Source: OHIO STATE UNIVERSITY submitted to
CHEMISTRY AND FUNCTIONALITY OF FLAVINOIDS: BIOLOGICAL ACTIVITY AND COMMERCIAL APPLICATIONS IN THE FOOD INDUSTRY
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0209069
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Nov 1, 2012
Project End Date
Sep 30, 2017
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
OHIO STATE UNIVERSITY
1680 MADISON AVENUE
WOOSTER,OH 44691
Performing Department
Food Science & Technology
Non Technical Summary
Consumption of fruits and vegetables has been associated with a reduced incidence of chronic diseases. Animal, clinical and in vitro studies provide compelling evidence that fruit and vegetables rich in antioxidants, particularly those known as polyphenolics (such as anthocyanins and isoflavones) may help protect against cancer, particularly in the gastro intestinal tract (GIT), including cancers of the mouth, esophagus and colon. The explosive growth of the functional food industry has led to many attempts to enhance the concentration of these health promoting compounds in specific foods. For that reason, optimizing health and performance through the diet has become one of the largest and most lucrative markets in the US. Polyphenols are very widely distribute in plants, and there are many different structures that occur naturally. One class of polyphenols, the flavonoids, exhibits more than 7000 structures present in nature, and close to 700 different structures among anthocyanins alone. Therefore, there is a need to increase our understanding of how the different molecular structures affect the functionality of those compounds. Flavonoids are believed to have potential health benefits, and these benefits are associated with their antioxidant and anti-inflammatory activity. In addition, some classes of flavonoids such as anthocyanins provide other unique characteristics to foods such as color. Anthocyanins are the pigments responsible for most of the re to purple to blue colors of plants. Our research goal is to increase the understanding of the role that polyphenolic compounds from fruits and vegetables play on health promotion. Of special interest are the flavonoids, including anthocyanins, proanthocyanins and isoflavones, and how horticultural and processing conditions affect their incidence in foods. Further knowledge is needed to investigate the absorption, excretion and biological activity of flavonoids as affected by their chemical structure. Our research will bring light on strategies to improve our diet. The information will be available to growers, to select plant materials with higher concentrations of compounds that can enhance health, or to determine growing conditions that favor the accumulation of certain compounds in the plant. This information will also help the food industry to use better ingredients and to understand how to preserve the integrity of the health promoting compounds during processing and storage. Our research will also provide information to the scientific community to better understand how to prevent chronic diseases and the role that some phytochemicals play in the health benefits of fruits and vegetables.
Animal Health Component
(N/A)
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
5022299200020%
7011499200020%
7015010200010%
5025010101020%
5025010309010%
7025010100010%
7025010200010%
Knowledge Area
702 - Requirements and Function of Nutrients and Other Food Components; 502 - New and Improved Food Products; 701 - Nutrient Composition of Food;

Subject Of Investigation
1499 - Vegetables, general/other; 2299 - Miscellaneous and new crops, general/other; 5010 - Food;

Field Of Science
3090 - Sensory science (human senses); 1010 - Nutrition and metabolism; 2000 - Chemistry; 1000 - Biochemistry and biophysics;
Goals / Objectives
Our overall goal is to increase the understanding of the role that non-volatile compounds from fruits and vegetables play on health promotion. Of special interest are the flavonoids, including anthocyanins, proanthocyanins and isoflavones, and how horticultural and processing conditions affect their incidence in foods. Further knowledge is needed to investigate the absorption, excretion and biological activity of flavonoids as affected by their chemical structure. Specific objectives 1. Increase knowledge on human exposure to flavonoids. 2. Evaluate anthocyanins as natural alternatives to the use of synthetic dyes and their incorporation into foods as value added ingredients 3. Evaluate the relationship between flavonoid chemical structure and functionality including stability in the food matrix, stability in the gastrointestinal tract, bioavailability and bioactivity. In the past we have worked on the transformation of anthocyanins in the intestines and colon. In the future we will generate more information of their transformations and interactions with the oral cavity an stomach. We expect to determine the contributions of anthocyanins and anthocyanin metabolites to their biological activity and help determine if they act better when combined or if isolated. Protein / flavonoid interactions are of great concern because of two main reasons: their impact on food quality an the interactions that occur in-vivo between dietary components and proteins in the body. We will provide clear alternatives to the use of synthetic colorants for the food industry. Our research has great potential for practical applications in addition to strong basic science components. Proanthocyanidins, catechins, and isoflavones will also be compounds investigated. Expected outputs include presentations of this work in at least 3-4 international meetings per year, publications in refereed journals, and invention disclosures. I also expect to bring external funding from government and the food industry to support our research efforts.
Project Methods
Flavonoid extraction Two approaches will be used: acetone and chloroform (great recoveries, and separates non-polar compounds such as carotenoids and chlorophyll from the more polar flavonoids), and methanol/water or acetonitrile/water mixtures (simple, rapid procedure). We have optimized methodologies to extract flavonoids from cells or animal tissues. Isolation of flavonoids Solid phase extraction will be used for purification or fractionation of different phenolics from plant mixtures, including C-18 cartridges, the most widely used for the analysis of phenolic compounds, and MCX cartridges, based on cation exchange and non-polar interactions. We developed a method to fractionate anthocyanins from other phenolics using MCX cartridges and a patent is pending. Chromatographic and spectroscopic characterization of flavonoids Spectral characteristics of flavonoids provide a great amount of information about their chemical structure. Quantitation of anthocyanins and total phenolics can be achieved by the use of colorimetric methods: pH-differential and Folin Ciocalteau method with aid of a spectrophotometer. HPLC chromatography constitutes a very powerful tool for characterization of flavonoids. Analytical and semi-preparative High Performance Liquid Chromatographers equipped with autosampler, photo-diode array, fluorescence, refractive index and mass detectors will be used. Color analysis Color will be evaluated by using a Hunter ColorQuest XE spectrophotometer with an integrating sphere with capability to measure color of both solid and liquid samples (reflectance and transmittance). Different types of illuminants to reproduce the typical lighting on a store shelve or during consumption. In vitro studies Cell lines derived from human carcinoma, or non-carcinogenic GIT tissues will be used to test the chemoprotective effects of different flavonoids. Parameters such as cell proliferation and GI50 (concentration required to inhibit 50% growth) will be monitored. Ex vivo studies Biological fluid or materials from animals or humans can be collected for later use in a variety of ex-vivo tests. Saliva from selected individuals will be collected and incubated with compounds of interest to determine their selected stability, degradation products and contribution of different saliva components. These studies also allow evaluating subject to subject variability as the source of the sample can be easily tracked. In-vivo studies In vivo studies will allow testing of the finding obtained in our in-vitro tests. In addition, in-vivo tests allow us to determine absorption into the plasma, and excretion into either urine or feces. Collaborations with the human nutrition department and the Medical School will allow access to animal and human samples as well.

Progress 11/01/12 to 09/30/17

Outputs
Target Audience:Our audience has included the following groups: - Our peers in the scientific community. We have presented our results at local, national and international scientific meetings, published articles in peer reviewed journals and edited booksthat will reach a large number of researchers in the USA and around the world. - Our stakeholders in the food industry. We have work with food companies transitioning to producing healthier products using more naturalingredients, as well as with ingredient manufacturers.We have also served on advisory committees for the food coloring industry, and given presentations specifically targetet for the foodindustry, at industry sponsored meetings. - Students in our field: the scope of our research has been presented to students in food science, training them about the importance of basic and applied research as well as the many possibilities to innovate through science and discoveries. - legistators: we have worked with other university experts, a color company and the USP to develop recommendations to improve the safety of color additives, affecting regulations. Main target has been students at our institution, however we have also offered workshops and classes abroad. - Local science school teachers. We have worked with the Ohio Department of Education helping train the teachers to use food science tools to deliver science content to their students. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?- We have trained students in our lab, our instiution, and abroad. This has been donethrough courses at Ohio State and at different institutions, including the Ohio State University, Mexico, and Peru. - We have trained teachers, throught workshops organized in collaboration with the Ohio Department of Education - We have trained other scientists, throught numerous presentations around the world. In addition, we offered a workshop on natural colorants for a team of researchers from the Philippines at Ohio State. - We continue to provide training to the food industry through conferences, visits, and presentation tailored to the food industry. This year we also presented at the Annual Meeting of the American Association of Color Manufacturers. We also published a white paper with recommendation for proactive safety measures that the colorant industry can take in order to prevent hazards. How have the results been disseminated to communities of interest?- Presentations at scientific meetings in 2017: in New Zealand, at the International Workshop on Anthocyanins, at the meeting of the AOAC in Arlington VA, at the Institute of Food Technologists meeting in Chicago, IL, at the Anthocyanin Meeting at the SISSA University in Italy, workshop on spectroscopy and colorimetry in Lima, Peru. - Publications in peer-reviewed journals, book chapters and books. - Visits to companies and other stakeholders. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Progress on Goal 1. Increase knowledge on human exposure to flavonoids. Human exposure to flavonoids is mainly atributed to the dietary intake of such compounds. We have strengthened the information regarding anthocyanins composition of different plant materials as well as in different food products. However, we have also demosntrated that human exposure to flavonoids can occur through the skin. Small and large polyphenolic structures are able to cross the skin barrier. The goal would be to take advantage of the health promoting properties of flavonoids not only throught the diet but also localized in the skin. Progress onGoal 2. Anthocyanins as Natural Alternatives to Synthetic Dyes. Over the last several years we showed many novel sources and applications of anthocyanins as food colorants. In particular, last year we placed more focus on anthocyanin-based blue colors, and on trying to reproduce the conditions found in nature that allow anthocyanins to express blue coloration. We have showen that the spatial configuration of the molecule is as important or even more than the type of units that make us the molecule. Anthocynins with sugars substitutions in positions 3 and 5 of the aglycone are able to produce blue colorations, either under neutral or alkaline coditions, or even under low acidic conditions when combined with metal ions.We have also made significant progress on working with pyranoanthocyanin formation. These pigments are derived from the condensation of anthocyanins with other reagents, such as pyruvic acid or some phenolic acids, and show higher stability to ascorbic acid or bisulfite bleaching. Formation of these pigments also depends on pH, temperature and chemical structure. Once again, spatial conformation played a more important role in this reaction than previously anticipated. Progress on Goal3. Evaluate the relationshipbetween flavonoid chemical structure and functionality including stability in the food matrix, andthe gastrointestinal tract.Over the last several years we have made great progress understanding the link between flavonoid chemical structure and function. We have worked on different areas of the digestive tract as well as on the skin, using both, in vitro and in vivo models. The last year of the project we were able to present and publish our major contributions on the impact of pH in the stomach for the absorption of anthocyanins. We demonstrated that anthocyanin absorption is improved under acidic conditions of the stomach, and that chemical structure plays an important role on transport and update by the gastric tissues. This work has been published in the International Journal of Molecular Science as well as in the Journal of Food Composition and Analyses. In addition, we have continued our work on anthocynin penetration into the skin. The major findings were published in the Journal of cosmetic science, showing that anthocyanins can penetrate the skin barrier and protect the skin from stressors such as UV-light, and oxidation, and can as well reduce formation of dark spots in the skin.

Publications

  • Type: Book Chapters Status: Published Year Published: 2017 Citation: Giusti, MM; Wrolstad, RE; Smith, DE. (2017). Calculation of CIE Color Specifications from Reflectance or Transmittance Spectra. (Suzanne Nielsen)(Eds.), Food Analysis Laboratory Manual (pp. 219-224). Location: Springer, New York.
  • Type: Book Chapters Status: Published Year Published: 2017 Citation: Schwartz, SJ; Cooperston, J; Cichon, M; Von Elbe, JH; Giusti, MM. (2017). Colorants. (S Damodaran, K Parkin)(Eds.), Fennemas Food chemistry (pp. 681-752). Location: CRC Press, Boca Raton.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Sigurdson, GT; Robbins, RJ; Collins, TM; Giusti, MM, "Effects of Hydroxycinnamic Acids on Blue Color Expression of Cyanidin Derivatives and their Metal Chelates". Food Chemistry. Vol. 234, 131-138. 2017. (Published) ( IF: 4.052 ) 10.1016/j.foodchem.2017.04.127.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Lao, F; Giusti, MM, "The Effect of Pigment Matrix, Temperature and Amount of Carrier on the Yield and Final Color Properties of Spray Dried Purple Corn (Zea mays L.) Cob Anthocyanin Powders". Food Chemistry. Vol. 227, 376-382. 2017.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Westfall, A; Giusti, MM, "Color Profiles and Stability of Lipstick Formulations Containing Anthocyanins Pigments: A Viable Alternative to Synthetic Colorants". Journal of Cosmetic Science. Vol. 68, no. 3: 1-12. 2017.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Castada, HZ; Mirondo, R; Sigurdson, GT; Giusti, MM and Barringer, S., "Deodorization of Garlic Odor by Spearmint, Peppermint, and Chocolate Mint Leaves and Rosmarinic Acid". LWT - Food Science and Technology. Vol. 84, 160-167. 2017.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Sigurdson, GT; Robbins, RJ; Collins, TM; Giusti, MM, "Spectral and Colorimetric Characteristics of Metal Chelates of Acylated Cyanidin Derivatives". Food Chemistry. Vol. 221, 1088-1095. 2017
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Lao, F; Sigurdson, GT; Giusti, MM, "Health Bene?ts of Purple Corn (Zea mays L.) Phenolic Compounds". Comprehensive Reviews in Food Science and Food Safety. Vol. 16, no. 2: 234-246. 2017.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Atnip, A; Sigurdson, GT; Bomser, J; Giusti, MM, "Time, Concentration, and pH-dependent Transport and Uptake of Anthocyanins in a Human Gastric Epithelial (NCI-N87) Cell Line". International Journal of Molecular Science. Vol. 18, no. 2: 446-457. 2017.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Sigurdson, GT; Tang, P; Giusti, MM, "Natural Colorants: Food Colorants from Natural Sources". Annual Review of Food Science and Technology. Vol. 8, 261-280. 2017
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Ayvaz, H; Bozdoganb, A; Giusti, MM; Mortasd, M; Tay, D; Manrique, I; Rodriguez-Saona, LE, "Improving the Screening of Potato Breeding Lines for Specific Nutritional Traits Using Portable Mid-Infrared (MIR) Spectroscopy and Multivariate Analysis". Food Chemistry. Vol. 211, 374-382. 2016
  • Type: Journal Articles Status: Under Review Year Published: 2018 Citation: Farr, JE; Srivastava, A; Machiraju, R; Giusti, MM, "ColorBySpectra: an application to automate conversion of spectral data to color spaces.". SoftwareX.
  • Type: Journal Articles Status: Under Review Year Published: 2018 Citation: Hernandez-Cumplido, J; Giusti, MM; Zhou, Y; Kyryczenko-Roth, V; Chen, YH; Rodriguez-Saona, C, "Testing the 'plant domestication-reduced defense' hypothesis in blueberries: The role of herbivore identity". Arthropod-Plant Interactions.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Simon, J; Decker, E; Ferruzzi, M; Giusti, MM; Mejia, C; Talcott, S., "Establishing Standards on Colors from Natural Sources". Journal of Food Science, Vol 88 (11): 2539-2553.


Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Our audience has included the following groups: - the scientific community. We have presented at scientific meetings, published articles in peer reviewed journals and worked on book chapter that will reach a large number of our peers in the USA and around the world. - the food industry. We work with different companies that produce food products, and are interested in our work as it applies to their bussinesses. We have also worked on advisory committees for the food coloring industry, and given industry focused presentations. - students in our field: the scope of our research has been presented to students in food science, training them about the importance of basic and applied research as well as the many possibilities to innovate through science and discoveries. - legistators: we have worked with other university experts, a color company and the USP to develop recommendations to improve the safety of color additives, affecting regulations. - local science school teachers. We have worked with the Ohio Department of Education helping train the teachers to use food science tools to deliver science content to their students. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During 2016 we have had the opportunity to train graduate students, undergraduates and high school students on basic and applied concepts of food science, food quality and the impact of food on human health. We have tought methologies and procedures to evaluate polyphenols, food quality, color communication and research procedures in our laboratories and laboratory meetings. We have also helped on the training of other professionals in our discipline through participation at different conferences by sharing our experiencesin different projects and workshops for professional development while learning from other researchers in similar areas. During 2015 I participated in an Expert Panel to evaluate the safety of colorants from natural sources. This expert committee came up with recommendations to the food colorant industry to improve safety monitoring of our food colorants. We have developed a white paper that is under evaluation for publication. We hope that the recommendations from the committee will help shape regulations in the field. I was invited to present the recommendation from the panel at the annual meeting of the International Association of Color Manufacturers. We hope this white paper can become a tool for training professional in the field of natural colorants. In addition, we worked together with the Ohio Department of Education to train high school science teachers on the use of food examples and matrices to teach important science concepts including chemistry, biology, processing, safety. We offered a 2 hour workshop during the Hands-On-Training offered by ODE, and followed up with a day long workshop Science and Technology of Food Course Professional Development. How have the results been disseminated to communities of interest?The results of our work has been disseminated to our different audiences in a number of different ways. - 2 book chapter was published during 2016and 2 more has been accepted for publication. - 6 peer-reviewed manuscripts were published and 3more have beenaccepted for publication. Other 3 manuscripts have beensubmitted for publication and are awaiting review. - Our work has also been communicated to peers through presentations at professional meetings, national and international. Our work was presentedin 2016 at the Institute of Food Technologists in Chicago, at regional ACS meeting in Grand Rapids, at a Colorants meeting in Trieste, Italy, at the Pigments in Foods in Romania, at the Workshop on Bioactive Compounds & Food Quality in Brazil, and at the Universidad Nacional Agraria, in Peru. - I also disseminated the work through courses on Colorants in Universidad Agraria, Peru, and a course on the Health Benefits of Polyphenols at the Universidad Nacional de Bogota, Colombia. What do you plan to do during the next reporting period to accomplish the goals?Next year we plan to continue to publish a lot of the work onanthocyanin transport through the gastric tissues, particularly as it pertains to inflamation, as well as our work with anthocyanin penetration through the skin. We will continue to pursue publication of the work on anthocyanin absorption throught the skin, that was delayed by trying to identify the right journal, as they do not fit the scope of our usual journals on food science. We plan to target high impact factor peer-reviewed journals. We will also continue exploring the anthocyanin stabilization through copigmentation with other phenolic acids, flavonols, and flavonoid mixtures. We will explore how copigments may enhance anthocyanin color and which one can positively affect stability. Our preliminary data suggests that compounds enhancing the color the most may not be the ones that result on the best stability of the color. We will also continue exploring pyranoanthocyaning, and plan to explore the condensation reaction between anthocyanins, phenols and ascorbic acid throught the use of NMR. We have also received a grant from the Ohio Soybean Association to further explore the copigmentation of anthocyanins and isoflavones, the estrogenic compounds in soy. We beleive that the position of sugars attached to the anthocyanin chromophore may play a more important role that previously reported. We also plan to continue disseminating the results of our work at different meetings of international impact. In 2017 I plan to attend the International Workshop on Anthocyanins, in New Zeland. I will make sure to present our work at the meeting and to facilitate the exchange of information with other laboratories around the world that work in similar areas.

Impacts
What was accomplished under these goals? GOAL 1. Increase knowledge on human exposure to flavonoids. Last year we completed a set of experiments to determine the impact of pH and anthocyanin structure on its absorption through the gastric tissues. This year we have continued with the data analyses and preparation of the manuscript for publication. Acidic environment of the stomach seems to favor anthocyanin transport through the gastric cells. And this absorption seems to be particularly efficient when the concentration of anthocyanins is low. This suggests that the consumption of the anthocyanins from the regular diet, usually present in low amounts, may help optimize their uptake. In addition, we evaluated the use of anthocyanins as bioactive colorants in lipstick formulations. Successful formulations showed similar shades to commercial lipsticks, high color stability to accelerated environmental testing, high free radical scavenging antioxidant ability, UV protection, and inhibition of melanin production. A study completed in 2015 using pig skin model systems was followed by a clinical trial where we were able to prove that anthocyanins can penetrate the human skin in concentrations high enought to be of biological relevance. We also obtained clear evidence of transversal difusion of the pigments. This suggests that exposure to anthocyanins through the skin can be also achieved through topical application, and that the tissues adjacent to the place of application may also benefit from their health protecting properties. Goal 2. We continue to evalutethe use of anthocyanins as natural alternatives to the use of synthetic. This year we continued to trying to reproduce the mechanisms used by flower to produce blue colors using anthocyanin pigments. In flowers, anthocyanins and metal ions are cobined in particualr ratios and environments. In foods, we can reproduce some of those conditions to produce similar hues. Acylated anthocyanins were more effective producing blue colors than other non-acylated anthocyanins. However, this year we discovered that sugars, previously believed to have little impact on color expression of anthocyanins, may play a critical role on the expression of blue colors when they are attached at particular positions of the cromophore. We have also continued our explorations of the interactions between anthocyanins and ascorbic acid. Ascorbic acid is an important vitamin commonly added to foods that degrades quickly and accelerate the degradation of anthocyanins. Bleaching of anthocyanins in the presence of ascorbic acid is believed to occur through condensation at position C4 of the anthocyanin. To better understand the mechanism of interaction, we have chemically sythesized pyranoanthocyanins derived from simple anthocyanin pigments. Pyranoanthocyanins have the C4 position unavailable for chemical reactions, and if the hypothesis is right, then pyranoanthcoyanis should be resistant to the bleaching in the presence of ascorbic acid. Our preliminare results suggest that the C4 position is a major site for the reaction but not the only site. In addition, we have initiated a new project to evaluate copigmentation. We are evaluating a large number of phenolic acids and flavonoid mixtures for their ability of enhance anthocyanin color. GOAL 3. Evaluate the relationship between flavonoid chemical structure and functionality including stability in the food matrix, stability in the gastrointestinal tract, bioavailability and bioactivity. We have new evidence of the interaction between anthocyanins and proteins. These interactions occur in foods, and could potentially occur also in the body. We believe that in foods anthocyanin-protein interactions may enhance the stability of the colors. However, depending on the environment, anthocyanin-protein complexes can become insoluble and precipitate. We will continue to explore these interactions as they affect the food quality and may give light to help elucidate anthocyanin-protein interaction in vivo.

Publications

  • Type: Book Chapters Status: Published Year Published: 2016 Citation: Rodriguez-Saona, LE; Giusti, MM; Shotts, M, "Advances in infrared spectroscopy for food authenticity testing". In "Advances in Food Authenticity Testing". Edited by:Downey, G. Duxford: Elsevier, Woodhead Publishing. 71-107. 2016.
  • Type: Book Chapters Status: Published Year Published: 2016 Citation: Rodriguez-Saona, LE; Plans Pujolras, M; Giusti, MM, "Targeted and Non-Targeted Analysis". In "Analytical Separation Science: Sample preparation, method validation, and applications". Vol. 5 Edited by:Pino, V; Anderson, JL. New York: Wiley & Sons, Inc.. 1401-1436. 2016.
  • Type: Book Chapters Status: Accepted Year Published: 2017 Citation: Schwartz, SJ; Cooperston, J; Cichon, M; Von Elbe, JH; Giusti, MM, "Colorants". In "Fennemas Food chemistry". Edited by:S Damodaran, K Parkin, and OR Fennema.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Ayvaz, H; Bozdoganb, A; Giusti, MM; Mortasd, M; Tay, D; Manrique, I; Rodriguez-Saona, LE, "Improving the Screening of Potato Breeding Lines for Specific Nutritional Traits Using Portable Mid-Infrared (MIR) Spectroscopy and Multivariate Analysis". Food Chemistry. Vol. 2016, 374-382. 2016.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Sigurdson, GT; Robbins, RJ; Collins, TM; Giusti, MM, "Evaluating the Role of Metal Ions in the Bathochromic and Hyperchromic Responses of Cyanidin Derivatives in Acidic and Alkaline pH". Food Chemistry. Vol. 208, 26-34. 2016.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Dhakal, S; Giusti, MM; Balasubramaniam, VM, "Effect of high pressure processing on dispersive and aggregative properties of almond milk". Journal of The Science of Food and Agriculture. Vol. 96, 3821-3830. 2016
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Zhou, Y; Giusti, MM; Parker, J; Salamanca, J; Rodriguez-Saona, C., "Frugivory by Brown Marmorated Stink Bug (Hemiptera: Pentatomidae) Alters Blueberry Fruit Chemistry and Preference by Conspecifics". Environmental Entomology. Vol. 0, 1-8. 2016.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Lao, F; Giusti, MM, "Quantification of Purple Corn (Zea Mays L.) Anthocyanins Using Spectrophotometric and HPLC Approaches: Method Comparison and Correlation". Food Analytical Methods. Vol. 9, 1367 - 1380. 2016.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Ahmadiani, N; Robbins, R; Collins, T; Giusti, MM, "Molar Absorptivity (?) and Spectral Characteristics of Cyanidin-Based Anthocyanins from Red Cabbage". Journal of Agricultural and Food Chemistry. Vol. 197, 900-906. 2016
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2017 Citation: Sigurdson, GT; Tang, P; Giusti, MM, "Natural Colorants: Food Colorants from Natural Sources". Annual Review of Food Science and Technology.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2017 Citation: Sigurdson, GT; Robbins, RJ; Collins, TM; Giusti, MM, "Spectral and Colorimetric Characteristics of Metal Chelates of Acylated Cyanidin Derivatives". Food Chemistry.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2016 Citation: Giusti MM. Anthocyanin Pigments Absorption, Degradation and Bioactivity in the Gastrointestinal Tract and the Skin. Workshop on Bioactive Compounds & Food Quality. Santa Maria, Brazil. October, 2016.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2016 Citation: Giusti, MM. Anthocyanin chemical structure affects their stability, bioavailability, and functionality in the human body. 8th International Congress on Pigments in Food. Romania. June, 2016.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2016 Citation: Giusti, MM. The Colorful World of Anthocyanins: Learning From Nature 091 Natural Colors: Challenges and Chances in a Global Perspective. Institute of Food Technologists Annual Meeting and Food Expo. Chicago, IL, June, 2016.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2016 Citation: Giusti, MM; Simon, JE; Decker, EA; Ferruzzi, MG; Mejia, CD; Talcott, ST. Establishing Standards on Colors from Natural Sources: Recommendations from an Expert Panel. International Association of Color Manufacturer, Natural Colorants Meeting. Alexandria, VA, November, 2016.


