Progress 09/01/11 to 08/31/16

Outputs
Target Audience:With this project we sought to reach the general public, food processors, and regulatory agencies. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Two graduate students worked on these projects. The graduate student who worked on high pressure assisted infusion of quercetin in cranberries completed her PhD in the year 2014; and the graduate student who worked on high pressure assisted infusion of calcium in baby carrots completed her MS in the year 2016. How have the results been disseminated to communities of interest?Results from this work were presented by the respective graduate students in their final PhD and MS theses defenses, in July 2014 and April 2016, respectively, which areopen to public. The results were also presented at national and international conferences. Two research papers were published in a scientific journal in the year 2015. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? During the course of this study, our group found that enhanced infusion of quercetin in both fresh and frozen-thawed cranberries was achieved under high-pressure (100 MPa - 551 MPa), when compared to infusion at ambient conditions. The amount of quercetin infused in frozen-thawed cranberries was found to be two to three times that of the amount infused by vacuum or atmospheric pressure infusion. The amount of quercetin infused did not significantly change beyond 100 MPa; and even low pressures up to 20 MPa were found to infuse twice the amount of quercetin when compared to vacuum or atmospheric pressure. High pressure processing was found to result in 144 times faster infusion compared to atmospheric pressure diffusion. Pressure cycling (2 cycles and 5 cycles) was observed to significantly increase the amount of infused quercetin. Experiments were performed to evaluate possible mass transfer mechanisms under pressure. Two systems were used for this purpose - fresh cranberries, cells of which were intact before high pressure processing, and frozen-thawed cranberries, cell of which were permeabilized by the freeze-thawing process before high pressure processing. The extent of cell permeabilization (quantified as a cell permeability index, zp) caused by freeze-thawing was similar to that of high pressure processed fresh cranberries. Moreover, the frozen-thawed cranberries showed no additional cell permeabilization after HPP. It was observed that the fresh cranberries had about two times the amount of infused quercetin than in the frozen-thawed cranberries, when procssed under similar high pressure conditions. If cell rupture were the only cause of infusion, then the amount infused into both, the fresh and the frozen-thawed cranberries, should have been similar. Our results suggested that the enhanced infusion under high pressure may be a combination of the effect of concentration driven diffusion due to cell permeabilization and the instantaneous pressure-driven mass transport. We estimatedthat the pressure-driven transport contributed to 60 % of the infusion while concentration-driven diffusion (because of cell permeabilization) contributed to 40 % of the infusion. Based on our above observation of enhanced infusion of quercetin in cranberries under high pressure, the study was extended to evaluate the effect of high pressure processing on enhanced infusion of an important micronutrient, calcium, in baby carrots. Baby carrots treated with pectinmethylesterase (PME) enzyme solution at 37 °C for 45 min, were infused with calcium using calcium lactate gluconate (CLG) solution under high pressure at various pressure-time combinations. A Box-Behnken design of experiments was used to study the effects of high pressure (150 MPa - 550 MPa), hold time (5 min - 15 min), and CLG concentration (3 % - 9 % w/v) on the amount of calcium infused (mg per serving size of 85 g).Unprocessed baby carrots served as the control. Processed baby carrots were also analyzed for the effect of high pressure assisted calcium infusion on the texture, color, and beta-carotene extractability. Baby carrots infused with calcium using CLG solution (3 % - 9 % w/v) using vacuum and atmospheric infusion for 15 min, were also used as controls. The individual processing parameters of pressure, time, and CLG concentration, as well as their interactions were observed to significantly affect the amount of calcium infusion. We observed that moderate pressures (up to 350 MPa) - less hold time (5 min) conditions infused up to 100 mg/serving of baby carrots (equivalent to 10 % RDI of calcium), when 9 % CLG solution was used. Along with increased infusion of calcium, the processed baby carrots were observed to have a higher amount of extractable beta-carotene (4 - 5 times compared to the unprocessed control), along with higher hardness values for texture and a darker color. Pressure cycling experiments (1 cycles, 3 cycles, and 5 cycles) were performed such that the hold time of high pressure processing was constant. Increasing the number of pressure cycles significantly increased the amount of infused calcium. Preliminary shelf-life analysis showed that the baby carrots infused with calcium at 350 MPa for 15 min using 9 % CLG solution are microbiologically safe up to 4 days. A lab scale sensory evaluation showed that the taste of the calcium infused baby carrots was not altered when compared to the unprocessed baby carrots. Thus, through these projects we demonstrated enhanced infusion of important biomolecules under high pressure in both a fruit system (cranberries) and a vegetable system (baby carrots), with little to no change in their texture, color, and sensory characteristics. This research can help develop process design guidelines that will be useful to the food industry and can be implemented for a broad class of snacks based on fruits and vegetables. The process design guidelines developed through this research will impact the food industry as well as consumers so as to develop and avail a broad class of nutrient infused healthy snacks based on fruits and vegetables. Developing new, convenient, affordable, and minimally processed products will provide an alternative snack choice for a population susceptible to vitamin deficiencies.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Gosavi, N.S., Salvi, D., Karwe, M. V. (2016). High Pressure Assisted Infusion of Calcium in PME Treated Baby Carrots. Conference on Food Engineering, Columbus, Ohio, September 12-14, 2016.
• Type: Theses/Dissertations Status: Published Year Published: 2014 Citation: Mahadevan, S. (2014). High pressure assisted infusion of phytochemical antioxidants into fruits: Influence of process parameters and mechanistic insights. Rutgers The State University of New Jersey-New Brunswick
• Type: Theses/Dissertations Status: Published Year Published: 2016 Citation: Gosavi, N. S. (2016). High Pressure Assisted Infusion of Calcium in PME Treated Baby Carrots. Rutgers, The State University of New Jersey, NJ, USA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Gosavi, N.S., Salvi, D., Karwe, M. V. (2016). High Pressure Assisted Infusion of Calcium in PME Treated Baby Carrots. IFT Annual Meeting, Chicago, July 16-19, 2016.

