AGFD 1

Chemistry and analysis of polyphenols in food and human samples

Elke Richling1, richling@chemie.uni-kl.de, Markus Schantz2, Dolores Mueller3, Denise Scherbl2, Thomas Erk3, Hannah Bergmann2. (1) Food Chemistry, Universtity of Kaiserslautern, Kaiserslautern, Germany (2) Division of Food Chemistry and Toxicology, Molecular Nutrition, University of Kaiserslautern, Kaiserslautern, Germany

Polyphenols are a class of phytonutrients, which are present in many plants. Especially food such as fruits and vegetables are a rich source of these compounds. Polyphenols act as antioxidants and seem to prevent the human body from degenerative diseases such as cardiovascular diseases or cancer. This presentation will provide an overview about the analytical methodologies to determine various polyphenol classes in food e.g. apples, coffee, and red berries. Additionally, information about methods to investigate the metabolism and body distribution of these classes of polyphenols in human samples after their consumption via food or extracts will be provided.

AGFD 2

Anthocyanins in the blood: Where are they going and how do they get there?

Joe A. Vinson1, joe.vinson@scranton.edu, Ibtehaj Alshdoukhi2. (1) Chemisry, University of Scranton, Dalton, Pennsylvania, United States (2) Department of Chemistry, University of Scranton, Scranton, Pennsylvania, United States

Consumption of a wide variety of berries and their juices have been shown to have beneficial effects. Anthocyanins are the major colored compounds in berries and they have been found in human plasma and urine although bioavailability is very low. They have been found in multiple organs in animal studies including the brain. Little is known about anthocyanins (AC) with respect to their transport in the blood to the cells where their effects will manifested. We have examined three AC in the chloride form; cyanidin (C), malvidin (M) and malvidin-3-glucose (MG) spiked at 20 µM in our ex vivo pig blood model. This model with selective extractions provides two plasma AC fractions (separated as HDL + albumin, and LDL + VLDL), intracellular fraction and surface-bound AC fraction (see figure below). The most hydrophilic compound MG had the greatest % recovered in the plasma and C and M had similar % plasma. All compounds had the majority of the plasma anthocyanin in the LDL+ VLDL compartment rather than in albumin the most prevalent plasma protein. M, the most lipophilic AC, had the greatest intracellular %, and C had the greatest surface-bound %. The chemical structure of the AC determines the compartmentalization in this blood/cell model.

AGFD 3

Understanding factors that influence the bioavailability and kinetic profile of strawberry anthocyanins: A focus on meal timing and fasted-fed state status

Aman Sandhu1, asandhu@iit.edu, Indika Edirisinghe1, Britt Burton-Freeman2. (1) Dept. of Food Science and Nutrition, Illinois Institute of Technology, Summit Argo, Illinois, United States (2) Institute for Food Safety and Health, Bedford Park, Illinois, United States

Strawberries are a rich source of polyphenols, especially anthocyanins, which not only impart red color to the strawberries but also provide health promoting effects for chronic disease prevention. However, if their health promoting action is dependent on bioavailability and or achieving a certain blood concentration or exposure time to impart biological activity then understanding the factors that influence these variables are critical. The bioavailability of anthocyanins is very low and previous work has indicated that food matrix will impact kinetic absorption and clearance profiles but not bioavailability. Foods are consumed under varying conditions throughout a day. The impact of when dietary anthocyanins are consumed in a day (under fasting conditions, with a meal or for a snack between meals) has not been studied. This presentation will provide a brief overview of known/published factors influencing strawberry anthocyanin bioavailability, their metabolites and kinetic profile and provide new data expanding the field to understand the role of meal timing and fasted-fed state status. The relevance of these profiles to metabolic and inflammatory responses will also be discussed.

AGFD 4

Absorption, distribution, metabolism, and excretion of orange juice flavanones in humans

Alan Crozier, alan.crozier@glasgow.ac.uk. University of Glasgow, Glasgow, United Kingdom

A diet rich in fruit and vegetables is a major factor in the maintenance of good health and in part, these effects are mediated plant derived (poly)phenolic compounds. Understanding the fate of these compounds within the body is key to elucidating their mode of action in reducing the incidence of chronic diseases. This talk will focus on the absorption, distribution, metabolism and excretion (ADME) of flavanones after the ingestion of orange juice by healthy human volunteers with emphasis on events occurring in the proximal and distal gastrointestinal tract.

AGFD 5

Cocoa flavanols – chemical nature, analysis, and fate during processing

Xianli Wu, xwu@hersheys.com. Hershey Company, Hershey, Pennsylvania, United States

Cocoa beans are rich in flavanols, which are major bioactive compounds in cocoa and chocolates. The link between flavanols and the health benefits of the consumption of cocoa/chocolate has been supported by numerous epidemiological and clinical studies. However, due to the extreme complexity of the chemical structures and rather confusing nomenclature, it is still a challenging for many researchers to fully understand the chemical natures and the analysis of this important group of compounds. The first objective of this presentation is to provide an overview of the chemical nature of cocoa flavanols, and to clarify the terminology and nomenclature. Some newly discovered A-type procyanidins are also discussed. Based on the understanding of the chemical structures, different analytical strategies of cocoa flavanols are discussed and compared, with the focus on the most commonly used HPLC-MS-FLD method. In addition, cocoa flavanols are known to be degraded during food processing. Flavanols are lost in almost every single step of chocolate making, from bean harvest to finished product, including fermentation, drying, roasting and Dutching (alkalization). In order to better understand the health benefits of cocoa flavanols, it is critical to understand their fate and structure changes during processing. But the underlying mechanisms of the flavanol degradation in cocoa beans through processing are still poorly understood. The last objective of this presentation is to summarize the current knowledge and provide some insights regarding this topic and for future studies.

AGFD 6

Comparison of polyphenolics and secoiridoids in California-style black ripe olives and dry salt-cured olives using UHPLC/MS/MS

Jerry Zweigenbaum1, j_zweigenbaum@agilent.com, Eleni Melliou3, Alyson E. Mitchell2. (1) Agilent Technologies, Wilmington, Delaware, United States (2) Food Science Technology, UC Davis, Davis, California, United States (3) 1Department of Pharmacognosy and Natural Products Chemistry, University of Athens, Greece, Greece

Abstract: Polyphenolics and secoiridoids have been associated with positive health effects and are components of the olive fruit. Their distribution in processed olives is depending on variety and the type of processing. This study examines the concentrations of 12 of these compounds in three types of black olives: California-style black ripe processed Manzanilla olives and two dry salt-cured olive varieties, Mission from California and Throuba Thassos from Greece. An ultrahigh pressure liquid chromatography/tandem mass spectrometry (UHPLC/MS/MS) method using a triple quadrupole mass spectrometer was used to detect and quantify the targeted polyphenols and secoiridoids. The methodology used including sample preparation along with the results of the distribution of these compounds will be described in this presentation.

AGFD 7

Phenolic acid profiles of Fuji, Golden Delicious, Granny Smith, and Pink Lady apples

Alyson E. Mitchell, aemitchell@ucdavis.edu. Food Science Technology, UC Davis, Davis, California, United States

Apples (Malus Domestica) are the second most consumed fruit in the United States and are the primary source of phenolic acids in the Western diet. Herein, phenolic acid profiles were measured in the flesh and peels of 4 commercially important varieties of apples using UHPLC-(ESI)QTOF MS/MS. Twenty four phenolic acids were identified and include: 9 hydroxybenzoic acid derivatives, 8 hydroxycinnamic acid derivatives, 6 hydroxyphenylacetic acid derivatives, and 1 hydoxyphenylpropanoic acid derivative. Of these, 11 were tentatively identified for the first time in apples and include: two methyl gallate isomers, two ethyl gallate isomers, hydroxy phenyl acetic acid, three phenylacetic acid isomers, 3-(4-hydroxyphenyl)propionic acid, and two dimethoxyphenylacetic acid isomers. Significant varietal differences (p < 0.05) were observed and the levels of total phenolic acids were higher in the flesh as compared to peel (dry weight basis) in all varieties examined. Apple peel contained significantly higher concentrations of 3-(4-hydroxyphenyl) propionic acid, vanillic acid, p-coumaric acid, protocatechuic acid, benzoic acid, hydroxy phenyl acetic acid, rosmarinic acid, and t-ferulic acid than apple flesh. The major phenolic acids identified include chlorogenic acid, caffeic acid, p-coumaric acid, and protocatechuic acid regardless of variety. Chlorogenic acid and caffeic acid predominated. Golden Delicious and Fuji apples contained the highest concentrations of total phenolic acids in the flesh (1,129.5 ± 45.3 and 1,065.9 ± 3.3 mg/kg dry weight (DW), respectively) whereas Fuji contained the highest concentration in the peel (736.7 ± 22.1 mg/kg DW) as compared to other varieties. Levels of chlorogenic and especially caffeic acid were lowest in the Granny Smith apples.

AGFD 8

Influence of different deposition forms of carotenoids in plant foods on their bioavailability

Ralf Schweiggert1, rswg@uni-hohenheim.de, Reinhold Carle1,2. (1) Institute of Food Science and Biotechnology, Chair of Plant Food Technology and Analysis, University of Hohenheim, Stuttgart, Deutschland, Germany (2) Biological Science Department, King Abdulaziz University, Jeddah, Saudi Arabia

Carotenoids are responsible for the yellow, orange, and red hues of numerous fruits and vegetables. They similarly occur at high concentrations in green plant foods, although their bright colors are being masked by chlorophylls. The most prominent bioactivity of many dietary carotenoids is their post-prandial conversion into vitamin A. In addition, a carotenoid-rich diet has been associated with numerous further health benefits, e.g., the prevention of specific cancer types and other chronic diseases. In order to exert these health effects, carotenoids need to be liberated from the food matrix and, subsequently, be absorbed to the human blood stream. Factors governing their liberation have been discussed comprehensively and, in particular, their physical deposition form within the plant’s chromoplasts appears of major importance. Thus, a complete overview of the

natural array of these deposition forms is of crucial importance for estimating or even technologically modulating their bioavailability. Well-known examples are the large solid-crystalline aggregates in carrots and tomatoes and the protein-complexed carotenoids in green plant foods. Being scarcely reported previously, truly lipid-dissolved forms in plant foods will be presented, such as, e.g., tangerine tomatoes and peach palm fruit (Bactris gasipaes Kunth). In addition, liquid-crystalline forms within so-called tubular chromoplasts will be discussed, although they were often overlooked in the past despite their presence in numerous fruits (e.g., papaya, mango, and bell pepper). Therefore, latest knowledge about the varying deposition forms and current findings from related human intervention studies will be presented. Ultimately, an overview of technological and dietary strategies to modulate carotenoid intake from food sources with different deposition forms will be provided.

AGFD 9

Application of different analytical systems for the characterization of food bioactives

Daniele Giuffrida2, dgiuffrida@unime.it, Francesco Cacciola2, Margita Utczas3, Marco Beccaria3, Paola Donato2, Paola Dugo3,4, Luigi Mondello1,4. (1) “Scienze del Farmaco e Prodotti per la Salute” , Viale Annunziata, 98168, University of Messina, Italy, Messina, Italy (2) "Scienze dell'Ambiente, della Sicurezza, del Territorio, degli Alimenti e della Salute", Viale F. Stagno d'Alcontres 31, 98166 , University of Messina, Italy, Messina, Italy (3) “Scienze del Farmaco e Prodotti per la Salute”, Viale Annunziata, 98168 – , University of Messina, Italy, Messina, Italy (4) University Campus Bio-Medico, Via Álvaro del Portillo, 21 - 00128 , University Campus Bio-Medico of Rome, Rome, Italy

Bioactive molecules are naturally occurring in plants and animal products, with a variety of physiological functions for promoting general body development and disease prevention. Food fractions with identified bioactives with proven health benefits can be incorporated into nutraceuticals and supplements. Functional foods are playing an increasingly important role in our community’s health and wellbeing, and scientists are developing the technologies to incorporate bioactives into foods so that their beneficial properties are maximised without reducing the sensory or taste attributes and quality of foods. Carotenoids are an important kind of natural pigment that can be widely found in plants and plant-derived food and products, with health-promoting properties. One-dimensional liquid chromatographic techniques are widely applied to the analysis of real world samples in several fields. However, such separation methods sometimes do not provide sufficient resolving power for the separation of target components in many complex samples. Comprehensive two-dimensional liquid chromatography (LC×LC) is a relatively novel technique involving two independent LC separation processes with orthogonal selectivities. In this presentation, the application of different chromatographic systems for the characterization of carotenoids in food products, both from the tropical and the Mediterranean areas will be shown.

AGFD 10

Effects of thermal and enzymatic treatment on polyphenol profiles during bilberry juice production

Fabian Weber1, Fabian.Weber@uni-bonn.de, Peter Heffels1, Franziska Bührle1, Daniela Kaumans1, Andreas Schieber2. (1) Department of Nutritional and Food Sciences - Food Technology and Food Biotechnology, University of Bonn, Bonn, Germany (2) Dept of Nutrition Food Science, University of Bonn, Bonn, Germany

Berry fruits have experienced a growing market share, since they contain large amounts of secondary plant metabolites, which are associated with numerous health promoting effects. Bilberries (Vaccinium myrtillus L.) contain large quantities of polyphenols, in particular anthocyanins. During fruit juice production, different steps are conducted to increase the overall yield and also to enhance the extraction of these beneficial compounds. However, processing inevitably leads to changes or even losses of polyphenols. We determined the influence of thermal and enzymatic treatment on the content and the profile of polyphenols in bilberry juice. High temperatures are widely applied to inactivate the polyphenol oxidases and thus prevent browning. In our experiments, temperatures below 80 °C proved to be insufficient to inhibit enzymatic oxidations, whereas higher temperatures led to a considerable increase in anthocyanin content. The polyphenol content as determined by the Folin Ciocalteau assay was not adversely affected. Noteworthy, the increase in pigments was different for the various anthocyanins. We also examined the effects of several commercial pectinolytic enzymes on the polyphenol content and profiles. Since these enzyme preparations usually exhibit a broad range of side activities, we assessed these activities in model solutions as well as in real fruit juice samples. Contrary to our expectations, we did not observe any correlation of the beta-glucosidase activity and the decrease in anthocyanin glucosides. The results provide valuable insights in the effects of thermal and enzymatic treatment on the polyphenol profiles of bilberry fruit juices and may help to optimize processing parameters in berry juice production.

AGFD 11

Biological effects of anthocyanins from fruits

Elke Richling, richling@chemie.uni-kl.de, Markus Schantz, Matthias Baum, Tamara Bakuradze, Dolores Mueller. Food Chemistry, Universtity of Kaiserslautern, Kaiserslautern, Germany

Anthocyanins are responsible for attractive color of red fruits and are attributed with beneficial effects. Especially bilberries have a broad spectrum of anthocyanins. In our recent in vivo study with human volunteers, we were able to show that (1) the stabilization of anthocyanins leads to a higher systemic bioavailability, (2) colon as adsorption site contributed significantly to the bioavailability of these compounds and (3) the anthocyanin degradation products might be responsible for the biological effects

observed. Recently, we could show that the consumption of anthocyanin rich bilberry extract and fruit juice rich in anthocyanins reduced the DNA damages in white blood cells (WBC) measured by the so called comet assay. This might be due to the induction of the ARE/Nrf2 pathway leading to an increased formation of antioxidative enzymes. The studies were supported by the German Ministry of Economics and Technology (via AiF) and the FEI (Forschungskreis der Ernaehrungsindustrie e.V., Bonn, Germany), cluster project no. 15614 N, and project no. 17039N/18068 N.

AGFD 12

Transport and uptake of anthocyanins in gastric tissue and their effect on the gastric inflammatory response: Developing an in vitro model using the NCI-N87 gastric cell line

Allison Atnip1, allison.atnip@gmail.com, M. Monica Giusti2, Joshua Bomser 3. (1) Rapid Prototyping, Abbott Nutrition, Columbus, Ohio, United States (2) Food Science and Technology, The Ohio State University, Columbus, Ohio, United States (3) Human Nutrition, The Ohio State Universtiy, Columbus, Ohio, United States

Anthocyanins are polyphenolic pigment compounds found in many commonly consumed fruits, which also have potential health benefits, including antioxidant and anti-inflammatory capabilities. In order to understand these benefits, the behavior of anthocyanins throughout the GI tract must be elucidated. Anthocyanins appear rapidly in plasma after ingestion, suggesting a role of the stomach on the in vivo activity of anthocyanins. Effects of the gastric environment and chemical structure on the transport and uptake of anthocyanins in the stomach have not been well characterized. Physiological pH used in cell culture is not ideal to study the acidic environment of the stomach, however the NCI-N87 line is an acid-stable model of the gastric epithelium, and here is investigated as a model to study transport and uptake of anthocyanins in the stomach. Cells formed a stable monolayer (18-32 days) post-confluence. The pHs of apical chambers were adjusted to 3.0, 5.0, or 7.4, with basolateral pH of 7.4, with minimal effects on monolayer integrity. Transport and uptake of anthocyanins from chokeberry was dependent on time, initial concentration, and apical pH. Anthocyanin recovery was higher at apical pH 3.0 than 5.0 and 7.4. Over time and increasing apical concentrations, the basolateral concentration of anthocyanins increased. The rate of transport increased over time, suggesting that the transport was not by passive diffusion alone. Apical pH of 3.0 showed higher transport, and pH 5.0 the lowest. There was an effect of anthocyanin structure on the transport and uptake from chokeberry (varying monoglycoside attachments to cyanidin) and red grape (6 anthocyanins attached to monoglucoside). Chokeberry anthocyanins inhibited IL-8 production in response to IL-1β induced inflammation of NCI-N87 cells, suggesting anti-inflammatory activity in the stomach, in this acute model. This work is further evidence that the stomach may play a significant role in the in vivo fate of anthocyanins.

AGFD 13

Optimized and validated method for the characterization and quantification of bioactive ellagitannins in pomegranate and other fruits and nuts

Francisco Tomas-Barberan1, fatomas@cebas.csic.es, Rocio Garcia-Villalba1, Kjersti Aaby2, Tuuli Koivumäki3, Marina Heinonen3, Ebru Pelvan4, Cesarettin Alasalvar4, Griet Jacobs5, Shikha Saha6, Juan Carlos Espin1, Paul Kroon6. (1) CEBAS-CSIC, Murcia, Spain (2) NOFIMA, Oslo, Norway (3) Food Chemistry, Helsinki University, Helsinki, Finland (4) TUBITAK, Istambul, Turkey (5) VITO, Mol, Belgium (6) IFR, Norwich, United Kingdom

Pomegranate bioactivity is mainly associated to its high content in polyphenols and particularly to ellagitannins. Despite the number of studies aiming to the evaluation of the health benefits of pomegranate polyphenols and their mechanisms of action, reliable data on their contents in pomegranates and derived extracts and food products is lacking, as it is usually underestimated due to their complexity, diversity and lack of standards. Here we describe a new method for the analysis of the ellagitannin content based on the quantification of the hydrolysis products that include ellagic acid, gallic acid, valoneic acid dilactone and gallagic acid dilactone, after acid hydrolysis of pomegranate samples. Method optimization was developed using pomegranate peel extracts and finally direct acid hydrolysis of the freeze dried samples with 4 M HCL in water at 90 ºC for 24h and additional extraction of the pellet with DMSO/MeOH (50/50, v/v) was selected. The method was validated with an intra and inter-laboratory trial, showing high repeatability and reproducibility across six European laboratories with relative standard deviations (RSD) less than 15 %. The study shows that the previously published methods for quantification of pomegranate polyphenols have underestimated the ellagitannin content, and have neglected the occurrence of ellagitannin C-glycosides and ellagitannin oligomers. Then its applicability was demonstrated in different pomegranate parts including husk, peel, arils and juices. The method was also adapted to the analysis of ellagitannins in other fruits and nuts including strawberry, raspberry, blackberry and walnuts. The results show that ellagitannins are fruit bioactives that can be quantitatively more relevant than previously expected. Acknowledgments: This work has been funded by the European Commission FP-7, Project BACCHUS (Grant Agreement, 312090).

AGFD 14

Olive oil authenticity and adulteration: Analytical tools and standards

Richard Cantrill, richard.cantrill@aocs.org. AOCS, Urbana, Illinois, United States

Olive oil has been listed by the European Union as one of the most regularly misbranded, misidentified and misassigned foods on the market. This is not a new problem, since traceability systems were designed to combat fraud in Roman times. A number of national and international standards have been developed in the last 50

years to address olive oil quality and authenticity. The International Olive Council (IOC), based in Madrid, has been a leading organization in the development and definition of olive oil grades and suitable testing methods to support specifications. More recently, similar standards have been adopted by the European Union and Codex Alimentarius. The United States Department of Agriculture also published its own standard modeled on the Codex Alimentarius standard but allowing for variations in the characteristics of domestic production. In the last year, the Olive Oil Commission of California has also published a standard for olive oil that covers oils produced in that state. Again, this standard also makes allowances for variations in composition found in domestic production and also included methods of analysis not included in other standards. Methods address quality parameters, such as free fattyacid, peroxide value, UV spectral properites and authenticity parameters such as fatty acid composition, sterol composition, triglyceride structure. Shelf-life and freshness may also be addressed by diacylglycerol composition and chlorophyll breakdown products.

AGFD 15

USP skim milk powder advisory group: The development of a toolbox of methods to detect food adulteration

Robert L. Magaletta1, bob.magaletta@mdlz.com, Jeffrey C. Moore2. (1) Mondelez International, East Hanover, New Jersey, United States (2) US Pharmacopeia, Silver Spring, Maryland, United States

Economically-motivated food adulteration is an important issue facing food producers. A major contamination incident in a particular food product or ingredient not only affects the company(s) directly involved, but undermines the public’s faith in all similar food materials, thus affecting all companies who use those ingredients. Those who adulterate food ingredients will use whatever substance is on hand that will do the job, and will move on to a different adulterant once they find out that the authorities have a method to detect the current adulterant. This underscores the need for a non-targeted approach to screening and detection of adulterants, a method that can answer the question, "is the ingredient the same as always or is it somehow different". Food safety is widely seen as a pre-competitive endeavor, and an independent standards-setting organization such as the United States Pharmacopoeia (USP) is an ideal forum for industry-wide cooperation to develop the tools needed to successfully detect economically-motivated adulteration. Additionally, it is very difficult for a single company to procure a wide-enough variety of “good” ingredient samples and potential adulterants to build a database representative of the natural variation of an ingredient, which is essential for a non-targeted approach to detection; a reputable and neutral third party such as USP can however be trusted to take inputs from many companies in confidentiality and develop the methods and databases essential to the detection of economic adulteration. This talk will discuss the progress of the USP Skim Milk Powder Advisory Group in developing a toolbox of methods suitable for the rapid detection of economically-

motivated food adulteration, using a variety of techniques ranging from wet chemistry to sophisticated instrumentation.

AGFD 16

Development of field screening methods using surface enhanced Raman spectroscopy (SERS)

Laura C. Pogue1, laura.pogue@fda.hhs.gov, Naimish P. Sardesai2, Betsy J. Yakes3, Steve Barcelo4, Mineo Yamakawa4, Anita Rogacs4, Zhiyong Li4, Abdur-Rafay Shareef2. (1) Division of Pharmaceutical Analysis, US Food & Drug Administration, St. Louis, Missouri, United States (2) Winchester Engineering Analytical Center, US Food & Drug Administration, Winchester, Massachusetts, United States (3) Center for Food Safety and Applied Nutrition, US Food & Drug Administration, College Park, Maryland, United States (4) HP Labs, Palo Alto, California, United States

The FDA is responsible for managing a complex global food and pharmaceutical supply chain with somewhat limited resources. Ideally, a FDA field investigator could quickly screen large numbers of collected samples and dedicated laboratories would receive “flagged” samples for confirmatory analysis, thus greatly increasing coverage of the U.S. food and drug supply. This new paradigm has recently been implemented and is currently being evaluated. To add to the Agency’s screening repertoire, we are now investigating the use of a portable Raman system utilizing pre-fabricated surface enhanced Raman spectroscopy (SERS) substrates for the detection of nitrogen containing species in both food and pharmaceutical products. Currently, the protein content of some foods and pharmaceutical excipients, as measured via nitrogen determination, may be artificially enhanced by the addition of nitrogen rich, low molecular weight compounds. Recent events, including the use of melamine for adulteration of Chinese infant formula and milk in 2008, highlight the need for screening a multitude of consumer products. As a rapid and highly sensitive technique, surface enhanced Raman spectroscopy is positioned as an ideal method for field applications. We will present our initial method development focused on milk, milk products, lactose and bulk protein products in order to detect the presence of compounds potentially used in economic adulteration. We will describe sample preparation which utilizes size exclusion chromatography to extract nitrogen containing compounds followed by the deposition of the sample onto the SERS substrate prior to insertion into the Raman spectrometer. Both melamine and dicyandiamide have been successfully isolated from spiked milk and infant formula samples. Future method development activities will broaden to include additional FDA regulated products.

AGFD 17

Meat fraud and speciation: From vulnerability assessment to analytical methods

Geoffrey Cottenet, geoffrey.cottenet@rdls.nestle.com. Food Safety & Quality, Nestlé Research Center, Lausanne, Switzerland

Europe was faced in 2013 with a case of food fraud with major coverage and lost of consumer confidence: beef adulterated with horse meat. The impact for businesses can be enormous and in some cases lead to bankruptcy. Fighting against food fraud and adulteration is an endless activity which requires the contribution from all the value chain including procurement, regulatory, specification management, analytical sciences. Vulnerability assessment of meat raw materials takes into consideration various parameters such as price variation of the commodity, previous cases of adulteration and level of processing of the raw material purchased. Meat powder is indeed more vulnerable to food fraud than real meat pieces. In order to identify meat species and detect potential meat substitution, analytical methods are needed. Considering also the latest Chinese cases of meat substitution where fox and rat meat were detected instead of donkey and lamb, there is a strong need to expand the range of detectable species. Althoug DNA sequencing is considered as a reference method to identify (meat) species, its capability to detect potential mixtures is limited. Another approach based on DNA macro-array was evaluated and allowed a simultaneous screening of 32 meat species with a detection of meat mixtures ≤ 1% (m/m) as required by the European legislation. Easy to use, fast, and cost efficient, this Meat LCD Array can be easily applied to monitor meat raw materials. Both DNA Barcoding and Meat LCD Array are DNA-based methods which allow the specific identification of meat species as required by food labelling regulations and maintain consumer trust.

AGFD 18

DNA-based species identification of seafood

Anne Eischeid2, Anne.Eischeid@fda.hhs.gov, Sarah Stadig 2, Sara Handy3, Fred S. Fry1, Jonathan Deeds2. (1) FDA HFS-707, FDA, College Park, Maryland, United States (2) Center for Food Safety and Applied Nutrition , U.S. Food and Drug Administration, College Park, Maryland, United States (3) US FDA, College Park, Maryland, United States

Seafood is among the most highly traded commodities in the world, and it is an increasingly important protein source in the U.S. diet. Proper labeling of seafood is necessary not only to prevent fraud, but also to address risks inherent to particular species, such as chemical and microbial contamination or the presence of allergens. Due to the incredible diversity of seafood products now available to U.S. consumers, accurate labeling requires reliable methods of species identification. FDA has recently adopted a DNA sequencing based method for species identification of seafood known as DNA barcoding. DNA barcoding involves amplification and sequencing of a short standardized segment of an animal’s genome with identification based on comparison to a library of reference sequences. DNA barcoding has proven to be a highly useful

tool in the regulatory arena for several reasons. The method is feasible, practical, and scientifically rigorous. Current DNA barcoding work builds on previous seafood identification work carried out at FDA. Information generated through FDA’s work on DNA barcoding has been disseminated via the public release of reference sequences, procedures, and other information through FDA’s website, and also through the scientific literature. FDA’s barcoding work has led to collaborations with academia and other government agencies and has resulted in initiatives to educate the industry.

AGFD 19

Honey adulteration: Methods currently applied in the routine control of commercial samples, analytical challenges, legal and regulatory aspects

Lutz Elflein, lutz.elflein@intertek.com. Intertek Food Services, Bremen, Germany, Bremen, Bremen, Germany

Honey trade is a global business, with high import rates into the EU and USA, and high export rates from Asia, South and Middle America, South and Eastern Europe. Honey is a natural and mostly untreated food produced by bees. The industrial processing is usually minimized to liquefaction, homogenization and filtration of raw honeys prior to packaging for retail. Therefore, the consumers esteem honey as a natural, pure authentic, healthy and valuable product. There are numerous honey varieties of different botanical and geographical origins available on the international market. Additionally, there are certain specialty honeys with particular properties like Manuka honey from New Zealand with its characteristic antibacterial activity. Honey is not only sold as a mono product, e.g. as breakfast spread or as sweetener for tea, but is also of major importance for the food industry as an ingredient to substitute corn, cane or beet sugar. As consumers become more ecosensitive and health-conscious the demand for natural and organic food is growing and people are willing to pay more money for that compared to conventional food. For this reason, it is very important to control raw and finished food products labeled with such properties in order to ensure their quality and purity and protect consumers from fraud. Honey is listed as one of the top ten food items particularly prone to fraud, because the demand is high and production volumes are limited. Furthermore, legal regulations for honey and food surveillance measures are not internationally harmonized, so that there are enough chances to gain profit with adulterated honey. This presentation will give an overview of the existing and currently applied analytical methods to detect honey adulteration, e.g. isotopic screening methods (EA/LC-IRMS) or specific methods to detect marker molecules of sugar syrups in honey, e.g. by LC-MS/MS, ICP-MS, HPLC or enzymatic methods, discuss their benefits and limitations, point out the future analytical challenges (e.g. NMR) and address particular problems with the analytical assessment for international trade and official food control.

AGFD 20

Is volunteer stratification necessary in clinical trials with phenolic phytochemicals?

Francisco Tomas-Barberan, fatomas@cebas.csic.es. CEBAS-CSIC, Murcia, Spain

Inter-individual variability in the metabolism and absorption of polyphenols can explain differences in the biological responsiveness observed in clinical trials that often reduces their statistical significance. In these trials, however, some volunteers respond to the treatment, while other can be labelled as non-responders. Several determinants of this interindividual variability have been identified, and among them gut microbiota composition has been considered a relevant player. Gut microbiota metabolism of polyphenols leads to metabolites that can modulate the biological effects and the bioavailability of the original compounds found in food. Depending on the gut microbial composition volunteers can be stratified in different phenotypes or different bioavailability/excretion levels. This has already been shown for soybean isoflavones, orange juice flavanones, high bran cereal lignans, and pomegranate ellagitannins. Correlations between phenotypes and health/biological effects have been demonstrated in some cases but other still wait to be studied. Volunteer stratification regarding gut microbiota metabolism needs to be taken into consideration in clinical trials with polyphenol-rich foods and nutraceuticals in order to understand the biological response observed.

AGFD 21

Predicting the mechanism of anthocyanin-induced insulin sensitization with molecular modeling

David Minh, dminh@iit.edu. Chemistry Division, Biological & Chemical Sciences , Illinois Institute of Technology, Chicago, Illinois, United States

The health benefits of flavonoids have commonly been attributed to their antioxidant properties. Anthocyanins, a subset of flavonoids that are particularly common in berries, appear to have more specific effects on insulin sensitivity. We used molecular docking to predict the binding pose and affinity of polyphenols to a series of prospective molecular targets. Among the peroxisome proliferator-activated receptors (PPARs), we predict gamma to be selective for anthocyanins and alpha and delta to not be. Estrogen receptors, the estrogen-related receptor alpha, and the AMP-activated protein kinase are also predicted to be selective for anthocyanins. However, the estrogen-related receptor gamma, glucose transporter type 4, and alpha-glucosidase are not predicted to be selective. We also plan to present structural rationales for selectivity and updated results based on a novel binding affinity estimation method.