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

Outputs
Target Audience:Our audience has included the following groups: - the scientific community. We have presented at scientific meetings, and published articles in peer reviewed journals that reach a large number of our peers in the USA and around the world. - the food industry. We workwith different companies that produce food products, and are interested in our work as it applies to their bussinesses. - students in our field: the scope of our research has been presented to students in food science, training them about the importance of basic and applied research as well as the many possibilities to innovate through science and discoveries. - legistators: we have worked with other university experts, a color company and the USP to develop recommendations to improve the safety of color additives, affecting regulations. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Graduate students, and high school students have been trained on methodologies and procedures to evaluate polyphenols, food quality, color communication and research procedures in our laboratories and laboratory meetings. Participation at different conferences give opportunities to all students and scientists in the projects for professional development and learning from other researchers in similar areas. We hosted during 2015 two visiting scholars, one student from Brazil and a researcher from Spain. Both of them were trained on laboratory procedues and techniques. I also participated in an Expert Panel to evaluate the safety of colorants from natural sources. This expert committee has come up with recommendations to the food colorant industry to improve safety monitoring of our food colorants. We hope that the white paper generated by the committee will help shape regulations in the field. In addition, we worked together with the department of education to train high school science teachers on the use of food examples and matrices to teach important science concepts including chemistry, biology, processing, safety. This was done through a day long workshopScience and Technology of Food Course Professional Development. How have the results been disseminated to communities of interest?The results of our work has been disseminated to our different audiences in a number of different ways. - 4peer-reviewed publications were published 2 more accepted for publication and an additional 5 manuscripts were submitted for publication in scientifichigh impact peer-reviewed journals. - 1 book chapter was published during 2015 and 2 more were submitted and accepted for publication. - Our work has also been communicated to peers through presentations at professional meetings, national and international. Our work has been presented in 2015 at the Americal Chemical Society annual meeting in Boston, the Institute of Food Technologists in Chicago, a regional ACS meeting in Grand Rapids, at a Colorants meeting in Trieste, Italy, and at the Universidad Nacional Agraria, in Peru. - We hosted two visiting scholars, one student from Brazil and a researcher from Spain. Both of them were trained on laboratory procedues and techniques. They have returned to their respective institutions to further disseminate their newly acquired knowledge. What do you plan to do during the next reporting period to accomplish the goals?Next year we plan to publish a large portion of our work onanthocyanin transport throughin the gastric tissues, particularly as it pertains to inflamation, as well as our work with anthocyanin penetration through the skin. We plan to target high impact factor peer-reviewed journals. We will also continue exploring the anthocyanin stabilization through copigmentation with other phenolic compounds, particularly in the presence of ascorbic acid, but also under other conditions that favor anthocyanin degradation. One new aspect of investigation will be the copigmentation of anthocyanins and isoflavones, the estrogenic compounds in soy.We wil also explore further the intramolecular copigmentation of different acylated cyanidin derivatives. We beleive that the position of the acylating group may be critical for the tinctorial strength and the color caracteristics of the pigments and plan to explore this issue further. We also plan to continue disseminating the results of our work at different meetings of international impact. In 2016 I will be a co-organizer and member of the scienfic commitee of the Pigments in Food Congress, that will take place in Romania. I will make sure to present our work at the meeting and to facilitate the exchange of information with other laboratories around the world that work in similar areas.

Impacts
What was accomplished under these goals? GOAL 1. Increase knowledge on human exposure to flavonoids. This year we completed a set of experiments to determine the impact of pH and anthocyanin structure on its absorption through the gastric tissues. We determined that the acidic environment of the stomach favors anthocyanin trasnport through the gastric cells. Transport of anthocyanins was also observed at pH close to natural, but to a lower extent, with the lowest absorption happening under pH close to 5. The rate of anthocyanin transport increased over time, suggesting that the transport was not by passive diffusion alone. Anthocyanin structure also affected the transport through the gastric tissue, with anthocyanin-arabinosides being bettertransported than anthocyaningalactosides, and cyanidin transported more efficiently thanmalvidin.Finally, anthocyanins showed anti-inflamatory effects byinhibiting the increased IL-8 production in response to the IL-1β induced inflammation of NCI-N87 cells. This suggests a potential anti-inflammatory activity of anthocyanins in the stomach, at least with respect to this acute inflammation model. In addition, we evaluated the use of anthocyanins as bioactive colorants in lipstick formulations.Successful formulations showed similar shades to commercial lipsticks, high colorstability to accelerated environmental testing, high free radical scavenging antioxidantability, UV protection, and inhibition of melanin production. In a pig skin model system as well as in a clinical trial, we demonstrated that anthocyanins can penetrate different layers of the skin, to reach areas of the skin where the UV-protection and anti-aging effects could be relevant. Therefore, exposure to anthocyanins through the skin can be also achieved through topical application. Goal 2. We evaluted the use of anthocyanins as natural alternatives to the use of synthetic. Our search for natural alternatives to synthetic colorants continues. Natural colorants behave in different ways depending on their chemical structure and their interaction with the matrix. We continue working to imitate the chemical conditions produced in flowers that result on blue coloration in the plant, in order to reproduce similar colors in food matrices. Ph, metal complexation and copigmentation with other polyphenols are some of the factors that have been shown to play a major role in the production of anthocyanin-based blue colors. We are evaluating now the role of acylating groups attached to the anthocyanin molecules for their ability to intereact with the metal ions and produce blue colors in low acidity environments. We have also continued our explorations of the interactions between anthocyanins and ascorbic acid. Ascorbic acid is an important vitamin commonly added to foods that degrades quickly and accelerate the degradation of anthocyanins. We have shown that addition of catechins can result on a slow down of the degradation rate of anthocyanins in the presence of ascorbic acid. We are evaluating the optimum ratios of the different ingredients to maximize vitamin activity and color stability, and exploring the underlaying mechanisms of their interactions. GOAL3. Evaluate the relationship between flavonoid chemical structure and functionality including stability in the food matrix, stability in the gastrointestinal tract, bioavailability and bioactivity. Anthocyanin structure also affected the transport through the gastric tissue, with anthocyanin-arabinosides being bettertransported than anthocyaningalactosides, and cyanidin transported more efficiently thanmalvidin.

Publications

  • Type: Book Chapters Status: Awaiting Publication Year Published: 2016 Citation: Schwartz, SJ; Cooperston, J; Cichon, M; Von Elbe, JH; Giusti, MM, "Colorants". In "Fennemas Food chemistry". Edited by:S Damodaran, K Parkin, and OR Fennema.
  • Type: Book Chapters Status: Awaiting Publication Year Published: 2016 Citation: Rodriguez-Saona, LE; Shotts, M; Giusti, MM, "Advances in infrared spectroscopy for food authenticity testing". In "Advances in Food Authenticity Testing". Edited by:Downey, G.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Bochi, VC; Godoy, HT; Giusti, MM, "Anthocyanin and Other Phenolic Compounds in Ceylon gooseberry (Dovyalis hebecarpa) fruits". Food Chemisty. Vol. 176, 234-243. 2015
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Jing, P; Qian, B; Zhao, S; Qi, X; Ye, L; Giusti, MM; Wang, X, "Effect of glycosylation patterns of Chinese eggplant anthocyanins and other derivatives on antioxidant effectiveness in human colon cell lines". Food Chemistry. Vol. 172, 183-189. 2015.
  • Type: Book Chapters Status: Published Year Published: 2015 Citation: Giusti, MM; Ahmadiani, N; Tang, P.; Ottinger, MA., "Isoflavone and Flavonoid Supplemented Eggs in Health". In "Handbook of Egg in Promotion of Health". Edited by:Watson, RR; De Meester, F.. Wageningen: Wageningen Academic Publishers. 333-364. 2015.
  • Type: Book Chapters Status: Published Year Published: 2016 Citation: Rodriguez-Saona, LE; Plans Pujolras, M; Giusti, MM, "Targeted and Non-Targeted Analysis". In "Analytical Separation Science: Sample preparation, method validation, and applications". Edited by:Pino, V; Anderson, JL. 2016
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Nangle, EJ; Gardner, DS. Metzger, JD; Rodriguez-Saona, LE; Giusti, MM; Dannebergerb, TK;Petrella, DP, "Pigment changes in cool-season turfgrasses in response to ultraviolet-B light irradiance". Agronomy Journal. Vol. 107, no. 1: 41-50. 2015.
  • Type: Journal Articles Status: Accepted Year Published: 2016 Citation: Ahmadiani, N; Robbins, R; Collins, T; Giusti, MM, "Molar Absorptivity (?) and Spectral Characteristics of Cyanidin-Based Anthocyanins from Red Cabbage". Journal of Agricultural and Food Chemistry.
  • Type: Journal Articles Status: Accepted Year Published: 2016 Citation: Lao, F; Giusti, MM, "Quantification of Purple Corn (Zea Mays L.) Anthocyanins Using Spectrophotometric and HPLC Approaches: Method Comparison and Correlation". Food Analytical Methods.
  • Type: Journal Articles Status: Under Review Year Published: 2016 Citation: Dhakal, S; Giusti, MM; Balasubramaniam, VM, "Effect of high pressure processing on dispersive and aggregative properties of almond milk". Journal of The Science of Food and Agriculture.
  • Type: Journal Articles Status: Under Review Year Published: 2016 Citation: Simon, J; Decker, E; Ferruzzi, M; Giusti, MM; Mejia, C; Talcott, S., "Establishing Standards on Colors from Natural Sources: An Expert Committee Report" Concise Reviews and Hypotheses in Food Science, IFT.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Wallace, TC and Giusti, MM, "Anthocyanins". Review of NUTRITION INFORMATION BRIEFS on Anthocyanins, by State of research knowledge. Advances in Nutrition. Vol. 6, 620-622. (2015)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Ahmadiani, N; Giusti, MM; Robbins, R; Collins, M. "Effect of Sinapic Acid Acylation on the Spectral Characteristics and Stability of Cyanidin-Based Anthocyanins From Red Cabbage". IFT Annual Meeting Abstracts. Vol. 098-047, Chicago: Institute of Food Technologists.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Chen, LA; Giusti, MM. "Effect of Catechin Copigmentation and Ascorbic Acid Fortification on Color Stability of Black Carrot and Chokeberry Anthocyanins Under Heat Treatment". IFT Annual Meeting Abstracts. Vol. 046-048, Chicago: Institute of Food Technologists.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Sigurdson, GT; Robbins, R; Collins, T; Giusti, MM. "Evaluating the Spectral Characteristics and Color of Acylated Anthocyanins Induced by Metal Chelation". IFT Annual Meeting Abstracts. Vol. 098-068, Chicago: Institute of Food Technologists.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Giusti, MM. "The multiple colors of anthocyanins: Learning from nature". JGLCRM ACS Abstracts. Grand Rapids: ACS JGLCRM.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Guo, J; Farr J; Giusti, MM; Kaletunc, G. "Comparison of Diffusion Characteristics of Dry and Wet Hydrogel Particles by Fluorescence". IFT Annual Meeting Abstracts. Vol. 031-024, Chicago: Institute of Food Technologists.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Lao, F; Rodriguez-Saona, LE; Giusti, MM. "Rapid Quantitation of Protein Content in an Anthocyanin-Rich System Using Fourier Transformed Mid Infrared Spectroscopy". IFT Annual Meeting Abstracts. Vol. 098-129, Chicago: Institute of Food Technologists.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Tang, P; Giusti, MM. "Acylation of Anthocyanin Influences Its Co-Pigmentation with Isoflavone". IFT Annual Meeting Abstracts. Vol. 098-004, Chicago: Institute of Food Technologists.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Westfall, A; Giusti, MM. "Evaluation of the Efficacy of Anthocyanins as Biologically Active Ingredients in Cosmetic Formulations". IFT Annual Meeting Abstracts. Vol. 067-046, Chicago: Institute of Food Technologists.
  • Type: Other Status: Published Year Published: 2015 Citation: Giusti, MM, Presenter. "Intramolecular and intermolecular copigmentation of acylated cyaniding derivatives from Red Cabbage for the production of blue colors." Presented at MARS Anthocyanin Research Meeting at the Scuola Internazionale Superiore di Studi Avanzati (SISSA). Trieste, Italy.


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

Outputs
Target Audience: Our audience has included the following groups: - the scientific community. We have presented at scientific meetings, and published articles in peer reviewed journals that reach a large number of our peers in the USA and around the world. - the food industry. We have worked with different companies that produce food products, and are interested in our work as it applies to their bussinesses. - students in our field: the scope of our research has been presented to students in food science, training them about the importance of basic and applied research as well as the many possibilities to innovate through science and discoveries. - legistators: the highlights of our research were presented to legislators in Ohio and around the country, sent by OARDC - Alumi from our university. Our work was highlighted in the magazine that is distributed to all OSU alumni, showing the relevance of our work to science and to the food industry. Changes/Problems: No major changes have been made. What opportunities for training and professional development has the project provided? Working in this project we have trained a team of graduate students, working directly in the project. We also used examples taken from this study in the classroom setting where we teach undergraduate students, and use the phenolic research to illustrate important concepts such as antioxidant activity, chromatography and colorimetry. In addition, we have had opportunity to train other professionals in our field through seminars, invited lectures and other presentations. How have the results been disseminated to communities of interest? The results have been disseminated in the following modalities: - a book was published on the health benefits of anthocyanins - 8 peer-reviewed publications were generated and published in high impact peer-reviewed journals - we presented our work at the Annual Meeting of the Institute of Food Technologists in New Orleans, at the American Chemical Society Regional Meeting in Pensilvania, at the 2nd International Congress on Food Technology, in Turkey, and at many other smaller meetings. - in addition, our work has been disseminated to the community through OSU bulletins and magazines highlighting the innovative character of our work. What do you plan to do during the next reporting period to accomplish the goals? We will continue to explore the effects of anthocyanins in the gastric tissues, particularly as it pertains to inflamation. We also plan to publish this work in high impact factor peer-reviewed journals. We will also continue exploring the anthocyanin stabilization through copigmentation with other phenolic compounds, particularly in the presence of ascorbic acid, but also under other conditions that favor anthocyanin degradation. We wil also explore further the intramolecular copigmentation of different acylated cyanidin derivatives. We beleive that the position of the acylating group may be critical for the tinctorial strength and the color caracteristics of the pigments and plan to explore this issue further. We also plan to continue disseminating the results of our work at different meetings of international impact.