Progress 10/01/14 to 09/30/15

Outputs
Target Audience:General Public, Food Processors, and Regulatory Agencies. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?A graduate student, Noopur Gosavi, has been involved in this study How have the results been disseminated to communities of interest?The results were presented at an international conference. What do you plan to do during the next reporting period to accomplish the goals?Future work for the infusion of calcium in baby carrots study includes, Measurement of cell permeability and correlation to infusion of calcium Measurement of extractability of beta carotene from the processed and unprocessed baby carrots Shelf life analysis of high pressure infused baby carrots

Impacts
What was accomplished under these goals? We found that infusion was enhanced under high-pressure (100 - 551 MPa), when compared to infusion at ambient conditions, in both fresh and frozen-thawed cranberries. While infusion was enhanced, frozen-thawed cranberries showed no additional cell permeabilization after HPP, suggesting that cell permeabilization may not be the only cause of enhanced infusion. Additionally, though both fresh and frozen-thawed cranberries showed similar cell permeability after HPP, the fresh cranberries had about two times the amount of infused quercetin than in the frozen-thawed cranberries, under similar conditions. If cell rupture were the only cause of infusion, then the amount infused into both, fresh and frozen-thawed cranberries, should have been comparable. Our results suggest that enhanced infusion by HPP may be caused by concomitant factors in addition to cell-membrane rupture. Based on developed model system of infusion under high pressure in cranberry with quercetin the study was extended to undestand infusion of calcium in baby carrots.A Box-Behenken Design (BBD) of experiments was employed with pressure (MPa), time (min), and CLG (infusate) concentration (% w/v) as variables. The pressure was varied from 150 MPa to 550 MPa, time from 5 min to 15 min, and CLG concentration from 3 % to 9 %. Each variable was varied at three levels, and a total of 15 experiments were performed. All the samples (untreated and treated) were evaluated for the texture (hardness), color and amount of calcium infused (mg calcium/serving). Results and progress on each objective are discussed bellow, Effect of process parameters on the amount of calcium infused in baby carrots: Calcium infusion of up to 150 mg/serving of carrots (equivalent to 15 % RDI) was achieved with HPP. The amount of calcium infused with HPP was significantly higher than that infused by vacuum infusion and osmotic infusion. Vacuum and osmosis infused samples had calcium content close to the untreated sample, highlighting that HPP accelerates the infusion of calcium in baby carrots. It was found that the pressure (MPa) and infusate (CLG) concentration had a significant linear effect on the infusion of calcium in baby carrots. Effect of high pressure processing on the texture and color of baby carrots High pressure processing did not significantly affect/alter the texture and color of baby carrots. The texture of processed baby carrots was similar to the untreated ones. Effect of high pressure processing on beta carotene extractability HPLC analysis is currently in progress to measure the amount of beta carotene extracted from processed samples. It is hypothesized that HPP would increase the extractability of beta carotene from baby carrots, which may indicate towards increased bioavailability of beta carotene from high pressure processed carrots. Effect of pressure cycling Pressure cycling did not have a significant effect on infusion of calcium in baby carrots. In conclusion, we were able to achieve infusion of calcium up to 15 % RDI in one serving of carrots without significantly affecting their texture and color. Thus, we were able to verify that HPP enhances the infusion of calcium in baby carrots without altering its sensory attributes of texture and color. HPP can therefore be effectively used as an infusion technique to infuse fruits and vegetables with nutrients and micronutrients they inherently lack.