AGFD 22

Grape seed extract authentication

Mark A. Kelm, mark.kelm@cbrands.com, Steve Kupina, Anil Shrikhande. R&D, Constellation Brands, Fresno, California, United States

As the popularity of grape seed extract (GSE) as a dietary supplement for health benefits has increased, so has the number of marketers of said products. Similar to other functional ingredients, GSEs are no exception when it comes to deviations in quality. As such, we endeavored to better delineate GSE quality from those producers that rely on the introduction of non-grape components. Ordinarily, B-type procyanidins are reported in GSEs, with high levels of catechin and epicatechin. However, recent studies indicated adulteration of grape seed extracts with A-type procyanidins which typify peanut skin extracts. Herein, we investigated the chemistry of commercially available GSEs for the presence of adulterants by comparing diol HPLC and MS data with a sample of confirmed source. In several GSEs, propelargonidins were characterized. In another extract, A-type procyanidin dimers and trimers were characterized; suggesting adulteration by peanut skins. Several grape seed extracts were found to have lower than typical levels of catechin and epicatechin. This work is currently ongoing.

AGFD 23

Proanthocyanidins and gut barrier function

Jess D. Reed1, jdreed@wisc.edu, Christian G. Krueger2. (1) Univ of Wisconsin Madison, Madison, Wisconsin, United States (2) University Wisconsin Madison, Madison, Wisconsin, United States

The central hypothesis of our research is that the effects of food proanthocyanidins (PAC) on health are mediated through interactions with gut digesta and microbiota, gut epithelium, and gut associated lymphoid tissue. Therefore, the low “bioavailability” of PAC is not relevant to their beneficial role in the diet. Two examples of our research to test this hypothesis will be discussed; 1) effects of cranberry PAC on the invasion of gut epithelial cells by intestinal and extra intestinal pathogenic bacteria, and 2) effects of cranberry PAC on gut barrier function. Cranberry PAC inhibit the invasion of intestinal epithelial cells by extra intestinal pathogenic Escherichia coli and Salmonella enterica serovar agona in a dose dependent manner and at concentrations that are obtainable in the gut lumen at normal levels of consumption of cranberry products. Cranberry PAC attenuate the negative effects of elemental enteral nutrition on gut barrier function in mice and restores immune and non-immune function in this model of gut barrier dysfunction. The implications of these results for the effects of dietary PAC on human health will be presented.

AGFD 24

Anthocyanin metabolism and transport across the blood brain barrier

Paul E. Milbury, paul.milbury@tufts.edu. Gerald J. and Dorothy R. Friedman School of Nutrition Science and Policy, Tufts University, Ipswich, Massachusetts, United States

Evidence is growing which suggests anthocyanins play beneficial roles in reversing age related cognitive impairment and protect against neurodegenerative disorders. The plausible mechanisms of anthocyanin bioactivity in brain tissue are dependent on their bioavailability to the brain. A bioavailability study where pigs were fed 2% whole freeze-dried, powdered blueberry in the diet for 8 weeks showed anthocyanin and anthocyanin glucuronides can pass the blood brain barrier. Individual anthocyanins and their glucuronides were found in all portions of the brain in the ranges of femtomoles per gram of fresh weight of tissue at 18 h postprandial, after anthocyanins had been removed from the blood by xenobiotic metabolism. At these concentrations plausible mechanism of neuroprotective action are more likely via modulation of signal transduction processes and/or gene expression in brain tissue rather than by direct antioxidant radical quenching. Anthocyanin stability, transport into brain tissue, and potential modes of action will be discussed.

AGFD 25

Bioactive compounds for cancer prevention and health aging

Rui H. Liu, rl23@cornell.edu. Food Science, Cornell University, Ithaca, New York, United States

More evidence suggests that a healthy eating pattern with increased consumption of fruits, vegetables, whole grains, and other plant-based foods has been negatively correlated to the risk of developing chronic diseases, such as heart disease, cancer, stroke, diabetes, Alzheimer’s disease, cataracts and age-related diseases. Phytochemicals in fruits, vegetables, and whole grains have been suggested to be responsible for their health benefits. Our group proposed that the major health effects of a diet rich in fruits, vegetables, and whole grains is attributed to the additive and synergistic interactions of bioactive compounds and other food components in whole foods by targeting multiple signal transduction pathways. This presentation will cover our current research on fruit phytochemicals for cancer prevention and health aging, and the molecular targets of bioactive food components in regulating signal transduction pathways.

AGFD 26

Citrus flavonones decreases oxidative stress in the liver and blood serum caused by high-fat diet feeding in C57BL/6J mice

Paula S. Ferreira1, paullasf@gmail.com, Ana Lucia M. Nasser1, Danielle Gonçalves1, Luis Carlos Spolidorio3, John A. Manthey2, Thais B. Cesar1. (1) Food and Nutrition, Sao Paulo State University, Araraquara, Sao Paulo, Brazil (2) USDA ARS, Fort Pierce,

Florida, United States (3) Physiology and Pathology, Sao Paulo State University, Araraquara, Sao Paulo, Brazil

High-fat diet and adipose tissue accumulation commonly leads to an increased oxidative stress and inflammation, contributing to the development of chronic diseases observed in obesity, such as diabetes mellitus and cardiovascular diseases. Citrus flavonoids have biological properties capable of attenuating oxidative stress, protecting against metabolic disorders resulting from obesity. In the present work we studied the effect of the citrus flavonones hesperidin, eriocitrin and eriodictyol (100, 200 and 200 mg/kg body weight, respectively) over oxidative stress in the blood serum and liver of C57BL/6J male mice fed high-fat diet (45% kcal from saturated fat) for 30 days. Mice fed high-fat diet showed visceral fat accumulation and increased TBARS levels in the blood serum (80%) and in the liver (57%), while hesperidin, eriocitrin and eriodictyol increased the serum total antioxidant capacity (7%, 9% and 6%, respectively) and decreased the TBARS levels in the liver (50 %, 57 % and 64 %, respectively). Eriocitrin and eriodictyol also decreased the blood serum TBARS levels in 48% and 47%, respectively. In addition, the liver fat accumulation and damaged hepatocytes (observed by histology analyses) was efficiently reduced by hesperidin and eriodictyol, and the serum levels of LDL-C was reduced in 28% by hesperidin. These results showed that hesperidin, eriocitrin and eriodictiol have protective effect against oxidative stress caused by high-fat diet feeding in mice, as indicated by reduced liver damage, liver fat accumulation, and TBARS levels in the liver or serum, making them potential candidates to prevent tissue damage and the development of metabolic dysfunction associated to chronic diseases caused by obesity.

AGFD 27

Using untargeted metabolomics to profile tomato products intended for clinical trials

Morgan J. Cichon, mjcichon@verizon.net, Ken M. Riedl, Steven Schwartz. Food Science & Technology, The Ohio State University, Columbus, Ohio, United States

Research has shown a correlation between increased consumption of tomato products and a decreased incidence of certain chronic diseases, such as prostate cancer. However, the mechanism behind this protective effect remains unclear. The carotenoid lycopene is believed to be one of the bioactive compounds present in tomatoes. The tangerine tomato has a unique carotenoid profile compared to the traditional red tomato, which translates into a difference in bioavailability and potentially a difference in biological activity between the two tomato varieties. For this reason, red and tangerine tomatoes were grown and processed into tomato juices at the Ohio State University for use in clinical trials with prostate cancer patients. In order to better understand any clinical effects of these juices, it is important to first study how they differ in their chemical composition beyond the known carotenoid differences. Untargeted LC/MS-based metabolomics was used to chemically profile the polar and non-polar compounds present in red and tangerine tomato juices intended for clinical interventions. Of the over

4000 compounds detected, the red and tangerine tomato juices were found to differ significantly in a number of polar and non-polar phytochemicals, including several lipid classes. Many of these compounds have been shown to possess biological activity and may also contribute to the protective effects associated with tomatoes.

AGFD 28

Cranberry oligosaccharides decrease biofilm formation by uropathogenic Escherichia coli

Jiadong Sun2, jiadong_sun@my.uri.edu, Johannes P. Marais1, Christina Khoo1, Navindra P. Seeram2, Kerry Laplante3,4, David C. Rowley2. (1) Research Sciences, Ocean Spray Cranberries, Inc., Lakeville-Middleboro, Massachusetts, United States (2) Biomedical and Pharmaceutical Sciences, College of Pharmacy, University of Rhode Island, Kingston, Rhode Island, United States (3) Pharmacy Practice, College of Pharmacy, University of Rhode Island, Kingston, Rhode Island, United States (4) Veterans Affairs Medical Center, Providence, Rhode Island, United States

Urinary tract infections (UTI) commonly occur in the kidney and bladder. UTI patients often experience frequent recurrence and increasing susceptibility to the drug resistant uropathogens. Over 80% of the UTIs are associated with Escherichia coli, which can form biofilms on the bladder wall that provide protection against antibiotic treatment. Previous studies have shown evidence that consumption of cranberry juice can decrease the presence of bacteria in the urine and reduce UTI symptoms. Phenolic compounds from cranberry have been extensively studied for their anti-microbial properties. However, the carbohydrate constituents from cranberry have received far less attention. In this investigation, we purified phenolic-free carbohydrate fractions from cranberry (Vaccinium macrocarpon) using ion exchange chromatography and size exclusion chromatography. Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry revealed constituents consistent with oligosaccharides possessing various degrees of polymerization. Further structural analysis revealed that these constituents are mainly composed of xyloglucan and arabinan oligosaccharides. In antimicrobial assays, one enriched fraction reduced biofilm production by the uropathogenic Escherichia coli CFT073 by over 50% at 1.25 mg/mL with no adverse effects on bacterial growth. These results suggest that, in addition to phenolic compounds, oligosaccharide components of cranberry products should be further explored for their role in the prevention of urinary tract infections.

AGFD 29

Antioxidant and antiinflammatory activity of protein hydrolysates from germinated black bean cotyledons

Lidia Lopez-Barrios, lidia.lopez.barrios@gmail.com. Biotecnología y Alimentos, ITESM, Monterrey, Nuevo Leon, Mexico

Phaseolus vulgaris seeds, raw and cooked, have been recently described as potential sources of antihypertensive, antioxidant, anti-inflammatory and tumor cell inhibitory bioactive peptides. It is very well known that germination improves the nutritional properties of leguminous seeds but its effects on the antioxidant and anti-inflammatory activities of the peptides obtained from protein fractions have not been studied. Protein hydrolysates at 60, 90, 120 and 180 min, were produced with pepsin-pancreatin digestion of flours obtained from black bean cotyledons germinated up to five days. Antioxidant and anti-inflammatory activities were normalized for total soluble protein content. The extraction of proteins was doubled after germination. All hydrolysates showed antioxidant activity on a range from 298-400 TE/g of soluble protein. Antioxidant activity of protein hydrolysates was significantly different from non-germinated seeds only when the 2nd day germinated black bean flour was hydrolyzed for 180 minutes. The antioxidant potential of flours from germinated black bean cotyledons was significantly higher than the obtained from raw black beans. Hydrolysates inhibited above 90% of nitric oxide at 0.1mg/mL of soluble protein. Results obtained indicate that germinated cotyledons are a potential source of bioactive peptides with antioxidant and anti-inflammatory activities. More studies are required to validate these results in an in vivo model.

AGFD 30

Potential antimicrobial and anticarcinogenic properties of Rhoeo discolor (Tradescantia spathacea) extracts

REBECA GARCIA-VARELA, rbk_varela@hotmail.com. BIOTECNOLOGY, INSTITUTO TECNOLOGICO DE ESTUDIOS SUPERIORES MONTERREY, Monterrey, NUEVO LEON, Mexico

Traditional medicine has led to the discovery of important bioactive substances that have found uses in several health related areas. In the present work, we studied antimicrobial and anticancer properties of bioactive molecules in Rhoeo discolor extracts; a plant commonly used in folklore medicine in Mexico. For antimicrobial properties, we evaluated the in vitro activity via agar-well diffusion assay against marker microorganisms, for their importance in human health; namely, Gram-positive Listeria innocua and Streptococcus mutans; Gram-negative Escherichia coli and Pseudomonas aeruginosa, and the fungus Candida albicans. Ten different plant extracts were tested in eight different doses on all the microorganisms. Results demonstrated that P. aeruginosa was the least affected by the different extract exposure. However, low doses of these extracts did produce a statistical significant effect on the other assayed microorganisms. Some extracts were effective as bactericidal and others as bacteriostatic agents. These results suggest that the addition of certain extracts in human diet or sanitizing products could increase aseptic conditions. For the anticancer effect, different controlled doses of each extract were tested on three human cancer cell lines HT 29, PC-3, Hep G2, and a control fibroblast cell line NIH 3T3. Results statistically demonstrated a significant growth inhibition or antiproliferativo activities in cancer cell lines, especially HT 29, but very little damage on the fibroblast cell line.

These results provide meaningful data that suggest that phytochemicals contained in Rhoeo discolor extracts are bioactive components with potential applications as antimicrobials and anticarcinogen.

AGFD 31

Analytical puzzle of allergenic peanut and almond residues in spices: Was adulteration the root cause?

Stephen L. Taylor, staylor2@unl.edu, Joseph L. Baumert, Subhashinee Wijeratne. Dept. of Food Science & Technology, Univ Nebraska, Lincoln, Nebraska, United States

Beginning in late 2014, a spate of recalls were announced in the U.S., Canada, and EU associated with the presence of undeclared, but detectble, levels of potentially allergenic peanut and almond residues in several spices, notably cumin, paprika and spice/seasoning blends containing one or both of these spices. Initially, the undeclared peanut and almond residues were traced to ground spices imported from Turkey. Initially, the levels of undeclared peanut were quite high, up to 50,000 ppm (or 5%) as detected by enzyme-linked immunosorbent assays (ELISA) while lower levels of undeclared almond were detected. Subsequently, much lower levels of undeclared peanut were found in whole cumin seed imported from India and ground in the U.S. Additionally, paprika with undeclared almond, but not peanut, was found on the marketplace in England and other EU countries. Analytically, the detection of high levels of undeclared peanut in cumin was confirmed by several different commercial ELISAs and by SDS-PAGE followed by immunoblotting and detection of IgE-binding proteins using sera from peanut-allergic subjects. With almond, positive results were also obtained using several different commercial almond ELISAs but IgE-binding proteins were not observed by immunoblotting. Additionally no almond DNA was detected by polymerase chain reaction (PCR). Furthermore, "mahlab" (a spice made from cherry pits in Turkey) was found to be fully cross-reactive in several commercial almond ELISAs. Thus, the undeclared almond in cumin and paprika may actually be undeclared cherry pits although further confirmation is sought. With peanut, some samples of cumin and garlic powder with low levels of detectable peanut by ELISA did not test positive by PCR. Thus, some analytical uncertainty remains regarding the presence of undeclared, low-level peanut in certain spices.

AGFD 32

Using fraud history to inform food fraud vulnerability assessments

Jeffrey C. Moore1, creaton2@gmail.com, Karen Everstine2. (1) US Pharmacopeia, Silver Spring, Maryland, United States (2) National Center for Food Protection and Defense, University of Minnesota, Saint Paul, Minnesota, United States

As economic adulteration gains awareness globally, there is a need for new tools to assess and manage the risks of food fraud. Recently introduced vulnerability

assessment tools for food fraud utilize fraud history to anticipate potential vulnerabilities and to identify potential impacts of fraud. This presentation will review currently available information tools including the USP Food Fraud Database and NCFPD’s EMA Incidents Database. Several case studies will be presented for specific ingredients, and important consideration will be given to accurate interpretation of fraud history data to conduct informed vulnerability assessments. Lastly, opportunities for the development of new information tools beyond fraud history will be discussed.

AGFD 33

Food fraud mitigation framework for industry and regulators

Karen Everstine1, ever0152@umn.edu, Jeffrey Moore2. (1) National Center for Food Protection and Defense, University of Minnesota, Saint Paul, Minnesota, United States (2) United States Pharmacopeial Convention, Rockville, Maryland, United States

In 2014, a draft Guidance on Food Fraud Mitigation was released by the United States Pharmacopeial Convention (USP). This document, authored by the USP Expert Panel on Food Ingredient Intentional Adulterants, guides industry users as they assess the vulnerability of the food ingredients they source to food fraud, otherwise known as economically motivated adulteration (EMA). The framework described in the document can be tailored to the specific ingredients and supply chains of each user, and allows mitigation resources to be focused on the most vulnerable ingredients. It also guides an assessment of potential consequences of food fraud and implementation of mitigation efforts. This presentation will include an overview of the guidance document, as well as results and lessons learned during a food industry tabletop exercise hosted by USP in March 2015. Finally, a discussion of the potential for adaptation of this vulnerability assessment framework to use by regulatory agencies will be presented.

AGFD 34

You can't test your way to safety

Shaun Kennedy1,2, skennedy@foodsysteminstitute.com. (1) The Food System Institute, North Oaks, Minnesota, United States (2) Veterinary Population Medicine, The University of Minnesota, St. Paul, Minnesota, United States

Whether conventional foodborne illness outbreaks or cases like melamine where economically motivated adulteration results in illness, a common question comes up: why aren’t contaminants found before they sicken consumers? In the case of accidental food contamination with pathogens like Salmonella in poultry products, E. coli O157:H7 in produce or Listeria monocytogenes in ready-to-eat meat, the problem starts with the simple fact that zero risk is only possible if everything is tested for everything, and no realistic amount of testing can ensure that each and every food item is free from accidental or intentional contamination. The low frequency of contamination alone is a problem, and it becomes more complicated when considering specificity (correctly

identifying the contaminant), sensitivity (confirming lack of contamination), time to result and pre-analytical sample preparation. All of this becomes even harder in the case of economically motivated adulteration (EMA) as the perpetrators usually know what the testing strategy is and are intentionally trying to evade detection. While it is easy, retrospectively, to say that melamine as a protein substitute in wheat gluten or horsemeat contamination of ground beef and pork could have been anticipated, proactively testing for something that isn’t supposed to be there is difficult to justify from either a company or a public health agency standpoint. Deciding to test for Salmonella in chocolate when it has only rarely been a problem before is one thing, testing for formaldehyde in tofu or diethylene glycol in wine is another one all together. The challenges associated with utilizing testing as an intervention strategy to prevent economically motivated adulteration from reaching the consumer require new strategies to make testing more effective, recognizing that testing alone can’t assure safety.

AGFD 35

Impact of antioxidants on the formation of volatile secondary lipid oxidation products in oil-in-water emulsions

Erica Bakota, elaraine@gmail.com, Jill K. Winkler-Moser, Hong-Sik Hwang. U.S. Department of Agriculture, Agricultural Research Service, Peoria, Illinois, United States

Food emulsions are particularly susceptible to lipid oxidation, which leads to the formation of off-flavors and odors, and ultimately, shorter product shelf lives. Here we examine antioxidants for use in emulsions from a variety of different sources, including natural product extracts as well as rationally-designed synthetic antioxidants. Natural product extracts represent an attractive source of food antioxidants for several reasons: they appeal to consumers’ ever-increasing preference for “natural” ingredients, they enjoy GRAS status as a result of their origin, and finally, natural product extracts have the potential to be inexpensive when derived from agricultural byproducts such as grape pomace. We will discuss antioxidant extracts from Salvia officinalis (garden sage) as well as grape pomace extracts, and the impact of these antioxidants on volatile secondary oxidation products. An extract of garden sage was prepared using supercritical carbon dioxide (SC-CO2) extraction, followed by hot water extraction. The resulting extract was enriched in polyphenols, including rosmarinic acid, which has shown promising health benefits in animals. We also examined pomace from eight varieties of Midwestern hybrid grapes for phenolic content and antioxidant activity. Extracts produced from the pomace of each grape variety were added to bulk soybean oil and oil-in-water emulsions to determine antioxidant activity, and the effect on the formation of volatile secondary antioxidants was determined. Rational design of new antioxidants also offers the potential to create targeted, process-specific antioxidants. Peptides are a class of molecules that can bridge the gap between

natural and synthetic: millions of peptide products occur in nature already, and yet the modular nature of amino acids allows the construction of new peptides that have not yet been observed. At least one class of peptide antioxidants and their ability to suppress the formation of secondary lipid oxidation products will be discussed.

AGFD 36

Performance stability of nonmigratory metal chelating active packaging materials in model food systems

Maxine Roman3, maxine.roman@gmail.com, Fang Tian3, Yoshiko Ogiwara3, Eric A. Decker1, Julie M. Goddard2. (1) Univ of Massachusetts, Amherst, Massachusetts, United States (2) Food Science, University of Massachusetts Amherst, Amherst, Massachusetts, United States (3) University of Massachusetts, Amherst, Amherst, Massachusetts, United States

Trace metal promoted oxidative degradation reactions produce rancid off-flavors that impact quality and shelf-life of many packaged foods. Although addition of synthetic chelators is effective in controlling the reactivity of metals, increasing consumer demand for reduction of additives in product formulations has prompted research in alternate preservation methods. We have developed a non-migratory metal chelating active packaging material in which hydroxamate metal chelating moieties are grafted from the food contact packaging surface. In order for such non-migratory active packaging material to enable the removal of synthetic chelators from product formulations, it must be able to perform in true food systems, which have physical and chemical properties that may inhibit reactivity. The objective of this study was to evaluate the performance stability of a non-migratory iron chelating active packaging material under a variety of pH (3.0-5.0), counter-ion (Na+, Mg2+, Ca2+), and viscosity (1-104 mPa·s) conditions typically found in packaged food products. Iron chelation was quantified by material-bound iron content using colorimetry and inductively coupled plasma-optical emission spectrometry (ICP-OES). Magnesium and sodium had no effect on the material chelating activity, at pH 3.0 to 5.0. Film-bound chelators had a reduced effectiveness in the presence of calcium, only at pH 5.0, however retained ~25% iron chelating activity. Maximum iron chelation for chelating materials was reached in model food systems up to viscosity ~102 mPa·s. In more viscous models (up to 104 mPa·s), iron chelating materials retained at least 76% chelating activity and reached a maximum chelation within 2-3 d. Iron chelating materials inhibited lipid oxidation in oil-in-water emulsions at pH 3.0, even exceeding the antioxidant activity of EDTA. This work demonstrates the potential application of non-migratory iron chelating active packaging in liquid and semi-liquid foods to allow for the removal of synthetic chelators, while maintaining food quality.

AGFD 37

Unravelling chemical pathways for wine aging: Role of quinones as intermediaries on wine oxidation as “Strecker degradation reagents”

António César C. Silva Ferreira1,2, asferreira@porto.ucp.pt, Carla Maria Oliveira1,3, Ana Rita Monforte1, Artur M. S Silva3. (1) CBQF, Escola Superior de Biotecnologia- Universidade Catolica Portuguesa, Porto, Portugal (2) IWBT-DVO, Stellenbosch University, Stellenbosch, South Africa (3) Departamento de Química & QOPNA, Universidade de Aveiro, Aveiro, Portugal

The main non-enzymatic reactions occurring during wines processing and storage are the Maillard reaction and oxidation or reactions with reactive oxygen species (ROS), leading to browning and aroma/flavor formation. It has been demonstrated that some key odorants play a critical role in the perceived oxidized character of wines (1,2). Nevertheless, the mechanisms for these key odorants formation are not fully understood; only that temperature and mainly oxygen have a synergistic impact on their formation (3). Reactions of ROS with phenolics, alcohols, organic acids, even sugars, can interact with Maillard reaction (“Strecker degradation”). “Strecker degradation” has been suggested to play a switching role in directing the Maillard reaction towards aromagenic rather than chromogenic pathways (4). However, “Strecker degradation” of amino acids is not only produced by a-dicarbonyl or a-hydroxycarbonyl compounds formed by carbohydrate thermal degradation (Maillard reaction) (5). Carbonyl compounds are produced after wine alcoholic and malolatic fermentation, but can also be formed by the oxidation reaction of the hydroxyl radical (HO•) with organic acids or lipid-derived degradation (6). In the same way, the quinones produced by phenolics oxidation (7, 8) have also been described as significant producers of “Strecker aldehydes”, although this route has not been yet fully understood. All these assignments suggest that the production of color/flavor compounds may be the result of different interrelated reactions. In this work, and for the first time, 3-deoxyosone (3DG) a Maillard reaction product was quantified in white wines and red Port. Formation of “Strecker aldehydes” through quinones, as “Strecker degradation reagents”, was also evaluated at wine conditions. It was observed that “Strecker aldehydes” can be formed by quinones intermediates at wine pH. Follow by (+)-catechin, gallic acid was the phenolic that had promoted higher levels of phenylacetaldehyde. Finally development of tools such as network reconstruction provide considerable amount of information that helps to understand the kinetic contexts of the molecules present on a perturbed matrix. This network-driven approach proven to be a useful tool for identifying compounds of interest related with oxidation and Maillard reaction and to do mechanistic hypothesis generation which can subsequently be tested experimentally.

AGFD 38

Changes to oat secondary lipid oxidation products as a function of initial moisture content

Michael J. Morello2, mike.morello@pepsico.com, Todd Rakofsky1. (1) PepsiCo Beverages and Foods, Barrington, Illinois, United States (2) PepsiCo Global RD, Barrington, Illinois, United States

Changes to secondary lipid oxidation products that form in oat groats, cut groats, and flakes were investigated as a function of initial moisture and storage temperature. Sixteen secondary lipid oxidation products, consisting of n-aldehydes, enals, and dienals, were monitored by headspace-SPME-GC-MS/MS using a modified standard addition protocol. Results confirm that low initial moisture levels results in accelerated formation of the oxidation products at both ambient and elevated storage temperatures. Additionally, relatively minor changes to the secondary lipid oxidation products were observed in the oat groats and cut groats when produced at the higher initial moisture levels.

AGFD 39

Complications of analyzing acetaldehyde as a wine oxidation product

Andrew L. Waterhouse, alwaterhouse@ucdavis.edu, Ana Peterson. Viticulture and Enology, University of California, Davis, California, United States

Wine oxidation is complex from several perspectives, and there are two major classes of products, the quinones and the carbonyls. The latter are dominated by acetaldehyde, the Fenton oxidation product of ethanol. The immediate Fenton product, the ethoxyl radical is readily detected in wine exposed to oxygen by EPR studies using free radical traps. Acetaldehyde might be considered a good measure of oxidation, but it is fairly reactive and is consumed by at least 3 well known pathways. These products might be considered useful measures of oxidation, but, complicating matters further, it appears that these products can largely equilibrate back to acetaldehyde. I will present some data analyzing one major product pathway, the acetals of glycerol and ethanol using NMR spectroscopy, and discuss the issues interpreting acetaldehyde quantities in wine.

AGFD 40

Complex coacervation: Principles and simple theories

Renko de Vries, renko.devries@wur.nl. Lab Physical Chemistry and Colloid Science, Wageningen University , Wageningen, Netherlands

Over the years, our lab has focused on a wide range of experimental systems where complex coacervation, or more generally macroion complexation plays a key role: polymer micelles with complex coacervate cores, coacervates of proteins and polyelectrolytes, but also the design of novel self-assembling polypeptides, where we sometimes exploit macroion complexation to control self assembly. To make sense of the experimental observations, we have extensively made use of simple analytical theories and of coarse-grained computer simulations. In this talk I first briefly introduce

our experimental work, and this will serve as a motivation for the main part of the talk: a concise overview of theories for complex coacervation, and a discussion of their usefulness and limitations in interpreting experimental work on complex coacervation.

AGFD 41

Polyelectrolyte complex-coacervate continuum

Joseph B. Schlenoff, schlen@chem.fsu.edu, Qifeng Wang. Florida State Univ, Tallahassee, Florida, United States

Oppositely-charged polyelectrolytes are used to make a fascinating variety of soft matter, such as multilayers, complexes, coacervates and solutions. Coacervates have the properties of viscous liquids but they also show elasticity. They are of current interest as biological adhesives and encapsulants. While the similarities between multilayers and complexes have been appreciated for some time, coacervates of polyelectrolytes are usually placed in a separate category. In this talk, I will discuss our recent work linking the entire complex-coacervate-solution spectrum by water and salt content. Central to this composition/property continuum is the concept of doping by salt ions, which break “sticky” interaction points between polymers. Doping is site-specific, reversible, follows a Hofmeister series and controls all properties of associated polyelectrolytes.

Illustration of the effect of adding salt (increasing from left to right) on the microstucture of polyelectrolyte solid (undoped), solid (doped), coacervate and soution. All steps are reversible.

AGFD 42

Electrostatic complexes between (bio)polyelectrolytes and nanoparticles. Effect of the chain persistence length over particle diameter ratio

Francois Boue, francois.boue@cea.fr. Laboratoire Léon Brillouin CNRS/CEA, Gif-sur-Yvette, France

Scattering (SANS, using neutrons, or SAXS, using X Rays) is used to show that mixing a polyelectrolyte (PEL), with oppositely charged nanoparticles results in electrostatics complexes of different shapes and compacity, depending on the regions of the phase diagram, and on the ratio Lp/R between the persistence length (the rigidity parameter) and the nanoparticle size R. R sizes vary from several tenth of nanometers -silica, anionic or cationic, down to a few nanometer (3 nm). For the Au nanoparticles, cationic, for which specific original synthesis has been developed to get monodispersity in size, good spatial dispersion, and well controlled positive charge. Different (bio)-polyelectroytes have been used, of different charge, and most importantly with different persistence lengths : polycations, chitosan(Lp = 9 nm), and polylysine ( Lp = 1 nm), as well as polyanion, polystyrene sulfonate (Lp0 = 1 nm), and hyaluronan (Lp > 5 nm). Hence the ratio Lp/R varies from 0.1 up to more than 2. Depending whether the system is in a Monophasic domain with excess in PEL, a Diphasic domain II (intermediate ratio), or a Monophasic domain III (excess in NPs), the apparent fractal dimension is different. In particular, nanorod-like arrays have been observed (1.). Finally in some cases, such rather simple materials, under simple mixing, produce spontaneously, in coexistence with electrostatic complexes, more ordered crystalline organization. 1. Nanorods of well defined length and monodisperse cross-section from stable electrostatic complexation of nanoparticles and semiflexible biopolymers F. Carn, L. Shi, F. Boué, E. Buhler, ACS Macroletters 1 (7), 857–861, 2012 2. Control over the electrostatic self-assembly of nanoparticle semiflexible biopolyelectrolyte complexes, Li Shi, F. Carn, F. Boué, G. Mosser and E. Buhler, Soft Matter, 2013, 9, 5004-5015.