Impacts
What was accomplished under these goals? Goal 1. In order to increase our understanding of human exposure to flavonoids we focused this year on the evaluation of anthocyanin absorption in the stomach. We used a new gastric cell line that can be grown under acidic enironment typical of the stomach in vivo. This is very remarkable as most cell lines can only be grown in vitro under neutral pH, and these conditions are not representative of the stomach environment and are detrimental to anthocyanins, the bioactive compounds we are evaluating. For this, we first had to demonstrate the feasibility of using the cell line. We proved that the cells can grow at pH as low as 3.0 producing a monolayer of good integrity. Second, we tested the absorption and uptake of anthocyanins by these gastric cells, under different pH growing conditions. We were able to prove that the absorption and uptake of anthocyanins can happen in these gastric cells, and it is dependant on pH, and anthocyanin chemical structure.The absorption and uptake was also dose dependant and increased over time of exposure. Goal 2. We evaluted the use of anthocyanins as natural alternatives to the use of synthetic blue colorants. This year we focused our attention into the interaction of anthocyanins from cabbage (acylated cyanidin derivatives) and eggplant (acylated delphinidin derivatives) with metals. These are the typical structures that produce the beautiful blue colors in flowers and other blue plants. We have been able to produce intense blue colorations that resemble those found in nature at pH levels between 5 and 8. An issue that is still to be resolved is the limited solubility of the anthocyanin metal complexes. Interactions of these complexes with other phenolics will be explored next in search for improved solubility. In addition we also started exploring the stabilization of anthocyanin colors by the use of catechins. Ascorbic acid is frequently added to foods because of their nutritional value, but they can act as prooxidants in the presence of compounds such as anthocyanins. We are exploring if catechins from tea can be used to decrease the degradation of antocyanins in the presence of ascorbic acid. Our preliminary results are very promising, and the work will continue. Another mechanism to stabilize anthocyanin color is through microencapsulation. We have determined that anthocyanins can be microencapsulate in a cold setting gel that retain anthocyanins under acidic conditions but can release them under neutral conditions such as those found in the intestines, potentially improcing the bioavailability of these compounds in the intestines. 3. The relationship between anthocyanin chemical structure and functionality was explored in the gastric cells system. We have proven that both, the type of anthocyanidin and the type of glycosylation can affect the transport of anthocyanins through gastric cells, as well as the retention of the compounds inside the cells.

Publications

  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Sigurdson, GR; Giusti, MM. "Bathochromic and Hyperchromic Effects of Aluminum Salt Complexation by Anthocyanins from Edible Sources for Blue Color Development." J. Agri Food Chem. Vol. 62, no. 29. (Feb 2014): 69556965.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2014 Citation: Jing, P; Qian, B; Zhao, S; et al. "Effect of glycosylation patterns of Chinese eggplant anthocyanins and other derivatives on antioxidant effectiveness in human colon cell lines." Food Chemistry.
  • Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Nangle, E; Rodriguez-Saona, LE; Giusti, MM; et al. "Pigment changes in cool season turfgrasses in response to ultraviolet-B light irradiance." Agronomy Journal.
  • Type: Books Status: Published Year Published: 2013 Citation: Wallace, Taylor C; Giusti, M. Monica, ed(s). Anthocyanins in health and disease. Boca Raton: CRC Press, Taylor and Francis group, 2013.
  • Type: Book Chapters Status: Published Year Published: 2013 Citation: Pu, J. and Giusti, MM. "Analysis of Anthocyanins in Biological Samples." In Anthocyanins in Health and Disease.. Edited by Wallace, TC. and Giusti, MM. Boca Raton: CRC Press, 2013.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Bochi, VC, Barcia, M; Rodrigues, D; Giusti, MM; et al. "Polyphenol extraction optimization from Ceylon gooseberry (Dovyalis hebecarpa)." Food Chemisty. Vol. 164, (Dec 2014): 347-354.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Ahmadiani, N; Robbins, R; Collins, T; et al. "Anthocyanins contents, profiles and color characteristics of red cabbage extracts from different cultivars and maturity stages." Journal of Agricultural and Food Chemistry. Vol. 62, no. 30. (Jul 2014): 75247531.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Hummer K, Durst R, Zee F, Atnip A, Giusti MM. "Phytochemicals in fruits of Hawaiian wild cranberry relatives." J Sci Food Agric. Vol. 94, no. 8. (Jun 2014): 15301536.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Giusti, MM; Polit, M; Ayvaz, H; et al. "Characterization and quantitation of anthocyanins and other phenolics in native andean potatoes." J Agr Food Chem. Vol. 62, no. 19. (Apr 2014): 44084416.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Kamonpatana, K; Failla, M; Kumar, P; et al. "Anthocyanin structure determines susceptibility to microbial degradation and bioavailability to the buccal mucosa." Journal of Agricultural and Food Chemistry. Vol. 62, no. 29. (Mar 2014): 69036910.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Snyder, AB; Sweeney, CF; Rodriguez-Saona, LE; et al. "Rapid authentication of concord juice concentration in a grape juice blend using Fourier-Transform infrared spectroscopy and chemometric analysis." Food Chemistry. Vol. 147, (Mar 2014): 295-301.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Paudel, Liladhar; Wyzgoski, F; Giusti, MM; et al. "NMR-based Metabolomic Investigation of Bioactivity of Chemical Constituents in Black Raspberry (Rubus occidentalis L.) Fruit Extracts." Journal of Agricultural and Food Chemistry. Vol. 62, (Feb 2014): 1989-1998.


Progress 01/01/13 to 09/30/13

Outputs
Target Audience: Our work is targeted to help the food industry find practical natural alternatives to the use of synthetic dyes. The scientific community is the main target audience for our scientific publications in the chemistry and biological activity of flavonoids. In addition, the public in general is an additional target audience as we try to provide tools for consumers to make more informed decisions. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Interaction with the industry: We have worked closely with the industry to advance the knowledge in the field and to find practical solutions for practical problems. We have worked with a number of food processing companies to help them understand the effects of processing on the integrity of flavonoids, in terms of both their functionality in the food as well as their bioavailability. Companies served include large US food processors, small local food processors, and companies in other cuntries, particularly in Peru, where we have strong lines of collaboration. Training Students An important part of our work is conducted with students, that I train on the skills needed. This year I have had 10 students working under my supervision, who I have trained through classes as well as in my laboratory. I also train a larger number of students through the class Food Colors and Pigments were we train graduate students on the different aspects of color and natural pigments, structures and functionality. Students in the class are trained on the chemistry and analytical techniques for flavonoid analyses. How have the results been disseminated to communities of interest? Our results have been presented in both, national and international conferences and workshops. This year, I was an invied speaker at two major international conferences: Giusti, MM, Presenter. "Anthocyanin Activity and Metabolism Starts on the Oral Cavity." Presented at International Workshop on Anthocyanins, Porto, Portugal. (Sep 2013) Giusti, M.M., Presenter. "Can we make food healthier and appealing at the same time?." Presented at ACS Annual Meeting, New Orleans (Mar 2013) In addition, we have disseminated our results at other conferences: Allison Atnip, Joshua Bomser, M. Monica Giusti,2013. Characterization of the Stability and Uptake of Anthocyanins by Gastric Cells Using a Novel Cell Model. Comp. 14-05. Poster presented at IFT Annual Meeting, July 13 – 16, 2013, Chicago Illinois. Santosh Dhakal, M. Monica Giusti, Shridhar Sathe, V.M. Balasubramaniam, 2013. Influence of High Pressure Processing on Physicochemical Properties of Almond Milk. COMP13-05. Poster presented at IFT Annual Meeting, July 13 – 16, 2013, Chicago Illinois. Neda Ahmadiani, M. Monica Giusti, 2013. Relationship Between Cultivar and Maturity with Color Formation in Seven Different Red Cabbage Anthocyanin Extracts in Acidic and Neutral pH Values. 071-03. Poster presented at IFT Annual Meeting, July 13 – 16, 2013, Chicago Illinois. Fei Lao, M. Monica Giusti, 2013. Efficiently Extract Anthocyanin from Purple Corn (Zea mayL.) Cob Using Food-Friendly Solvents. 157-01. . Poster presented at IFT Annual Meeting, July 13 – 16, 2013, Chicago Illinois. Kom Kamonpatana, Purnima Kumar, M. Monica Giusti, Mark Failla, 2013. Anthocyanin Extracts Inhibit IL-8 Secretion by IL-1β-activated Oral Epithelial Cells. 312-36. Poster presented at IFT Annual Meeting, July 13 – 16, 2013, Chicago Illinois. Jing, P; Giusti, MM. 2013. Anthocyanin Structure-function relationships and their impact on cancer cells. Oral presentation. ACS annual meeting. September, 8-12. Indianapolis. Sigurdson, G; Giusti, MM. 2013. Developing blue food colors from anthocyanin-metal complexation.Presented at International Workshop on Anthocyanins, Porto, Portugal. (Sep 2013) What do you plan to do during the next reporting period to accomplish the goals? We will continue working on all objectives simultaneously. For objective 1, we will explore the bioavailability of anthocyanins in the stomach, and we plan to initiate a study to explore the impact of anthocyanins on appetite. For our second objective we will continue the work on anhocyanins as sources of blue colors. We will also continue exploring the impact of chemical structure on gel encapsulation of anthocyanins. We also want to explore the stability of color and pigment of gels produced with different anthocyanin chemical strcutures. For the last objective, we will continue to investigate interaction of anthocyanins with a variety of materials. We will have a special emphasis on the interactions of anthocyanins with other flavonoids, with proteins and with other phenolics. In addition we will continue investigating anthocyanins in the gastric environment.

Impacts
What was accomplished under these goals? 1. Progress has been made in this field. We were able to demonstrate that anthocyanin degradation starts in the oral cavity. Anthocyanins are susceptible to transformation and degradation in saliva, and the chemical sructure of the anthocyanins has an impact on its stability and ability to be taken up by the epithilium and mucus cells of the oral cavity. 2. Great progress and groundbreaking results were obtained in this area. This period we focused a lot of efforts on the production of anthocyanin-based blue colors. We determined that red cabbage anthocyanins have the righ composition to produce blue colors under alkaline pH that best resemble the color characteristics of the synthetic blue FD&C Blue 2. Even more, we determined that some of those pigments are more effective than others. All pigments in red cabbage are acylated cyanidin derivatives but the position and number of acylations vary among the close to 15 different pigments present. We have been able to differentiate which of those chemical structures produce better hues and higher color stability when placed under alkaline conditions. The next steps will be to investigate the possible reasons for these differences. We have also investigated many different varieties of red cabbage to determine the best variety and maturation for color production. These results are rather preliminary and will continue to be investigated. Another approach to obtaining blue color we are investigating is the use of anthocyanin-metal complexations. Most of the anthocyanin-based blue colors are produced by these anthocyanin-metal complexations. We have determined that acylated cyanidin and delphinidin derivatives respond best to the interaction with metals, to produce a variety of hues. The addition of excess amounts of salts to anthocyanin mixtures, particularly the acylated cyanidin and delphinidin derivatives, results on bathochromic and hyperchromic shifts, allowing for the production of blue colors under acidic pH environments. The blue colors produced with red cabbage only, required the use of alkaline pH, around 7-8. One additional approach has been the use of anthocyanin microencapsulation. We have investigated the impact of athocyanin size and chemical srtucture on our ability to trap them into a cold-setting gel. We determined that size of the anthocyanin was not an important factor to improve gel color stability. Our hypothesis that larger molecules would be better trapped did not hold true. It was rather the chemical interaction of the flavonoid with the gel components what seemed to be more critical. Further research will be conducted to determine what portion of the molecule is more critical. 3. Flavonoids can interact greatly with both the food matrix as well as the body after ingestion. Interaction of anthocyanins with saliva were explored. In addition we have started a new study to investigate the interaction of anthocyanins with the stomach environment. The hypothesis is that the acidic environment of the stomach will not only favor anthocyanin stability but that will also favor uptake of these compounds into gastric tissues and absorption into the plasma. This hypothesis is currently being investigated. Anthocyanin-metal complexes obtained are being evaluated in a variety of food matrices to determine what is the impact of the matrix composition on the color and stabilty characteristics. Another project that adressed flavonoid interaction with the matrix is dealing with purple corn anthocyanins. These pigments tend to form insoluble complexes, or polimerize during extraction and concentration. We suspect the interactions may be with proteins. These pigments are of great relevance to the world of natural colorants, particularly in Asia. These interaction can serve as a model to understand better how anthocyanins may interact with proteins in the body.

