, L.S. 2019. Encapsulation efficiency of Polycapprolactone. Poster Presentation at the FAS 50th Anniversary Golden Symposium. ARC Auditorium. Normal, Alabama, October 25.
• Type: Conference Papers and Presentations Status: Submitted Year Published: 2020 Citation: Al-Alam, M.A. and L.S. Kassama. 2019. Characterization and Comparative Analysis of Polylactic Acid vs Polylactic Co-glycolic Acid for Nanoencapsulation. Abstract (Submission ID: 26166) submitted for the IFT20 Annual Event ad Food Expo Conference, McCormick Place, Chicago, Illinois, July 12-15.

Progress 10/01/17 to 09/30/18

Outputs
Target Audience: Graduate minority students: will be trained to develop expertise in food process with emphasis on the in vitro simulation of the GIT; Undergraduate minority students: experiential learning program to develop skillsets for research; The information will be vital for the food industry, particularly the functional food manufacturers. Changes/Problems:The project was delayed due to the untimely recruitment of a graduate student. Finally, a graduate student was recruitment in August 2018, and he is making a significant progress. What opportunities for training and professional development has the project provided?Currently one graduate student was recruited and being trained on the design and development of the simulation system and also currently developing a doctoral dissertation proposal based on the project objectives How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period the following project goals, objectives and tasks is planned to be conducted, including the preparation of a doctoral dissertation proposal based on the project: Objective 1: Design and develop custom made simulation model with a diffusion stage of the GIT: Task I - Design of the simulation system: Design and development of a simulation system based on the upper and lower GIT. Task II: Special diffusion module will be deigned to the incorporated into the small intestine to simulate the intestinal wall. Take III: Developing a Standard Operating Procedure (SOP) Objective 2: Investigate the impact of conventional thermal, microwave, Pulse Electric Field processing on the leaded nanoparticles (LNP); Task I - Conventional Heat Treatment: Steam retort will be used for conventional heat treatment. Loaded nanoparticles will be incorporated in a starchy material comprising of corn starch and methyl cellulose will be used as the nanoemulsion (NAE). The NAE will be canned in the #300 can and sealed and subsequently retort. Task II - Microwave: The microwave treatment will be done using the Microwave Work Station-240 with FISO Commander Workstation software for sensors and result management (FISO Technologies Inc. Quebec, Canada). Task III- Pulse Electric Field (PEF): Samples (NAE) will be treated in a bench-scale, batch type, pulsed electric fields system (OSU-4J PEF, Ohio State University, Columbus, OH). The treatment module consisted of six co-fields flow, tubular chambers in series with diameters of 2.3 mm and electrode gaps of 2.92 mm. Each sample will be loaded into a sterile disposable 60 mL syringes. A bi-polar, square wave form electric pulse will be used for the experimental treatments. An oscilloscope will be used to measure the electric fields strength (30kV/cm), pulsed duration (2.9 to 485 μs), and pulse repetition rates.

Impacts
What was accomplished under these goals? Task Activity I - Simulation System Design: Design and development of a simulation system based on the concept illustrated in Figure 4. The system approach will factors in the upper and lower GIT. A graduate student was recruited during the 2018 fall Semester, hence the project is ongoing

Publications