AGFD 43

Multivalent counterion-induced bridging of polyelectrolyte chains

Blair K. Brettmann1, bbrettmann@uchicago.edu, Nicolas Laugel2, Philip Pincus2, Matthew V. Tirrell1. (1) Institute for Molecular Engineering, The University of Chicago, Chicago, Illinois, United States (2) The University of California, Santa Barbara, Santa Barbara, California, United States

To expand the breadth of applications for polyelectrolyte complex coacervates, the interaction of the polyelectrolytes and coacervates with additional components in the medium must be more completely understood. In particular, the interaction of closely-packed polyelectrolytes with multivalent ions, which are often present in water sources, is essential to understanding their behavior in complex systems. For our initial model

system, we use a single polyelectrolyte species grafted onto a planar surface, a system for which the interaction of the polyelectrolyte chain with monovalent ions is well-understood, and expand existing models to consider the bridging of the polyelectrolyte chains by multivalent ions. We use an energy balance approach, with the energy represented as a sum of mean-field terms for the electrostatic, polymer and entropic contributions. The multivalent ions induce attractive interactions between adjacent polyelectrolyte chains, bridging, and this is taken into account by a phenomenological mean-field term. The free energy is minimized with respect to the populations of counterions and the brush length. It is found that increasing the concentration of multivalent counterions leads to collapse of the polyelectrolyte brushes in agreement with experimental observations.

AGFD 44

New opportunities for complex coacervation control exposed by bridging the gap between two classical models

Charles E. Sing, cesing@illinois.edu, Mithun Radhakrishna. Chemical and Biomolecular Engineering, University of Illinois, Savoy, Illinois, United States

Polymeric complex coacervation occurs when solutions of two oppositely-charged polyions are mixed to form a liquid-liquid phase separation consisting of a polymer-rich phase and a polymer-deficient phase. Their utility in everything from encapsulants in foods to underwater adhesives to even biomedical scaffolds motivates their continued study. Conceptual understanding of this complex coacervation process is informed by two classical models: Voorn-Overbeek and counterion release. Current theories on coacervates are successors to one or the other of these models, however each model makes distinctly different statements about the physical driving force of coacervation. We explore how these models differ, and focus on their deficiencies. In particular, we consider how despite the strong assumptions in Voorn-Overbeek, matching to experimental data is possible. We have developed both a theoretical picture and subsequently a computational picture that spans the two theories (Voorn-Overbeek and counterion release). In doing so, we provide explanations for how Voorn-Overbeek maintains matching to experimental data via a cancellation of errors. This cancellation of errors is due to cooperative interactions between the charged groups in the polyions, as well as the corresponding excluded volume correlations. This allows us to suggest and probe new ways to control coacervation, such as the position of charged groups along the polymer chain or the flexibility of the polyions. These new insights will motivate new ways to use complex coacervation as a motif in the self-assembly of polymers in solution.

AGFD 45

Complex coacervates for enzyme encapsulation and stabilization

Bradley D. Olsen, bdolsen@mit.edu, Allie Obermeyer, Carolyn Mills, Xuehui Dong, Weichao Shi. Chemical Engineering, MIT, Cambridge, Massachusetts, United States

Coacervation of proteins with polyelectrolytes provides a powerful method to encapsulate enzymes, using the ionic environment within a coacervate domain to stabilize the protein against environmental stress. When block copolymers containing charged and uncharged blocks are used, the charged proteins interact preferentially with the charged polymer domains, providing a potential method for protein nanopatterning and control over nanostructure in both solution and solid-state materials. For example, any potential applications of biocatalytic chemistry occur in organic solvents, and complex coacervate core micelles may provide a method to disperse enzymes in these media to catalyze reactions. Using polymers composed of poly(oligoethyleneglycol acrylate-block-4-vinylpyridine), we have encapsulated the enzyme organophosphorous hydrolase (OPH) in aqueous solution and shown the ability to transfer the resulting coacervate core micelles into an organic solvent environment. In particular, the micelles may be transferred to a bulk organophosphate fluid, providing an important technology for the decontamination of pesticides and neurotoxins. Coacervate core micelles also offer significant processing advantages for the preparation of biofuncitonal coatings because they can be solution-cast in a single step to form layers of bioactive plastics. Using spin coating or flow coating, we have shown the ability to coat these materials on uniform substrates such as silicon and on textured substrates such as cloth. The coacervate domains prevent aggregation of the protein during coating and lead to the retention of a high degree of protein function, even after complete drying of the film. By engineering the properties of the block copolymer used for micelle formation, the nanostructure of the resulting film can also be controlled.

AGFD 46

Effect of multivalent ions on hydrated polyelectrolyte multilayers

Dariya Reid, reid1805@neo.tamu.edu, Avanti Kavarthapu, Jodie L. Lutkenhaus. Texas AM University, College Station, Texas, United States

Layer-by-layer (LbL) assemblies, formed from the alternate adsorption of oppositely charged (or complementary) species, form conformal coatings and are of interest for many applications. The LbL structure, where layers are highly interdigitated, is similar to that of a polyelectrolyte complex. Because the interactions are noncovalent, the structure and properties of the film are sensitive to salt, pH, and other factors. This presentation will present the effects of multivalent ions on the structure and properties of LbL assemblies comprised of model strong polyelectrolytes, poly(diallyldimethylammonium chloride) (PDAC) and poly(styrene sulfonate) (PSS). Specifically, we ask how multivalent cations affect the nature of the thermal transition of PDAC/PSS LbL assemblies, previously reported in the presence of monovalent salt (NaCl). The transition itself is related to the dehydration of the polyanion, in which it remains unclear if ions are major participants. Quartz crystal microbalance (QCM-D) is

used to monitor the film structure and modulated differential scanning calorimetry (MDSC) is used to assess the thermal transition as a function of salt type and concentration. Results are compared against the well-known monovalent salt case, where it is shown that transition persists even in the presence of multivalent ions. These results can be extended to complexes in which multivalent ions also strongly affect properties.

AGFD 47

Characterization of the activity of dietary organosulfides from vegetables as natural donors of hydrogen sulfide in cell line model

Dejian Huang1, ajianhuang@yahoo.com, Dong Liang1, Chenhui Wang1, Haixia Wu2, Restituto Tocmo1. (1) Food Science and Technology Programme, National University of Singapore, Singapore, Singapore, Singapore (2) Food Science and Technology Programme, National University of Singapore, Singapore, Singapore

We have developed a highly sensitive fluorescent probe, BCu, for quantification of H2S releasing capacity in cell line model. MCF-7 cells in 96 well plate were incubated with a H2S probe BCu for 3 h, then treated with dietary organosulfides, which are biotransformed in the cells to produce H2S. The H2S releasing activity was quantified by fluorescent turning on degree of the probe. A dose response between organosulphur concentration and fluorescence intensity was applied to quantify the H2S releasing capacity of a given sample. We measured the the H2S releasing capacity of ten organosulphur rich fruits and vegetables (garlic, red onion, yellow onion, scallion, shallot, leek, spring onion, Chinese chives, durian, and stinky beans) and found that, stinky beans (a vegetable from Southeast Asia) exhibits extraordinary H2S donating capacity. The structure and H2S releasing activity of the unique cyclic polysulfides are discussed. Our research shed some light on the potential action mechanisms organosulfide rich foods in promotion of cardiovascular health.

AGFD 48

Characterization of tomato volatiles by headspace-solid-phase micro extraction

Guddadarang Jayaprakasha1, gjayaprakasha@ag.tamu.edu, Bhimu Patil2. (1) Vegetable Fruit Improvement Center, Texas A & M University, College Station, Texas, United States (2) Vegetable and Fruit Improvement Center, College Station, Texas, United States

Fruits and vegetables provide essential micronutrients, vitamins, fiber, volatiles, carotenoids, and antioxidants. Tomato (Solanum lycopersicum) is one of the most widely grown vegetable in the world. Considering low levels of volatiles in tomato, the samples needs more optimized and sensitive methods to understand volatile impact constituents present in various tomato cultivars. In the present study, headspace solid-phase microextraction (HS-SPME) method for gas chromatography-mass spectrometry

(GC/MS) system has been employed for quantifying volatiles from different varieties of tomato. Five SPME fiber coatings including 50/30μm divinylbenzene- carboxen- polydimethylsiloxane (DVB-CAR-PDMS), 65μm PDMS-DVB, 85μm CAR-PDMS, 85μm polyacrylate, and 85μm PDMS were tested for their volatiles adsorption efficiencies. In addition, key parameters such as extraction and desorption time were optimized. The maximum number of volatiles was absorbed to DVB-CAR-PDMS coating on SPME fiber. More than 15 volatiles constituents were separated Zebron-1 capillary column and identified by comparing the mass spectral data, Kovats indices, and retention times of authentic standards. These conditions were used for the identification of volatiles from different plant varieties of tomato including Roma, Vene, Campari, Sweet golden sun, Zima sweet, Heavenly villagio marzano and Maroon dark red tomato. The developed HS-SPME coupled to GC/MS technique is simple, rapid, sensitive, solvent free automated method for the routine analysis of large number samples.

AGFD 49

Bioactives from berries and their by-products

Fereidoon Shahidi, fshahidi@gmail.com. Biochem Dept, Memorial Univ of Newfoundland, St. Johns, Canada

Berries are considered as small fruits and occasionally are referred to as super fruits due to their high antioxidant potential. we also find that berry processing by-products and co-products are a good source of antioxidants. Thus, berry seed meals, pomace and also leaves are a good source of antioxidants that may be used in different food and supplement applications. Asfaras the fruit is concerned, the efficacy of phenolic antioxidants is dictated primarily by their genotype as well as degree of ripeness. The presentation will provide examples to demonstrate the importance of differeet variables in the efficacy of such products as natural sources of antioxidants in food applications and as supplements to reduce the risk of communicable diseases.

AGFD 50

Establishing biochemical justification for the value of fruit pomace a path from discovery to application

John W. Finley, jfinley@agcenter.lsu.edu. Louisiana State University, Baton Rouge, Louisiana, United States

Food processing waste and by-products is a growing environmental problem which requires large amounts of water or energy for disposal. Fruit and berry processing results in production of pomace which is used as animal feed, put back on the field as field dressing or must be treated as municipal waste. We will discuss the use of processing waste as nutrient rich food ingredients. Pomace from various fruits and berries have been studied and are rich sources of polyphenolic compounds which are can be converted to bioactive components by colonic microflora. In addition fruit

pomace can be fermented to short chain fatty acids. The potential values of various fruit or berry by products will be demonstrated a model digestive system. Strategies will be discussed to maximize the value of fruit in a range of food products.

AGFD 51

Organic resveratrol: Natural occurrence and sunlight phototransformations

Andrei A. Gakh1,2, gakhaa@yahoo.com, Andrey Sosnov3. (1) The University of Virginia, Charlottesville, Virginia, United States (2) The Discovery Chemistry Project, Bethesda, Maryland, United States (3) OrCheMed, Moscow, Russian Federation

Driven by our interest in the proliferation-modulating properties of primary resveratrol matrix components (trans-resveratrol, cis-resveratrol, and dihydro-resveratrol) towards hormone-dependent cancers, we extended our ongoing effort aimed at the identification and quantification of these compounds in the worldwide library of wine samples, with particular attention towards red wines with elevated levels (>7 mg/L) of trans-resveratrol. While no significant correlations between the resveratrol content and common environmental variables (location, grape variety, climate, manufacturing technology) were observed in resveratrol-poor samples, resveratrol-rich samples demonstrated strong, easily detectable correlation patterns. Among other trends, we serendipitously discovered that 44% of resveratrol-rich red wines were produced from organic grapes which constitute only 8.2% of all analyzed products. Puzzled with these results, we explored in detail other available samples of organic wines. The analysis of the extended data set revealed that high levels of resveratrol content is a characteristic feature of organic grapes from a narrow set of predominantly European producers. This geographic specificity is likely a result of particular agricultural standards or climate conditions. An attempt was made to correlate the levels of another bioactive compound, quercetin, also produced in grapes via comparable phenylpropanoid pathway. However, unlike resveratrol, the content of quercetin is no different in organic wines than in regular wines. While performing this research, we observed persistent phototransformations in resveratrol-rich products under direct sunlight exposure. The reaction proceeds in several steps, starting with the reversible accumulation of cis-resveratrol in a photoequilibrated manner followed by the irreversible formation of highly fluorescent 2,4,6-trihydroxyphenanthrene and (E)-4-(6,8-dihydroxynaphthalen-2-yl)but-3-en-2-one. Only cis-resveratrol was detected in significant quantities in sun-protected wine samples, even though high level of direct sunlight radiation exposure is expected during the agricultural production. Low chemical and/or photochemical stability could explain the unexpectedly low natural abundance of 2,4,6-trihydroxyphenanthrene and (E)-4-(6,8-dihydroxynaphthalen-2-yl)but-3-en-2-one.

AGFD 52

Preservation of anthocyanins in solid lipid nanoparticles: Optimization of microemulsion dilution method by Placket Burman and Box Behnken design

RAHELEH RAVANFAR1,2, rravanfar@bsu.edu, Ali Mohammad Tamadon3, Mehrdad Niakousari2, niakosar@shirazu.ac.ir, Mahmoud Reza Moein4. (1) Chemistry, Ball State University, Muncie, Indiana, United States (2) Food Science and Technology, Shiraz University, Shiraz, Iran (the Islamic Republic of) (3) Nanoparmaceutics, Shiraz University of Medical Sciences, Shiraz, Iran (the Islamic Republic of) (4) Shiraz University of Medical Sciences, Shiraz, Iran (the Islamic Republic of)

Anthocyanins are the main polyphenol components in extract of red cabbage (Brassica oleracea L. Var. Capitata f. Rubra) with inherent antioxidant activity. The anthocyanins were effectively stable in the acidic gastric digestion conditions but the total recovery after simulated pancreatic digestion was around 25% compared to around 100% recovery of phenol content. To protect them against harsh environmental conditions (e.g. pH and temperature), solid lipid nanoparticles were prepared by dilution of w/o microemulsion containing anthocyanins in aqueous media. The formulations were characterized regarding particle size and encapsulation efficiency. The formulation parameters (e.g. % total lipid, volume of internal aqueous phase, homogenization time, % total surfactant, % stabilizer) were optimized by Placket-Burman and Box-Behnken experimental designs. The value of entrapment efficiency (89.2±0.3%) was obtained when mean particle size was 455±2 nm. Scanning Electron Microscopy study revealed a spherical morphology of the particles.

AGFD 53

DOPC liposomes doped with octadecylferulate

Kervin Evans1, Kervin.Evans@ars.usda.gov, David L. Compton3, Joseph A. Laszlo2. (1) USDA, Peoria, Illinois, United States (2) USDA ARS, Peoria, Illinois, United States (3) Ncaur-Ars-USDA, Peoria, Illinois, United States

Octadecyl ferulate, found in limit quantities in plants, is a natural phenolic derivative with the potential as a cosmeceautical, nutriceutical and/or pharmaceutical ingredient because of its lipophilic and antioxidant properties. The current work demonstrates our ability to chemically synthesize and purify octadecyl ferulate to a 42% yield using solid acid catalyst and supercritical fluid chromatograph. Fluorescence leakage demonstrates that octadecyl ferulate stabilized liposomes against loss of internal contents. Molecular modeling suggests that the saturated acyl chain possibly spans the bilayer. DSC measurements indicate a 74°C melting phase transition for octadecyl ferulate. And, AFM imaging show that 5-mole% present in a lipid bilayer induced domain formation.

AGFD 54

Reactive carbonyl species: Will they be the next food safety issue?

Chi-Tang Ho, ho@aesop.rutgers.edu. Food Science, Rutgers University, New Brunswick, New Jersey, United States

Hyperglycemia is the most important factor for the onset and progress of diabetic complications. A growing body of evidence indicated that the increase in reactive carbonyl intermediates such as methylglyoxal (MG) is a consequence of hyperglycemia in diabetes. Several studies have shown that higher levels of MG are present in diabetic patients’ plasma compared to non-diabetics. Glyoxal (GO) and MG, the two major a-dicarbonyl compounds found in humans, are very reactive and lead to nonenzymatic glycation in vivo. Glycation is a complex series of reactions between reducing sugars and amino compounds, and it will lead to the formation of advanced glycation end products (AGEs). AGEs and dicarbonyl species are both linked to possible clinical significance in chronic and age-related diseases. We have discovered that commercial carbonated beverages contain extremely high levels of MG. Due to the prevalence of Maillard reaction, it is highly possible thet GO and MGO may widely present in various process foods.

AGFD 55

Formation and reduction of furan in various food model systems

Jae-Young Her, Min Yeop Kim, Kwang G. Lee, kwglee@dongguk.edu. Dongguk University, Jung Gu Seoul, Korea (the Republic of)

The objective of this study is to find out its formation and develop a method to reduce the level of furan utilizing food additives in food model systems. To investigate the effect of the temperature on the formation of furan, Maillard model systems were conducted at 90, 121, and 150 °C. A total 15 models, including alanine (ALA), serine (SER), ribose (RIB), RIB/ALA, RIB/SER, glucose (GLU), GLU/ALA, GLU/SER, sucrose (SUC), SUC/ALA, SUC/SER, furoic acid (FUR), GLU/FUR, acetaldehyde (ACET), and GLU/ACET, were prepared. The maximum level of furan was detected in the GLU/SER and GLU/ALA models at a molar ratio of 0.5:0.5. The formation of furan was proportional to the temperature in all models. The RIB/SER model generated the greatest amount of furan among the 11 models under all temperature conditions. Among the precursor models, the FUR model formed the greatest amount of furan ranging from 1058.2 to 13 927.9 ng mL−1 at all temperatures. The effects of food additives (each 0.1 M) on the reduction of furan were measured. Furan model systems consisting of monosaccharides, amino acids and/or ascorbic acid were heated at 121 °C for 25 min. The level of furan in the model systems of glucose/serine and glucose/alanine increased 7–674% when food additives were added. In contrast, the level of furan decreased by 18–51% in the Maillard reaction model systems that included ribose and alanine/serine with food additives except zinc sulphate. The formation and reduction of furan using a soy sauce model system were investigated. The concentration of furan fermented up to 30 days increased by 211% after sterilization compared to without sterilization. Regarding fermentation temperature, furan level after 30 days’ fermentation was the highest at 30 (86.21 ng/mL). The furan

levels in the soy sauce fermentation at 20 and 40 were reduced by 45% and 88%, respectively compared to 30fermentation. The addition of metal ions such as magnesium sulfate and calcium sulfate reduced the furan concentration significantly by 36–90% and 27–91%, respectively in comparison to furan level in the control sample (p<0.05). Iron sulfate and ascorbic acid increased the furan level at 30 days’ fermentation in the soy sauce model system by 278% and 87%, respectively. In the case of the BHT and BHA, furan formation generally was reduced in the soy sauce model system by 84%, 56%, respectively.

AGFD 56

Influence of California-style black ripe olive processing methods on acrylamide formation

Alyson E. Mitchell, aemitchell@ucdavis.edu. Food Science Technology, UC Davis, Davis, California, United States

Methods used in processing California-style black ripe olives generate acrylamide. California-style black ripe olives contain higher levels of acrylamide (409.67 ± 42.60 - 511.91 ± 34.08 µg kg-1) as compared to California-style green ripe olives (44.02 ± 3.55-105.79 ± 22.01 µg kg-1), Greek olives (< 1.42 µg kg-1), and Spanish olives (not detected) and indicate that the higher temperatures used to sterilize the California-style green ripe and black ripe olives are required for acrylamide formation. Pre-processing brine storage influenced the formation of acrylamide in a time-dependent manner. Acrylamide increased during the first 30 days of storage. Longer brine storage times (> 30 days) result in lower acrylamide levels in the finished product. Air-oxidation and the neutralization of olives prior to sterilization significantly increase the formation of acrylamide whereas lye-processing decreases the levels of acrylamide in the final product. These results indicate that specific steps in the California-style black ripe olive processing may be manipulated to mitigate the formation of acrylamide in finished products.

AGFD 57

Free radical mediated 3-MCPD fatty acid ester formation and the potential catalytic effect of Fe

Zhongfei Zhang1, Boyan Gao2,1, raphaelgao1985@gmail.com, Xiaowei Zhang1, Haiming Shi1, Liangli L. Yu2,1. (1) Shanghai Jiao Tong University, Shanghai, China (2) Univ of Maryland, College Park, Maryland, United States

Formation mechanisms of 3-MCPD di- and mono-fatty acid esters from TSG and DSG will be discussed. TSG or DSG was reacted with inorganic chloride compounds including NaCl, KCl, FeCl2, CuCl2, ZnCl2, FeCl3 and dry HCl, or organic chlorine compound lindane at different temperatures. Electron spin resonance (ESR) determination of TSG or DSG and 5,5-dimethylpyrroline-N-oxide (DMPO) reactions with

and without Fe2(SO4)3 revealed a free radical mechanism and the potential of Fe ion in promoting free radical generations under the experimental conditions , whereas FT-IR examination indicated a possible involvement of a carbonyl group in 3-MCPD ester formation reaction. The possible mechanisms for generating 3-MCPD fatty acid di- and mono- esters from TSG and DSG were proposed and validated. In addition, chelating agent (EDTA-2Na) was added to the model reactions to examine the potential role of Fe in 3-MCPD ester formation. Finally, the results provided an explanation for the co-formation of glycidol esters with 3-MCPD esters. The results from this study may be useful for reducing the level of 3-MCPD esters and related toxicants in the refined edible oils and food products.

AGFD 58

Chemodiversity and biosynthesis of cereulide, the food-born emetic toxin of Bacillus cereus

Thomas Hofmann1, thomas.hofmann@wzw.tum.de, Sandra Marxen1, Timo D. Stark1, Andrea Rutschle2, Genia Luecking2, Elrike Frenzel3, Siegfried Scherer2, Monika Ehling-Schulz3. (1) Food Chemistry, TU Munich, Freising, Germany (2) TU München, Neufahrn, Germany (3) University of Veterinary Medicine Vienna, Vienna, Austria

Food-borne intoxications are increasingly caused by the dodecadepsipeptide cereulide, the emetic toxin produced by Bacillus cereus. As such intoxications pose a health risk to humans, a more detailed understanding on the chemodiversity and biosynthesis of this toxin is mandatory for the reliable risk assessment of B. cereus toxins in foods. A large variation of Bacillus cereus strains were discriminated into non, low, medium and high producer strains for their emetic toxin cereulide by means of UPLC-TOF-MS and a high-precision stable isotope dilution analysis (SIDA) using 13C6-cereulide as the internal standard. UPLC-TOF-MS screening of cereulide high-producers showed a series of at least 18 cereulide variants, among which the previously unknown isocereulide A-G were determined for the first time by means of MSn sequencing, 13C-labelling experiments, and post-hydrolytic dipeptide and enantioselective amino acid analysis. Most intriguingly, the isocereulides were found to differ widely in their cell toxicity correlating with their ionophoric properties. Moreober, the data demonstrate a high microheterogeniety in cereulide and show evidence for a relaxed proof reading function of the non-ribosomal cereulide peptide synthetase (NRPS). UPLC-TOF MS analysis of B. cereus strain extracts now revealed tetra-, octa-, and dodecapeptides of a previously unkown sequence, namely (L-O-Val-L-Val-D-O-Leu-D-Ala)1-3, as intermediates of cereulide biosynthesis. Surprisingly, also di-, hexa-, and decadepsipeptides were identified which, together with the structures of isocereulides E, F, and G, do not correlate to the current mechanism for cereulide biosynthesis and violate the canonical NRPS biosynthetic logic. UPLC-TOF MS metabolite analysis and bioinformatic gene cluster analysis highlighted dipeptides rather than single amino or hydroxy acids as the basic modules in tetradepsipeptide assembly and proposed the CesA C-terminal C* domain and the CesB C-terminal TE domain to function as a cooperative esterification and depsipeptide elongation center repeatedly recruiting the action of the C* domain to

oligomerize tetradepsipeptides prior to the release of cereulide from the TE domain by macrocyclization.

AGFD 59

Food phenolics, their bioactivities, and their metabolites

Fereidoon Shahidi, fshahidi@mun.ca. Biochem Dept, Memorial Univ of Newfoundland, St. Johns, Canada

Determination of food phenolics is often a first step in evaluation of their bioactivities . ASs phenolics are present in the free, soluble esters and glycosideds as well as insoluble-bound form, simple solvent extraction may not allow the extraction of the latter fraction, thus leading to underestimation of the results. In addition, bioavailability, bioaccessibility and metabolism of phenolics may be influenced by their chemical nature as well as other variables. Thus, metabilites may indeed be more effective than the starting material and structure modification may actually improve their efficacy, at least partly, due to these factors. Examples will be provided to demonstrate the importance of these variables in the efficacy of food phenolics and their role in health promotion and disease risk reduction.

AGFD 60

Interplays between microbiota and plant bioactives

C-Y O. Chen, oliver.chen@tufts.edu. JM USDA HNRCA, Tufts University , Boston, Massachusetts, United States

An intimate symbiotic relationship between human and microbiota mediates an array of essential functions of the host, e.g., biochemistries, physiology, immunity, detoxification, all of which play a crucial role in the wellbeing of the host. Metabolic activities provided by microbiota range from utilization of non-digestible carbohydrates to deconjugation and dehydroxylation of bile acids, to biosynthesis of vitamins, and to metabolism of amino acids and xenobiotics. These functions intertwine tightly with our truly indigenous metabolic processes, resulting in extensive transgenomic co-metabolism of many substrates originating from humans, microbes, and diets. Composition and diversity of intestinal microbiota are subject to the influence of diet. For example, plant bioactives, such as fibers and polyphenols, are not absorbed or poorly absorbed in the upper gastrointestinal tract and reach the colon, where they may have a reciprocal relationship with microbiota. Thus, these dietary constituents can potentially reshape the gut microbiota ecology, which in turn has a significant impact on health locally and/or systemically. In addition to subjecting to the modulating effects of plant bioactives on their composition and diversity, microbes contribute extensively to metabolism of unabsorbed polyphenolics to phenolic. To realize the full health potentials of plant bioactives, it is essential to examine their interactions with microbiota. In this presentation, these interactions will be discussed.

AGFD 61

2-Way interaction of dietary polyphenols with gut microbiota and effects on human health

Francisco Tomas-Barberan, fatomas@cebas.csic.es. CEBAS-CSIC, Murcia, Spain

Dietary polyphenols and their effects on human health have been the objective of active research over the last 25 years. The physiological relevance of the clinical trials has, however, being rather limited due to the large inter-individual variability observed. The absorption of these phytochemicals in the gastro-intestinal tract is limited and they reach the colon almost unaltered where they interact with the colon microbiota. The colon microorganisms have a two-way relationship with polyphenols, as these food constituents in one hand modulate the microbiota population, and on the other hand the microbiota transforms polyphenols producing metabolites that differ from the original dietary constituents. Dietary polyphenols activate the development of some bacterial groups while inhibit the growth of other. This can be associated with some health benefits. Colonic microbes can metabolize polyphenols leading to compounds that are better absorbed than the original compounds or show improved health effects. Therefore, depending on the composition of the gut microbiome, the bioavailability and biological effects of dietary polyphenols can be modulated. The identification of the bacteria responsible for the metabolic transformation of specific phenolics is an active area of research, and members of the Coriobacteriaceae and Lactobacilli and Biffidobacteria have been associated with specific metabolic transformations of polyphenols in the gut. The mechanisms through which these microbiota metabolites exert their biological effects are currently studied. This means that individuals can produce, absorb and excrete different polyphenol metabolites, and enjoy different biological effects due to polyphenols intake, depending on their microbiome, and this could partly explain the inter-individual variability observed in human intervention studies with polyphenols. This opens new opportunities for the development of drugs, nutraceuticals and functional foods. The discovery of the human enterotypes will eventually have future implications in the nutritional and medicinal plants treatments and in the development of specific drugs and food products for individuals with a specific enterotype within the field of personalized nutrition.

AGFD 62

Biotransformation of cranberry A-type proanthocyanidins: Influence on gut microbiota and immune function

Jason W. Soares1, jason.soares@us.army.mil, Kenneth Racicot1,3, Laurel A. Doherty1, Steven Arcidiacono1, Emmanouil Apostolidis2, C O. Chen3. (1) US Army NSRDEC, Natick, Massachusetts, United States (2) Framingham State University, Framingham, Massachusetts, United States (3) JM USDA HNRCA, Tufts University, Boston, Massachusetts, United States

The gut microbiome consists of bacteria that play a pivotal role in innate immune health. Our works centers on elucidating the systematic processes of colonic bacteria involved in the biotransformation of dietary components and the influence of the derived metabolites on inflammation and immunity. In vitro colonic fermentation was employed to investigate the bioconversion of cranberry A-type proanthocyanidins (PAC). Static and dynamic batch fermentations were performed utilizing fecal inocula, obtained from three individuals, in a nutrient-rich anaerobic media supplemented with purified cranberry PAC. Protein content analyses indicated that PAC supports bacterial growth in a dose-dependent manner. A linear increase in protein content was observed, resulting in ~200% increase in growth at 50 mgs relative to 1 mg suggesting that purified cranberry PAC may have a prebiotic effect on gut microbiota in vitro with potential effects in vivo. Influences of metabolic byproducts assessed through GC-FID and identification through 16S RNA sequencing of pivotal bacteria that play a role in biotransformation are ongoing. GC/MS identified two phenolic metabolites, (4-hydroxyphenyl) acetic acid and (3-(4-hydroxypheyl) propionic acid, that were introduced to an in vitro host model to assess influence of metabolites on inflammation and immunity. Lastly, studies related to the detoxification of PAC metabolites will be discussed. Our efforts toward understanding the gut microbiota metabolism of PAC and the influence on innate immune function could impact future dietary supplementation strategies for both the military and civilian sectors to enhance overall gut and immune health.

AGFD 63

Metabolism of oat avenanthramides by gut microbiota

Shengmin Sang, Pei Wang, wangpeijx@gmail.com, Huadong Chen. Center for Excellence in Post-Harvest Technol, North Carolina AT State University, Kannapolis, North Carolina, United States

Oats have been cultivated for two thousand years in various regions throughout the world. Oat is a multifunctional crop nutritionally superior to many other unfortified cereals. It is commonly consumed as whole grains and known to provide healthy nutrients to humans. Oats contain a unique type of compounds, avenanthramides (AVAs), which are a group of substituted N-cinnamoylanthranilic acids. They are all composed of an anthranilic acid part and a cinnamic acid part, where the substitution pattern on the two parts is what distinguishes the different AVAs from each other. AVAs have been reported to have anti-oxidative, anti-inflammatory, anti-itching, and anti-cancer activities. We recently studied the biotransformation of the three major AVAs in mice and by human gut microbiota. Our results indicated that reduction of the double bond in the cinnamic acid unit and the cleavage of the amide bond are the major metabolic pathways of AVAs in mice and by human gut microbiota. We also demonstrated that the major metabolites of AVAs remain bioactive against the growth of human colon cancer cells.

AGFD 64

Artificial cells in picoliter droplets

Wilhelm Huck, w.huck@science.ru.nl. Radboud University Nijmegen, Nijmegen, Netherlands

A cell is the common unit structure shared by all living organisms, but even ‘simple’ prokaryotic cells are extremely complex chemical reactors. Despite enormous progress in our understanding of the complex relationships between all the individual components in the cell, we do not understand the basic underlying physical chemistry of the cell. In this talk I will show initial results of our studies into new protocells formed by coacervation. Compartmentalization via lipid bilayers is considered essential for the emergence of cells,1 but there are alternative routes to membrane-free compartments arising from liquid-liquid phase transitions.2,3 However, it is unknown if and how coacervation of collections of proteins and other molecules can form compartments capable of executing complex sets of reactions. To study this, we developed a method based on picoliter water-in-oil droplets4 that allows us to induce coacervation in Escherichia coli cell lysate and follow in vitro transcription and translation under crowded and non-crowded conditions. Coacervation creates an artificial cell-like environment in which the rate of mRNA production is increased significantly. The effect of crowding on the kinetics of the fundamental machinery of gene expression has a direct impact on our understanding of biochemical networks in vivo. Moreover, our results show the intrinsic potential of cellular components to drive phase separation and to facilitate macromolecular organization into membrane-free compartments.5 1 Szostak, J. W., Bartel, D. P. & Luisi, P. L. Synthesizing life. Nature 409, 387-390 (2001). 2 Oparin, A. I. The Origin of Life. (Dover Publications, Inc, 1953). 3 Hyman, A. A. & Simons, K. Beyond Oil and Water-Phase Transitions in Cells. Science 337, 1047-1049 (2012). 4 Theberge, A. B. et al. Microdroplets in Microfluidics: An Evolving Platform for Discoveries in Chemistry and Biology. Angew Chem Int Edit 49, 5846-5868 (2010). 5 Sokolva et al. Enhanced transcription rates in membrane-free protocells formed by coacervation of cell lysate Proc. Nat. Acad. Sci. USA 110, 11692-11697 (2013).