Publications

• Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: Gosavi, N.S., Salvi, D. A., Karwe, M. V. 2015. High Pressure Assisted Infusion of Calcium in PME Pre-treated Baby Carrots. 12th International Congress on Engineering and Food (ICEF'12) - Poster session, Quebec City, Quebec, Canada. June 14 - 18, 2015.

Progress 10/01/13 to 09/30/14

Outputs
Target Audience: General Public, Food Processors, and Regulatory Agencies. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? The graduate student working on thisproject completed her Ph.D. A new graduate student has started working on this part of this project. How have the results been disseminated to communities of interest? Results were presented in a final Ph.D. thesis defense, which is open to public, in July 2014. One peer-reviewed manuscript has been accepted for publication in the year 2015 and another is under review process. What do you plan to do during the next reporting period to accomplish the goals? We plan to extend the knowledge and understanding generated through this project to other fruit and vegetable systems such as carrots.

Impacts
What was accomplished under these goals? In this study we investigated, using a model system, HHP driven infusion of quercetin into cranberries. The main objectives of the study were to test whether enhanced infusion of quercetin can be achieved using HHP and to test whether the commonly accepted mechanism of cell permeabilization is operative in this model system. Two systems were used in this study - fresh cranberries, cells of which are intact and frozen-thawed cranberries, cells of which get permeabilized during freeze-thawing process. Under HHP, infusion of quercetin was enhanced, compared to infusion at ambient conditions (control), in both fresh and frozen-thawed cranberries. While the amount of quercetin infused under pressure was 3-5 times that in control, it was independent of the applied pressure in a broad range (5-551 MPa). Low pressure of 5 MPa was sufficient to cause enhanced infusion in frozen-thawed cranberries. Furthermore, pressure cycling treatment (2 cycles and 5 cycles) significantly increased the amount of quercetin infused. Unlike other studies, no additional cell permeabilization was observed in frozen-thawed cranberries after HHP, although amount infused was higher. This result suggested that the commonly accepted mechanism of cell permeabilization may not be the only cause of enhanced infusion. Additionally, while both fresh and frozen-thawed cranberries showed similar cell-membrane permeability after HHP, fresh cranberries had about two times greater amount of infused-quercetin than in frozen-thawed cranberries after HHP. If cell-membrane rupture were the only mechanism of infusion, then the amount infused into both, fresh and frozen-thawed cranberries, should have been equal. These results suggest that HHP infusion is not just cell-permeabilization based but may be caused by pressure-driven flow (Darcy flow in porous media). Understanding the actual mechanisms of transport under pressure may enable the development of process guidelines that will help the food industry to develop value-added foods.

Publications

• Type: Journal Articles Status: Accepted Year Published: 2015 Citation: Mahadevan, S., Nitin, N., Salvi, D., and Karwe M. V. (2015). High Pressure Enhanced Infusion in Cranberries: Influence of process parameters. Journal of Food Process Engineering, Accepted for publication, December 2014
• Type: Journal Articles Status: Accepted Year Published: 2015 Citation: Mahadevan, S., Salvi, D., and Karwe M. V. (2015). High Pressure Enhanced Infusion in Fresh and Frozen-Thawed Cranberries: A Comparative Study. Journal of Food Process Engineering, Accepted for publication, January 2015

Progress 10/01/12 to 09/30/13

Outputs
Target Audience: Food Processors, Food Scientists, and General Public. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? The student got opportunties to present the results at scientific meetings. How have the results been disseminated to communities of interest? Yes.The results were presented at the Annual Meeting of the Institute of Food Technologists,Chicago, IL, (July 2013) What do you plan to do during the next reporting period to accomplish the goals? In order to gain further insights into high pressure assisted enhanced infusion experiments will be carried out to develop a predictive model to predict the amount of substance infused given the processing conditions of pressure, time and material properties.