Publications

  • Type: Books Status: Published Year Published: 2013 Citation: Wallace, TC; Giusti, MM. (editors). 2013. Anthocyanins in Health and Disease Prevention. Taylor & Francis Group. CRC Press, p. 355.
  • Type: Book Chapters Status: Published Year Published: 2013 Citation: Pu, J. and Giusti, MM, 2013. Analysis of Anthocyanins in Biological Samples. In "Anthocyanins in Health and Disease". Edited by TC Wallace and MM Giusti. Boca Raton: CRC Press.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Hummer, K; Durst, R; Zee, F; Atnip, A; Giusti, MM. 2013. Phytochemicals in fruits of Hawaiian wild cranberry relatives. J. Sci. Food Agric.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2013 Citation: Snyder, AB; Sweeney, CF; Rodriguez-Saona, LE; Giusti, MM. 2014. Rapid authentication of concord juice concentration in a grape juice blend using Fourier-Transform infrared spectroscopy and chemometric analysis. Food Chem. 147, 295301.
  • Type: Journal Articles Status: Submitted Year Published: 2013 Citation: Sigurdson, G.; Giusti, MM. 2014. Bathochromic and Hyperchromic Effects of Aluminum Chelation by Anthocyanins from Edible Sources for Blue Color Development. J. Ag Food Chem. Special Issue on Anthocyanins. Submitted.
  • Type: Journal Articles Status: Submitted Year Published: 2013 Citation: Kamonpatana, K; Failla, ML; Kumar, PS; Giusti, MM. 2014. Anthocyanin structure determines susceptibility to microbial degradation and bioavailability to the buccal mucosa. J. Ag Food Chem. Special Issue on Anthocyanins. Submitted.


Progress 01/01/12 to 12/31/12

Outputs
OUTPUTS: The year 2012 has been a year of international impact and of great relevance to the food industry. In addition, the relevance of our work to society has been confirmed by the support received by a grant from the USDA. We worked closely with the food industry to address a changing scenario for the food colorant industry, as a result of changing regulations. In addition, my laboratory and research team has hosted visiting scholars from Brazil and Rumania, fostering new collaboration between Universities in those countries and OSU. I was invited speaker at international conferences in the USA, Colombia, and Peru. Our work with universities in Peru has continued, specifically with the Universidad Nacional Agraria, La Molina, in Lima, and we have established new contacts in Peru with a leading company: Alicorp, one of the largest food processors in that country. This company is also trying to respond to the increasing demand for natural food colorants, and we are exploring potential collaborations. The results of our work were presented at several national and international meetings, and to industry or universities. Invited talked I presented in 2012 were: 1. The Relationship Between Anthocyanin Chemical Structure and Its Stability in Foods and the GI Tract. IFT Annual Meeting, Las Vegas, NV, June 25-28. 2. Chemoprotective Effects of Aronia (Chokeberry) Components. Annual Conference of the Midwest Aronia Association. Des Moines, Iowa. March 29-31. 3. Increasing Dietary Intake of Bioactive Flavonoids without Major Changes in Dietary Habits. ASA, CSSA, and SSSA Annual Meetings, Oct. 21-24. Symposium: Harvesting Knowledge from Model Crops to Enhance Crop Nutritional Value. Cincinnati, Ohio, October 21-24. 4. Polyphenol Compounds in Pharmaceutical and Food Industries. XV COIFFA & 1st Symposium of Pharmaceutical and Food Sciences, Medellin, Colombia. September 27-29. 5. Effects of Food Processing in Polyphenol Compounds. XV COIFFA & 1st Symposium of Pharmaceutical and Food Sciences, Medellin, Colombia. September 27-29. Giusti, M. Monica. 2012. Antocianinas: Color y Salud. Simposio Internacional de Innovacion Alimentaria (SIIA). Montana. Lima, Peru. September. Our work was also presented by my students at different local and international conferences, and shared with others, including high-school science teachers and students through workshops and short courses. In June, 2012, in collaboration with the Ohio Department of Education, Ohio Department of Agriculture we presented the workshop "Food Pigments Laboratory", as part of the Hands On Training (HOT) Conference for Agriculture Teachers, in Parker Food Science. I also mentored a team of high school students (David Glass and Audrey Hoey) in their science project about a comparison of the anthocyanins from fresh and frozen blueberries. The work done by these students from the Zane Trace Local Schools in Chillicothe, Ohio under my supervision, received third place and a gold rating at the National FFA Agriscience Fair in Indianapolis in fall 2012. They are moving forward with their project and continuing in 2013. PARTICIPANTS: M Monica Giusti, Principal investigator. Collaborators: Mark Failla (Human Nutrition), Purnima Kumar (College of Dentistry), Gonul Kaletunk (Food and Biological Engineering), Derek Hansford (Biomedical Engineering), Joshua Bomser (Human Nutrition), Robert Durst (Linus Pauling Institute), Luis Rodriguez-Saona (Food Science), Carmen Socaciu (Cluj-Napoca Agricultural University, Rumania). Visiting scholars: Vivian Bochi (Brazil) and Zorita Sconta (Rumania). Students: Neda Ahmadiani, Steven Simmons, Alison Atnip, Andrew Barry, Fei Lao, Kom Kamonpatana, Gregory Sigurdson, Abby Snyder. TARGET AUDIENCES: Scientific community, Food Industry, Consumers, and Regulatory Agencies. PROJECT MODIFICATIONS: The current project is a revision of the original project submitted. As a result, overal goals remain the same, but adjustments have been made to the approach based on findings from previous years.

Impacts
Health improvement and disease prevention through the diet are topics of great current interest. In particular, the world of colorants has received attention due to negative publicity of synthetic colorants and the need for suitable natural alternatives. The EU requires now a warning on foods colored with synthetics, and the FDA was close to follow. A lot of our efforts have been dedicated to facilitate the transition of the food industry into the use of colorants from natural sources. We worked with large US food manufacturers to identify natural alternatives to their synthetic colors. Anthocyanins can serve as alternatives to Red No. 40. This year we continued to explore better sources. In 2011 we developed a method to harvest high quality pigments from cull radishes. A company is currently working on implementing our technology, resulting on new jobs creation. Radishes previously discarded due to size, shape or other minor defects will now be utilized to produce a high quality added value product. Another challenge for the food colorant industry is the lack of availability of natural sources of blue colors. Our goal is to understand how the chemical structure of the pigment, its spatial configuration, the interaction with other compounds in the matrix (such as metals), and the pH of the environment affect the hue and intensity of the color obtained from an anthocyanin source. We identified 2 vegetable sources that can produce blue hues very close to that of blue No. 2, with stability suitable for some food applications. Another approach investigated in our laboratory to facilitate incorporation of natural pigments into foods is through microencapsulation. A multidisciplinary team from food science, food engineering, biomedical engineering and human nutrition is working together to increase the stability of the pigments, mask undesirable aromas or flavors and increase the delivery of these compounds to the target organs in the human body. Funding has been received from the USDA / NIFA program as well as from the Food Innovation Center to explore this approach. In addition, other plant materials are being investigated for total anthocyanins, and other phenolics, with particular interest on proanthocyanidins. Proanthocyanidins are responsible for the protection against urinary tract infections attributed to cranberries. Similar properties may be found in other plant materials to date not characterized. Studies have been done in collaboration with other institutions from Brazil, Rumania, and the Linus Pauling Institute, in Oregon. Lastly, we have investigated the stability and transformations of anthocyanins in the gastrointestinal tract. This year we focused in the oral cavity, where they are believed to exert chemoprotective and anti-inflammatory effects. We have determined that there is selective degradation of certain anthocyanin chemical structure in saliva, and that the mechanisms of degradation differ to those reported on other sites of the GIT. In addition, oral microbiota plays an important role and it is being investigated.

Publications

  • Kamonpatana, K; Giusti, MM; Chitchumroonchokchai, C; MorenoCruz, M; Riedl, KM; Kumar, P; Failla, ML, 2012. Susceptibility of anthocyanins to ex vivo degradation in human saliva. Food Chem. 135 (2) : 738-747.
  • Giusti, M. Monica. 2012. Polyphenol Compounds in Pharmaceutical and Food Industries. XV Reunion de COIFFA. Universidad de Antioquia, Medellin, Colombia. VITAE 19 (2), 19.
  • Giusti, M. Monica. 2012. Effects of Food Processing in Polyphenol Compounds. XV Reunion de COIFFA. Universidad de Antioquia, Medellin, Colombia. VITAE 19 (2), 41.
  • Kamonpatana, K., MorenoCruz, Maria, Kumar, P.; Giusti, M.M.; Failla, M. 2012. Metabolism and Uptake of Anthocyanins in Human Oral Cavity. IFT Annual Meeting Book of Abstracts. Abstract 084-28.
  • Ayvaz, Huseyin; Giusti, Maria M; Rodriguez-Saona, Luis. 2012. Application of Infrared Sensors for Screening Acrylamide Content in Snack Products. IFT Annual Meeting Book of Abstracts. Abstract 193-12
  • Barry, Andrew M; Kaletunc, Gonul; Giusti, Maria M. 2012. Encapsulation of Purple Corn (PC) Anthocyanins by Cold Setting Alginate and High-Methoxy Pectin Gel. IFT Annual Meeting Book of Abstracts. Abstract 144-24.
  • Atnip, Allison; Giusti, M. Monica. 2012. Characterization of the Proanthocyanidin (PAC) Content of 2 Species of Berry from the Genus Vaccinium by HPLC-Fluorescence-MS and DMAC Colorimetric Assay. IFT Annual Meeting Book of Abstracts. Abstract 275-01.
  • Giusti, M. Monica. 2012. The Relationship Between Anthocyanin Chemical Structure and Its Stability in Foods and the GI Tract. IFT Annual Meeting Book of Abstracts. Abstract 093-01.
  • Ahmadiani, Neda; Giusti, M. Monica. 2012. Factors Affecting the Color Quality of Some Anthocyanin Mixtures at Neutral and Alkaline pH Values. IFT Annual Meeting Book of Abstracts. Abstract 275-09.
  • Bochi, Vivian; Rodrigues, Daniele; Godoy, Helena T; Giusti, M. Monica. 2012. Optimization of Extraction Procedure for Anthocyanin and Phenolic Compounds in Ceylon Gooseberry (Dovyalis hebecarpa). IFT Annual Meeting Book of Abstracts. Abstract 275-14.
  • Bochi, Vivian; Godoy, Helena T; Giusti, M. Monica. 2012. Anthocyanin Pigment Composition and Major Phenolic Compounds of Ceylon Gooseberry (Dovyalis hebecarpa) at Different Ripening Stages. IFT Annual Meeting Book of Abstracts. Abstract 275-23.
  • Ahmadiani, Neda; Giusti, M. Monica. 2012. Factors Affecting the Color Quality of Some Anthocyanin Mixtures at Neutral and Alkaline pH Values. IFT Annual Meeting Book of Abstracts. Abstract 310-03.