AGFD 65

Biomimetic microcompartmentalization by aqueous phase separation

Christine D. Keating, keating@chem.psu.edu. Penn State University, University Park, Pennsylvania, United States

The thermodynamically nonideal aqueous solution chemistry of macromolecules offers an attractive possible means of molecular organization in a prebiotic milieu or in the interior of extant cells. Aqueous polymer solutions will form multiple coexisting thermodynamic phases under a variety of readily accessible conditions. When phases are present, solutes such as ions, small molecules, proteins or nucleic acids can be compartmentalized by partitioning. The extent of localization depends on solute identity

and can be tuned by compartment size or composition. Aqueous two-phase systems can also be prepared inside cell-sized lipid vesicles by encapsulating multiple polymers at several weight percent. The resulting structures are a primitive model of biological cells that can mimic several basic functions of biological cells: including protein relocalization in response to stimulus, symmetry-breaking, and asymmetric vesicle division. These seemingly complex phenomena are observed despite the absence of any genetic molecules, enzymes, or cellular machinery, and as such could provide clues to possible intermediates in the early evolution of cell-like assemblies. Aqueous phase systems also point to the possibility of “cytoplasm first” systems in which functional microcompartments can be maintained in the absence of an amphiphile bilayer membrane, and could themselves serve as a template for assembly of a permeable membrane-like coating.

AGFD 66

Directing the phase behavior of biopolyelectrolyte complexes

Lorraine Leon Gibbons2, lorraineleon@gmail.com, Sarah L. Perry4, Michael J. Lueckheide5, Jeffrey Vieregg3, Ryan A. Klein2, Naomi Pacalin2, Matthew V. Tirrell1. (1) IME, University of Chicago, Chicago, Illinois, United States (2) Institute for Molecular Engineering, University of Chicago, Chicago, Illinois, United States (3) Jones 222, University of Chicago, Chicago, Illinois, United States (4) Chemical Engineering, University of Massachusetts Amherst, Amherst, Massachusetts, United States

Electrostatically driven polymer self-assembly mechanisms are vastly underexplored compared to that of amphiphilic based assemblies, and yet offer unique opportunities for both encapsulation of charged therapeutics and controlled delivery via the tailoring of intermolecular interactions using pH and salt. This study investigates the effect of chirality on the resultant solid or liquid character of electrostatically self-assembled polyelectrolyte complexes formed from oppositely charged polypeptides. We demonstrate that the formation of dynamic, liquid polyelectrolyte complexes requires at least one racemic polyelectrolyte in order to disrupt backbone hydrogen bonding networks. Conversely, the use of homochiral polypeptides enables the collapse of the complexes into compact, fibrillar solids with a β-sheet structure. Molecular dynamics simulations support these results while enabling visualization of the molecular structure of these complexes. The ability to use polypeptide chirality as a means for controlling the liquid or solid character of the resultant polyelectrolyte complexes is a useful tool in developing new self-assembled materials based on polyelectrolyte complex formation without otherwise altering the chemical composition of the material. By designing peptides using chiral patterning we can tailor the amount of hydrogen bonding between oppositely charged peptides and thus create materials with tunable materials properties. Additionally, nanoscale stabilization of polyelectrolyte complex formation can be achieved by coupling the polyelectrolyte to a neutral yet hydrophilic block, forming nanometer-sized micelles with a polyelectrolyte complex core and a hydrophilic corona. In this work, we characterize the structure and stability of these polypeptide based model micellar systems using light scattering, electron microscopy, circular dichroism

and small angle x-ray scattering. Additionally, we create polyelectrolyte complexes that contain therapeutically relevant charged molecules such as nucleic acids and demonstrate that the choice of nucleic acid can affect the liquid or solid nature of the polyelectrolyte complex. Time permitting, we will explore the use of these therapeutic polyelectrolyte complexes as mechanisms to inhibit microRNA’s involved in atherogenesis, with the ultimate goal of creating a treatment for atherosclerosis.

AGFD 67

Design and construction of higher-order structure and function in coacervate-based protocells

Stephen Mann, s.mann@bris.ac.uk. University of Bristol, Bristol, United Kingdom

The design and construction of compartmentalized chemical ensembles for modelling complex biological systems, exploring the origin of life, and advancing future living technologies is attracting considerable interest in a wide range of research communities. Most of these studies are focused on approaches that involve the spontaneous assembly of membrane-bounded micro-compartments such as lipid vesicles [1], colloidosomes [2] or proteinosomes [3]. However, there is also a resurgence of interest in the studies of Oparin, who proposed that liquid micro-droplets in the form of complex coacervates were a plausible model for the origin of life [4]. Although these ideas have generally fallen out of favour, we have recently initiated a series of investigations into the use of coacervate micro-droplets as an alternative model for synthetic membrane-free protocells [5]. In this talk, I will discuss our recent results on the design and construction of higher-order structure and function in coacervate-based protocells. I will demonstrate that these molecularly crowded membrane-free micro-systems exhibit high levels of chemical enrichment [5], rapid uptake and selective sequestration of biomolecules (enzymes, DNA) [6], increased rates of enzymatic transformations [7], and surface properties compatible with facilitating the membrane assembly of fatty acid [8] or inorganic building blocks [9,10]. The latter systems also result in a dramatic re-organization of the coacervate medium to produce hybrid protocells with complex sub-divided micro-architectures. The impact of these studies on designing new types of chemical micro-ensembles, and their importance as a step towards minimal representations of synthetic cellularity will be discussed. References: [1]. Dzieciol A J and Mann S, , Chem. Soc. Rev. 41, 79-85 (2012). [2]. Li M et al.,. Nature Chem., 5, 529-536 (2013). [3]. Huang X, et al.,. Nature Commun. 4, 2239 (2013) DOI: 10.1038/ncomms 3239, 1-9 (2013). [4]. Oparin A I. The origin of life, Macmillan, New York 1938. [5]. Koga, S., et al., Nature Chem. 3, 720-724 (2011). [6]. Williams, D W et al., Soft Matter 8, 6004-6014 (2012).

[7]. Crosby J, et al., Chem. Commun., 48, 11832-11834 (2012). [8]. Tang T-Y D, et al., Nature Chem. 6, 527- 533 (2014). [9]. Williams, D S, Patil A J, Mann S, Small, 10, 1830-1840 (2014). [10]. Fothergill J, et al., Langmuir, 30, 14591-14596 (2014).

AGFD 68

Biomimetic effects on actin cytoskeletal filament growth

Sarah L. Perry1,2, perrys@engin.umass.edu, Patrick McCall3, Samanvaya Srivastava2, David Kovar4, Margaret Gardel3, Matthew V. Tirrell2,5. (1) Department of Chemical Engineering, University of Massachusetts Amherst, Amherst, Massachusetts, United States (2) Institute for Molecular Engineering, University of Chicago, Chicago, Illinois, United States (3) Department of Physics, University of Chicago, Chicago, Illinois, United States (4) Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, Illinois, United States (5) Argonne National Laboratory, Argonne, Illinois, United States

Living cells have evolved sophisticated intracellular organization strategies that are challenging to reproduce synthetically. Biomolecular function depends on both the structure of the molecule itself and the properties of the surrounding medium. The ability to simulate the in vivo environment and isolate biological networks for study in an artificial milieu without sacrificing the crowding, structure, and compartmentalization of a cellular environment, represent engineering challenges with tremendous potential to impact both biological studies and biomedical applications. Emerging experience has shown that polypeptide-based complex coacervation (electrostatically-driven liquid-liquid phase separation) produces a biomimetic microenvironment capable of tuning protein biochemical activity. We have investigated the effect of polypeptide-based coacervates on the dynamic self-assembly of cytoskeletal actin filaments. Coacervate materials are able to directly affect the nucleation and assembly dynamics. Utilizing fluorescence confocal microscopy and fluorescence-based filament growth assays, we have observed the impact of macromolecular crowding, polymer chain length, and chemical specificity of the encapsulating polypeptides-based coacervate phase. These results are supported by similar observations made using specific actin-binding proteins. Moving forward, we are looking to determine the potential impact of specific chemical interactions through the incorporation of sequence-specific motifs into our polypeptide-based coacervate materials. Such systems may prove particularly useful for biochemical assays because of the potential for free transport of molecules across the coacervate phase boundary. *This work is supported by the University of Chicago and the U.S. Department of Energy Office of Science program in Basic Energy Sciences and the Materials Sciences and Engineering Division, and the University of Chicago NSF-MRSEC.

AGFD 69

In vitro reconstitution of a nonmembrane-bound RNA-protein compartment

Shambaditya Saha, ssaha@mpi-cbg.de, Anthony A Hyman. Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany

It is well known that cells use membranes to spatially separate biological activities in specialized compartments e.g. nucleus, Golgi bodies, mitochondria. On the other hand, compartmentalization is also possible in the absence of a membrane separating the compartment from its surroundings. Such membrane-less compartmentalization is observed in cellular structures containing both proteins and RNA, for instance nucleolus, Cajal bodies, P bodies, promyelocytic leukaemia nuclear bodies, stress granules, P granules. How compartmentalization occurs and sustains in the absence of a surrounding membrane is poorly understood. In order to address this, we used P granules as a model, and reconstituted in vitro a P granule-like compartment from purified components. In the germ-line cells of the worm Caenorhabditis elegans, RNA and more than forty proteins concentrate in P granules. Surprisingly, we found that a single P granule protein is sufficient for phase separating in vitro into a non-membrane-bound compartment with many properties of P granules observed in vivo. These compartments behave like liquid-drops; they grow and fuse with each other and form bigger drops. These drops are dynamic; molecules of protein intermix spatially within drops, and also exchange between drops and the surroundings. When incubated with pre-formed drops, total RNA isolated from C. elegans preferentially concentrate within drops. We used structure-function analysis to address which parts of the protein are responsible for the properties of phase-separation and “liquidity”. We found that an N-terminal self-interaction domain mediates phase-separation into drops, while a domain located at the immediate C-terminus tunes the viscosity in drops. We conclude that a single P granule protein is sufficient for generating a liquid coacervate with many properties of P granules observed in vivo. It is an intriguing possibility that the non-membrane-bound RNA-protein compartments in cells are generated from only a few of its constituent proteins with “self-interaction” and “liquidity” domains, while rest of the constituents are recruited into the compartments.

AGFD 70

Coacervation of mussel-inspired zwitterionic adhesives

Herbert Waite, herbert.waite@lifesci.ucsb.edu, B. Kollbe Ahn. University of California Santa Barbara, Santa Barbara, California, United States

Adhesives capable of sticking to wet, salt-encrusted, fouled, and corroded surfaces are urgently needed for a broad range of environmentally and medically robust applications. The adhesive strategies of mussels and sandcastle worms have inspired the design of biomimetic synthetic platforms to address these needs, especially in three essential areas: fluid-fluid phase-separation, polymer adsorption to surfaces, and curing. Initial studies emphasized adoption of a single amino acid in the native adhesive proteins,

namely 3, 4-dihydroxyphenylalanine (Dopa), to create a family of catechol-functionalized polymers for energetic surface adsorption. Dopa (catechol) was then oxidized to provide quinone-mediated curing by protein cross-linking. Lysine and phosphoserine were then adopted, often to enhance adsorption but also to drive charge-dependent complex coacervation needed for the dense fluid phase-separation of adhesive polyelectrolytes. Although several of these bio-inspired formulations are beginning to show promise for biomedical applications, they now include so many components and steps that require optimization and stabilization, that there is growing concern that they will never be suitable for use in the field. Here, we sought to simplify and combine all motifs borrowed from adhesive mussel foot proteins (mfps) into a single, low molecular weight, one-component adhesive system. By taking a previously reported zwitterionic surfactant platform with charged quaternary amine and phosphate groups and two alkyl tails, we replaced one of the alkyls with a catechol, and varied the other to create a family of mfp-mimetic zwitterionic adhesive homologs (mass <500 Da), exhibiting spontaneous catechol-mediated adsorption to surfaces, stable phase-separation as one-component coacervates, and periodate-mediated curing. The measured underwater adhesion (or cohesion) energy (Wc ~ 47 mJ/m2) is the highest reported to date; 3.3 times greater than mfp-5, and likely to stimulate applications that require atomically smooth and thin (<4 nm) glue lines, e.g., electronics.

AGFD 71

Heavy metals and aflatoxins in various herbal medicines and health functional foods

Kwang G. Lee, kwglee@dongguk.edu. Dongguk University, Jung Gu Seoul, Korea (the Republic of)

Heavy metals such as lead (Pb), cadmium (Cd), total mercury (Hg) and methyl mercury (MeHg) and aflatoxins (AFs: B1, B2, G1 and G2) in functional health foods (FHFs) and herbal medicines (HMs) were analyzed by the inductively coupled plasma mass spectrometry and HPLC. A total of 672 samples were collected from 2,347 people (1015 adults, 557 students and 775 infants and children) who lived in Korea. Method validation was carried out using standard reference material (SRM), recovery rate and limits of detection and quantification. Recovery rates for Pb and Cd using three SRMs were 94.9–101.6% and 96.7–115.2%, respectively. Mean Pb values in FHFs and HMs were 0.146 and 0.349 mg kg-1, respectively. Mean Cd levels in FHFs and HMs were 0.035 and 0.056 mg kg-1, respectively. The levels of total Hg in 416 samples were lower than the limit of detection. The MeHg analysis was conducted in 45 samples and the ranges of MeHg levels in samples were N.D. – 38.92 ng g-1. Regarding to the analysis of AFs, Two analytical methods, trifluoroacetic acid and Kobra cell derivatization methods, were compared; The latter was selected based on high linearity and sensitivity. The limits of detection of AFs using the Kobra cell method were 0.07 – 0.32 ng g-1. Recoveries of AFs using various matrixes such as solid, semi-solid, liquid samples and CRM were 81.81 – 119.87%. The Z-score and linearities of calibration curves were 0.53 and

0.9996 – 0.9999, respectively. Among 241 samples, only Angelica gigas NAKAI extract products (2 products) were detected to have 7.93 and 5.70 ng g-1 of aflatoxin G2.

AGFD 72

Reactions between polyphenolic dietary supplements and other biomolecules dictate bioactivity, bioavailability and analysis

Ann E. Hagerman, hagermae@miamioh.edu. Chemistry & Biochemistry, Miami University, Oxford, Ohio, United States

Interactions between polyphenols and proteins have been studied for many decades, with a primary focus on the noncovalent, reversible complexes that form rapidly in aqueous solution. These interactions have long attracted the interest of food and beverage scientists, since they are factors in important properties including haze formation and astringency. Recent appreciation of the beneficial health effects of polyphenols has driven additional research into interactions with proteins that may impact bioactivity, bioavailability or analytical methodology. For example, in the blood stream, polyphenols may inhibit or exacerbate protein glycation, changing the physiological function of both protein and polyphenol. Interaction with protein in the gastrointestinal tract may dictate uptake of polyphenols, while interaction with serum albumin may prolong the lifetime of polyphenols after absorption. Reaction to form covalent complexes may mask the polyphenol, making it analytically inaccessible and changing its targets and bioactivities. Unanswered questions and directions for future research will be highlighted.

AGFD 73

Rosemary: From nature to table

Maria J. Jordan1, mariaj.jordan@carm.es, Cristina Martinez-Conesa1, Sancho Bañon2, Jose A. Sotomayor1. (1) Natural Resources, Murcia Institute of Agri-Food Research and Development (IMIDA), La Alberca, Murcia, Spain (2) Department of Food Science and Technology and Nutrition, Faculty of Veterinary Science, University of Murcia, Murcia, Murcia, Spain

Southeastern Spain is well known for its richness of spontaneous aromatic plants. Among the different species found, R officinalis L. should be highlighted, mainly because of its characteristic chemical profile, in which many active components have been shown to exhibit important biological activities. This is probably the main reason why rosemary is an important source of raw materials for several companies and industries throughout the world. The geographical area in which rosemary grows dramatically affects its chemical composition. Such variability needs to be taken into account, since the quality and biological activity of rosemary extracts will depend on their composition. Study of the chemical variability exhibited by individual plants from spontaneous

rosemary populations has allowed the correlation between the chemical composition and the biological properties to be determined. Thus, the presence of eucalyptol and α-pinene as the major components in the volatile fractions increases the bactericide capacity of oils against food borne pathogens. With regards to the polyphenolic fractions, higher bacteriostatic activities were observed when the concentration of carnosol was higher than that of its corresponding precursor, carnosic acid. Also related to the polyphenolic fraction in rosemary, it has been stated that at equal amounts of rosmarinic acid, the relative concentrations of carnosic acid and carnosol do not affect its antioxidant activity. These observations suggested that rosemary could be used as an additive in animal feeding as a strategy to confer added value to the meat. In this line, a lamb diet was supplemented with a typified rosemary extract during the fattening stage. The main goal was to evaluate the level of deposition of diterpenic metabolites in the Longissimus dorsi muscle and any consequent enhancement of the meat antioxidant status. The results confirmed the presence of a diterpenic metabolite (C19H22O3) in the muscle, which conferred an improvement to the antioxidant capacity of the lamb meat.

AGFD 74

Simple UPLC–MS to monitor the presence of pomegranate in pomegranate juices

Caroline Mathon2, carolinemathon@hotmail.fr, Andrew Green2, Cynthia K. Larive1. (1) Univ of California, Riverside, California, United States (2) Chemistry, University of California Riverside, Riverside, California, United States

Pomegranates (Punica granatum L.) – a delicious fruit once used in ayurvedic medicine, now is largely known for its antioxidant properties, as well as its health benefits against diseases like cancer and cardiovascular diseases. These beneficial effects are associated with the high content of tannins, which are present in many forms, including punicalagin, an ellagitannin, and anthocyanins, which are responsible for fruit and juice coloration. Pomegranate juice is a common constituent of juice blends, and a goal of this work is to quantify the composition of juice samples where the pomegranate is a minor component. A simple UPLC-MS method was developed to detect the punicalagin, which is specific to pomegranate, and correlate its concentration with the pomegranate composition of juice samples.

AGFD 75

New methods and antioxidants to prevent oxidation of omega-3 oil supplements

Matthew Fhaner2, Hong-Sik Hwang1, hongsik.hwang@ars.usda.gov, Jill K. Winkler-Moser1, Erica L. Bakota1, sean X. liu1. (1) USDA, ARS, NCAUR, Peoria, Illinois, United States (2) Chemistry, University of Michigan-Flint, Flint, Michigan, United States

Omega-3 oils have gained much attention recently due to their beneficial health effects. However, their polyunsaturated fats (PUFAs) are so prone to oxidation in the presence

of oxygen, heat, light, and metal ions that a strong antioxidant is needed to prevent oxidation during manufacturing processes, transportation, and storage. Oxidation of PUFAs occurs even in our body and, therefore, the intake of antioxidants along with omega-3 oil rich diet is suggested to overcome the oxidative stress caused by PUFAs. As our continuing effort to discover effective antioxidants for omega-3 oil, we have examined two natural antioxidants, sesamol and gamma-oryzanol for their antioxidant activities and compared with a synthetic antioxidant, butylated hydroxytoluene (BHT) and a commercial natural antioxidant, rosemary extract. While gamma-oryzanol did not show an impressive antioxidant effect, sesamol was found to be a very strong antioxidant, stronger than BHT. Compared to the commercial antioxidant, rosemary extract, sesamol showed higher antioxidant activity at 8.4 mM due to its higher solubility in oil. Another technique to prevent oxidation of omerga-3 oil in which flowability of oil is reduced by organogelation method will be discussed. Plant waxes added in oil as organogelators could effectively reduce oxidation rate of omega-3 oil.

AGFD 76

Facile synthesis and characterization of curcumin metformin adduct: Potentially important gama-secretase inhibitor for Alzheimer disease

Bishambar Dayal2,1, dayalbi@njms.rutgers.edu. (1) Microbiology, Biochemistry and Molecular Genetics, Rutgers University, Newark, New Jersey, United States (2) Medicine, Rutgers University, Newark, New Jersey, United States

NBA

AGFD 77

Coffee-based dietary supplements contain kaurane diterpenoid glycosides inhibiting adenine nucleotide translocase in mitochondria and reduce respiration

Roman Lang1,2, roman.lang@tum.de, Tobias Fromme1,3, Anja Beusch1,2, Tatjana Lang1,2, Martin Klingenspor1,3, Thomas Hofmann1,2. (1) Technische Universität München, Freising, Germany (2) Chair for Food Chemistry and Molecular Sensory Science, Freising, 85354, Germany (3) ZIEL Unit Molecular Nutritional Medicine, Freising, Germany

Dietary supplements containing raw coffee extract are advertised to the consumer as an easily available source rich in antioxidants as part of a healthy diet. In addition to phenolic antioxidants, raw coffee extracts were found to contain a series of kaurane diterpenoid glycosides that are structurally related to carboxyatractyloside, which is reported to block oxidative phosphorylation by effectively inhibiting the adenine nucleotide translocase (ANT) in mitochondria. After their isolation and purification from raw coffee extract, studies on the impact of the atractyligenin, 2-O-β-D-glucopyranosyl-atractyligenin, 3'-O-β-D-glucopyranosyl-2'-O-isovaleryl-2β-(2-desoxy-atractyligenin)-β-D-glucopyranoside, and 2-O-β-D-glucopyranosyl-carboxyatractyligenin on ANT activity

in isolated mouse mitochondria were performed. For the first time, the latter glucoside was found to strongly inhibit ANT activity leading to reduced respiration. Quantitative LC-MS/MS analysis of these bioactive phytochemicals in commercially available green coffee based dietary supplements, roasted coffees, and model roasting experiments demonstrated the thermal instability of these kaurane diterpenoid glycosides and revealed significant amounts of these compounds in dietary supplements depending on the way of manufacturing. Based on these data, the impact of raw coffee based dietary supplements on human health will be discussed.

AGFD 78

Multivitamin and mineral supplements: An overview of key product issues

E. T. Finocchiaro, terry.finocchiaro@pfizer.com. Pfizer Consumer Healthcare, Madison, New Jersey, United States

Vitamins and minerals (MVM) are essential micro-nutrients in foods that are needed in very small amounts for growth and maintenance of good health. In February 2015, the Dietary Guidelines Advisory Committee (DGAC) released their scientific report summarizing current status and trends in food and nutrient intakes. The report identified “short fall” nutrients that are under-consumed in the USA. Consequently MVM supplementation represents an option for consumers to assure a nutritionally-complete diet for populations that are not able to consume the recommended daily intake of micronutrients.Commercial U.S dietary supplements are regulated by the FDA under the Dietary Supplement Health and Education Act of 1994 (DSHEA). Unfortunately many smaller manufacturers may leverage claims language beyond what scientific evident can support. Consequently the supplement labeling can vary widely in the nutritional language applied to marketed products. Most reputable companies leverage claims language that meets DSHEA guidelines to assure that these claims are grounded in sound science. Like many food components, multivitamins include a complex array of chemical entities and, as such, a variety of analytical methodologies are required to assure the precise quantification of micronutrients throughout the product’s shelf life. In general, pharma-like delivery forms have less stability issues than the more complex food-like forms but certain labile vitamins none-the-less require careful formulation and handling to assure the true nutrient content(s) meets the package-claimed content. Further, particularly in food forms, control of chemical stability is critical to maintaining consistent and desirable visual and organoleptic attributes.

AGFD 79

Generation of reactive oxidative species during thermal and UV processing of sugars

Rohan V. Tikekar, rtikekar@umd.edu. Nutrition and Food Science, University of Maryland- College Park, College Park, Maryland, United States

Sugars, particularly fructose, are known to accelerate the degradation of food components such as anthocyanins and ascorbic acid during thermal and ultraviolet (UV, 254 nm) processing. Yet, the mechanism by which this occurs is not well understood. We showed that sugars generated diverse oxidative species (ROS) during these processing methods. Using the FOX assay, we demonstrated that hydrogen peroxide was generated from fructose during thermal and UV processing. The concentration of hydrogen peroxide increased with the duration of treatment, presence of oxygen and a high solution pH. In UV treatment and possiblly thermal treatment, hydrogen peroxide photolyzed further to produce hydroxyl radicals. Using a fluorescent singlet oxygen sensor green (SOSG) dye, we demonstrated that singlet oxygen was also formed during both thermal and UV treatment of fructose. In addition to ROS, acidic byproducts were also generated upon thermal and UV-treatment of fructose. A remarkable similarities of the intermediates and final products between the two processes suggest that fructose undergo similar degradation process. We expect this process to be an oxidation reaction. Glucose showed similar behavior as fructose during thermal treatment, while no effect was observed during UV processing. Sucrose showed no ability to generate ROS during thermal or UV processing. The results of this study may be useful in optimizing thermal and UV based processes for foods containing high amount of sugars.

AGFD 80

Lipid oxidation as a source of diverse food toxicants

Bruno E. De Meulenaer, bruno.demeulenaer@ugent.be. Food Quality and Food Safety, Ghent University, Zwijnaarde, Belgium

Lipid oxidation is the main cause of chemical deterioration of microbiologically stable foods. During lipid oxidation, a diversity of compounds is produced which are relevant with respect to food safety. In this presentation special attention will be given to the presence of a number of highly reactive seconday lipid oxidation products in foods such as malonaldehyde, 2-butenal, 4-OH-hexenal and 4-OH-nonenal. In adition the unexpected presence of epoxy fatty acids in relatively high concentrations in a diversity of foods will be highlighted. Topics covered include the analytics of these compounds, a survey of their presence in a variety of foods, combined with a dietery exposure assessment and risk assessement, their formation in a number of model systems and their reactivity in complex foods with other ingredients. From the data obatined it could be concluded that the reactivity of some of these compounds is so high that probably other molecular targets should be selected in order to evaluate their presence in foods.

AGFD 81

Evaluation of temperature effect on the concentration levels of polycyclic aromatic hydrocarbons (PAHs) in edible vegetable oil

Olatunde S. Olatunji, snf_olatunji@ymail.com, Beatrice O. Opeolu, Olalekan S. Fatoki, Bhekumusa J. Ximba. Department of Chemistry, Cape Peninsula University Of Technology, Cape Town Western Cape, South Africa

Introduction The consumption of foods processed in reuse or spent edible vegetable oil at high temperature portent a risk to human health. This is because heating of edible vegetable oil at high temperatures may result in the formation of probable carcinogenic polycyclic aromatic hydrocarbons(PAHs) and heterocyclic aromatic amines(HAA), vis-a-viz the de-hydro cyclization of olefins in the oil. In this study, levels of PAHs; phenanthrene(Phe), benzo[a]pyrene(BaP), and benzo[k]fluoranthene(BkF), was investigated in selected refined edible vegetable oils subjected to thermal treatments at different temperatures. Experimental Four brands of edible vegetable oils were purchased over the counter in some shops in Cape Town, South Africa. About 50 ml of the different oils were subjected to heat treatment on hot plate at temperatures 25, 100, 150 and 200 ͦ C for 15 min. Thereafter, PAHs was recovered from each of the thermally treated edible oil in a 1:4 mixture of dichloromethane and hexane, after alkali hydrolysis in 90% methanolic potassium hydroxide. The extracts were cleaned up using solid phase extraction, and each concentrated to about 1 ml under nitrogen stream. The PAHs were separated and detected using a Gas Chromatograph coupled to a Flame Ionization Detector (GC-FID). Results and Discussion Table 1 The mean concentrations of Phe, BaP and BkF in edible oils heated at 25, 100, 150 and 200 ͦ C treated were 0.4525, 0.6579, 1.1110 and 1.8660 µg/L; 0.9046, 1.0418, 1.0460, and 2.1298 µg/L; and 0.9045, 1.0452, 1.0459, and 1.9035 µg/L, respectively. Conclusion The concentrations of the examined PAHs increased with increase temperature, although the observed levels were lower than the recommended levels suggested for BaP in smoked food.

Table 1: Mean concentration levels of phenanthrene, benzo[a]pyrene and benzo[k]fluoranthene in edible vegetable oils heated at different temperature

AGFD 82

Effects of thermal and high pressure processing on chemical migration in food contact polymers

John L. Koontz1, John.Koontz@fda.hhs.gov, Yoonseok Song1, Yue Zhou2, Karthik Pillai1, Kun Zhao2, Rima O. Juskelis2. (1) Center for Food Safety and Applied Nutrition, US Food and Drug Administration, Bedford Park, Illinois, United States (2) Food Science and Nutrition, Illinois Institute of Technology, Bedford Park, Illinois, United States

Migration of chemical constituents from food packaging into foods is an important component of the safety assessment of new food contact materials. The effects of thermal and high pressure processing on migration of additives from food contact polymers into foods will be reviewed. The lack of comprehensive studies and unified results of the treatment effect of HPP on migration has led FDA to investigate this perceived knowledge gap. Our earlier migration studies with polypropylene showed that HPP at 700 MPa for 5 min at 71 °C decreased migration of low molecular weight surrogate compounds into Migylol 812 and 10% ethanol food simulants relative to equivalent thermal processing. Polyethylene terephthalate (PET) pellets were loaded with four low molecular weight surrogate compounds methyl salicylate (MS), phenyl cyclohexane (PC), methyl laurate (ML), and benzophenone (BP), followed by melt compounding at 275 °C and cast extrusion into films. PET films were extracted with methylene chloride for 24 hours at 40 °C followed by ultrasonication to quantify initial surrogate loading by GC/MS prior to HPP treatment and migration studies. The average concentrations (w/w) of MS, PC, ML, and BP in manufactured PET films were 607 ± 13, 509 ± 13, 729 ± 12, and 687 ± 13 μg/g, respectively. Percent retention of surrogate in PET film after melt blending was 69% BP, 58% ML, 34% PC, and 27% MS, which followed according to surrogate vapor pressure. Overlapping temperature profiles were developed to isolate the pressure effect of HPP at 700 MPa for 5 min at 121 °C from an equivalent thermal process at atmospheric pressure (0.1 MPa). Migration data acquired from HPP will confirm that the conditions of use of food contact polymers in recommended migration testing protocols are comprehensive and ensure a margin of safety for current processing technologies.