Impacts
What was accomplished under these goals? We found that infusion was enhanced under low pressure (5 to 100 MPa), when compared to infusion under ambient conditions (control) in frozen-thawed cranberries. Additionally, the amount infused was independent of applied pressure, suggesting that very low pressure (~ 5 MPa) is sufficient to cause enhanced infusion in frozen-thawed cranberries. While the amount infused was enhanced after pressure treatment compared to control, there was no significant difference in cranberry cell permeability before and after pressure treatment. These results suggest that cell permeabilization may not be the only cause for enhanced infusion under pressure. Furthermore, pressure cycling treatment (1, 2 and 5 cycles) significantly increased quercetin infused but did not change cranberry cell permeability with increase in the number of pressure cycles.This work demonstrates the potential of low pressure treatment as a low energy and low-cost process to develop nutraceutical enriched food products.

Publications

• Type: Conference Papers and Presentations Status: Accepted Year Published: 2013 Citation: Swetha Mahadevan and Mukund.V. Karwe, Enhanced Infusion under Low High Pressure with and without Induced Cell-Membrane Permeabilization, IFT Annual Meeting, Chicago, July 13-16, 2013.

Progress 10/01/11 to 09/30/12

Outputs
OUTPUTS: This study is undertaken to demonstrate and understand the mechanism of enhanced infusion of medium size (300-350 Da) phytochemical antioxidants into fruit substrates using high pressure processing (HPP). With quercetin-cranberry as the model antioxidant-fruit system, the objectives are to (a) test whether enhanced infusion can be achieved using HPP, (b) determine the dependence of enhanced infusion on process variables (pressure and time) and, (c) test whether the commonly accepted mechanism of cell permeabilization is in fact operative in this model system. Osmotic dehydration has been traditionally used in the food industry for infusion of substances into food materials. Since diffusion, the underlying phenomenon of osmotic dehydration, is a slow process, there is a need for methods that can accelerate infusion. High hydrostatic pressure processing (HPP) is one such alternative. High pressure processing (HPP) has been shown to infuse small size molecules into food materials much faster than diffusion-based processes. The accepted mechanism is that HPP permeabilizes cells and therefore increases their diffusivity. Hence, infusion is enhanced when foods are treated under high pressure. The enhanced infusion is explained in terms of increased effective diffusivity (Deff), which does not elucidate the actual mass transport mechanisms. We present here results of a study that tests this commonly accepted mechanism by conducting HPP infusion in two systems - fresh cranberries, cells of which are intact and frozen-thawed cranberries, cells of which are already permeabilized during freeze-thawing process. In our experiments, scarified-frozen-thawed and scarified-fresh cranberries were infused with quercetin, an antioxidant, at pressures between 100-551 MPa for 1-60 minutes at or near room temperature. Infused quercetin and cell-membrane permeabilization in processed cranberries were determined using RP-HPLC and impedance spectroscopy, respectively. Fruit microstructure was analyzed using optical microscopy. We found that infusion was enhanced under high-pressure (100 - 551 MPa), when compared to infusion at ambient conditions, in both fresh and frozen-thawed cranberries. While infusion was enhanced, frozen-thawed cranberries showed no additional cell permeabilization after HPP, suggesting that cell permeabilization may not be the only cause of enhanced infusion. Additionally, though both fresh and frozen-thawed cranberries showed similar cell permeability after HPP, the fresh cranberries had about two times the amount of infused quercetin than in the frozen-thawed cranberries, under similar conditions. If cell rupture were the only cause of infusion, then the amount infused into both, fresh and frozen-thawed cranberries, should have been comparable. Our results suggest that enhanced infusion by HPP may be caused by concomitant factors in addition to cell-membrane rupture. Our results suggest that enhanced infusion by HPP may be caused by additional factors, other than cell-membrane permeabilization, and needs further investigation. PARTICIPANTS: S. Mahadevan, M.V. Karwe TARGET AUDIENCES: USFDA, USDA, Food Processors, U.S. Army PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
This research elucidates important aspects of the science of pressure-enhanced infusion. In addition, this work demonstrates the potential of HPP to develop nutraceutical-enriched food products. This understanding can be used to develop design guidelines that will be useful to the food industry and can be implemented for a broad class of fruits and infusing molecules to develop value-added nutrient rich foods.

Publications

• No publications reported this period