Progress 01/01/11 to 12/31/11

Outputs
OUTPUTS: We have worked closely with the food industry to address current challenges faced. In addition, we have established new collaborations with other researchers at OSU as well as at other institutions, including universities in Brasil and Peru. I am hosting a visiting scholar from Brazil, and another one from Mexico, and have visited the Universidad Nacional Agraria, La Molina, in Lima, and the Universidad San Antonio Abad de Cusco in Cusco, Peru. The results of our work were presented at several national and international meetings, and to industry or universities. Our work was presented at: Giusti, MM. 2011. Anthocyanin Colors and Chemistry. MARS Science and Technology Workshop - Natural Colors. Davis, California. Giusti, MM. 2011. Bioactive Berry Pigments: Uptake and Transformations in the Gastrointestinal Tract. Berry Health Benefits Symposium. Westlake, California. Giusti, MM. 2011. Incorporando salud y color en la dieta diaria (Incorporating health and color in the daily diet). VIII Iberoamerican Congress in Food Engineering (CIBIA) 2011. Miraflores, Lima, Peru. (October) Giusti, MM. 2011. Anthocyanins for Color and Health. Presented at Winter 2011 HortScience Department Seminar Series. Columbus, Ohio. Kamonpatana, K; Chitchumroonchokchai, C; Giusti, MM; Failla, M. 2011. Ex vivo degradation of chokeberry anthocyanins in human saliva. IFT Annual Meeting and Food Exposition (AMFE), New Orleans. Kamonpatana, K; Giusti, MM; MorenoCruz, M; Kumar, P; Failla, M. 2011. Differential susceptibility of anthocyanins to degradation in human saliva. IFT AMFE, New Orleans. Kottman, S; Giusti, MM. 2011. Production of a red radish juice concentrate for use as a food colorant. IFT AMFE, New Orleans. Witarsa, SY; Giusti, MM. 2011. Comparative study of phytonutrients stability in functional beverages stored under different environment conditions. IFT AMFE, New Orleans. Giusti, MM; Kamonpatana, K; Failla, ML; Kumar, P. 2011. Anthocyanin Degradation in Human Saliva. International Worshop on Anthocyanins. North Carolina, USA. (September). Rinaldi, PL; Paudel, L; Scheerens, JC; Wyzgoski, FJ; Reese, RN; Giusti, MM; Johnson, J; Najjar, NW; Bomser, J; Hardy, JK; Chanon, AM; Smiljanic, D; Wesdemiotis, C. 2011. Identification of Black Raspberry Polyphenols by High Field NMR and HPLC-ESI-MS/MS. International NMR Conference. (September). Our work was also shared with other students and researchers as well as with high-school science teachers through workshops or short courses: 6/2011 Food Pigments. Ohio Department of Education, Ohio Department of Agriculture. HOT Conference - Agriculture Teachers Conference, Parker Bldg, OSU. 8/2011, II Curso Internacional. Analisis Moderno de Alimentos y Productos Naturales. San Antonio Abad de Cusco. Aplicaciones Cromatograficas y Espectroscopicas para la Caracterizacion de Productos Naturales, Cusco, Peru. 8/2011 Aplicaciones Cromatograficas y Espectroscopicas para la Caracterizacion de Productos Naturales, Monitoreo de Autenticidad y Calidad de Alimentos. Universidad Nacional Agraria La Molina (Lima, Peru). PARTICIPANTS: M Monica Giusti, Principal investigator. Collaborators: Mark Failla (Human Nutrition), Purnima Kumar (College of Dentistry), Gonul Kaletunk (Food and Biological Engineering), Derek Hansford (Biomedical Engineering), Joshua Bomser (Human Nutrition), Robert Durst (Linus Pauling Institute). Visiting scholars: Obed Osorio (Mexico) and Vivian Bochi (Brazil). Students: Shirley Witarsa, Scott Kottman, Neda Ahmadiani, Steven Simmons, Alison Atnip, Andrew Barry, Fei Lao, Kom Kamonpatana, Abby Snyder. TARGET AUDIENCES: Scientific community, Food Industry, Consumers, and Regulatory Agencies. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
This has been an important year for natural colorants. FDA announced last March that the safety of synthetic food colorants would be re-evaluated. A lot of our efforts have been dedicated to facilitate the transition of the food industry into the use of colorants from natural sources. A vegetable juice concentrate was produced from cull radishes to be used as a food colorant. Different methods were evaluated to extract the highly stable radish pigments while minimizing the extraction and formation of aroma and flavor components. This work was sponsored by a company that is currently working to commercialize the colorant using our newly developed technology. Radishes that were previously discarded due to size, shape or other minor defects will now be utilized to produce a high quality added value product. Being a new line of product for the company, this will create jobs and growth for that company. Another challenge for the food colorant industry is the lack of availability of natural sources of blue colors. Last year we also focused on identifying potential anthocyanin-based sources of natural blue colors. We have determined that different anthocyanin sources under exactly the same conditions - concentration, matrix, pH - can produce very different colors, particularly at neutral to alkaline pH. We have identified 2 vegetable sources that can produce blue hues very close to that of blue No. 2, with stability suitable for some food applications. Drying of the product significantly enhanced the stability of the color. Another approach being used to facilitate incorporation of these natural pigments into food matrices is the use of microencapsulation. A multidisciplinary team from food science, food engineering, biomedical engineering and human nutrition is working together to increase the stability of the pigments, mask any undesirable aromas or flavors associated with the starting material and increase the delivery of these compounds to the color, where anthocyanins have been found to exert chemoprotective effects. Funding has been received from the USDA / NIFA program as well as from the Food Innovation Center to explore this approach. In addition, other plant materials are being investigated for total anthocyanins, and other phenolics, with particular interest on proanthocyanidins. Proanthocyanidins are responsible for the protection against urinary tract infections attributed to cranberries. Similar properties may be found in other plant materials to date not characterized. This work has been done in collaboration with other institutions from Brazil, Mexico, as well as with the Linus Paulin Institute, in Oregon. Lastly, we have investigated the stability and transformations of anthocyanins in the gastrointestinal tract. This year we focused in the oral cavity, where they are believed to exert chemoprotective effects. We have determined that there is selective degradation of certain anthocyanin chemical structure in saliva, and that the mechanisms of degradation differ to those reported on other sites of the GIT. In addition, oral microbiota plays an important role and it is being investigated.

Publications

  • Osorio-Esquivel, O; Ortiz-Moreno, A; Alvarez, VB; Dorantes-Alvarez, L; Giusti, MM. 2011. Phenolics, betacyanins and antioxidant activity in Opuntia joconostle fruits. Food Research International. Vol. 44, no. 7. : 2160-2168.
  • Wallace, TC; Giusti, MM. 2011. Selective Removal of the Violet Color Produced by Anthocyanins in Procyanidin-Rich Unfermented Cocoa Extracts. Journal of Food Science. Vol. 76. : C1010-C1017.
  • Del Carpio Jimenez, C; Serrano Flores, C; He, J; Tian, Q, Schwartz, SJ; Giusti, MM. 2011. Characterization and preliminary bioactivity determination of Berberis boliviana Lechler fruits anthocyanins. Food Chemistry. Vol. 128. : 717-724.
  • He, J; Giusti, MM. 2011. High-purity Isolation of Anthocyanins Mixtures from Fruits and Vegetables - a Novel Solid-Phase Extraction Method Using Mixed Mode Cation-exchange Chromatography. Journal of Chromatography A. Vol. 1218, no. 44. : 7914-7922.
  • Johnson, JL; Bomser, JA; Scheerens, JC; Giusti, MM. 2011. Effect of Black Raspberry (Rubus occidentalis L.) Extract Variation Conditioned by Cultivar, Production Site, and Fruit Maturity Stage on Colon Cancer Cell Proliferation. Journal of Agricultural and Food Chemistry. Vol. 5, no. 59. : 1638-1645.
  • Kamonpatana, K; Chitchumroonchokchai, C; Giusti, MM; Failla, M. 2011. Ex vivo degradation of chokeberry anthocyanins in human saliva [Abstract]. IFT Annual Meeting Book of Abstracts: Abstract 098-14. Kamonpatana, K; Giusti, MM; MorenoCruz, M; Kumar, P; Failla, M. 2011. Differential susceptibility of anthocyanins to degradation in human saliva [Abstract]. IFT Annual Meeting Book of Abstracts: Abstract 098-18.
  • Kottman, S; Giusti, MM. 2011. Production of a red radish juice concentrate for use as a food colorant [Abstract]. IFT Annual Meeting Book of Abstracts: Abstract 155-01.
  • Witarsa, SY; Giusti, MM. 2011. Comparative study of phytonutrients stability in functional beverages stored under different environment conditions [Abstract]. IFT Annual Meeting Book of Abstracts. Abstract 150-08.
  • Giusti, MM; Kamonpatana, K; Failla, ML; Kumar, P. 2011. Anthocyanin Degradation in Human Saliva [Abstract]. Proceedings of the 2011 International Worshop on Anthocyanins. North Carolina, USA. (September).
  • Rinaldi, PL; Paudel, L; Scheerens, JC; Wyzgoski, FJ; Reese, RN; Giusti, MM; Johnson, J; Najjar, NW; Bomser, J; Hardy, JK; Chanon, AM; Smiljanic, D; Wesdemiotis, C. 2011. Identification of Black Raspberry Polyphenols by High Field NMR and HPLC-ESI-MS/MS [Abstract]. Book of Abstracts of the International NMR Conference. (September).
  • Giusti, MM; Atnip, A; Sweeney, C; Rodriguez-Saona, LE. 2011. Rapid Authentication of Fruit Juices by Infrared Spectroscopic Techniques. In Progress in Authentication of Food and Wine. Edited by Ebeler, SE; Takeoka, GR; Winterhalter, P. Washington, DC, USA: ACS Symposium series.
  • Jing P, Giusti MM. 2011. Contribution of Berry Anthocyanins to Their Chemopreventive Properties. In Berries and Cancer Prevention. Edited by G.D. Stoner, N.P. Seeram. New York, NY, USA: Springer Science+Business Media. 3-40.


Progress 01/01/10 to 12/31/10

Outputs
OUTPUTS: This year we have worked closely with the food industry on bringing science and phytonutrients into regularly consumed products. Among the products we have improved and / or evaluated are power drinks - to improve stability of health enhancing components, cranberry extracts rich in flavonoids and cull radishes, to extract anthocyanin pigments from the skins to produce added value ingredients from waste materials. In addition, we have initiated a clinical study to evaluate the metabolism of anthocyanins in the oral cavity and the role of saliva and oral microbiota in anthocyanin degradation. Anthocyanins have been linked to oral cancer protective effects and it is important to understand what chemical forms are more biologically active, and how these compounds change after they come in contact with the mouth. This work was done in collaboration with Mark Failla from the Human Nutrition department and Purnima Kumar of the Dental School, and with OARDC funding. The results of our work was presented at several international meetings in the USA, Chile, Brasil, and Peru. I was an invited speaker at the American Chemical society annual meeting to share our work with authenticity of grape juices. In Chile, it was CREAS (Regional Center for Research on Healthy Foods) that requested our contribution in the field of flavonoids, and important contacts were established that may initiate new collaborations. I was an invited speaker at the XXII Brazilian Congress of Food Science and Technology, where I shared our results on the transfer of isoflavones into poultry tissues. In addition, we continue our collaborations with the Universidad Nacional Agraria La Molina, the International Potato Center, and the Universidad San Antonio Abad de Cusco, in Peru. I visited all those institutions in June/July and had the opportunity to exchanged ideas and samples with faculty and researchers, and interacted with graduate students. My laboratory hosted a student from the Instituto Politecnico Nacional, Mexico working in the experimental portion of his thesis. He investigated the phenolic and pigment composition of Opuntia joconostle,a fruit of high antioxidant power. We initiated a collaboration with Gonul Kaletunc, department of food agriculture and biological engineering to encapsulate anthocyanins. This could enhance stability of the pigments in foods while enhancing their bioavailability in the intestines. A new company, AnthoScyantific, has been created to commercialize a technology developed in my laboratory, patent pending. This technology allows for purification of large quantities of anthocyanins with high purity with a rapid method. The material produced has a cost much lower than any comparable product in the market, making animal and clinical trials with pure anthocyanins possible. PARTICIPANTS: M Monica Giusti, Principal investigator. Collaborators: Mark Failla, Purnima Kumar, Gonul Kaletunk, Alicia Ortiz Moreno, Luis E. Rodriguez Saona, Anthony Buffington. Students: Shirley Witarsa, Scott Kottman, Neda Ahmadiani, Steven Simmons, Alison Atnip, Obed Osorio Esquivel, Abby Snyder. TARGET AUDIENCES: Scientific community Food Industry Consumers Regulatory Agencies in the USA as well as in countries visited, including Brasil, Chile and Peru. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Cranberries are a major source of anthocyanins and procyanidins, potent phytonutrients believed to benefits in the vascular system and other age / obesity related disorders. Dimeric and trimeric procyanidins may prevent urinary tract infections. We optimized and applied a protocol for the quantification of monomeric anthocyanins and procyanidins in 8 cranberry extracts. Optimal extraction procedures for the quantification of anthocyanins and procyanidins in commercial cranberry extracts with varying matrices and processing techniques were evaluated and protocols developed. Flavonoids of all cranberry extracts, including those containing inert binding agents, were best extracted using an acetone / water technique versus an acid / alkaline extraction. Procyanidin extracts were semi-purified using Sephadex LH-20 to remove interfering compounds. Cranberry anthocyanins were characterized and quantified using the pH Differential method and reverse phase HPLC-PDA-MS analysis. Procyanidins were characterized and quantified using the DMAC method and normal phase HPLC-Fluorescence-MS analysis, a catechin standard curve, and a Diol stationary phase column. The more efficient extraction techniques and increased HPLC resolution of cranberry anthocyanins / procyanidins enabled us to more accurately quantify these phytonutrients and to advance protocols for use by the cranberry industry. Opuntia joconostle is a cactus grown in Mexico since pre-Hispanic times, yielding more than 10,000 tons per year. Xonocostle, the fruit of O. joconostle is consumed fresh in salads, sauces and other food products, and in traditional medicine as treatment for diabetes, hypertension and obesity. Our team characterized for the first time the biocompounds of the fruit. Samples were obtained from San Juan Teotihuacan, Mexico. Fruit extracts were obtained from peel, pulp, and seed by using 70% aqueous acetone with 0.01% HCl followed by chloroform partition, and purified by solid-phase extraction with a C18 cartridge. Total phenolics, anthocyanins, and betalains were analyzed spectrophotometrically. Pigments were identified by HPLC-PDA-MS, and the antioxidant activity was determined by the DPPH+. The results showed that xoconostle is an important source of phenolic compounds; the highest concentration was in the peel (346.09 mg gallic acid equivalents (GAE) /100 g fresh fruit), followed by the seed and the pulp (142.00 and 102.12 mg GAE/100 g fresh fruit, respectively). No anthocyanins were detected. The color of the peel and pulp was mainly due to the presence of betacyanins (5.94 and 0.85 mg betanin/100 g fresh sample in pulp and peel, respectively) including betanin, isobetanin, betanidin, and isobetanidin. Crude extracts of xonocostle had high antioxidant activity mainly in the peel (68.62 expressed as % inhibition of DPPH+), followed by pulp (54.74) and seed (20.05). It was concluded that xonocostle is a rich source of antioxidant compounds that can be marketed as a functional food. A vegetable juice concentrate was produced from cull radishes to be used as a food colorant. Different methods are being evaluated to remove aroma and flavor components from the pigments.