AGFD 83

FDA update on acrylamide, furan, and other processing toxicants

Lauren Jackson, lauren.jackson@fda.hhs.gov. FDA, Bedford Park, Illinois, United States

Although processing generally improves the quality and safety of foods, it can decrease nutrient levels and bioavailability and produce chemical and physical changes that result in the formation of toxic compounds. Process-induced toxic compounds such as acrylamide and furan have gained international media attention over the past decade. In April 2002, researchers in Sweden reported finding acrylamide at levels over 1000 µg/kg in a wide range of heated, carbohydrate-rich foods such as fried, baked and roasted potato products, breakfast cereals, breads and crackers, as well as coffee. Soon after the discovery of acrylamide in food, scientists at the U.S. Food and Drug Administration (FDA) found furan in a wide range of heat-processed foods at levels greater than were previously reported. After the discovery of the two heat-induced toxicants in food, FDA initiated a broad range of activities on acrylamide and furan. FDA’s accomplishments include development of analytical methods for both compounds; analysis of a wide variety of foods for furan and acrylamide; exposure assessments; research on toxicology, formation and mitigation; and participation in international activities. FDA participated in assessments of acrylamide and furan by the WHO/FAO Joint Expert Committee on Food Additives, which will help guide future risk management efforts. In 2013, FDA also issued a Draft Guidance for Industry: Acrylamide in Foods which provides information to help growers, manufacturers, and food service operators reduce acrylamide in certain foods. This presentation will provide an update on FDA’s activities on acrylamide, furan and other processing toxicants.

AGFD 84

Metabolic and colonic microbiota transformation may alter the bioactivities of dietary food bioactives

Chi-Tang Ho3, ho@aesop.rutgers.edu, Min-Hsiung Pan2, Fereidoon Shahidi1. (1) Biochem Dept, Memorial Univ of Newfoundland, St. Johns, Canada (2) National Taiwan University, Taipei, Taiwan (3) Food Science, Rutgers University, New Brunswick, New Jersey, United States

Epidemiological evidences throughout the years have indicated that consumption of phytochemicals may play important functions in the regulation ofpathological and normal biological processes. Growing information has indicated that the bioactive compounds from foods may exert beneficial effects in part by their metabolites. The bioactive metabolites were converted by the gut microflora, liver microsomes and hepatocytes, and identified in intestinal, plasma, feces, and urine after dietary ingestion. Surprisingly, recent studies suggest that many metabolites possess more active biological functions than their precursors. In order to explore the possibilities of metabolites in food bioactive compounds, more clear understanding of the metabolic pathways and the molecular targets responsible for health promotion and disease prevention are needed. Several examples from our own research as well as from recent literature will be discussed.

AGFD 85

Metabolites of wheat phytochemicals as the exposure biomarkers of whole grain wheat intake

Yingdong Zhu, yzhu1@ncat.edu, Shengmin Sang. Center for Excellence in Post-Harvest Technol, North Carolina AT State University, Kannapolis, North Carolina, United States

Increased whole grain consumption has been inversely associated with the risk for developing some diet-related disorders, such as type 2 diabetes, obesity, cancer, and cardiovascular diseases. Many epidemiological studies, however, have failed to generate consistent results on this topic due to a lack of accurate tools to assess dietary intake of whole grains. In order to better understand the beneficial health effects of whole grains/cereal bran, biomarkers for their exposure and effects are needed. The bran fraction of the whole grain contains important bioactive phytochemicals and is the major source of cereal fiber. The metabolism of these phytochemicals may reflect inter-individual differences. This presentation will discuss the potential of different wheat phytochemicals and their metabolites as the exposure biomarkers to reflect the intake of whole grain wheat.

AGFD 86

Gastrointestinal biotransformation of resveratrol and pterostilbene in mice

Yue Sun, Minyue Song, Fang Li, Yong Cao, Hang Xiao, hangxiao@foodsci.umass.edu. Food Science, University of Massachusetts, Amherst, Massachusetts, United States

Resveratrol and pterostilbene are members of the hydroxystilbene family of compounds commonly found in fruits such as blueberry and grapes. They have been associated with a broad range of potential health benefits. In order to understand the molecular mechanisms underlying the beneficial effects of resveratrol and pterostilbene, we determined their gastrointestinal (GI) biotransformation pathways in mice. Resveratrol and pterostilbene were fed to male CD-1 mice for 35 days to achieve a stable metabolic profile in the GI tract. The major phase I metabolites of resveratrol and pterostilbene were oxyresveratrol and pinostilbene, respectively, while the major phase II metabolites are trans-resveratrol-sulfate and trans-pterostilbene-sulfate, respectively. It was also found that resveratrol and pterostilbene generated corresponding glycoside metabolites, e.g. resveratrol-O-glucopyranoside and pterostilbene-O-glucopyranoside, in the GI tract. Most interestingly, the major form of metabolites in small intestine were conjugates generated by phase II metabolism, whereas these conjugated were converted to their free forms, e.g. oxyresveratrol and pinostilbene in the colon by microbiome. Our results further demonstrated that oxyresveratrol and pinostilbene had considerable inhibitory effects against colon cancer cells. Overall, our results provide important information on the identities of metabolites potentially responsible for the biological effects of resveratrol and pterostilbene in the GI tract.

AGFD 87

Biological importance of fucoxanthin and its metabolites

Kazuo Miyashita1, kmiya@fish.hokudai.ac.jp, Masashi Hosokawa1, Nana Mikami2, Yasuo Kokai2. (1) Hokkaido Univ Fisheries Scis, Hakodate Hokkaido, Japan (2) Department of Biomedical Engineering, Sapporo Medical University School of Medicine, Sapporo, Japan

Dietary carotenoids are considered to play a role in the prevention of common chronic diseases. Carotenoids have been implicated as important dietary nutrients having antioxidant potential. There is little doubt that, under the right conditions, carotenoids can show their health beneficial effects by protecting cells, tissues and other structures such as lipoproteins against oxidative damage. On the other hand, we can understand that not all of the biological activities ascribed to carotenoids must be necessarily linked to their ability to scavenge free radicals and reactive oxygen species. Other mechanisms of action that are independent of the antioxidant activity are likely to be also important. For the better understanding the physiological effect of carotenoids, efforts have been sometimes required to make clear the modulation effect of carotenoids or their metabolites on specific gene and protein expressions in biological systems. The effect is usually based on the interaction between a carotenoid or its metabolite and biological key molecule such as receptor protein and co-activator, where specific chemical structure of carotenoid may be essential for binding to these biomolecules. From this viewpoint, unique mechanism for the anti-obesity effect of brown seaweed carotenoid, fucoxanthin, is interesting, since this effect of fucoxanthin is based on the induction of UCP1 in abdominal white adipose tissue (WAT) mitochondria to lead fatty acid oxidation and heat production in the WAT. Fucoxanthin can also improve through the down-regulation of adipokines such as TNFa, MCP-1, IL-6, and PAI-1 in WAT and up-regulation of glucose transporter 4 (GLUT4) in skeletal muscle and the promotion of its translocation to the cell membrane. The present study reports the metabolites of fucoxanthin in human and its biological activities with their detail molecular mechanisms.

AGFD 88

Are anthocyanins PPARα agonists?

Agnes M. Rimando1, agnes.rimando@ars.usda.gov, Shabana Khan2, Cassia Mizuno3, Guang Ren4, Suresh Mathews5, Hyunsook Kim6, Wallace Yokoyama7. (1) USDA ARS, University, Mississippi, United States (2) University of Mississippi, National Center for Natural Product Research, University, Mississippi, United States (3) University of New England, College of Pharmacy, Portland, Maine, United States (4) University of Alabama at Birmingham School of Medicine, Division of Endocrinology, Diabetes & Metabolism, Birmingham, Alabama, United States (5) Auburn University, Department of Nutrition, Dietetics and Hospitality Management, Auburn, Alabama, United States (6) Konkuk University, College of Veterinary Medicine, Seoul, Korea (the Democratic People's Republic of) (7) USDA ARS, WRRC, Albany, California, United States

High-fat diet supplemented with 2% blueberry powder has been reported to reduce abdominal fat in rats, and was associated with increased PPARα activity. Our group also previously demonstrated an up-regulation of hepatic PPARα mRNA expression in hamsters fed diet supplemented with 2% blueberry peel extract. It became of interest to determine whether anthocyanins activate PPARα since anthocyanins are the most abundant polyphenolic compounds in blueberries. Anthocyanins, together with catechin, epicatechin, resveratrol and pterostilbene were evaluated for the activation of PPARα in H4IIEC3 cells at concentrations ranging from 3.12 µM to 25.00 µM. Pterostilbene exhibited a dose-dependent activation of PPARα similar to those of ciprofibrate and Wy-14643. The responses of resveratrol were lower than those of pterostilbene, while the anthocyanins and catechins did not activate PPARα. Additionally, real-time PCR analysis also revealed that pterostilbene significantly and dose-dependently (at 10, 20 and 50 µM) increased PPARα gene expression, and exhibited greater induction of PPARα than 100 and 200 µM of fenofibrate, in H4IIEC3 cells. While low bioavailability of anthocyanins due to quick metabolism maybe considered a factor, studies have shown that anthocyanins are found in the blood after oral administration of berries or berry extracts, with maximum concentration being reached between 0.5 to 2 hours. Anthocyanins could therefore reach target sites for PPARα activation; however, our present study suggests anthocyanins are not PPARα ligands.

AGFD 89

Effect of supercharging on coacervation between proteins and polyelectrolytes

Allie Obermeyer, acober@mit.edu, Carolyn Mills, Xuehui Dong, Bradley D. Olsen. Chemical Engineering, MIT, Cambridge, Massachusetts, United States

The encapsulation of proteins in complex coacervate core micelles serves as a promising technology for protein drug delivery and patterning. However, a large number of proteins do not form complex coacervate core micelles with ionic-neutral block copolymers. Unlike DNA or RNA, which possess a uniform negative charge, proteins have a relatively low charge density, a “patchy” charged surface, and a specific, globular fold. Chemical modification of proteins to generate a more uniformly supercharged species provides a simple strategy to generate protein based complex coacervate core micelles. However, the fundamental understanding of the principles of coacervation between proteins and polymers is underdeveloped. Using mass spectrometry, we were able to quantify the formal charge distribution of modified proteins. A panel of proteins with varying charge densities was synthesized and their propensity to form coacervates with model polyelectrolytes was measured. While the model proteins studied do not readily form coacervates in their native state, the proteins all formed coacervates once they surpassed a critical ratio of negatively to positively charged residues. Additionally, as the protein charge density increased they formed coacervates over an increasing range of polymer ratios. Finally, the charge ratio at maximum coacervation indicates a strong induced charging effect of the protein in the coacervate state. These bulk coacervation studies were translated to the design of

complex coacervate core micelles. The micelles were characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM).

AGFD 90

Effect of charge patterning on polypeptide-based complex coacervation

Li-Wei Chang, liweichang@umass.edu, Sarah L. Perry. Chemical Engineering, University of Massachusetts Amherst, Amherst, Massachusetts, United States

Complex coacervation is an electrostatically-driven liquid-liquid phase separation that occurs when aqueous solutions of oppositely-charged polyelectrolytes are mixed. Complex coacervates find application in many fields, such as cosmetics, process food and microencapsulates for drug and gene delivery, and is affected by many factors such as the polycation/polyanion ratio, total polymer concentration, pH, temperature, and pattern of charges present. Among these factors, the effect of charge patterning has been rarely studied, due to the difficulty of synthesizing polyelectrolytes with the equal chain length and charge density, but different distributions of charge. Polypeptides represent a model platform for the synthesis and study of polyelectrolytes with precisely controlled charge patterning at molecular level. We examine the effect of charge density and charge patterning on the formation, stability, and overall material properties of complex coacervates formed from polycations and polyanions with matched and mismatched sequences of charged residues. This systematic investigation of the effects of charge patterning will help to elucidate design rules to facilitate the tailored creation of complex coacervate-based materials with defined properties for a wide range of applications.

AGFD 91

Liquid-to-solid transitions in polyelectrolyte complexes

Yalin Liu, yalinliu@umass.edu, H. H. Winter, Sarah L. Perry. Chemical Engineering, University of Massachusetts Amherst, Amherst, Massachusetts, United States

Polyelectrolyte complexation has long been known to result in both liquid and solid complexes. However, the exact nature of the liquid-to-solid transition remains an open question. To seek answers explaining these phenomena, we will use rheology to study the nature of the liquid-to-solid transition from the liquid side. Advanced analysis methods will enable unambiguous determination of the mechanism for this phase transition (e.g., gelation vs. glass transition). In examining the nature of liquid-to-solid phase transitions in polyelectrolyte complexes, we will study the effects of two orthogonal chemical functionalities: electrostatic interactions and hydrogen bonding. Salt has been shown to facilitate a solid-to-liquid transition in polyelectrolyte complexes by acting as a plasticizing agent and softening intermolecular electrostatic interactions between oppositely-charged polyelectrolytes.

Separately from the electrostatic effects that drive polyelectrolyte complex formation, interpolymer hydrogen bonding has been shown to stabilize the formation of solid complexes. In a manner similar to that of salt-based plasticization, the addition of urea has been shown to disrupt these stabilizing interactions and facilitate the transition to a liquid state. Modulation of these orthogonal chemistries has the potential to enable independent strategies for controlling the state of polyelectrolyte materials, and facilitate their use in a wide range of applications.

AGFD 92

Polypeptide complexation: From bulk coacervates to nanoscale assemblies

Dimitrios Priftis, jim.priftis@gmail.com, Lorraine Leon, Khatcher O. Margossian, Anna Tropnikova, Matthew V. Tirrell. IME, University of Chicago, Chicago, Illinois, United States

A variety of materials with diverse structures and properties can form as a result of electrostatic interactions between oppositely charged macromolecules. Under defined conditions, complexation can lead to a phase separation phenomenon, referred to as complex coacervation. This talk will focus on complex coacervation of a specific category of polyelectrolytes, polypeptides. Using polypeptides as a model system we were able to identify a number of parameters that affect the complexation (e.g. salt concentration, chirality) as well as the thermodynamics involved. Building on this work, we designed a series of self-assembled materials including coacervate core micelles. For the preparation of these materials copolymers consisting of polypeptide and poly(ethylene glycol) blocks were used. The key to this design strategy is the microphase separation of coacervate domains from the aqueous solution, when the copolymers are mixed. The response of these assemblies to external parameters will be discussed in detail with the use of a variety of experimental techniques such as TEM, SAXS and light scattering.

AGFD 93

Polyelectrolyte complex formation in acetone-water mixture

Handan Acar1, hacar@uchicago.edu, Samanvaya Srivastava1, Dimitrios Priftis1, Joao Cabaral2, Matthew V. Tirrell1. (1) Institute for Molecular Engineering, The University of Chicago, Chicago, Illinois, United States (2) Department of Chemical Engineering, Imperial College London, London, United Kingdom

Polyelectrolyte complexation is a phase separation phenomena that occurs upon mixing of solutions of oppositely charged and sufficiently long polyelectrolytes. These polyelectrolyte complexes can vary from a liquid-like phase with high water content to a solid phase with negligible water content depending on the strength of the interactions among the polyelectrolytes. Effects of a range of parameters that influence polyelectrolyte complex formation, such as the ionic strength, salt concentration,

polycation/polyanion ratio, pH, molecular weight of the polyelectrolytes, and temperature, have all been studied in detail. Polyelectrolyte complexes retain large amounts of water, and have extremely low interfacial tension between the coacervate phase and the coexisting supernatant phase. All the theoretical calculations are based on water as solvent, so far. In this study, we investigated polyelectrolyte complex formation in various organic solvents and their mixtures with water. Intriguing trends in complex formation in acetone-water mixtures were observed, including persistence of the complex phase at high salt concentrations, and are ascribed to significant differences in the dielectric constant of the two solvents. Polyelectrolyte conformation in different solvents, as explored via small angle X-ray scattering, is also presented and aids in unraveling the underlying physics of the complexation phenomena. We also reconstruct the bimodal salt concentration/polymer concentration phase diagram for the new acetone-water solvent.

AGFD 94

Chirality-selected phase behavior in complexes of ionic polypeptides

Matthew V. Tirrell, mtirrell@uchicago.edu. IME, University of Chicago, Chicago, Illinois, United States

Polyelectrolyte complexes present new opportunities for self-assembled soft matter. Factors determining whether the phase of the complex is solid or liquid remain unclear. Ionic polypeptides enable examination of the effects of stereochemistry on complex formation. Here, we demonstrate that chirality determines the state of polyelectrolyte complexes, formed from mixing dilute solutions of oppositely charged polypeptides, via a combination of electrostatic and hydrogen bonding interactions. Fluid complexes occur when at least one of the polypeptides in the mixture is racemic, which disrupts backbone hydrogen bonding networks. Pairs of purely chiral polypeptides, of any sense, form compact, fibrillar solids with a β-sheet structure. Analogous behavior occurs in micelles formed from polypeptide block copolymers with polyethylene oxide, where assembly into aggregates with either solid or fluid cores, and eventually into ordered phases at high concentrations, is possible. Chirality is an exploitable tool for manipulating material properties in polyelectrolyte complexation. In collaboration with: Sarah L. Perry, Lorraine Leon, Kyle Q. Hoffmann, Matthew J. Kade, Dimitrios Priftis, Katie A. Black, Derek Wong, Ryan A. Klein, Charles F. Pierce III, Khatcher O. Margossian, Jonathan K. Whitmer, Jian Qin,<span style="font-size:11px; line-height:17px"> and</span> Juan J. de Pablo.

AGFD 95

Marine sandcastle worm-inspired medical adhesives based on liquid-liquid phase separation

Russell Stewart1, russell.stewart@utah.edu, Monika Sima1, Ryan O'Hara2. (1) Department of Bioengineering, University of Utah, Salt Lake City, Utah, United States (2) Department of Radiology, University of Utah, Salt Lake City, Utah, United States

There remains much room for improvement of adhesives for bonding living tissue. In an approach inspired by marine sandcastle worms, we are developing water-borne adhesives by associative macrophase separation (complex coacervation) of synthetic copolyelectrolytes (co-PEs) that mimic the adhesive proteins of the natural sandcastle worm glue. The water-borne, injectable adhesives can be applied to and bond fully-submerged substrates, including tissue. Several in situ setting and curing mechanisms have been developed to harden the glue after application to tissues, including photo- and chemically initiated covalent crosslinking. The crosslinked adhesives swell less than 1% after 30 days in physiological saline, a critical feature for adhesives intended for tissue repair. Pre-clinical testing of the adhesives is in progress for applications in fetal membrane repair following in utero surgery and as embolic agents to control blood flow.

AGFD 96

Self-assembled nanostructures from block copolymers for biomedical application

Yasutaka Anraku, anraku@bmw.t.u-tokyo.ac.jp. Department of Materials Engineering, The University of Tokyo, Tokyo, Japan

Nanotechnology-based medicine (Nanomedicine) has been receiving progressive interest for the treatment of intractable diseases, such as cancer, as well as for non-invasive diagnosis through various imaging modalities. Engineered polymeric nanostructures with smart functions play a key role in nanomedicine, including drug carriers, gene vectors, and imaging probes. Nano-structures with 10-100 nm in size can be prepared by programmed self-assembly of block copolymers in aqueous entity. Most typical example is the polymeric micelles with distinctive core-shell architecture. Hydrophilic shell provides polymeric micelles with stealthiness against foreign body recognition system in the body, while inner core works as a nano-reservoir of various cargo compounds. Furthermore, smart functionalities as nano-structures to executing the structural change responding to external and internal signals can be installed in the polymeric micelles, thereby enabling smart therapy and diagnosis at the target site in the body. Additionally, we have developed the first example of polyion complex vesicles (PICsomes) with a unilamellar PIC membrane obtained by the simple mixing of water-soluble and oppositely charged block copolymers. Many advantages of them have been revealed so far, such as easy encapsulation of water-soluble macromolecules, semipermeability of the vesicle wall, size tunability, etc. Another important aspect of PICsomes is facile cross-linking of PIC layers using an amide coupling reagent. In fact, significantly prolonged blood circulation of crosslinked PICsomes was demonstrated irrespective of their sizes. PICsomes are expected to be a versatile platform to deliver enzymes to tumor for enzyme prodrug therapy, utilizing their features, such as encapsulation of enzymes and substrate-

permeability. This presentation focuses on the development of self-assembled nano-structures from block copolymers for targeted therapy of intractable disease.

AGFD 97

Beyond elastin: New peptide olymers that exhibit aqueous coacervation

Ashutosh Chilkoti, chilkoti@duke.edu. Duke Univ, Durham, North Carolina, United States

Elastin like polypeptides (ELPs), composed of repeats of VPGXG pentapeptides that recur in all tropoelastin sequences, are the best studied class of peptide polymers that exhibit aqueous coacervation behavior above their cloud point, leading to the formation of a polymer rich phase and a second, largely aqueous phase. These coacervates are thermally reversible with little to no hysteresis and show no internal microstructure. To test the hypothesis that coacervation behavior requires a largely unstructured polymer that is capable of temperature-triggered dehydration, we synthesized a set of ELPs that contain periodic (Ala)25 domains that are interspersed within the VPGXG repeats in the range of 0-50% of the total chain length. The inclusion of ordered helical oligoalanine domains led to several unexpected consequences on their coacervation behavior. First, these systems show a tunable degree of hysteresis in their cloud point depending on the fraction of Ala residues. Second, the coacervate forms a fractal-like network structure that is absent in the control ELPs. Third, these polymers show syneresis at high temperatures (>80ºC) that the control polymer does not exhibit. Fourth, when these oligoalanine-containing ELPs were fused to a more hydrophobic ELP segment that does not contain oligoalanines, the diblock copolymers, unexpectedly, form vesicles at lower temperatures (~20 ºC), which transition into spherical micelles at higher temperature (40ºC). This system hence progresses from soluble unimers to vesicles to spherical micelles to a coacervate as the temperature is raised. In contrast, a control dibock polymer that is missing these oligoalanine domains does not show an intermediate vesicle phase, suggesting that the phase diagram of these oligoalanine containing ELP diblocks is considerably different from prototypical ELPs that do not contain these oligoalanine segments. In a second line of investigation into the origins of coacervation in ELPs, we used bioinformatics to guide the synthesis of a large family of peptide polymers that are predicted to exhibit LCST phase behavior. Analysis of the LCST phase behavior of these polymers provides sequence heuristics to encode LCST phase behavior in intrinsically disordered peptide polymers.

AGFD 98

Directing encapsulated stem cell fate via in situ forming, growth factor-loaded coacervate microparticle-embedded hydrogels

Eben Alsberg, eben.alsberg@case.edu, Oju Jeon. Biomedical Engineering, Case Western Reserve University, Cleveland, Ohio, United States

Microencapsulation by complex coacervation, which can be spontaneously formed upon mixing oppositely charged polyelectrolytes in aqueous solution, has been widely studied for food science, drug delivery and biomedical applications. However, there are no known examples of cytocompatible complex coacervate systems as potential three-dimensional biomaterials for cell transplantation and tissue engineering applications, due to the harsh physicochemical conditions necessary to typically form them (i.e., low and narrow pH range and high temperature). Here, we report the spontaneous formation and properties of coacervate microdroplets and/or coacervate-laden photocrosslinked hydrogels derived from the simple mixing of photocrosslinkable oxidized, methacrylated alginate (OMA) and methacrylated gelatin (GelMA) over a wide pH range at room temperature. This system enables simultaneous creation of drug-laden microdroplets and encapsulation of stem cells in photopolymerized coacervate hydrogels under physiological conditions, which can be utilized as a novel platform for in situ formation of localized, sustained bioactive molecule delivery to encapsulated stem cells for therapeutic applications.

AGFD 99

Coacervates of ionic polysaccharides for tissue engineering

Ozge Karabiyik2, Elif Kilic Iyilik1, Gamze Kose2, Ayse B. Kayitmazer1, bkayitmazer@gmail.com. (1) Department of Chemistry, Bogazici University, Istanbul, Turkey (2) Genetics and Bioengineering Department, Yeditepe University , Istanbul, Turkey

Oppositely charged macroions may result as liquid-liquid phase separation, i.e. coacervation, under certain conditions. For coacervates of hyaluronic acid (HA) and chitosan (CH), these conditions are pH, ionic strength, order of addition, degrees of acetylation of chitosan, HA molecular weight, and charge ratio of the two biopolymers. In the first part of this study, we will present our light scattering results about the effect of ionic strength (I) on the size of the particles. Despite a maximum in turbidity versus I was observed for this system, radius of gyration didnot follow the same relationship. Zeta potential experiments were also done to understand the relationship with the charge ratio of the two biopolymers in the coacervates. It was confirmed that coacervation was non-stoichiometric for the HA-CH system. In the second part of our study, HA-CH coacervates will be applied to cell encapsulation, and the cell viability will be shown at different experimental conditions.

AGFD 100

Complex coacervates as protein delivery vehicles: Preserved activity, controlled release rate, and in vivo efficacy

Noah Johnson, william chen, Yadong Wang, yaw20@pitt.edu. University of Pittsburgh, Pittsburgh, Pennsylvania, United States

Clinical translation of protein therapies faces multiples challenges; the most significant is how to maintain bioactivity. My lab uses heparin and biocompatible polycations to form an injectable coacervate that preserves the bioactivities of heparin-binding proteins. This includes many growth factors and morphogens. I designed the delivery vehicle to mimic the interaction among fibroblast growth factor-2 (FGF2), heparin and FGF receptor. The effectiveness of the coacervate delivery system is demonstrated using multiple proteins (FGF2, VEGF, HGF, HB-EGF, IL10, Shh, PDGF, NGF among others) in multiple animal models (mice, rat, pig). The coacervate provides spatial and temporal control of the release of heparin-binding proteins. I will discuss three applications: angiogenesis, skin wound healing, and cardiac repair post- infarction. Promising data in large animals and diabetic animal models suggest the potential of clinical translation. Moving forward, there are challenges that need to be addressed, these will be discussed as well.

AGFD 101

Using real time measurement of galvanic electrode potentials to clock the course of Maillard reactions

George P. Rizzi2,1, georgerizzi@yahoo.com. (1) 542 Blossomhill Lane, Current Address, Cincinnati, Ohio, United States (2) Chemistry, Miami University of Ohio, Middletown, Ohio, United States

The Maillard reaction of amino acids and carbohydrates has received widespread attention because of its relavence to food flavor and the complex biochemistry of cell ageing. In Maillard systems redox intermediates like deoxyosones can define an electrochemical cell capable of producing measurable voltages between a Pt and a Ag/AgCl (reference) electrode. Changes in Galvanic (zero load) potential measurements are known to correlate with redox activity and can thereby provide a useful , non-invasive measurement to follow the course of the reaction. In this study real time changes in galvanic potential were measured to continuously follow the course of model Maillard reactions at 70 and 100o C. Data are presented for reactions of glycine and α- and β-alanine with typical hexoses and pentoses in phosphate buffer at pH 7.0.

AGFD 102

Different reaction pathways generate aroma-active amino acid degradation products during fermentation, roasting and eating of cocoa

Peter H. Schieberle, peter.schieberle@lrz.tum.de. Technical Univ of Munich, Freising, Germany

During thermal treatment of foods, free amino acids are easily degraded and some of them are well-known precursors of aroma-active aldehydes, acids and amines. Commonly, their formation is suggested via the so-called Strecker reaction starting with a Schiff’s base formed from the amino group and an alpha dicarbonyl compound, which

usually stems from carbohydrate degradation at higher temperatures. Our recent findings indicated that Strecker aldehydes are already formed in quite high amounts during cocoa fermentation, prompting us to investigate the formation of the aldehydes in more detail at lower temperatures. For this purpose, model studies with polyphenol oxidase were performed, and the yields of aroma active compounds as well as reaction products from a catechin template will be presented. In addition, results on the role of Amadori products in the generation of Strecker compounds will be discussed, and the importance of both, amino acids and Amadori compounds, as aldehyde and amine precursors will be compared to the role of the recently found oxazolines, another group of important Strecker intermediates These compounds are able to liberate aroma active Strecker aldehydes during eating of cocoa or chocolate, respectively. This in-mouth chemistry will be demonstrated for a few other thermally processed foods, such as malt or potato chips, respectively.

AGFD 103

On the role of Amadori-rearragement products as precursors of aroma-active Strecker aldehydes in cocoa

Sandra Hartmann, sandra.hartmann@lrz.tu-muenchen.de, Peter Schieberle. German Research Centre for Food Chemistry, Freising, Germany

The unique aroma of cocoa is a result of the fermentation process followed by roasting. While roasting, a complex cascade of reactions takes place among which the so-called Maillard reaction between reducing carbohydrates and amino compounds plays an important role. 1-Amino-1-desoxyketoses, known as Amadori rearrangement products (ARPs) have often been proposed as key intermediates in the formation of Strecker aldehydes, such as methylpropanal, 3-methylbutanal or phenylacetaldehyde. To study their role as precursors of cocoa aroma compounds, first six ARPs and their respective isotopically labeled homologues were synthesized and then an assay for their reliable quantitation was developed based on LC/MS/MS. Quantitation of the ARP formation during cocoa fermentation and a correlation with the amounts of Strecker aldehydes generated after roasting showed a good correlation between ARP formation/degradation and Strecker aldehyde formation. Interestingly, the ARPs were already formed during the fermentation of cocoa beans. To elucidate the role of ARPs as precursors of the Strecker aldehydes in “competition” with the free amino acids, an isotope enrichment experiment was performed. The obtained data clearly suggested the ARPs as effective precursors of Strecker reaction products. Furthermore, inappropriately fermented beans (slaty beans) showed a clearly lower amount of ARPs compared to brown beans, and thus, a lower amount of Strecker aldehydes.

AGFD 104

Formation of Strecker aldehydes and biogenic amines as a consequence of carbonyl-amine reactions initiated by oxidized lipids

Rosario Zamora, M. Mercedes Leon, Francisco J. Hidalgo, fhidalgo@ig.csic.es. Instituto De La Grasa, Sevilla, Spain

Nonenzymatic browning reactions are produced during food processing and have important consequences on nutritional and sensory properties of foods. Many of these consequences are related to amino acid degradations. Thus, amino acids can suffer a Strecker degradation to produce many flavors, including Strecker aldehydes, pyrazines, pyridines, pyrroles, and oxazoles, among other compounds. On the other hand, amino acids can also suffer a decarboxylation to produce biogenic amines in a first step, which can be then converted into vinylogous derivatives such as styrene or acrylamide. The objective of this study was to determine the reaction conditions favoring the formation of either Strecker aldehydes or biogenic amines. Studies were carried out with phenylalanine, as a model amino acid, which was heated in the presence of different primary, secondary, and tertiary lipid oxidation products. The formation of phenylacetaldehyde and phenylethylamine was studied as a function of pH, concentration of reactants, water activity, atmosphere composition, time, temperature, and the presence of other amino acids as potential competitors. The obtained results showed that it is possible to promote either the oxidative decarboxylation of the amino acid, to produce flavors, or its simple decarboxylation, to produce biogenic amines, as a function of reaction conditions. The reaction conditions that favor the different alternative pathways were identified.

AGFD 105

Food-borne taste modulators from mother nature and culinary art

Thomas Hofmann, thomas.hofmann@wzw.tum.de. TU München, Neufahrn, Germany

Varying widely across the world, reflecting unique environmental, economic, and cultural traditions, various drying, fermentation, cooking and roasting procedures have been empirically developed during the last millenniums and, since then, the alluring flavor of the dishes prepared do attract consumers on a global scale. In particular, the food manufacturing techniques leading to the most premium tastes promise to contain essential taste compounds and/or taste modulators generated from sensory inactive precursors upon processing of the raw materials. This evolutionary refinement of food manufacturing procedures is, therefore, expected to open an interesting avenue towards the discovery of natural taste (modulating) compounds, which might be applied as natural solutions to overcome flavor challenges associated with the production of, in particular, sugar, salt or fat-reduced products. The presentation will highlight analytical strategies to identify key taste compounds and taste modulators in processed food by means of a Sensomics approach.

AGFD 106

Improved detection methods for food toxin with nanotechnology

Bosoon Park1, bosoon.park@ars.usda.gov, Bin Wang2, Bingqian Xu3. (1) USDA, ARS, Athens, Georgia, United States (2) Biological and Agricultural Engineering, University of Georgia, Athens, Georgia, United States (3) Coll of Engr Chemistry, University of Georgia, Athens, Georgia, United States

Food safety is a complex issue that has an impact on all segments of society. Although most food poisoning is caused by bacteria, viruses and parasites rather than toxic substances in the food, some cases of food poisoning is link to natural or chemical toxins. In the United States, an estimated 48 million foodborne illnesses occur each year. More than 250 pathogens and toxins are known to cause foodborne illness. Traditional detection and identification methods for toxins in food typically involve time consuming and expensive bioassays. Most of the rapid tests currently available require some sort of extraction and purification procedure before they can detect toxins in food matrices. Recently, concern about the possibility of terrorist attack with food toxins has prompted the development of commercial test kits for bio-defense applications. However, the methods developed previously could be used for rapid screening of food samples with limited sensitivity. Therefore, improved sensing platforms that can detect food toxins rapidly with high sensitivity and specificity are required for food industry. In this research advanced nanotechnology was used for the label-free detection of ricin. Atomic force microscopy (AFM) has the ability to detect single molecule in samples, which provide highest sensitivity and the useful information for toxin surface properties in terms of binding with anti-ricin aptamers as well as antibody. The AFM dynamic force spectroscopy (DFS) was used to measure the single-molecule interactions between antibody or aptamer and ricin protein in buffer solution, which maintains the physiochemical activity of ricin in bulk solution. Also, surface plasmon resonance (SPR) was used for fast detection of ricin. Both AFM and SPR detection required special surface modification with aptamers to capture food toxins, so that surface modification and ricin deposition protocols will be presented. The results of this study will facilitate further development of label-free biosensor for the detection of both natural and chemical toxins in foods.

AGFD 107

Lipid rafts may involve in TFA-induced apoptosis and inflammation of human umbilical vein endothelial cells

Zeyuan Deng1,2, zeyuandeng@outlook.com, Huan Rao2, Bin Qiu1, Benxin Liu1, Jing Li1. (1) State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, Jiangxi, China (2) School of Food Science and Technology, Nanchang University, Nanchang, Jiangxi, China

There is sufficient data showing that TFA do not provide any known nutritional benefits. Rather, some adverse health effects including cardiovascular events, diabetes, compromised fetal growth and child development, and cancers have been acknowledged. This study was to explore how trans-fatty acids activate the caspase pathway, whether

there exist specific receptors induced apoptosis by comparing normal cells and non-rafts cells treated with elaidic acid (9t18:1), linolelaidic acid (9t12t18:2) and oleic acid (9c18:1), respectively. Compared to normal cells treated with 9t18:1 and 9t12t18:2, the cell viability increased and the number of apoptotic cells decreased in non-rafts cells treated with 9t18:1 or 9t12t18:2 (p < 0.05), and the expression levels of pro-apoptotic proteins such as caspase-3, -8, -9, Bax, and Bid decreased, and expression of antiapoptotic protein Bcl-2 increased (p < 0.05). In addition, Fas/FasL expression in cell membrane decreased significantly (p < 0.05). In conclusion, the lipid rafts may involve in TFA-induced apoptosis of human umbilical vein endothelial cells.

AGFD 108

3-MCPD 1-monopalmitate induces apoptosis in NRK-52E cells via activation of p53-JNK pathway

Guoren Huang1, huangguoren@sjtu.edu.cn, Man Liu1, Haiming Shi1, Xiangjun Sun1, xjsun@sjtu.edu.cn, Liangli L. Yu2,1, lyu5@umd.edu. (1) Shanghai Jiao Tong University, Shanghai, China (2) Univ of Maryland, College Park, Maryland, United States

3-MCPD fatty acid esters are reported to be nephrotoxic. However, mechanisms underlying the kidney toxicity induced by 3-MCPD esters are still unclear. Here, we reported that 1-palmitoyl-3-chloropropanediol (3-MCPD 1-monopalmitate) inhibited the growth of NRK-52E rat kidney cells in a dose and time dependent manner. The concentrations required for 50% reduction of cells (IC50 ) were 230.1 μM for 24 h and 151.7 μM for 48 h treatments respectively. This is first demonstration that 3-MCPD 1-monopalmitate-mediated NRK-52E cell apoptosis may involve the activation of ERK, JNK and p53, leading to the up-regulation of caspase 3 gene and bax/bcl2 ratio. In addition, the knockdown of p53 via shRNA transfection significantly decreased the protein levels of p-JNK, p-c-Jun, caspase 3 and bax/bcl2 ratio, indicating the important role of p53 in the 3-MCPD 1-monopalmitate induced apoptosis. In conclusion, the present study provided evidences that 3-MCPD 1-monopalmitate may induce apoptosis in NRK-52E cells via activation of p53-JNK pathway.

AGFD 109

Stable isotope labeling experiments - a useful tool to identify formation pathways of food-borne toxicants

Michael Granvogl, michael.granvogl@ch.tum.de. Technical University of Munich, Garching, Germany

In the past, many studies have been undertaken to mitigate the formation of the so-called “food-borne toxicants” during food processing. But, sometimes, an established pathway is not known or well understood, leading to approaches on the basis of "trial and error". Thus, beside the development of accurate, reliable, sensitive and selective quantitation methods mostly using stable isotopically labeled standards, the elucidation

of formation pathways and parameters influencing the generation of these toxicants is very important. This knowledge at hand enables systematic minimization strategies. The lecture will demonstrate that the use of labeled compounds is a useful tool to get deeper insights into the formation of food-borne toxicants. Thereby, the labeled compounds are not used as internal standards for quantitation but as precursors or intermediates to follow their reactions in model systems or real food by mass spectrometry experiments, e.g., GC-MS or LC-MS/MS. In detail, the proof of formation mechanisms of acrylamide, acrolein, and furan will be presented. Further, the development of quick and simple quantitation methods on the basis of headspace GC-MS analysis will be presented. Their robustness and reliability was proven in comparison to more time-consuming derivatization methods, exemplarily shown for the (E)-2-alkenals acrolein and crotonaldehyde by stable isotope dilution analysis (SIDA) using [13C3]-acrolein and synthesized [13C4]-crotonaldehyde.

AGFD 110

Chemoprotection effect of catechins on detoxcity of dietary acrylamide in mercapturic acid adduct level in rats

Yu Zhang, yuzhangzju@163.com, Jun Cheng, Qiao Wang, Xinyu Chen. Department of Food Science and Nutrition, Zhejiang University, Hangzhou, Zhejiang, China

Many research outcomes regarding analysis, formation and reduction mechanism have been documented since acrylamide contaminant was found in heat processing foods. However, few studies focus on the control of acrylamide in vivo. This study systematically investigated the chemoprotection effect of catechins on acrylamide in rats based on biomarker level of urine mercapturic acid adducts. Results of toxicokinetic studies showed that the half-life period of acrylamide mercapturic acid adducts ranged within 12 hours for both female and male rats with different gavaged doses. The molar ratio of cumulative excretion of adducts in urine to the gavaged dose level of acrylamide and the molar ratio of N-acetyl-S-(2-carbamoyl-2-hydroxyethyl)- L-cysteine (GAMA) to N-acetyl-S-(2-carbamoylethyl)-L-cysteine (AAMA) in high gavaged dose (50 mg/kg bw) was lower than corresponding molar ratios in low gavaged dose (1 mg/kg bw). Results of chemoprotection studies demonstrated that the molar ratio of cumulative excretion of acrylamide mercapturic acid adducts in urine to the gavaged dose of acrylamide in the tea polyphenols, epigallocatechin gallate (EGCG) and epicatchin (EC) intervention groups increased by 24.6%, 14.7% and 22.0%, respectively. Our study revealed that EC mainly contributes to the excretion of acrylamide via its reduction pathway while tea polyphenols mainly contributes to the excretion of acrylamide excretion via its oxidation pathway. This finding indicated that the 3-hydroxyl functional group of flavanols may play an important role in the chemoprotection of acrylamide in vivo.

AGFD 111

Utilization of agricultural by-products in healthful food products: Organogelators, antioxidants, and spreadable products

Hong-Sik Hwang, hongsik.hwang@ars.usda.gov, Jill K. Winkler-Moser, Erica L. Bakota, sean X. liu. USDA, ARS, NCAUR, Peoria, Illinois, United States

It was found that several agricultural by-products could be utilized for healthful food products. Three major applications that our research group has been focusing on will be discussed: 1) plant waxes for trans-fat free, low saturated fat-containing margarine and spread products, 2) extracts of corn distillers dried grains with solubles (DDGS) as antioxidants for frying oil, and 3) rice bran oil extracts as spreads and antioxidants. Plant waxes are obtained as by-products during the oil refining process of vegetable oils such as rice bran oil and sunflower oil. They showed strong gelation ability towards edible oils. They can be added into healthful vegetable oils to form organogels, which are incorporated in margarine and spread products. In these products, solid fats containing high contents of saturated fats and occasionally trans-fats, too are replaced with a small amount of a plant wax and therefore, the major ingredient in the product is the healthy vegetable oil. Corn distillers dried grains with solubles (DDGS) are co-products of dry-milled ethanol production. We have found that corn steryl ferulates extracted from DDGS showed strong antioxidant activity in frying oil and, therefore, they could be utilized as antioxidants. We also found that fractions separated from crude rice bran oil at low temperatures could create a series of spreads showing promise as functional ingredients and as alternatives to peanut butter. A rice bran oil derived spread can also extend the lifetime of frying oil and can also be incorporated into baked goods such as bread and granola as a trans-fat free alternative to butter or shortening.

AGFD 112

Extraction and analysis of high-value compounds in agricultural and forest byproducts using water, ethanol, and carbon dioxide at elevated temperature and pressure as solvents

Victor Abrahamsson, Said Al-Hamimi, Firas Jumaah, Jiayin Liu, Merichel Plaza, Mingzhe Sun, Margareta Sandahl, Charlotta Turner, charlotta.turner@chem.lu.se. Department of Chemistry, Lund University, Lund, Sweden

Water, ethanol and carbon dioxide at elevated temperature and pressure are considered as “green and clean” solvents used in extraction and chromatography processes. In our research, we have developed sample preparation and chromatography methods based on the use of these solvents as extractant/mobile phase [1-3]. In particular, extraction kinetics for lipids and phenolic compounds have been studied and described by mathematical models [3-5]. In this lecture, a background about the fundamentals of pressurized hot water, ethanol and carbon dioxide, and mixtures of these, will be presented, and results from our latest research in sample preparation and analysis of complex samples will be given. Especially, applications in the area of polyphenols in food/forest/agricultural industry byproducts and fat-soluble

compounds in plants and microalgae will be considered. Finally, some aspects on what makes a method or solvent “green” will be discussed [6-7]. References 1. V. Abrahamsson, et al., J. Chromatogr. A., 2012, 1250, 63-68. 2. M. Plaza, et al.,, J. Agric. Food Chem, 2013, 61, 5500-5510. 3. J. Liu, et al., Anal. Bioanal. Chem. (ABC), 2014, 406, 441-445. 4. E. V. Petersson, et al., Anal. Chim Acta, 2010, 663, 27-32. 5. V. Abrahamsson, et al., Anal. Chim. Acta, 2015, 853, 320-327. 6. C. Turner, J. Pure Appl. Chem., 2013, 85, 2217-2229. 7. A. Ekman, et al., J. Cleaner Prod., 2013, 57, 46-58.

AGFD 113

Components responsible for the functional properties of corn fiber gum

Madhav P. Yadav, madhav.yadav@ars.usda.gov. USDA, ARS, ERRC, Wyndmoor, Pennsylvania, United States

Corn fiber gum (CFG) is a hemicellulose (arabinoxylan)-enriched fraction obtained by the extraction of corn bran/fiber using a mild alkaline hydrogen peroxide process. The unique polysaccharide, CFG, with its low solution viscosity has been proposed as a stabilizer for oil-in-water emulsions. We have verified that in some model systems, CFG can out-perform the "gold standard" emulsifier, gum arabic. Our results have also shown that "pure" CFG fractions and the fraction adsorbed on the oil droplets contain considerable amounts of associated lipids, phenolic acids and proteins which contribute to its emulsifying properties. The extraction of CFG with alkaline hydrogen peroxide was investigated using different combinations of alkali concentration and time to identify the optimum extraction condition to retain its functional groups (protein, lipids and phenolic acids). The pure CFG prepared by this process was hydrolyzed with 1.5 N methanolic KOH at 70 °C for one hour to release hydroxycinnamic acids (p-coumaric and ferulic) and lipids. The released phenolic acids and lipids were identified and quantified using HPLC with detection by both UV and evaporative light-scattering detection (ELSD). The presence of these phenolic acids, lipids and protein in CFG may contribute to its excellent emulsifying properties and may combine to give other chemical, physical, and even nutritional properties. Understanding these critical structural elements required for optimal emulsification properties will allow future commercial producers of CFG to provide consistent quality and functionality in their products.

AGFD 114

Hemp waste as a potential source of valuable chemicals

Andrew Hunt1, ajh134@york.ac.uk, Thomas Attard1, Camille Bainier1, Marine Reinaud1, Alexandra Lanot2, Simon McQueen-Mason2, James Clark1. (1) Green Chemistry Centre of Excellence, University of York, York, United Kingdom (2) Centre for Novel Agricultural Products, University of York, York, United Kingdom

Hemp (Cannabis sativa L.) is a C3 plant that has its origins in Central North-East Asia. Hemp fibres have always been known to be of very high quality and can be used in a number of industries such as paper-manufacturing, textiles industries, automotive industries and bio-building industries. During the processing of hemp for fibre production, large amounts of hemp dust are generated, which could be a potential source of valuable chemicals and add value to this otherwise waste residue. Up to 33% of the total hemp in processing can result in dust, which is left unutilised or burnt for energy. Herein, it is demonstrated that dust residues from fibre processing contain significant quantities of high value lipophilic molecules including fatty acids, policosanols (fatty alcohols), fatty aldehydes, hydrocarbons, sterols, triterpenoids and cannabinoids (Cannabidiol (CBD)). Interestingly, dust collected from different stages of the mechanical process gave rise to lipophilic extractives that have significantly different amounts of these hydrophobic components. Of particular interest is CBD, which has low toxicity and is non-psychotropic. CBD has attracted much attention for clinical-level studies on its therapeutic efficacy (alone or combined with various cannabinoids) in the treatment of a variety of central nervous system (CNS) and peripheral disorders. Supercritical carbon dioxide (ScCO2) extraction was conducted on dust samples and optimisation of the process was carried out using the factorial experimental design. Interestingly, it was found that conditions of 400 bar and 65 oC gave rise to the highest % yield of crude wax, in-depth analysis on the concentrations of each family of compounds showed that 350 bar and 50 oC gave rise to the highest yields of the majority of compounds (with the exception of fatty acids). ScCO2 has been demonstrated as a promising pre-treatment step in an integrated biorefinery and as such this may open new doors in hemp waste utilisation. The authors would like to acknowledge the MultiHemp project (agreement number 311849) of the European FP7 framework for financial contribution.

AGFD 115

Pressurized liquid sequential and direct extraction of phytochemicals from Dancy tangerines for their comphrensive characterization by LC–DAD–ESI-HR-MS

Guddadarang Jayaprakasha1, Bhimu Patil2, bpatil@ag.tamu.edu. (1) Vegetable Fruit Improvement Center, Texas A&M University, College Station, Texas, United States (2) Vegetable and Fruit Improvement Center, Texas A&M University, College Station, Texas, United States

Most of the present conventional extraction methods require large amount of solvents, longer extraction time and highly labor-intensive for the extraction of phytochemicals. Additionally, extraction efficiency and reproducibility of current methods are unsatisfactory. Recently, pressurized liquid extraction (PLE) has been emerged as rapid and reproducible promising technique for the isolation of health promoting compounds

from natural sources. This technique uses single or gradient solvents at high pressure with variable temperature above their boiling point to extract different phytochemicals. In the present study, lyophilized Dancy tangerine fruit powder was extracted in both sequential and directly using PLE at various temperature (40, 80, 120 and 160ºC) by different solvents including hexane, ethyl acetate, acetone, methanol and water. All extracts were analyzed for total phenolics, radical scavenging activity and the chemical constituents were identified by liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry (LC-ESI-QTOF-MS). A database was constructed by recording HRMS and MS/MS spectra of tangerine phytochemicals, utilizing both positive and negative ionization modes. PLE provided the highest extraction efficiency within the shortest extraction time (<30min) compared to conventional methods. The developed LC-MS methodology was useful for the dereplication of targeted phytochemicals within 10min. This is the first report on combining PLE with LC–DAD–ESI/MS for the extraction and quantification of limonoids, flavonoids and coumarins from Dancy tangerines. This project is based upon work supported by USDA-NIFA No. 2010-34402-20875, “Designing Foods for Health,” through the Vegetable & Fruit Improvement Center.

AGFD 116

Orange peel by-products as a source of bioactive compounds

John A. Manthey, John.Manthey@ars.usda.gov, Randall G. Cameron. USDA-ARS-USHLR, Fort Pierce, Florida, United States

As a close approximation half of the Florida citrus crop constitutes juice processing by-products. Citrus peel composition has been extensively studied with emphasis on pectin and the polyphenolic antioxidants, mainly flavanones, polymethoxylated flavones, and hydroxycinnamates. The Florida citrus crop can produce upwards to 5000 tons of bioflavonoids. Other value-added byproducts such as limonene occur in peel oil recovered during juice processing. Nonvolatile peel oil constituents are rich in phytosterols as well as the polymethoxylated flavones. The peel flavonoids have been widely studied for their cholesterol-lowering properties and anti-inflammatory actions, and are targets for health-based food supplements. Research is underway to characterize the active blood serum metabolites of the citrus flavonoids, and identification of the biologically active chemical agents. New sources of peel occur with the premature fruit drop caused by the citrus greening disease currently plaguing the Florida citrus crop. Joint recoveries of pectin, limonene, and soluble flavonoids are being explored by the use of steam-explosion technology to extract these components from the main bulk peel, and improve the economy of recovery.

AGFD 117

Interaction/coacervation between food proteins: Mechanisms and potential application

Saïd Bouhallab1,2, said.bouhallab@rennes.inra.fr, Guilherme Tavares1,2, Anne-Laure Chapeau1,2, Pascaline Hamon1,2, Thomas Croguennec1,2. (1) INRA, Rennes, France (2) Agrocampus-Ouest, Rennes, France

The application of fundamental physicochemical concepts for rational design of functional assemblies from food proteins constitute a response to the growing trend toward the development of new and innovative food products and also an opportunity to generate new protein-based supramolecular structures with new applications. Because of their omnipresence in food systems and their biodegradability, proteins are the focus of many attempts for their use as building blocks for such supramolecular structures. Controlled self- co-assembly of proteins can generate a variety of supramolecular structures that vary in shape, size and density (fibrils, spherulites, nanotubes, etc). For instance, well-defined microspheres called coacervates can be formed by control mixing of oppositely charged proteins.1 The objective of our research is to understand the mechanisms behind such spontaneous coacervation process from molecular interaction to micro-scale characterization. In this presentation, we will summarize the results obtained on several binary protein systems and will show that co-assembly of proteins into coacervates (Figure 1) is a generic process that is, de facto, independent of the amino acid composition. We will report on the requirements that drive such spontaneous co-assembly: protein conformational state and flexibility, molar stoichiometry, total protein concentration, charge anisotropy, etc. The research challenges and the promising uses of these supramolecular structures in food and non-food sectors (encapsulation of bioactives, design of edible films) will be discussed. [1]: S., Bouhallab and T., Croguennec. Adv. Polym. Sci. 2014,256, 67–102

AGFD 118

Assembly of protein/polysaccharide complexes-based Pickering emulsions for nutraceutical delivery

Qingrong Huang, qhuang@aesop.rutgers.edu. Food Science, Rutgers Univ, New Brunswick, New Jersey, United States

Carboxymethyl Dextran (CMD) has broad applications due to its intrinsic merit in chemical structure and universal compatibility with other biomacromolecules. Understanding the microstructure, the interaction in CMD solutions and its complexes with globular proteins is crucial for rational development of CMD based products. In this paper, small angle X-ray scattering and turbidimetric titration were applied to investigate CMD solutions and CMD complexes with beta-lactoglobulin (BLG) as a function of CMD concentration. CMD solutions behave like typical polyelectrolyte solutions where chains become collapsed due to the screening of electrostatic repulsions with concentration increase. In the CMD and BLG mixed solutions, proteins tend to distribute along CMD chains, and the sizes of CMD/BLG complexes were determined by the combined effects of the microstructure of CMD solution, the globular electrostatic interaction and the total charge neutrality. The complexes formed by CMD and BLG were further used to form

Pickering emulsions and encapsulate polyphenols. Their microstructures, rheological properties and digestion profiles will also be discussed.

AGFD 119

Formation of a coacervate film across the oil-water interface: Stabilization of emulsions

Hélène Monteillet, Mieke Kleijn, mieke.kleijn@wur.nl, Frans Leermakers, Joris Sprakel. Physical Chemistry and Soft Matter, Wageningen University, Wageningen, Netherlands

Using two oppositely charged polymers of which one is soluble in oil and the other in water, it is possible to obtain an electrostatic complex at the oil-water interface. We demonstrate this with a new oil-soluble polyanion, poly-(fluorene-co-benzothiadiazole-co-benzoic acid), in combination with various water-soluble polycations. The polyelectrolytes on their own are not interfacial active, as shown by interfacial tension measurements; however, when both types of polyelectrolytes are present, in the oil and water phases respectively, the interfacial tension rapidly decreases. Confocal fluorescence microscopy verifies the presence of both polyelectrolytes at the interface. Owing to the purely electrostatic nature of the interfacial complex, it is very sensitive to ionic strength and pH. Emulsions can be stabilized for several weeks; using the sensitivity of the complex to pH, it is possible to reversibly make and break emulsions. Bright field images of pendent water drops in oil during coacervation show an interesting phenomenon: overtime, the droplets become less transparent, indicating the slow formation of the coacervate “skin” and suggesting that this is inhomogeneous on optical length scales. This motivated us to do a theoretical study using the Scheutjens-Fleer self-consistent field theory. Although far from the practical situation we first focus on a model which is fully symmetric with respect to chain architectures, dielectric permittivities, excluded volume interactions and concentrations of all components. When the counter ions are non-selective (no preference for the oil or the water phase), we find that the theory does appropriately predicts the coacervate formation at the oil-water interface. From calculated adsorption isotherms we conclude that the system follows a pseudo partial wetting scenario, characteristic for systems that feature interactions on different length scales. The partial wet state implies lateral inhomogeneities along the coacervate film and this possibly explains the observed loss of transparency of the pendent water drops.

AGFD 120

Complex coacervation in heteroprotein systems: A special form of macromolecular liquid-liquid phase separation

Daniel Seeman1, dseeman@chem.umass.edu, Paul L. Dubin2. (1) UMass-Amherst, Amherst, Massachusetts, United States (2) Dept. of Chemistry, UMass-Amherst, Amherst, Massachusetts, United States

There has been a resurgence of interest in complex coacervation, a form of liquid-liquid phase separation (LLPS) in systems of oppositely charged macroions, but very few reports describe the somewhat anomalous coacervation between acidic and basic proteins, which occurs under very narrow ranges of conditions. Lactoferrin (LF) and beta-lactoglobulin (BLG) are among the protein pairs that exhibit heteroprotein coacervation, a unique and relatively unexamined type of liquid–liquid phase separation (LLPS). It was previously found that LF and BLG undergo coacervation at highly constrained conditions of pH, ionic strength and protein stoichiometry. The precise balance of repulsive and attractive forces among these units, thought to stabilize the coacervate, is achieved only at limited conditions of pH and I. Our purpose here is to define the process by which such structural units form, and to elucidate the forces among them that lead to the long-range order found in equilibrium coacervates, requiring a very precise balance of repulsive and attractive forces.

AGFD 121

Complex coacervation with oppositely charged polymer and surfactant: Determination factor in the morphology of coacervated complexes during the dilution process

Miyuki Miyake, miyamiyu@lion.co.jp. Fubctional Materials Sciencece research laboratories, Lion Corporation, Edogawa-ku, Tokyo, Japan

The complex coacervation system is quite useful as a surface modification in the field of cosmetics and toiletries, especially that of coacervates with cationic polymer and anionic surfactant, which affect the hair texture during rinsing of conditioning shampoo. The mechanism in which the coacervated complexes change the hair texture was investigated by phase diagrams and light scattering to solubilized states and rheology and morphology observation to coacervated complexes. The high charges of cationic polymer and anionic micelle and the flexibility of polymers conducted complexes to be densely packed. The low charges of these and the stiffness of polymers formed looser morphology of complexes which caused hair texture to become smoother.

AGFD 122

Optimization of milk protein–native gum (tragacanth and Persian gums) interactions: Complex coacervation and soluble complexes

Soleiman Abbasi, mabbasifood@hotmail.com, Fatemeh Azarikia. Food Science, TMU, Tehran, Tehran, Iran (the Islamic Republic of)

Aggregation and precipitation of proteins at isoelectric pH, can be prevented by their interaction with negatively charged polysaccharides. However, electrostatic interaction between these negatively charged biopolymers either may lead to formation of stable colloidal system or cause phase separation via complex coacervation. Therefore, in the present study, the efficiency of soluble fraction (SF) of Iranian native gums [gum

tragacanth (GT) and Persian gum (PG)] on formation of complex coacervation and soluble complexes with milk proteins [sodium caseinate (CN) and whey protein isolate (WPI)] over a wide range of pH (2–7) was investigated. Furthermore, the capability of soluble complexes on stabilization of emulsions was examined. Based on visual observation, optical density, and zeta potential measurements, soluble complexes of CN–SFPG, CN–SFGT, WPI–SFPG, and WPI–SFGT were formed at 0.4:0.6, 0.4:0.6, 0.4:2 and 0.4:1wt% ratios over a wide range of pH (2–7), respectively. Whereas, complex coacervation of CN–SFPG (0.4:0.2 and 0.4), CN–SFGT (0.4:0.2), WPI–SFPG (0.4:0.2 to 0.8), and WPI–SFGT (0.4:0.2 to 0.8) was only occurred at narrow pH range (2.6–3.2). Based on zeta potential measurements, electrostatic repulsion had important role on the stability of emulsions; although, steric repulsion and high viscosity of systems were effective, as well. With regard to our achievements, we believe that soluble complexes and complex coacervates can potentially be used in formulation of emulsions to enhance their stability against environmental stresses as a novel emulsifiers as well as microencapsulation of functional ingredients, respectively.

AGFD 123

Study of complex coacervation of gelatin A with sodium carboxymethyl cellulose/sodium alginate/carrageenan: Formation of smart microparticles and encapsulation

Nirmala Devi1, nirmaladevi2040@gmail.com, Tarun K. Maji2, Dilip K. Kakati3. (1) Chemistry, Indian Institute of Technology Guwahati, Assam, India, Guwahati, India (2) Department of Chemical Sciences, Tezpur University, Napaam, India, Tezpur, Assam, India (3) Chemistry, Department of Chemistry, Gauhati University, Guwahati, India, Guwahati, Assam, India

Complex coacervation behaviour in some of the naturally occurring biopolymer systems was studied. Complex coacervation between gelatin A and SA/SCMC/carrageenan was carried out for the synthesis of microparticles and encapsulation of active agents.The efficiency of complex coacervate formation was followed by measuring viscosity, coacervate yield and turbidity of the polymer test solutions. Maximum complex coacervation for gelatin/SCMC was found to occur at pH=3.5 and SCMC/gelatin ratio of 1:2.33 in the mixture. For SA/gelatin, effective complex coacervation was realized at pH= 3.7 and when the ratio of the two polymers was 1:3.5. Whereas gelatin/carrageenan system showed maximum complex coacervation at pH=3.5 and gelatin/carrageenan ratio of 2:1 in the mixture. Microparticles containing different solid and liquid active agents such as isoniazid, ascorbic acid and oil were prepared by using the complex coacervation technique at the optimized conditions.The encapsulation efficiency of active agent was dependent on the amount of crosslinker, active agent loading and polymer concentration. Scanning electron micrographs (Fig.1) showed the formation of free flowing spherical microparticles on loading the active agents. The formation and the size of the microparticles were also dependent on the amount of surfactant, crosslinker and stirring speed. The microparticles were found to be pH responsive. The microparticles were further characterized by fourier transform infrared

spectroscopy (FTIR), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC) and X-ray diffraction (XRD) study.

Fig.1: Representative SEM images of the complex coacervate microparticles

AGFD 124

Variation in Maillard reaction product formation in oats from 13 cultivars

Michael J. Morello, mike.morello@pepsico.com, Bret C. Vastano. PepsiCo Global RD, Barrington, Illinois, United States

Oats typically undergo a thermal process, kilning, prior to their use in consumer products. This thermal processing induces formation of Maillard Reaction products, which can influence flavor. As with many small grains multiple cultivars are grown and co-mingled throughout any crop year. To assess potential variation in formation of Maillard Reaction Products across a growing season, Maillard Reaction Products from 13 cultivars grown in three locations underwent thermal processing in a pilot system designed to mimic kiln processing, and were analyzed by HS-SPME-GC-NPD. Results

demonstrating the relative variation in formation of Maillard Reaction Products across the various cultivars will be presented. Additionally, results will be presented that demonstrate growing location had a smaller impact on formation of Maillard Reaction Products than cultivar.

AGFD 125

Characterization of key aroma-active compounds in raw and roasted mustard seeds (Sinapis alba L.)

Michael Granvogl, michael.granvogl@ch.tum.de, Eva Ortner, Peter H. Schieberle. Technical University of Munich, Garching, Germany

Thermal processing of food leads to the formation of different desired quality aspects, mostly accompanied by characteristic changes in aroma, taste, and color. Thereby, the differences in the overall aroma of the raw and heat-processed product is based on interactions of odorants, which are either formed by reaction of different compounds or by release from specific precursors during heat-treatment on the one hand. But, on the other hand, it might also be the case that important compounds for the overall aroma of the raw food are degraded. The lecture will present these characteristic changes, exemplified for the seeds of white mustard (Sinapis alba L.; Brassicaceae), using the Molecular Sensory Science Concept consisting of aroma extract dilution analysis (AEDA), identification experiments, stable isotope dilution analysis (SIDA), calculation of odor activity values (OAVs), and, finally, recombination experiments. After identification and quantitation of 33 aroma-active compounds in raw mustard seeds eliciting a green, cabbage-like aroma and 40 odorants in roasted seeds having a popcorn-like, coffee-like aroma, the respective OAVs were calculated. Thereby, dimethyl sulfide (OAV = 327; asparagus-like aroma), methanethiol (186; cabbage-like, sulfury), 2-isopropyl-3-methoxypyrazine (21; earthy, green bell pepper), 3-(methylthio)propanal (31; cooked potato-like), and 1,8-Cineol (16; eucalyptus-like) were shown to be important aroma compounds for the raw seeds. In roasted seeds, 28 key odorants had an OAV > 1 revealing 2-furfurylthiol (OAV = 1870; coffee-like, burnt) 3-methylbutanal (73; malty), and 2-acetyl-1-pyrroline (290, popcorn-like) as important contributors to the overall aroma. Further, remarkably concentration changes were analyzed for, e.g., 4-hydroxy-2,5-dimethylfuran-3(2H)-one (caramel-like) increasing from 27.7 µg/kg (raw) to 2010 µg/kg (roasted), or the formation of new compounds like 2-ethyl-3,6-dimethylpyrazine (roasty, earthy) or 2,3-pentandione (butter-like) was proven. Possible formation mechanisms of single odorants will finally be presented and discussed.