Publications

  • Johnson, JL; Bomser, JA; Scheerens, JC; Giusti, MM. 2010. Effect of Black Raspberry (Rubus occidentalis L.) Extract Variation Conditioned by Cultivar, Production Site, and Fruit Maturity Stage on Colon Cancer Cell Proliferation. J Agr Food Chem. In Press.
  • Wallace, TC; Giusti, MM. 2010. Evaluation of Parameters that Affect the 4-Dimethylaminocinnamaldehyde Assay for Flavanols and Proanthocyanidins. J Food Sci. 75 (7), C619-C625.
  • Wallace, TC; Giusti, MM. 2010. Extraction and Normal Phase HPLC Fluorescence Electrospray MS Characterization and Quantification of Procyanidins in Cranberry Extracts. J Food Sci. 75 (8), C690-C696.
  • He, J; Giusti, MM. 2010. Anthocyanins: Natural colorants with health promoting properties. Annual Review of Food Science and Technology . 1 (1), 163-187.
  • Giusti, MM; Wrolstad, RE; Smith, DE. 2010. Calculation of CIE Color Specifications from Reflectance or Transmittance Spectra. In Food Analysis Laboratory Manual. Second ed. Edited by Suzanne Nielsen. New York, New York, USA: Springer Science+Business media. 171-177.
  • Giusti, MM; Rodriguez-Saona, LE; He, J; Sweeney, C. 2010. Authentication of fruit juices by spectroscopic techniques [Abstract]. American Chemical Society, 239th Annual Meeting and Exposition [Peer Reviewed] (Published)
  • Giusti, MM. 2010. Chemoprotection, bioavailability and metabolism of anthocyanins from small fruits [Abstract]. IFT Annual Meeting Book of Abstracts [Peer Reviewed] (Published)
  • Osorio-Esquivel, O; Alvarez, V; Ortiz-Moreno, A; Ji, T; Dorantes-Alvarez, L; Giusti, MM. 2010. Phenolic content, antioxidant activity and identification of pigments of sour prickly pear from Opuntia joconostle [Abstract]. IFT Annual Meeting Book of Abstracts [Peer Reviewed] (Published)
  • Wallace, TC; Giusti, MM. 2010. Assessment of normal and reverse phase HPLC-MS and fluorescence/photodiode array detection methods for characterization and quantification of procyanidins and anthocyanins in cranberry extracts [Abstract].I FT Annual Meeting Book of Abstracts [Peer Reviewed] (Published)
  • Giusti, MM. 2010. Increasing the dietary intake of polyphenolics without changing dietary habits. Pacifichem, The International Chemical Congress of Pacific Basin Societies. Value-Added Food Products from Fruits and Vegetables, (Symposium #162)-Area- Agrochemistry. Book of Abstracts,


Progress 01/01/09 to 12/31/09

Outputs
OUTPUTS: We have continued making great progress on the evaluation of different plant materials, the evaluation of improved analytical techniques, food applications and the inhibition of colon cancer cell growth of anthocyanins and anthocyanin-rich extracts. New materials evaluated include plant materials provided by the International Potato Center, in Peru, strengthening international collaborations. Native potato varieties were evaluated for enhanced polyphenolic content, with the goal of using this staple commodity to enhance nutrition. I participated in national and international meetings to enhance our collaborations and exchange information with other researchers working in this field of research. Early in January, I visited the Universidad Nacional Agraria La Molina and the International Potato Center in Peru, where we exchanged ideas and samples for chemical characterization of phenolics. We also have joined the graduate program as Visiting Faculty. International contacts were also established with Colombia, where I presented a seminar and served as external examiner for 2 dissertations dealing with functional foods. Other opportunity to disseminate the results from our research was at the Annual Meeting and Food Exposition of the Institute of Food Technologists (IFT), in Anaheim CA, as an invited speaker sharing our results on the protective effects of anthocyanin-rich commodities, and through 8 other presentation by students working in the research. Additional presentations were at the Americal Chemical Society. Our research on Enhancing Nutrition with Phenolic-Rich Potatoes was highlighted as a keynote talk at the BioPotato Network, in Canada. In March 2009 I was chair of the organizing committee for the first Ohio Valley IFT Symposium: Functional Foods for the Health Conscious Consumer. This project has involved the participation and effort of several investigators as well as several students, mainly graduate students. This year 5 graduate students obtained their degree after working in this project. PARTICIPANTS: M Monica Giusti, Principal investigator Collaborators: Steven Schwartz Joshua Bomser, Bernadene Magnuson, Mark Failla, Luis E. Rodriguez Saona Pu Jing. Students: Jian He Taylor Wallace Matthew Bunce, Maria Fernanda Polit, dante Vargas, Jennifer Willig, Christian Sweeney, Jodee Johnson. TARGET AUDIENCES: Scientific community Food Industry Consumers Regulatory Agencies PARTICIPANTS: PARTICIPANTS: M Monica Giusti, Principal investigator Collaborators: Steven Schwartz Joshua Bomser, Bernadene Magnuson, Mark Failla, Luis E. Rodriguez Saona Pu Jing. Students: Jian He, Taylor Wallace, Matthew Bunce, Maria Fernanda Polit, Dante Vargas, Jennifer Willig, Christian Sweeney, Jodee Johnson, Nuryati Pangestu, Kom Komanpatana. We also received support and collaboration from industry and institutions including: Universidad Nacional Agraria - La Molina, Internacional Potato Center, Universidad San Antonio Abad de Cusco, Welch's, MM Mars Botanicals, Cliffstar, Tropical Traditions. TARGET AUDIENCES: Scientific community, Food Industry, Consumers, and Regulatory Agencies PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Our laboratory has characterized new plant materials, and determined content and profiles of anthocyanins and other phenolics. We have found different varieties of potato with enhanced concentration of polyphenolic compounds. Potatoes are considered a staple commodity and they are the 5th most important crop worldwide. Delivering nutrition through a staple commodity has great potential for impacting large population groups. Our in vitro and in vivo studies are providing compelling evidence of the chemoprotective effects of anthocyanin-rich extracts against colon cancer. Anthocyanin rich extracts (AREs) from different plant materiasl were evaluated. Purple corn, chokeberry, bilberry, purple carrot, grape, radish, and elderberry were tested for growth inhibition using a human colorectal adenocarcinoma (HT29) cell line. All AREs suppressed HT29 cell growth to varying degrees. Purple corn ARE showed the highest growth inhibition (GI50~14 microg/mL as cy-3-glucoside equivalent), followed by chokeberry and bilberry. Elderberry and radish AREs showed the lowest growth inhibition (GI50:~130 microg/mL; ~108 microg/mL). ACN played a major role in AREs' chemoprotection and exerted an additive interaction with OPF. AREs at these concentration ranges and under similar conditions had little effect on NCM460 normal colon cells. Statistical analyses demonstrated that type of anthocyanin aglycone, glycosylation, and/or acylations affected growth inhibition activity. In addition, we evaluated a number of black raspberry varieties grown at different locations and at different maturity stages. All these factors can have an effect of the chemoprotective effects of the anthocyanin extract. Our results suggest that not only anthocyanins by also precursors of anthocyanins can increase the chemoprotective effects of black raspberry extracts in a colon cancer model system. These findings should be considered for development of anthocyanin applications in functional foods and/or during crop and cultivar selection. Anthocyanin-based colorants used by the food industry owe their color to the presence of acylated anthocyanins. We also investigated the impact of the acylation on the chemoprotective effect of anthocyanins. We determined that acylatin groups attached to the pigments do not decrease their chemoprotective effects, they may in some cases increase their activity. These results are extremely relevant to the food industry as they show that pigments used in food application can also have protective effects against colon cancer.

Publications

  • Rubio-Diaz DE; Pozza ME; Dimitrakov J; Gilleran JP; Giusti MM; Stella JL; Rodriguez-Saona LE; Buffington CA. 2009. A candidate serum biomarker for bladder pain syndrome/interstitial cystitis. The Analyst. Vol. 134, no. 6. (June 1): 1133-1137. He, J; Wallace, TC; Keatley, KE; Failla, ML; Giusti, MM. 2009. Stability of black raspberry anthocyanins in the digestive tract lumen and transport efficiency into gastric and small intestinal tissues in the rat. Journal of Agricultural and Food Chemistry. Vol. 57, no. 8: 3141-3148. Giusti MM and Wallace TC. 2009. Flavonoids as natural pigments. In Handbook of Natural Colorants. Wiley Series in Renewable Sources. Edited by T Bechtold and RAM Mussak. West Sussex, UK: John Wiley and Sons. 257-275. Giusti, MM, Bomser JA, Schwartz SJ, Magnuson BA. 2009. "Expanding the market of anthocyanin-rich commodities through the characterization of components with biological activity, Abstract 239-01." Abstract. Annual Meeting of the IFT Book of Abstracts. Johnson JL, Bomser JA, Giusti MM, Scheerens JC. 2009. "Effects of different black raspberry extracts on colon cancer cell proliferation in an in vitro cell system, Abstract 126-18." Abstract. Annual Meeting of the IFT Book of Abstracts. Johnson JL, Bomser JA, Scheerens JC, Giusti, MM. 2009. "Effects of black raspberry extracts on colon cancer cell proliferation, Abstract AGFD 135." Abstract. American Chemical Society, 238th Annual Meeting and Exposition. Polit MF, Shiroma-Kian C, Rodriguez-Saona LE, Tay D, Manrique L, Giusti, MM. 2009. "Characterization and quantification of anthocyanins and other phenolics in Native Andean potatoes, Abstract 204-33." Abstract. Annual Meeting of the IFT Book of Abstracts. Sweeney CF, Giusti MM, Rodriguez-Saona LE. 2009. "Rapid determination of Concord grape content in commercial grape juices by infrared spectroscopy, Abstract 057-14." Abstract. Annual Meeting of the IFT Book of Abstracts. Vargas DM, Giusti, MM. 2009. "Identification and quantification of soy isoflavones and the isoflavone metabolite equol in commercial eggs, Abstract 061-06." Abstract. Annual Meeting of the IFT Book of Abstracts. Vargas DM, Latshaw D, Giusti MM. 2009. "Quantification of soy isoflavones and equol in hen egg and tissues: Determination of isoflavone transfer from the feed, equol production and total accumulation in eggs and specific tissues, Abstract 061-33." Abstract. Annual Meeting of the IFT Book of Abstracts. Wallace TC, Giusti, MM. 2009. "Optimization of the 4-dimethylaminocinnamaldehye assay for flavan-3-ols., Abstract AGFD 158." Abstract. American Chemical Society, 238th Annual Meeting and Exposition. Willig JA, Jing P, Bomser JA, Giusti, MM. 2009. "The effect of acylation on the antiproliferative effects of anthocyanin pigments using a human colon cancer cell line, Abstract 257-18." Abstract. Annual Meeting of the IFT Book of Abstracts.


Progress 01/01/08 to 12/31/08

Outputs
OUTPUTS: This year I worked with 9 graduate students focusing on 4 main areas: the evaluation of new plant materials, the evaluation of improved analytical techniques, food applications and the inhibition of colon cancer cell growth of anthocyanins and anthocyanin-rich extracts. In Spring, I offered the course: "Food colors and Pigments" to training students on the importance and potential health benefits of natural pigments. I participated in national and international meetings to enhance our collaborations and exchange information with other researchers working in this field of research. Early in January, I visited UNALM where we exchanged ideas and samples for chemical characterization of phenolics. We have worked with a native berry from the Andes, and evaluated its potential as natural colorant. During the same trip, I visited Montana, a colorants company in Peru, where I shared our results about the advantages and potential health benefits of anthocyanins. Later on, that same January, I presented our work at the DSM 2008 Juice Workshop in Orlando, FL. This meeting was sponsored by DSM Food Specialties USA, Inc. and was targeted mainly to representatives of the food industry. This was an excellent opportunity to interact with industry representatives, confirm that their interest in fruit and vegetable products is in the raise, and exchange ideas about the production of healthier food products. Other opportunity to disseminate the results from our research was in Mexico (Chiapas, March) at the V Congreso Internacional de Ingenieria Bioquimica. The audience included students, faculty and other researchers from South, Central and North America from the fields of food science and technology, chemistry, biochemistry and biochemical engineering. A presentation in Finland, gave us the chance to interact with a much more specialized audience, at the 5th International Congress on Pigments in Foods, Helsinki (August). This meeting brought together the main researchers in the areas of natural pigments in foods. We presented our work on the bioavailability of anthocyanin compounds, and learned about the latest related research. Dissemination of this project also was done through presentations by my graduate students at the annual IFT meeting in New Orleans. I accepted an invitation to teach a short course / workshop in Cusco, Peru, in October this year about the use of chromatographic techniques for the analyses of natural products. This course took place during the latinamerican meeting for chemists (Congreso Iberoamericano de Quimica). During this event, I visited the Universidad Nacional San Antonio Abad de Cusco, and interacted with faculty from the chemistry, natural products, and biology programs. Collaborations with this institution are expected to continue in the future. New collaborations have been established with the department of Animal Sciences to study the transfer of isoflavones, potent phytoestrogens, from animal feed into animal tissues. This investigation is under way. A provisional patent application has been filed for commercialization of a new technology developed in our lab to fractionate phenolics with expected biological activity. PARTICIPANTS: PARTICIPANTS: M Monica Giusti, Principal investigator Collaborators: Steven Schwartz Joshua Bomser, Luis E. Rodriguez-Saona, Joe Scheerens, Faith Wyzgoski, Peter Rinaldi at Ohio State University. Collaborators: Pu Jing, at Jiangsu University. Collaborators in Peru: David Campos, Francisco Salas, Carla del Carpio. Students: Jian He, Taylor Wallace, Matthew Bunce, Maria Fernanda Polit, Jennifer Willig, Jodee Johnson, Christian Sweeney, Dante Vargas. TARGET AUDIENCES: Scientific community, Food Industry Consumers, Regulatory Agencies PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Our laboratory has continued the evaluation of new plant materials, including qualitative and quantitative evaluation of total phenolics, anthocyanins, and other flavonoids. Important berries evaluated include Berberis boliviana, a Peruvian native berry with unusually high pigment content, and Sambucus peruviana, a close relative of elderberries that had not been well characterized before. We also evaluated these plants as potential sources of colorants for food applications. Berries were incorporated into foods typically colored with synthetic dyes, and representing challenging applications for natural pigments: yogurt and candy. They were also used in combination with other phenolics considered copigments, compounds that can enhance the color and stability of the pigments, in a juice model system. We showed that these foods can be successfully enriched with polyphenolics with acceptable stability for commercial applications. Our in vitro and in vivo studies are providing compelling evidence of the chemoprotective effects of anthocyanin-rich extracts against colon cancer. In addition, we have shown that the chemical structure of anthocyanins plays a major role on their ability to slow down colon cancer cell growth. Anthocyanins have been extracted from vegetables containing acylated pigments, and their activity has been compared to that of the same anthocyanins with no acylation. Anthocyanin acylation is known to increase pigment stability, and we hypothesize that it will also have an impact on the bioavailability and bioactivity of the pigments. All AREs suppressed HT29 cell growth to varying degrees. Statistical analyses demonstrated that type of anthocyanin aglycone, glycosylation, and/or acylations affected growth inhibition activity. These findings should be considered for development of anthocyanin applications in functional foods and/or during crop and cultivar selection. A study has been conducted to evaluate unexpected sources of isoflavones, potent phytoestrogens, in our diet. The main sources of dietary isoflavones are soy beans which are mainly used in the USA for animal feed. The isoflavones from animal feed may transfer and accumulate in animal tissues. First, isoflavones were found and quantified in commercial eggs. Then, an experiment was designed in collaboration with the animal sciences department to test the modulation of isoflavone concentration in hen eggs and tissues by the use of varying degrees of soy isoflavones in the animal feed. This will provide valuable information regarding the human exposure through poultry to these potent compounds. One student completed her MS degree and 1 completed his dissertation. Both have joined the food industry and continue to work in areas related to functional foods.