AGFD 126

Defining mechanisms of flavor development in foodstuffs

Devin G. Peterson1, dgp@umn.edu, Liyun Zhang1, Smaro Kokkinidou2. (1) 225 FScN, Univ Minnesota, Saint Paul, Minnesota, United States (2) Food Science and Nutrition, University of Minnesota, Saint Paul, Minnesota, United States

This presentation will focus on the characterization of flavors developed during the browning reactions of foodstuffs throughout thermal processing and storage. Emphasis will be placed on aroma and taste development in food products such as cereal, milk, cocoa, wheat, potato and fruit. Discovery of novel compounds with flavor activity and modulating attributes as well as mechanisms of flavor generation will be discussed. Analytical approaches such as targeted sensory-guided fractionation techniques, mapping of key intermediates, and isotopic labeling methods will be presented. An improved mechanistic understanding of flavor generation from food ingredients provides the ability to develop higher flavor quality from process optimization rather than by the addition of flavoring materials.

AGFD 127

Formation of reactive fragmentation products during Maillard degradation of higher sugars

Margus A. Glomb, marcus.glomb@chemie.uni-halle.de, Mareen Smuda, Christian Henning. Institute of Chemistry - Food Chemistry, Martin-Luther-University Halle-Wittenberg, Halle, Germany

The amin induced fragmentation of higher sugars plays a pivotal role during the formation of browned flavors. The shorter-chained products are reactive intermediates in the further reaction cascades leading to taste or odor active, or browned compounds, or they are flavor active as such. The three mechanisms of fragmentation are retro-aldol-reactions, hydrolytic β-dicarbonyl cleavage and hydrolytic oxidative α-dicarbonyl cleavage. A fourth mechanism, namely hydrolytic α-dicarbonyl cleavage, present in the older Maillard literature was outruled as void. The mechanisms are discussed in detail for the degradation of ascorbic acid. Here, the amin catalyzed breakdown was comprehensively investigated on the basis of isotopic labeling experiments with 13C-ascorbic acid isotopomers and 18O2 atmosphere. As a result a multitude of carbonyl compounds, carboxylic acids and carboxylic amides was established explaining more than 75% of the breakdown. These results can also be transferred to other reducing sugars present in foods.

AGFD 128

Characterization of color formation in juice products

Laurianne Paravisini2, lparavis@umn.edu, Devin G. Peterson1. (1) 225 FScN, Univ Minnesota, Saint Paul, Minnesota, United States (2) 225 FScN, University of Minnesota, Saint Paul, Minnesota, United States

In juice products, non-enzymatic browning is the most important reaction responsible for quality and color changes during prolonged storage. Thus far, it has been well established that ascorbic acid degradation and the Maillard reaction (MR) are both involved in browning. The decomposition of ascorbic acid is reported to be the major degradation reaction occurring during the storage of citrus products. Numerous studies have shown a strong correlation between the loss of ascorbic acid and an increase in browning over time. However, the mechanisms of browning and the contribution of the precursors still remain unclear. The main objective of this study was to characterize the mechanisms of browning by focusing on the intermediate reactive carbonyl compounds (RCS) involved in both ascorbic acid degradation and MR pathways. The RCS composition of apple, orange and lemon juices were characterized and monitored over 10 weeks of storage under ambient (22oC), elevated temperature (35oC) and refrigerated (4oC) conditions. The correlation between the evolution of RCSs concentrations and the color development was then evaluated. Finally, precursors of RCSs were investigated by isotope-labeling techniques in model systems, allowing for the identification of the potential precursors of browning in juices. The RCS composition over time appeared to be highly dependent on the juice system and involved the degradation of both sugars and ascorbic acid. For example, both threosone and 3-deoxyglucone were highly correlated with brown color development in all of the juice systems. In summary, RCS intermediates from both sugar and ascorbic acid degradation were highly correlated to non-enzymatic browning in juice. This work will provide the foundation to develop novel technology to control color formation in juice.

AGFD 129

Development and applications of surface-enhanced Raman spectroscopy in food science

Lili He, lilihe@foodsci.umass.edu. food science, University of Massachusetts-Amherst, Belchertown, Massachusetts, United States

In this oral presentation, Dr. Lili He will introduce the development and applications of advanced and innovative surface-enhanced Raman scattering (SERS) techniques in three different areas in food science: food safety, food chemistry and food bioscience. In the food safety application, a single aptamer based SERS method for rapid detection of multiple pesticides in food will be introduced. This method is able to rapidly detect and discriminate four pesticides simultaneously in a complex food matrix (i.e. apple juice). In the food chemistry application, Dr. Lili He will talk about the fabrication of novel lipophilic and amphiphilic gold nanoparticles to study food lipids and emulsion interfaces, respectively. In the food bioscience application, a label-free imaging approach to study the interactions between polymethoxyflavones and cancer cells will be introduced.

AGFD 130

Primary expectations of secondary metabolites

Jungmin Lee, jungmin.lee@ars.usda.gov. USDA-ARS-HCRU worksite, Parma, Idaho, United States

Plant secondary metabolites (e.g., phenolics) are important for human health, in addition to the organoleptic properties they impart to fresh and processed foods. Consumer expectations such as appearance, taste, or texture influence their purchasing decisions. Thorough identification of phenolic compounds is key to discerning how cultivar selection, breeding strategies, environment, post-harvest conditions, and processing methods ultimately affect food quality. Highlights on the elucidation and quantification of phenolics in grapes, berries, culinary and medicinal herbs, along with dietary supplements will be presented; data can improve production and processing. Discussion will include how (1) canopy, cover crop, and nutrient management alter metabolites in wine grapes; (2) winemaking conditions influence wine structure; (3) characterizing cultivar variability in polyphenolics aiding enhanced germplasm selection in berries; (4) processing methods alter efficiency of phenolic extraction in final products; and (5) utilizing phenolic profiles to detect adulteration.

AGFD 131

Concentration of propolis extract using hydrophobic membrane

Choe Peng Leo, chcpleo@eng.usm.my. School of Chem Eng-Engineering Campus, Universiti Sains Malaysia, Nibong Tebal Penang, Malaysia

Unlike honey, propolis cannot be consumed without solvent extraction. The solvent in propolis extracts must be reduced or eliminated to produce concentrated extract. Traditionally, vacuum distillation, lyophilization or evaporation is used to concentrate propolis extract. The mentioned techniques are high in energy consumption, sometimes they also involve high temperature which may cause the degradation of bioactive compounds especially flavonoid and phenolic compounds. Hence, the potential of membrane distillation in the concentrate of propolis extract was evaluated in this work. A hydrophobic membrane works as the barrier for liquid phase, allowing only the vapor (water or solvent) to be transported through the membrane pores under vacuum. The driving force of the process is the partial vapor pressure difference which was commonly controlled through temperature difference. The propolis extract was successfully concentrated using membrane distillation. However, fouling remained to be the main obstacle in this separation technique. The fouled membrane was examined to understand the interaction between propolis components and hydrophobic membrane.

AGFD 132

Utilization of in vitro and in vivo gastrointestinal models in the production of optimum functional food formulations

Yuwen Ting1,3, pam.yt.ting@gmail.com, Yike Jiang1, Chi-Tang Ho1, Qingrong Huang2,3. (1) Food Science, Rutgers University, Edison, New Jersey, United States (2) Food

Science, Rutgers Univ, New Brunswick, New Jersey, United States (3) School of Food Science and Technology, Wuhan Polytechnic University, Wuhan, China

Nutraceuticals are bioactives found in many dietary sources. Numerous publications reported their ability to prevent the development of degenerative diseases through modulation of the physiological and physiochemical processes. Having sufficient concentration at the target site of action is the critical factor for them to deliver the health benefits. For consumers, it is most accepted to ingest the bioactive components through oral route since it is convenient, cost-efficient, and allows flexible dosing schedule. Thus, it is important to develop methods that can rapidly evaluate the effects of food formulations and processing conditions on bioavailability of those bioactive compounds. To predict the oral bioavailability of nutraceuticals, many in vitro and in vivo models have been used to select for the optimum system and to predict the pharmacokinetic process in human. Application of those screening models will greatly benefit the investigator when developing functional food products giving higher absorption rate of bioactive compounds.

AGFD 133

High-resolution mass spectrometry for the exploration of novel plant sterol conjugates

Laura Nystroem, laura.nystroem@hest.ethz.ch. ETH Zurich, Zurich, Switzerland

Plant sterols, which are commonly applied as cholesterol lowering ingredients in functional foods, have recently been discovered to possess also several additional technological and health benefits, like surface activity and emulsifying capacity, as well as immunomodulating and anticarginogenic activities of steryl glycosides and steryl phenolates. The functions of these polar steryl conjugates in plants and their metabolism in humans are still poorly understood, partially due to limitations in their analysis. We have developed new, high-resolution mass spectrometry (HR-MS/MS) based methods for the detailed analysis of sterol conjugates in plant based foods and biological samples, and have successfully applied them to identify new compounds and collect detailed information on the contents and profiles of steryl glycosides and steryl phenolates in plants. Furthermore, we can enzymatically produce these conjugates in high yields, thus facilitating the production of pure steryl conjugates for use as analytical standards and multifunctional ingredients in functional foods.

AGFD 134

Integrated advanced multi-omics strategies: A new paradigm for agricultural and medicinal plant biotechnology

Joaquim Marques2, jmarques@wsu.edu, Doralyn S. Dalisay2, Michael Costa2, Barri Herman2, Laurence B. Davin1, Norman G. Lewis2. (1) Inst of Biological Chem,

Washington State University, Pullman, Washington, United States (2) Institute of Biological Chemistry, Washington State University, Pullman, Washington, United States

The advancement and incisive application of big-data “omic” technologies are occurring at breakneck speed to resolve long-standing challenges/bottlenecks in agricultural/medicinal plant biotechnology. Two examples are described here. First, poplar lines, genetically engineered and purposed for their potential as biofactories to produce fine/specialty/commodity chemicals, were subjected to multi-omics analyses. In one example, a biochemical pathway was introduced to produce the main component of rose oil, 2-phenylethanol. In this way, we can now readily meet the world’s demand for this natural product widely used as an aroma/flavor/fragrance. Untargeted metabolomics analyses provided new insight into the efficacy of these genetic manipulations. Secondly, combination of multi-omics datasets, MALDI imaging mass spectrometry/laser capture microdissection enabled identification of a new step in podophyllotoxin biosynthesis (used extensively in chemotherapy) in Podophyllum species, and unveiled an unknown biochemical pathway leading to an anxiolytic alkaloid biosynthetic pathway. These examples are used to demonstrate the potential of these rapidly advancing technologies.

AGFD 135

Recovery and development of value-added applications of fruit juice processing byproducts

Yanyun Zhao, yanyun.zhao@oregonstate.edu. Food Science & Technology, Oregon State University, Corvallis, Oregon, United States

Fruit juice processing results in significant amount of bio-residuals, named “fruit pomace”. Fruit pomace contain a large amount of bioactive compounds including polyphenolics, dietary fibers, smaller amounts of proteins and minerals, and some remaining juices with acids, sugars, and other soluble substances. Disposal of pomace has been a burden for the juice industry since the direct disposal of pomace into soil or landfills can pose potential environmental problem, thus inhibited. Fruit pomace may be utilized for various value-added applications, including as functional food ingredient in a wide range of food products for promoting human health and extending product shelf-life, as fermentation substrates, extraction of the bioactive compounds, and making into biodegradable packaging materials. Through technology innovation, the tons of waste product from being a burden can be transformed to becoming a valuable source of revenue for the juice and wine companies. This presentation will report our studies in the recovery of the bioactive compounds from fruit pomace, and provide an overview on the bioactive compounds in various fruit pomace, economically feasible methods to dry wet pomace for long-term storage, and examples of utilizing pomace in different food products (from bakery goods to processed meat products) and as bulk material for developing biodegradable packaging materials.

AGFD 136

Eco-innovative polyphenol extraction using subcritical water from red and white pomace, coupled with purification by membrane processes

Sami YAMMINE1, sami.yammine@u-bordeaux.fr, Xavier Vitrac2, Robin Rabagliato3,4, Martine Mietton Peuchot3,4, Rémy GHIDOSSI3,4. (1) Université de Bordeaux USC 1219 Œnologie INRA/IPB/UBS EA 4577, Unité de recherche Oenologie, Villenave d'Ornon, France (2) Laboratoire PHENOBIO SAS, Centre Montesquieu, Martillac, France (3) Université de Bordeaux, Institut des Sciences de la Vigne et du Vin, EA 4577, Unité de recherche œnologie, Villenave d'Ornon, France (4) INRA, ISVV, USC 1366 Œnologie, 210 Chemin de Leysotte, CS 50008, F-33882 Villenave d'Ornon, France, Villenave d'Ornon, France

Grape by-products are rich in health promoting polyphenols. Their extraction from winery waste and their consequent purification are of special interest to produce extracts with high added value compounds. Meanwhile, the growing concern over environmental problems associated with economic constraints, require the development of environmentally sustainable extraction technologies. The extraction using semi-continuous subcritical water, as a natural solvent at high temperature and pressure is a promising “green” technology. The suitable feature of subcritical water as leaching agent is its capacity to decrease dielectric constant as a function of increase in temperature, allowing a better solubility for the compounds of interest. In our study subcritical water extraction of polyphenols from the red and white grape pomace of Dunkelfelder, Cabernet Franc, Merlot, and Chardonnay was performed. Semi-continuous extraction leads to crude extracts rich in different families of polyphenols. A purification step prior to industrial usage is essential. Coupling subcritical water extraction with membrane processes, due to large array of flexibility, offers a solution for the purification and fractioning of the crude extracts. The combined effects of extraction temperature (from 60 to 200 °C), pressure (from 25 to 100 bar), flow rate (1 to 10 ml/min), sample mass (5, 70, 1000g) were investigated and compared to traditional solvent extraction (1:1 ethanol/water). Optimal extraction conditions were found to be 150°C and 6ml/min irrelevant of the pressure used. These conditions produced crude extracts containing 268 mg/100g Dry Weight (DW) of anthocyanins (+23% compared to traditional methods of extraction) and 3406 mg/100g DW procyanidins (+34%). Subsequently, to complete the purification of the crude extract, several organic membranes having differential molecular weight cut off points ranging from 200 Da up to 0.45 μm were tested. The results allow evaluating both the permeate flux through the membranes and the rejection rates of the major compounds found in the crude extract. The tested membranes have differential selectivity for polysaccharides, proteins, and different families of phenolic compounds with high purity. Confirming the applicability of membrane separation for the fractionation and purification of pomace extracts. More

research is needed to validate the industrial up scaling and the intended application of the produced extracts.

AGFD 137

Incorporation of pressurized fluid technology in the recovery of bioactive constituents from pomace processing wastes

Jerry W. King2, kingjw100@hotmail.com, Luke Howard1. (1) Univ of Arkansas, Fayetteville, Arkansas, United States (2) Department of Food Science , University of Arkansas, Fayetteville, Arkansas, United States

Further utilization of food processing wastes is highly desirable and also has the potential for producing economically-viable, value-added products. This presentation focuses on the use of pressurized fluids, i.e. such as compressed carbon dioxides and water, to extract and enrich in particular naturally-occurring antioxidants, such as polyphenolic laden constituents, from grape and berry pomaces, in an environmentally-safe and sustainable manner. In the case of the polar polyphenolic constituents, the use of subcritical water versus supercritical carbon dioxide will be examined based on solubility parameter - molecular structure correlations. Fruit wastes contain both lipophilic constituents and aqueous soluble components; the former group having solubility parameters (δ) in the range of 15-18 MPa1/2 while the latter group embraces constituents having δ of 30-37 MPa1/2. Hence without the use of co-solvents, SC-CO2 is not the optimal solvent for extracting polyphenolic-based antioxidants. Optimization when using one or both of the above media for extraction- and fractionation-based processing will be described in terms of the following parameters: (a) pressure, temperature and solvent/feed (S/F) ratio; (b) the time of extraction, space velocity of the fluid, reactivity of the targeted solutes to yield processing times in the range of 10-20 min. or lower, and (c) modeling of the global extraction rate curves to assist in minimizing extraction times at S/F’s < 10-20. The potential of ultrahigh pressure SFE to enhance the extraction of more polar constituents existing at low concentration levels in waste pomace will also be noted thereby avoiding the use of an organic co-solvent. Scaling up of these critical fluid-based processing options will also be described – particularly for the use of continuous feed processing, the hot-cold extraction option, and semi-continuous batch extraction. This will a description of companies that re now producing products containing the above constituents, the resultant products, and the integration of the critical fluid processing platforms into existing food processing operations. Finally, the cost of manufacturing (COM) of these extracts from food/agricultural waste materials using critical fluids, will be characterized using the formula of Turton.

AGFD 138

Physicochemical challenges to recover polyphenolics from Concord grapes skins

Steve Talcott, stalcott@tamu.edu. Nutrition and Food Science, Texas AM University, College Station, Texas, United States

Polyphenolic consumption from grapes has a positive health association, especially against inflammation and cardiovascular disease. The polyphenolic content of Concord grapes (Vitis labrusca) is a mixture of anthocyanins, flavan-3-ols and their polymers, phenolic acids, and flavonols. Concord grapes are abundant in the US and therefore a significant source of phytochemicals, yet relatively few value-added by-products are commercially available. Concord finds its primary market in fruit juice and puree, leaving a residual biomass that is high in sugars and demands an immediate processing solution to limit microbial growth. The amount of recoverable polyphenolics in grape skins is largely dependent on the method of extraction, with aqueous systems less expensive and more universal than alcoholic solvents. Variable extraction methods were evaluated including solvent selection, acidity, temperature, and physical conditions such as the presence of grape seeds and use of cell wall degrading enzymes. Polyphenolics were then evaluated before and after drying and following desorption from solid phase supports such as Amberlite FPX-66 resin to remove sugars. Although alcoholic solvent extraction was superior in total polyphenolic recovery compared to acidified water for each phytochemical class, stability of the solvent extracts were lower (45% loss) than water extract (73% loss) under oxidizing conditions. The incorporation of either seed tannins or soluble proteins was exceedingly detrimental to both anthocyanins and total phenolic recovery with differences ranging from 33-50% compared to a control due to hydrogen bonding and polymerization reactions. Acidified hot water added directly to grape pomace was deemed the most viable extraction process due to its low cost and reduced interactions among polyphenolics and proteins. However, the commercial applications for Concord grape by-products may dictate actual processing conditions ranging from dietary supplements to water-soluble food ingredients.

AGFD 139

Comparison between antioxidant potentials of extracts from black chokeberry pomace and walnut husk using supercritical carbon dioxide and ethanol

Jonathan Wenzel3, jwenzel@kettering.edu, Thomas Dixon3, Evan Tucker3, Laron Burrows3, Nate Dwarshuis3, Elyse Hossink1, Lihua Wang2, Michelle Ammerman1, Cheryl Samaniego1. (1) Applied Biology, Kettering University, Flint, Michigan, United States (2) Chemistry, Kettering University, Flint, Michigan, United States (3) Chemical Engineering, Ketteirng University, Flint, Michigan, United States

During processing of berries and nuts, a considerable amount of material such as pomace, husks, and shells is produced and often discarded as compost or landfill. In the case of berries, such as chokeberries, cranberries, and grapes, the skins and seeds that make up the pomace have higher amounts of bioactive polyphenolic compounds than the pulp which contains most of the juice. Chokeberries are shown to contain a higher total polyphenol content and anthocyanin content than other commercially grown

fruits. Health benefits that have been reported from chokeberries include inflammation reduction, antimicrobial properties, as well as improvements in vascular health. In producing walnuts, the husks are often discarded. Walnut husks contain tannins and other compounds and the husk has been used for treatment of intestinal parasites and fungal infections. Supercritical fluids, due to their high diffusivity are excellent solvents for extracting chemical species from plant matter. Supercritical carbon dioxide, by itself has a relatively low dielectric constant, and ethanol can be added as a modifier to help improve extraction. This work evaluates the effects of pressure, temperature, and ethanol content of supercritical carbon dioxide with ethanol upon the extraction of walnut husks and chokeberry pomace. The quality of the two plants extracts are compared and ideal extraction conditions will be discussed. The antioxidant potential was evaluated using the total phenolic content (TPC) assay and the ferric reducing ability of plasma (FRAP) assay. DNA protection abilities of the extracts, as well as chemical composition, will also be presented.

AGFD 140

Acrylamide – a challenge to food scientists in industry and academia

Donald S. Mottram, d.s.mottram@reading.ac.uk. Department of Food and Nutritional Sciences, University of Reading, Reading, United Kingdom

The enormous improvements in the quantity and quality of food available to the consumer since the 1950s have been possible because of tremendous advances in agricultural and food science. Multidisciplinary research and collaboration between researchers in academia, industry and government has been key to so many of these advances. Throughout his career Andy Taylor has sought to promote such collaborations. One of the best examples of cooperation is way in which the major problem of acrylamide in food has been tackled. The discovery in 2002 of acrylamide, a possible carcinogen, in fried and baked potato and cereal products caused one of the biggest challenges ever to hit the food industry. Investigations in Europe and the USA established quickly that acrylamide derives, during the high temperature cooking of cereal- and potato-based foods, from the Maillard reaction between the amino acid asparagine and reducing sugars. Establishing the detailed mechanism of formation proved more difficult but by far the greatest challenge has been to mitigate acrylamide formation during cooking while maintaining the characteristic flavors and colors of fried and baked foods that are also formed in the Maillard reaction. While complete elimination of acrylamide from food has proved to be unachievable, significant advances have been made in reducing its level though modification to processing methods, plant breeding and agronomy. This has necessitated multidisciplinary research involving food chemists, food technologists, biochemists, agriculturalists and plant geneticists. Acrylamide research has seen unprecedented levels of collaboration between industry and academia, especially in Europe, and food companies have widely shared knowledge so the levels of acrylamide foods are kept as low as is practically attainable. The next 25 years will see increasing pressures on the agric-food industries to deliver more food for the growing world population while reducing adverse impact on the

environment. Similar levels of collaborations between academia, industry and government are going to be necessary to achieve these new goals.

AGFD 141

2,5-Diketopiperazines – interesting markers of reaction or compounds with sensory and bioactive properties?

Neil C. Da Costa, neil.dacosta@iff.com, Michael Z. Chen. R&D, IFF, Union Beach, New Jersey, United States

2,5-Diketopiperazines are a class of semi-volatiles compounds ever increasingly found and reported in foods, beverages and ingredients containing thermally processed proteins and peptides. Derived from the cyclization of two amino acids moieties, they are generally stable compounds first reported in cocoa, beer, beef and coffee and often described as bitter tasting with few apparent applications. The combinations of cyclic dipeptides and their occurrence has increased greatly in recent years. However in the pharmaceutical industry these compounds have been known for several years as having potential bioactive properties, such as antitumor, antiviral, antifungal and antibacterial. This presentation is an overview of these compounds, occurrence and reported properties.

AGFD 142

On-line aroma monitoring with mass spectrometry and link to flavor release and flavor perception

Jean-Luc Le Quere, jllequere@dijon.inra.fr. Center for Taste and Feeding Behavior, INRA, Dijon, France

On-line monitoring of aroma compounds has benefitted from the development of direct injection mass spectrometry (MS) techniques that add the time dimension to the precision, sensitivity and robustness of MS in the analyses of volatile organic compounds. Thus proton transfer techniques such as atmospheric pressure chemical ionization (APCI), proton transfer reaction (PTR) or more recently selected ion flow tube (SIFT) coupled to MS have been used quite extensively for the last 10 years in the food area. These direct injection MS techniques accommodate the necessary instrumental constraints in terms of fragmentation, speed and response (linearity and limit of detection) compatible with in vitro aroma analyses (aroma release function of food matrix effects, process monitoring) as well as in vivo analysis, where volatiles to be analyzed are present in the breath of human subjects. The pioneering work of Linforth and Taylor (1) in developing the so-called ‘MS-nose’ system based on APCI-MS has allowed the development of breath-by-breath analyses also called the ‘nosespace’ technique. Starting with this pioneering work, the presentation will review the different MS

techniques available for on-line monitoring of aroma compounds with a special focus on the latest developments. Then some in vitro and in vivo applications using APCI-MS and PTR-MS will be presented. Using a PTR-Time of Flight-MS as a MS-based electronic nose, an efficient method of classification of brandy samples according to their origin will be described. On-line monitoring of odorants metabolism using ex-vivo olfactory mucosa of rats will illustrate the metabolism capacity of xenobiotic metabolizing enzymes present in high amount in the olfactory mucosa. Finally, some in vivo flavor release studies using model foods and real foods will be discussed in relation with flavor perception with a special focus on the latest developments using dynamic sensory evaluation techniques such as temporal dominance of sensations. 1. Taylor, A. J.; Linforth, R. S. T.; Harvey, B. A.; Blake, A., Atmospheric pressure chemical ionisation mass spectrometry for in vivo analysis of volatile flavour release. Food Chemistry 2000, 71, 327-338.

AGFD 143

Separation and concentration of trace high-impact odorants using multidimensional gas chromatography-mass spectrometry-olfactometry with integrated preparative fraction collection

Lewis Jones1, Lewis.Jones@eu.effem.com, Kathleen Chu1, Bryan White2, Andrew Ward2. (1) Mars Petcare, WALTHAM Centre for Pet Nutrition, Waltham-on-the-Wolds, Leicestershire, United Kingdom (2) JSB, Maidenhead, Berkshire, United Kingdom

Volatile extracts of food are complex mixtures of compounds and present challenges when analysed using Gas Chromatography-Odour Port (GC-O) techniques. Some compounds can be detected reliably on the Odour Port but are below the limits of detection (LOD) of the GC detectors and therefore, cannot be identified. Increasing the amount injected onto the GC column to improve detection can exceed peak capacity and decrease resolution of other compounds. Multidimensional (MD) GC has been used for many years to increase peak capacity and resolve co-eluting peaks [1], while preparative (p) GC has been used to collect fractions so as to concentrate specific analyses before re-injection into a GC [2]. Instruments previously designed to combine both MDGC and pGC have required the manual removal of the collected material and reinjection by an automated thermal desorption system [3]. Here we describe a MDGC system with integrated, automated preparative fraction collection and desorption. The system comprises an Agilent 7890A GC with two low thermal mass (LTM) ovens, which can be used for one dimensional GC-Mass Spectrometry (MS) and Olfactometry (O), to identify odour-active areas of the chromatogram. To identify and quantify co-eluting peaks or analytes below LOD, an Agilent capillary flow technology (CFT) Deans switch is used to heart-cut the peak or area of interest to a SIM Ice Cube cryo-trap. The cryo-trap has a large volume, and multiple heart cuts can be collected before thermal desorption onto the second column and detection at MS and O.

The presentation will show the concentration effects (and reproducibility) of the system using test mixtures of volatile compounds, as well as application to real food systems. [1] Marriott, P.J., et al., Trends in Analytical Chemistry 34 (2012) 34 1 [2] Kim, L. and Marriott, P .J. Preparative Gas Chromatography. In Gas Chromatography, Poole C., Ed., Elsevier: Amsterdam (2012) 395. [3] Ochiai, N. and Sasamoto K., Journal of Chromatography A 21 (2011) 3180.

AGFD 144

Encapsulation, multimodal perception, and its applications

Gary Reineccius, greinecc@umn.edu. Food Science and Nutrition, University of Minnesota, St Paul, Minnesota, United States

Despite many years of research, the food and flavor industries still struggle with improving the shelf-life of oxidizable food components (e.g. flavorings, polyunsaturated triglycerides, natural pigments, vitamins, etc.). Microencapsulation has been a common means of offering some protection to these ingredients. Among a variety of microencapsulation methods, spray drying is the most widely used process in the food and beverage industries for encapsulation: cost, availability and flexibility of the process make it a good choice. Recently new tools have become available to optimize the formulation used in spray drying and the processing parameters. Substantial understanding has been gained using these new tools that translates into manufacturing products with longer shelf-lives. This presentation will provide an overview of global research focused on optimizing spray drying of both flavorings and fish oils. The presentation will end with where I believe we have opportunity to further improve the performance of this process.

AGFD 145

Modelling mass transfer under simulated nasal conditions – innovative laboratory experimental systems

Masayuki Yabuki1, yabuki.masayuki@kao.co.jp, David Scott3, Loïc Briand4, Andrew J. Taylor2. (1) Kao Corporation, Tokyo, Japan (2) Mars Petcare, Leicestershire, United Kingdom (3) University of Nottingham, Leicestershire, United Kingdom (4) Center for Taste and Feeding Behaviour, Dijon, France

An in vitro experimental system was developed to investigate uptake, retention and release of odorants by the human olfactory mucus layer including Odorant Binding Proteins (OBPs). To mimic the situation in the human nose, (when odorants in the gas phase pass over the olfactory epithelium and odorants transfer between the gas phase and the liquid phase and finally to the olfactory receptors (ORs)), selected gaseous odorants were introduced into a 100-mm section of a polar, deactivated capillary in which aqueous rat-OBP3 films (about 1 µm) were formed, and the change in the gas-

phase concentration of the outflow gas was monitored in real time using Atmospheric Pressure Chemical Ionization - Mass Spectrometry (APCI-MS). Since functional human OBPs are not yet available, the well-characterised rat-OBP3 was used. Benzaldehyde, 2-methylpyrazine, 2-isobutyl-3-methoxypyrazine, 2-isobutylthiazole and 2,4,5-trimethylthiazole were chosen because they exhibited a broad range of dissociation constants with rat-OBP3 and because they were amenable to detection by on-line APCI-MS. All the odorants were quantitatively bound by rat OBP3, which resulted in an effective concentration of the odorants in the aqueous layer (about 50,000-fold). Odorant release from the rat-OBP3-odorant complex into the gas phase showed that odorant release was governed by the dissociation constant of the complex and the flow rate of odorant-free air. When two odorants were introduced into the system, odorant uptake and release were influenced by the method of introduction and their relative affinities for the protein. Because rat-OBP3 exhibits typical odorant binding characteristics, the results not only provide fundamental information on the kinetics of odorant mass transfer induced by the presence of OBPs in the olfactory mucus layer but also support the possibility that vertebrate OBPs may facilitate the accumulation of odorants in the olfactory epithelium.