Publications

  • Shiroma, C., Tay, D., Manrique, I., Giusti, M.M., and Rodriguez-Saona, L.E. 2008. Improving the screening process for selection of potato breeding lines with enhanced nutritional traits. J. Agric. Food Chem. 56, 9835-9842.
  • Wallace, T., and Giusti MM. 2008. Determination of color, pigment, and phenolic stability in yogurt systems colored with non-acylated anthocyanins from Berberis boliviana L. as compared to other natural/synthetic colorants. J. Food Science. 73, 241-248.
  • Giusti MM and Wallace T. 2009. Flavonoids as Natural Pigments. Chapter 16 in Handbook of colors. MA Mussak (Ed.) CRC Press. In Press.
  • Jing, P; Bomser, JA; Schwartz, SJ; He, J; Magnuson, BA; Giusti, MM. 2008. Structure-function relationships of anthocyanins from various anthocyanins-rich extracts on the inhibition of colon cancer cell growth. J Agr Food Chem. 56, 9391-9398.
  • He, J., Wallace, T.C., Keatley, K.E., Failla, M.L., and Giusti, M.M. 2008. A Kinetic Study of Black Raspberry Anthocyanins in Rat Stomach and Intestine. 5th Pigments in Food Congress - for Quality and Health. Heinonen M. (Ed). Helsinki, Finland. Pages 36-37.
  • Giusti, M.M. and Wrolstad, R.E. 2008. Color Quality of Maraschino Cherries. Chapter 4 in Color Quality of Fresh and Processed Fruits and Vegetables. Eds. Wrolstad and Culbert. ACS publications. Pp 43-53.
  • Rodriguez-Saona, L.E., Giusti, M.M. and Wrolstad, R.E. 2008. Expanding the Potato Industry: Exotic Colored-Fleshed Tubers. Chapter 9 in Color Quality of Fresh and Processed Fruits and Vegetables. Eds. Wrolstad and Culbert. ACS publications. Pp 114-130.


Progress 01/01/07 to 12/31/07

Outputs
OUTPUTS: Consumption of fruits and vegetables has been associated with reduced incidence of chronic disease. Animal, clinical and in vitro studies provide compelling evidence that commodities rich in polyphenolics (such as anthocyanins and isoflavones) are powerful antioxidants and may be chemoprotective. With more than 5000 flavonoid structure present in nature, and close to 600 different structures among anthocyanins alone, there is a need to increase understanding structure/function relationships. Our efforts have been focused on increasing the understanding of the role that non-volatile compounds from fruits and vegetables play on health promotion. Of special interest are the flavonoids, including anthocyanins and isoflavones, and how horticultural and processing conditions affect their incidence in foods. This year our projects have involved evaluation of new plant materials, development of new analytical techniques, applications in food systems and evaluation of bioactivity of polyphenolics. I have also developed a course: "Food colors and Pigments" targeted to training students on the importance and potential health benefits of natural pigments. We have participated in national and international meetings to enhance our collaborations and exchange information with other researchers working in this field of research. In August I presented our research at the American Chemical Society national meeting in Boston. Our research was chosen by the media for an associated press release, and featured in more than 10 different countries, and in different languages. We also participated on a collaborative effort sponsored by the USDA and the PR China Ministry of Agriculture. We visited China, including Beijing, Shanghai, Nanjing and other cities in the Jiangsu province. In all different cities we visited Universitites and / or research institutions where we disseminated results from our research and exchanged ideas for future collaborations. A collaboration with Jiangsu University has been established to investigate radish cultivars as sources of natural colorants for food applications. Dissemination of this project also was done through presentations by the students at the annual IFT meeting, as well as by presentations by the PI at research institutions in Peru (International Potato Center, Universidad Nacional Agraria), as well as to the food industry at companies in USA, Peru and Bolivia. A novel method has been developed for the isolation of anthocyanin compounds from plants. Many biological studies in the past have tried to evaluate bioactivity of these compounds using crude extracts of mixtures of anthocyanins and other phenolics. Use of such mixtures include many confounding factors that complicate data interpretation as other compounds present in those extract might exert biological effects. Use of pure anthocyanins had been prohibitive in the past due to the high cost and limited availability of most anthocyanins. Our new methodology uses a novel system of solid phase extraction that combines cation exchange and hydrophobic interactions. A provisional patent application will be filed to explore commercialization of this new technology. PARTICIPANTS: M Monica Giusti, Principal investigator Collaborators: Steven Schwartz Joshua Bomser Bernadene Magnuson Luis E. Rodriguez Saona Pu Jing Students Jian He Taylor Wallace Matthew Bunce TARGET AUDIENCES: Scientific community Food Industry Consumers Regulatory Agencies

Impacts
Our laboratory has characterized new plant materials, and determined content and profiles of anthocyanins and other phenolics. We have found a new berry, Berberis boliviana, with a pigment content comparable to that of commercial color extracts. A berry with such high concentration offers great potential to the food industry as it would not require color extraction to be applied into food systems. Berries were incorporated into foods typically colored with synthetic dyes, and representing challenging applications for natural pigments: yogurt and candy. We showed that these foods can be successfully enriched with polyphenolics with acceptable stability for commercial applications. Our in vitro and in vivo studies are providing compelling evidence of the chemoprotective effects of anthocyanin-rich extracts against colon cancer. Anthocyanin rich extracts (AREs) from different plant materiasl were evaluated. Purple corn, chokeberry, bilberry, purple carrot, grape, radish, and elderberry were tested for growth inhibition using a human colorectal adenocarcinoma (HT29) cell line. All AREs suppressed HT29 cell growth to varying degrees. Purple corn ARE showed the highest growth inhibition (GI50~14 microg/mL as cy-3-glucoside equivalent), followed by chokeberry and bilberry. Elderberry and radish AREs showed the lowest growth inhibition (GI50:~130 microg/mL; ~108 microg/mL). ACN played a major role in AREs' chemoprotection and exerted an additive interaction with OPF. AREs at these concentration ranges and under similar conditions had little effect on NCM460 normal colon cells. Statistical analyses demonstrated that type of anthocyanin aglycone, glycosylation, and/or acylations affected growth inhibition activity. These findings should be considered for development of anthocyanin applications in functional foods and/or during crop and cultivar selection. Also, the effect of chemical structure of different anthocyanins was evaluated on their chemoprotection in this system. Additive, synergistic or antagonistic effects between anthocyanins with other phenols were also evaluated. We believe that it is possible to increase consumption of polyphenolics with potential health benefits at the same time that we increase the appeal of food products. Anthocyanins provide an excellent source of polyphenols, and can improve the visual appeal of foods being an alternative to the use of synthetic dyes. We want to increase the understanding of how their chemical structure will affect their color characteristics, stability and bioavailability, particularly when incorporated into food systems.

Publications

  • Wallace, T., and Giusti MM. 2008. Berberis boliviana pigments as source of natural colorants. Accepted J. Food Science.
  • Jing, P., Noriega, V.; Schwartz, S.J. and Giusti, M.M. 2007. Effects of growing conditions on Purple Corncob (Zea mays L) anthocyanins. J. Agr Food Chem 55, 8625-8629.
  • Jing, P. and Giusti, M.M. 2007. Effects of extraction conditions on improving the yield and quality of an anthocyanin-rich purple (Zea Mays L.) corn color extract. J. Food Sci. 72, 1517-1522.
  • Singletary, K.W.; Jung, K.J. and Giusti, M.M. 2007. Anthocyanin-rich grape extract blocks breast cell DNA damage. J Med Food, 10 (2) 2007, 244-251.
  • He, J.; Rodriguez-Saona, L.E.; Giusti, M.M. 2007. Mid-Infrared Spectroscopy for Juice Authentication - Rapid Differentiation of Commercial Juices. J Agr Food Chem, 55, 4443-4452.
  • Giusti, M.M. and Wrolstad, R.E. 2008. Color Quality of Maraschino Cherries. Chapter in Color quality of fresh and processed fruits and vegetables. Eds. Wrolstad and Culbert. ACS publications. In Press.
  • Rodriguez-Saona, L.E., Giusti, M.M. and Wrolstad, R.E. 2008. Expanding the Potato Industry: Exotic Colored-Fleshed Tubers. Chapter in Color quality of fresh and processed fruits and vegetables. Eds. Wrolstad and Culbert. ACS publications. In Press.
  • Schwartz, S.J., Von Elbe, J.H., and Giusti, M.M. 2007. Colorants. Ch. 9 In Fennema's Food Chemistry. S. Damodaran, K.L. Parkin, O.R. Fennema (Eds.), 4th edition. CRC Press. Pp 571-638.
  • GIUSTI, M.M. Jing, P. 2007. Analysis of Pigments and Colorants: Analysis of Anthocyanins. Ch 6.3 In Food Colorants: Chemical and Functional Properties, C. Socaciu (Ed). CRC Press. Pp. 479-506.
  • GIUSTI, M.M. Jing, P. 2007. Natural pigments of berries: functionality and applications. Chapter 4 in Berry fruit: Value-added products for health promotion, Yanyun Zhao (Ed.). CRC Press. Pp 105-146.
  • Anonymous. 2007. Color Esperanza: Antocianinas y cancer al colon. Suplemento Somos, diario El Comercio, August 26, 2007. Article highlight study in Dr. Giusti's lab on chemoprotective effects of anthocyanin rich commodities.
  • Anonymous. 2007. Study: dark colored fruits and veggies fight cancer. Fox News, Tuesday August 21, 2007. www.foxnews.com.
  • Anonymous. 2007. Darker fruits could fight cancer. BBC News. Monday august 20, 2007. www.news.bbc.co.uk.
  • Bunce, MG, S.J. Schwartz and M.M. Giusti. 2008. Hard candy enriched with anthocyanin and tea extracts to increase visual appeal and total phenolics. To be submitted to J. Food Science.
  • Jing, P., Joshua Bomser, M. Monica Giusti, 2007. Intrinsic and extrinsic factors affecting anthocyanin chemoprotection. Accepted for presentation at the Institute of Food Technologists Annual Meeting and Food EXPO. Chicago, IL, July.
  • Wallace, TC, M. Monica Giusti. 2007. Berberis boliviana anthocyanins as natural colorants for yogurt Accepted for presentation at the Institute of Food Technologists Annual Meeting and Food EXPO. Chicago, IL, July.
  • He, J and MM Giusti. 2007. High-purity Fractionation of Anthocyanins from Fruit and Vegetables - Advantages of Ion-exchange Techniques over Conventional Methods. Accepted for presentation at the Institute of Food Technologists Annual Meeting and Food EXPO. Chicago, IL, July.
  • He, J., X Zhang, A Broughton, and MM Giusti. 2007. The Shelf-life Extension of Cottage Cheese - The Effect of Vacuum Packaging and the Addition of Naturally Occurring Antimicrobial Phenolics from Berry Extracts. Accepted for presentation at the Institute of Food Technologists Annual Meeting and Food EXPO. Chicago, IL, July.
  • Keatley, KE, J He, R Klein, MM Giusti, LA Kresty, ML Failla. 2007. Inhibition of proliferation of human bladder epithelial cancer cell lines by anthocyanin-rich fraction from black raspberry. Accepted for presentation at the Institute of Food Technologists Annual Meeting and Food EXPO. Chicago, IL, July.
  • GIUSTI, MM. 2007. Effect of glycosylation and acylation on the chemoprotective effects of anthocyanins. American Chemical Society, Division of Agricultural & Food Chemistry. 234th National Meeting & Exposition, Boston, MA August 19-23