AGFD 146

Biology of taste: Studies of the order Carnivora

Gary K. Beauchamp, beauchamp@monell.org, Peihua Jiang. Monell Chemical Senses Center, Philadelphia, Pennsylvania, United States

Each animal species lives in a separate sensory world that is coordinated with its chemical ecology. A dramatic example of this occurs for the chemical sense of taste where the interrelationships between sensory perception and diet choice are clearly evident. The fundamental underlying hypothesis for the existence of a small number of basic or primary taste qualities (sweet, bitter, sour, salty, umami and perhaps a few others) is that these qualities evolved to detect and to guide adaptive responses to critical nutrients and potential poisons. We have studied members of the order Carnivora with the goal of understanding how taste receptors and taste perception in these species are related to different feeding ecologies with a particular focus on sweet and bitter compounds. This order is instructive because some Carnivora species are obligate carnivores (e.g. cats), others are more omnivorous (e.g. dogs), whereas others are strictly herbivorous (e.g. giant pandas). Many years ago, we demonstrated that domestic and wild cats (Felis and Panthera) are indifferent to all sweeteners tested but are highly responsive to certain amino acids. Later we demonstrated that the cat’s indifference to sweeteners can be explained by the pseudogenization of the Tas1r2 gene which encodes the T1R2 sweet receptor. From this, we reasoned that other exclusively meat-eating species might also have an inactive form of this gene. Indeed, sequencing of the entire coding region of Tas1r2 from twelve carnivore species revealed that seven of these species, all exclusive meat eaters, had independently fixed a defective Tas1r2 allele. More recently, we have initiated investigations of bitter taste in Carnivora with special attention to cats. We have been testing the prediction that

obligate carnivores should have reduced responsiveness to bitter compounds since it is presumed that bitter taste perception functions primarily to insure avoidance of plant-based toxins. In addition to advancing mechanistic understanding of how taste perception is coordinated with diet at the molecular level, these studies also may help in the development of novel screening tools to enhance and improve the diet and health of companion animals such as domestic cats. This work was supported by NIH-NIDCD grants DC0101842 and 1P30DC011735 and by Monell Institutional funds.

AGFD 147

Understanding colloidal stability and thermal transitions in polyelectrolyte complexes

Yanpu Zhang1, yanpuzhang@tamu.edu, Dariya Reid1, Erol Yildirim2, Hanne S. Antila2, Ran Zhang2, Maria Sammalkorpi2, Jodie L. Lutkenhaus1. (1) Chemical Engineering, Texas A&M University, College Station, Texas, United States (2) Department of Chemistry, Aalto University, Espoo, Finland

Polyelectrolyte complexes (PECs) form by mixing polycation and polyanion solutions together. One challenge for PEC processing and application is that their physical properties are often time-dependent. In this work, we examine the influences of polycation/polyanion mixing ratio and ionic strength on the colloidal stability of PECs made from strong polyelectrolytes poly(diallyldimethylammonium chloride) (PDAC) and poly(sodium 4-styrenesulfonate) (PSS). Physical properties such as turbidity, hydrodynamic size, and zeta potential are investigated as a function of time. The results are used to deduce various critical values that mark the boundary at which a PEC suspension is either colloidally stable or unstable. We discuss these results in relation to the stepwise aggregation model that depicts the aggregation of primary PEC particles into larger secondary particles. Finally, we perform detailed molecular dynamics simulations to examine the structure and effective charge distribution of the PECs at varying mixing ratios and salt concentrations to support the experimental findings. These results emphasize the time-dependent, non-equilibrium nature of PECs and provide useful boundaries for future processing approaches. Thermal properties are another import aspect in understanding the nature of PECs. Polyelectrolyte multilayers (PEMs), which are also formed from oppositely charged polymers share many similarities in structure and properties with PECs. The thermal properties of PEMs have been studied in various systems and the results show that they undergo a thermal transition in which both water and monovalent counter-ions are involved. The transition to-date has only been demonstrated with PEMs and the question remains as to whether the transition is universal to PECs. Here, we will present our investigation of the thermal transition in PECs and compare them to that of PEMs. From the results, we draw conclusions about the nature of the transition and its relationship to the overall structure of the PEM or PEC.

AGFD 148

Thermal transition in polyelectrolyte complexes via LCST mechanism

Erol Yildirim1, Yanpu Zhang2, Ran Zhang3, Jodie L. Lutkenhaus4, Maria Sammalkorpi5, maria.sammalkorpi@aalto.fi. (1) Department of Chemistry, Aalto University, Helsinki, Finland (2) Texas A&M University, College Station, Texas, United States

Hydrated polyelectrolyte (PE) complexes and multilayers undergo a well-defined thermal transition that bears resemblance to a glass transition. Even though its origins are not well understood, this transition of the complex coacervate phase is widely used in, e.g., thermo-responsive drug delivery and in the emerging field of saloplastics. Here, we probe via molecular simulations interconnected with modulated differential scanning calorimetry (MDSC) and quartz crystal microbalance with dissipation (QCM-D) experiments the fundamental nature of this thermal transition. Our findings show the diffusion behavior, hydrogen bond formation, and bridging capacity of water molecules plasticizing the complex experiences a sudden change at the transition temperature. We quantify the behavior and map its chemistry specificity through comparison of strongly and weakly charged PE complexes, as well as, different ionic strengths and species in the system. We conclude the transition is actually via a lower critical solution temperature (LCST) type mechanism and not a glass transition as previously supposed.

AGFD 149

Polyelectrolyte complexes of DNA and polypeptides

Michael J. Lueckheide1,2, milueckheide@gmail.com, Lorraine Leon2, Jeffrey Vieregg2, Matthew V. Tirrell2. (1) Physical Chemistry, University of Chicago, Chicago, Illinois, United States (2) Institute for Molecular Engineering, University of Chicago, Chicago, Illinois, United States

Polyelectrolyte complexes form upon mixing oppositely charged polymers in solution. The complexes formed include liquid-like complexes called coacervates, and solid complexes called precipitates. In the case of coacervates, the initial complexation of the polymers produces coacervate droplets that can coalesce to form a distinct macroscopic phase. Tethering a neutral, hydrophilic polymer to the polycation or the polyanion forms a diblock copolymer; diblock copolymers can form core-shell structures called micelles. Polyelectrolyte complexes can be formed using negatively charged DNA or other therapeutic nucleic acids for applications in gene therapy; these complexes are often called polyplexes. In this work, DNA is the anionic component of the complexes of interest, and biocompatible polypeptides, like poly-L-lysine, serve as the cations. Many variables have been shown to influence the complexation process; these include salt concentration, polymer mixing ratio, polymer chain length, and polymer charge density. We hypothesize that the hybridization state of the DNA will also have an impact. Our use of precisely defined, monodisperse DNA strands and polypeptides allows us to easily vary polymer lengths and charge densities to understand the conditions at which DNA-polypeptide complexes phase separate, and what types of complex are produced. We use methods including turbidity and optical

microscopy to characterize solutions of DNA and polypeptides while varying solution salt concentration; polymer lengths, mixing ratios, and charge densities; and DNA hybridization state. The results of this study may also provide insight into polyelectrolyte complex micelles formed from DNA and a block-copolymer. The inner cores of the micelles form via the same electrostatic interactions that create polyelectrolyte complexes, so we expect them both to be influenced by the same variables.

AGFD 150

Complexation of linear poly(ethylene imine)/poly(acrylic acid) and branched poly(ethylene imine)/metal ions: The effect of ionic strength, molar ratio, and pH

Huan Zhang, hz29@zips.uakron.edu, Nicole Zacharia. Dept. of Polymer Engineering, University of Akron, Akron, Ohio, United States

Polyelectrolyte complexes (PECs) have attracted considerable interest in a range of applications. This includes polyelectrolyte multilayers (PEMs), which are thermodynamically very similar to complexes, which present a wide range of applications, such as sensors, separation membrances, coatings, and barrier materials. Therefore the investigation of thermodynamics and kinetics of complexation of polyelectrolyte becomes important. For weak polyelectrolytes, complexation is readily affected by surrounding environment (i.e. ionic strength, pH, solvent, additives, and temperature). In this work, the complexation of linear poly(ethylene imine) (LPEI) (as polycation)/poly(acrylic acid) (PAA) (as polyanion) was investigated in aqueous solutions under different pH, concentrations of polyelectrolytes and salts, and the molar ratios of LPEI/PAA. The stability of LPEI/PAA complexes was studied as well. Considering coordination bonds can be formed between poly(ethylene imine) (amine group) and metal ions, which can affect the properties of PECs, eight different metal ions were used to study the interactions with branched poly(ethylene imine) (BPEI), which are alkali metal ions (KCl, KNO3), alkaline earth metal ion (MgCl2), transition metal ions (Fe(NO3)3, AgNO3, Cu(NO3)2, ZnCl2), and lanthanide (LaCl3), respectively. Dynamic light scattering (DLS) and phase analysis light scattering (PALS) were employed to analysis the particle size and zeta potential of complexes. Fourier transform infrared (FTIR) and X-ray photoelectron spectroscopy (XPS) were mainly used to study the interactions between BPEI and metal ions.

AGFD 151

Hydrogen bonded polymer complexes with hydrophobic associations

Yuanqing Gu2, Robert A. Weiss2, Nicole Zacharia1, nzacharia@uakron.edu. (1) Dept. of Polymer Engineering, University of Akron, Akron, Ohio, United States (2) Polymer Engineering, University of Akron, Akron, Ohio, United States

Complexes formed between polymers that are capable of hydrogen bonding are well known to be materials with many uses including drug delivery. Such interpolymer

complexes are capable of encapsulating hydrophobic small molecules and therefore dispersing them in aqueous solutions. While these materials have interesting solutions properties, when dried they are often brittle. One of the goals of our work is to improve mechanical properties of these materials by incorporating different types of secondary interactions. In this work, copolymers of acrylamide and a fluorinated group, 2-(N-ethylperfluorooctance sulfonamide), are complexed with polyacrylic acid. These fluorinated copolymers have been shown in other work to be able to be used to make toughened hydrogels. Acrylic acid and acrylamide groups hydrogen bond with one another, and then the fluorinated repeat units are able to form hydrophobic associations which can act to toughen the complex. In this work, interpolymer complexes are formed between polyacrylic acid and homopolymers of polydmethylacrylamide and then between polyacrylic acid and the fluorinated copolymers. Characterization of these materials is done with FTIR, DSC, and rheology to show the diferences in properties between the complexes with and without the hydrophobic associations. Several properties are noticed in the fluorinated materials; they are more tough than the complexes without the fluorinated groups and there are changes in Tg as observed by DSC.

AGFD 152

Mineral nutrient profile of orange juice

Murat Azik, muratazik@msn.com, Dorothy McLean. Scientific Research Department, Florida Department of Citrus, Lake Alfred, Florida, United States

Citrus fruits and juices contain organic compounds and inorganic minerals. The major mineral nutrients in orange juice are potassium, phosphorus, calcium, and magnesium. Orange juice is also known having very low concentration of sodium that is accepted as zero by FDA label rule. In addition, there are trace levels of minerals in orange juices, like manganese, copper, iron and zinc that are essential not only for orange trees but also for the human body at certain quantities. It is expected that commercially available orange juice types; not from concentrate (NFC), from concentrate (CON) and reconstitute (RCO) should have similar levels of mineral constituents at 11.8 degree Brix. In this study, commercial orange juice samples; NFC, CON and RCO collected all around the year from the stores in Florida were monitored and compared with each other and to the USDA nutrient database for their mineral nutrient composition. All the samples were microwave digested with nitric acid and concentrations of Na, Mg, P, K, Ca, Mn, Cu, Fe and Zn were measured utilizing Inductively-Coupled Plasma Mass Spectrometer (ICP-MS) with Collision Cell and Kinetic Energy Discrepancy mode. The data showed that the average concentrations, amount per serving, of Na and Ca were increasing consecutively by juice type, NFC < CON < RCO as 1.43 ± 1.03; 23.7 ± 3.10 < 2.51 ± 1.67; 26.7 ± 3.43 < 7.67 ± 4.09; 34.0 ± 3.90 mg/240 mL for the samples collected in 2012 – 2013. The quality of water used for juice reconstitution may be a major factor for the higher values of Na and Ca in the concentrated orange juices (CON and RCO) than that of not from concentrate (NFC) ones. These results suggest that orange juice types should be evaluated and classified separately for their nutrients.

AGFD 153

Residual effects of low and/or high temperature treatment at mature green stage on volatile production of tomatoes at following ripeness stages

Libin Wang1,2, Baldwin Elizabeth1, Anne Plotto1, Jeffrey Brecht3, Zhifang Yu2, Jinhe Bai1, jinhe.bai@ars.usda.gov. (1) USDA, ARS, HRL, Fort Pierce, Florida, United States (2) Nanjing Agricultural University, Nanjing, China (3) Horticultural Sciences Department, University of Florida, Gainesville, Florida, United States

Mature green ‘FL 47’ tomatoes were exposed to heat (52 °C water for 5 min) and/or cold (5 °C for 4 d) stress and the residual effects on volatile production during ripening at 20 °C were determined. Fruit were sampled directly after each treatment, and then at breaker, turning, pink and red stages. Results showed that chilling, although it did not cause visual injury, substantially slowed the ripening process, and suppressed ethylene production and respiration rate. When fruit turned red, 8 out of 11 important flavor volatiles in ‘FL 47’ tomatoes suffered a significant reduction in chilled fruit. Although it advanced the ripening process, heating had less impact on volatile production at the red-ripe stage in agreement with sensory panel results. On the other hand, heating prior to cold storage alleviated the chilling-caused reduction of ethylene during ripening. Meanwhile, 6-methyl-5-hepten-2-one, 2-methylbutanal and 2-phenylethanol were higher in heating + chilling treated fruit at the red-ripe stage, which was consistent with sensory panel results in that panelists perceived more tomato flavor in heating + chilling treated fruit than fruit that were chilled only.

AGFD 154

Use of foliar fungicide spray for control of HLB-related pre-harvest fruit drop

Wei Zhao1, Jinhe Bai1, jinhe.bai@ars.usda.gov, Greg McCollum1, Tim Gottwald1, Anne Plotto1, Baldwin Elizabeth2. (1) USDA, ARS, HRL, Fort Pierce, Florida, United States (2) Horticultural Research Laboratory, USDA-ARS , Ft. Pierce, Florida, United States

In a recent study, high incidence of Diplodia colonization (Diplodia natalensis or Lasiodiplodia theobromae) was found in HLB-symptomatic orange (presumed pathogen, Candidatus Liberibacter asiaticus, Clas) calyx abscission zones (AZ-C), and the fungal infection was correlated with fruit ethylene production (R2 = 0.70) and reduction in fruit detachment force (FDF, R2 = 0.73) along with an increase in postharvest stem end rot. This suggested presumptive involvement of the fungus in the excessive fruit drop. Therefore, in this study, the effect of fungicide application (Quadris Top, Syngenta Canada Inc.), on fruit drop was evaluated in a HLB-affected grove on five citrus cultivars (Navel Orange, Early Gold Orange, Midsweet Orange, Murcott Tangor, and Ruby Red Grapefruit). Samples were collected at multiple time points for FDF measurement and Diplodia titer detection by qPCR. Fruit drop count and number of fruit remaining on the tree were recorded periodically and the fruit drop ratio was calculated. Quadris Top spray significantly reduced the fruit drop ratio on ‘Early Gold’

(P<0.05), ‘Midsweet’ (P<0.05) and ‘Murcott’ (p<0.001) over the season, and reduced fruit drop by a cumulative 45%, 30% and 46%, respectively. Consistent with this, FDF for the sprayed group was significantly higher than for controls. Quadris Top did not have an effect on ‘Ruby Red’ grapefruit for either FDF or fruit drop ratio; and no effect for fruit drop ratio for ‘Navel’, although sprayed ‘Navel’ fruit had a higher FDF. The average Diplodia Ct values were higher in sprayed groups for ‘Early Gold,’ ‘Midsweet’ and ‘Murcott’ (indicates lower titers) than in controls, but not for ‘Navel’ or ‘Ruby Red’ Grapefruit. However, Diplodia titers in AZ-C increased in the both groups as the season progressed, indicating that the foliar spray of Quadris Top was unable to totally prevent Diplodia from invading fruit tissues. In another experiment, samples of AZ-C were taken from Hamlin fruit that dropped when trees were shaken for comparison to fruit that remained on the tree Analysis by qPCR for Diplodia and Clas DNA showed that the “dropped” fruit AZ-C had lower Ct values (higher titers) for both organisms which was most significant for Diplodia. Optimization of the formula, dosage and timing of application is needed to improve the efficacy of the fungicide foliar spray and avoid resistance development by the fungal pathogen in control of HLB-related fruit drop.

AGFD 155

Self-assembly of two-way nanotubes by proteolysis of wheat bran albumins with protease V8, in presence of calcium ions

Guadalupe Chaquilla-Quilca1, Rene Renato R. Balandran-Quintana1, rbalandran@ciad.mx, Jose A. Azamar-Barrios2, Gabriela Ramos Clamont-Montfort3, Ana M. Mendoza-Wilson1, Jorge N. Mercado-Ruiz1, Tomas J. Madera-Santana1, Yolanda L. Lopez-Franco4. (1) CTAOV, Centro de Investigacion en Alimentacion y desarrollo, AC, Hermosillo, Sonora, Mexico (2) Fisica Aplicada, CINVESTAV-Merida, Merida, Yucatan, Mexico (3) Ciencias de los Alimentos, Centro de Investigacion en Alimentacion y Desarrollo, AC, Hermosillo, Sonora, Mexico (4) Coordinacion de Tecnologia de Alimentos de Origen Animal, Centro de Investigacion en Alimentacion y Desarrollo, AC, Hermosillo, Sonora, Mexico

Wheat bran contains more than 16% w/w of proteins that are underutilized, which causes an interest for alternative uses, such as nanoparticle production. Among the proteins of wheat bran are included the albumins, which are easily extracted with water. These are globular proteins with significant amounts of acidic, hydrophobic and aromatic amino acids, predicting a wide range of non-covalent intra- and inter molecular interactions. The latter is important for the formation of protein nanoparticles by self-assembly processes. For the self-assembly to occur molecular blocks are required, which can be obtained by proteolysis with specific proteases. Furthermore, the presence of divalent ions provides additional possibilities of intermolecular electrostatic interactions, so that the potential range of structures to be formed is wide. Considering the above, in the present work it was hydrolyzed the albumin fraction of wheat bran with the V8 protease. Calcium chloride was added to the reaction mixture for assaying a range of molar ratios of protein:protease:Ca2+. The hydrodynamic size and morphology of the reaction products were characterized by dynamic light scattering and scanning

electron microscopy, respectively. In general, it was observed that proteolysis resulted in the formation of lobular shaped nanostructures, imbibed into a matrix; the higher the protein to protease ratio the better the definition. The addition of CaCl2, in an increasing molar ratio, transformed gradually the matrix into strands, so that for the largest molar ratio were observed strands of 200 nm in diameter and several microns in length. Under these conditions, what previously was seen as lobular shape nanoparticles it seems to be short branches of the strands. After ultrasonication and subsequent visualization by electronic scanning microscopy in STEM mode, it was found that the strands are, in fact, two-way nanotubes, 100 nm internal diameter, 200 nm outer diameter and more than 35 mm length.

AGFD 156

Synthesis of wheat bran albumin nanoparticles by a cold gelation/desolvation method

Jesus G. Luna-Valdez1, Rene Renato R. Balandran-Quintana1, rbalandran@ciad.mx, Jose A. Azamar-Barrios2, Gabriela Ramos Clamont-Montfort3, Ana M. Mendoza-Wilson1, Jorge N. Mercado-Ruiz1, Tomas J. Madera-Santana1, Agustin Rascon-Chu1. (1) Coordinacion de Tecnologia de Alimentos de Origen Vegetal, Centro de Investigacion en Alimentacion y Desarrollo, Hermosillo, Sonora, Mexico (2) Fisica Aplicada, CINVESTAV-Merida, Merida, Yucatan, Mexico (3) Ciencias de los Alimentos, Centro de Investigacion en Alimentacion y Desarrollo, AC, Hermosillo, Sonora, Mexico

The water soluble proteins of wheat bran, i.e. the albumin fraction, is easily extracted and has a significant content of Asp and Glu, which could make such proteins be feasible for the fabrication of nanoparticles through adaptations to the method of cold gelation. This method includes a pre-heating step to a protein suspension, at basic pH, to denaturate the protein molecules and to expose negative charges to subsequently be crosslinked with a divalent cation, usually Ca2+. The method has proven to be successful for fabricating firm gels from protein isolates, but in cases such as the wheat bran albumin fraction, where other components are co-extracted and further purification of proteins is not attractive because of economical reasons, the gel matrix is disrupted so that gels are turned weak. However, this circumstance and the dual role of the calcium as agent of both desolvation and crosslinking, could be exploited to fabricate nanoparticles. In the present work was assayed the cold gelation/desolvation of the albumin fraction of wheat bran with CaCl2. Aggregates >5 mm were formed after addition of CaCl2 to preheated wheat bran albumin suspensions, as detected by dynamic light scattering. Scanning electron microscopy showed that such aggregates consisted of spherical nanoparticles with diameters between 80 and 100 nm, immersed into a matrix composed mainly by polysaccharides. This matrix was disrupted by ultra-sonication and the individual nanoparticles were visible by scanning electron microscopy in transmission mode. This is the first report on the formation of spherical nanoparticles from wheat bran albumins. The main features of the cold gelation/desolvation method is that takes advantage of other non-protein components present in the albumin fraction and avoids the use of ethanol/acetone and

glutaraldehyde as desolvation and crosslinking agents, respectively, as described currently in the protocols found in literature about the albumin nanoparticles fabrication.

AGFD 157

Improvements in the measurement of chlorophylloids in soybean oil

Avrom C. Litin, alitin@comcast.net, David D. Brooks. Innovation Center, Oil-Dri Corporation of America, Chicago, Illinois, United States

An improved procedure using HPLC with fluorescence detection for the measurement of chlorophylloids in refined bleached soybean oil was developed. These improvements allow a simple and rapid method for the analysis of chlorophylloids in products including chlorophyll, pheophytin and pyropheophytin in refined bleached soybean oil. By exchanging the standard photomultiplier tube in the fluorescence detector with a red-sensitive Hamamatsu R928HA PMT, a 300 fold increase in sensitivity was achieved. A direct analysis of diluted soybean oil was performed without further sample preparation. Calibration of the analysis was performed by quantification of purified chlorophylloids by UV/VIS spectroscopy.

AGFD 158

Impact of food preparation on total phenolic contents and anti-oxidant capacities of regularly consumed botanicals

Lu Yu1, yulu0640514@gmail.com, Boyan Gao1, raphaelgao1985@gmail.com, Thomas T. Wang2, Liangli L. Yu1, lyu5@umd.edu. (1) Univ of Maryland, College Park, Maryland, United States (2) Beltsville Human Nutrition Research Center, ARS, U.S. Department of Agriculture, Diet, Genomics and Immunology Laboratory, Beltsville, Maryland, United States

This study investigated the effects of different home-used food preparation methods on total phenolic contents and anti-oxidant capacities of the selected common vegetables and fruits. The total phenolic contents (TPC), DPPH radical scavenging capacity (DPPH) and oxygen radical scavenging capacity (ORAC) were assayed to evaluate the anti-oxidant capacities of kale, broccoli, carrot, and blueberry processed by blending, microwaving, and chopping. The data suggested that the availability of anti-oxidants and anti-oxidant capacities of selected botanicals were strongly impacted by different food preparation approaches. Factors such as particle size of plants, enzymes from plants and saliva, thermal energy applied and glycosylation level of phytochemicals altered the availabilities of nutraceuticals and the anti-oxidant capacities of these vegetables and fruits. These findings suggest that choosing an appropriate food processing method for each type of botanical might be critical to obtain desirable health beneficial effects.

AGFD 159

Effects of home-based preparation approaches in determining the release of bioactivity compounds in fruits and vegetables

Boyan Gao1, raphaelgao1985@gmail.com, Lu Yu1, Thomas T. Wang2, Liangli L. Yu1, lyu5@umd.edu. (1) Univ of Maryland, College Park, Maryland, United States (2) Beltsville Human Nutrition Research Center, ARS, U.S. Department of Agriculture, Diet, Genomics and Immunology Laboratory, Beltsville, Maryland, United States

The effect(s) of home-based preparation approaches on the availability of nutritional components of blueberry, kale, broccoli and carrot were evaluated and compared using high performance liquid chromatography (HPLC). The results indicated that home-based preparation approaches could effectively change the particle size of fruits and vegetables, inactivate the enzymes from botanical and saliva, heat process the foods, and thus impact the release of bioactivity compounds from those foods. These data represent the relationship among particle size, enzyme activity, and thermal process with the release of nutrients in foods. All the results above indicate the importance in selecting appropriate home-based preparation approaches in consuming those fruits and vegetables during the daily life.

AGFD 160

Determination of the heavy metals in the health functional foods by inductively-coupled plasma/atomic emission spectrometry

Jee Eun Hong1, hje7682@korea.kr, Chae Hyung Lim1, Young-Mi Chang1, Chul Joo Lim1, Tae-seok Kang2. (1) Center for food and drug analysis, Gyeongin regional food and drug administration, Ministry of Food and Drug Safety(Rep. of Korea), Incheon, Korea (the Republic of) (2) Nutrition and functional food research team, Ministry of food and drug safety, Cheongju, Korea (the Republic of)

As the consumption of health functional foods is increased, the intake of heavy metals is one of the important problems for the human health. Determination of heavy metals such as Pb, Cd, As and Hg was performed by inductively coupled plasma-atomic emission spectroscopy (ICP-AES) and direct mercury analyser. Microwave digestion with HNO3-H2O2 and dry ashing procedures were compared for the recovery and precision. In order to validate the quantitative analysis, several parameters have been taken into account and evaluated; linearity, accuracy, detection limit and repeatability. We have performed the monitoring of heavy metals in the coenzyme Q10, saw palmetto extract, lecithin, green tea extract, lutein, chitosan and chito-oligosaccharide samples. The result of this study might be used as a valuable data for the safety control of the health functional food in Korea.

AGFD 161

Multiresidue analysis of pesticides in commercial agricultural products using LC-MS/MS

Se Ra Won, serawon@korea.kr, Seong Cheol Kim, Nam Suk Kang, Yun Jung Kang, Do Hyeong Kim, Ho Won Chang, Dong Eon Kim, Il Yong Jung, Seung Ho Woo, Sang Yub Kim, Ju Young Park, Hae Seong Yoon. Ministry of Food and Drug Safety, Busan, Korea (the Republic of)

This study investigated 141 kinds of pesticide residue levels using a multiresidue screening method by LC-MS/MS in agricultural products. The purpose of this study was to promote consumer safety by excluding the food containing pesticide residues from markets. 90 samples from 5 major cities were collected and composed of vegetables, fruits, cereals, seed, etc. For the validation of multiresidue screening methods, the items of recoveries, linearity and limit of quantitation(LOQ) were analyzed. Most pesticides were recovered in the range of 60~130% at spike levels of 0.05, 0.1 and 0.5 mg/kg. Linearity showed excellent correlation over the study with a correlation coefficient of 0.9900 ~ 0.9996. LOQ ranged from 4 to 10 μg/kg by LC-MS/MS. In the results, pesticide residues were found in 10 samples. Azoxystrobin was detected frequently and the levels of detected pesticide residues were not exceeded the Koreans maximum residue limits(MRLs). Violations of Korean MRLs were not observed in all the samples.

AGFD 162

Antimicrobial peptide segments from soy protein for use in food safety

Ning Xiang1, Yuan Lyu1, Arun Bhunia2, Ganesan Narsimhan1, narsimha@purdue.edu. (1) Dept of Agric Biological Eng, Purdue University, West Lafayette, Indiana, United States (2) Food Science, Purdue University, West Lafayette, Indiana, United States

Antimicrobial peptides (AMPs) kill microbial cells through insertion and damage/permeabilization of the cytoplasmic cell membranes and has applications in food safety. Soy protein may be such an attractive, cost-saving candidate for commercial consideration because the protein subunits have amino acid sequences that contain several α-helix or 3-10 helix domains which possess characteristics of AMPs. A methodology for identification of AMPs from soy protein is proposed. They were identified from soy β-conglycinin (7S) and glycinin (11S) based on (i) number of amino acids, (ii) positive charge, (iii) hydrophobicity and (iv) hydrophobic moment. Explicit solvent molecular Dynamics (MD) simulation was employed to assess the secondary conformation of these peptides in POPC/POPG bilayers to mimic their permeation action on the cell membrane of microorganism. The effects of number of peptides, their orientation and hydrophobic moment on the deformation of membrane and formation of water channel were investigated. A mathematical model for the prediction of energy barrier for deactivation (antimicrobial activity) was proposed accounting for free energy of formation of a pore by an aggregate of peptides. The antimicrobial activity of three synthetic selected peptides against Listeria monocytogenes and E. Coli was demonstrated.

AGFD 163

Flavonol glycosides in wild and cultivated berries of two major subspecies of sea buckthorn and influence of growth sites

XUEYING MA, xueyingnice@gmail.com, Oskar Laaksonen, Heikki Kallio, Baoru Yang. Department of Biochemistry and Food Chemistry, University of Turku, Turku, Finland

Flavonol glycosides in samples (n = 127 × 3) of wild berries (Hippophaë rhamnoides ssp. sinensis, RS) from China and cultivated berries (Hippophaë rhamnoides ssp. mongolica, RM) from Finland and Canada were identified and quantified by HPLC-DAD-ESI-MS/MS and HPLC-DAD, respectively. Twenty-six flavonol glycosides were found. Isorhamnetin and quercetin were the major aglycons. The contents of flavonol glycosides ranged from 23 to 252 mg per 100 g fresh sea buckthorn berries (SBB).There was significant difference (p < 0.05) in the contents of 26 compounds among berries of different sources. The independent-sample t test showed that the content of flavonol glycosides was significantly higher in the RS SBB than that RM SBB (p < 0.01). Isorhamnetin-pentoside-rhamnoside and isorhamnetin glycoside I were detected only in the wild berries, but isorhamnetin-hexoside-rhamnoside and isorhamnetin-rhamnoside were detected just in the cultivated berries of RM. The contents of total flavonol glycosides in berries from different origins followed the order: SC > QH > SX > IM > HB > HLJ > Kittilä > Québec > Sammalmäki. The influence of different altitude on the content of flavonol glycosides was studied in wild berries collected from nine natural growth sites in China. Increasing trends were shown in the contents of most of the compounds as the altitude increased.

AGFD 164

Flavonol glycosides in leaves of different varieties of black currant, green currant, red currant, white currant and changes of growing season, growth location, leaf position

WEI YANG, yaungwei@gmail.com, Heikki Kallio, Baoru Yang. University of Turku, Turku, Finland

Currant leaves contain many bioactive compounds with potential benefits for human health such as flavonol glycosides. In this study, 27 flavonol glycosides were identified in the leaf extracts of three black currant cultivars, a green currant cultivar, a red currant cultivar, and a white currant cultivar. Among them, four compounds were identified for the first time in currant leaves by NMR. This study also investigated flavonol glycosides variation relative to cultivar, growth location, growing season and the position of the leaves in the branches. The variety ‘Mikael’ differed from ‘Mortti’ and ‘Jaloste n:o 15’. The white currant leaves showed a clearly different profile of flavonol glycosides compared with others. The leaves grown at lower latitudes (60°23'N, Piikkiö, Finland) contained higher levels of flavonol glycosides than those grown at higher latitudes (66°34'N, Apukka, Finland). The contents of flavonol glycosides reached the highest values at the end of July at the lower latitude (Piikkiö) and at mid-August or early September at higher latitude (Apukka). For green and white currants, the total content of

flavonol glycosides in the leaves collected randomly from different sites of the bushes were higher than in the leaves collected in the middle position of the branches; the situation was vice versa in the red currant leaves. This systematic report of flavonol glycosides composition in currant leaves and variation relative to cultivar, location, growing season, leaf position provided important information for utilization of currant leaves as potential raw material of nutraceuticals and functional foods.

AGFD 165

Pilot-scale bioreactor production and long term stability of feruloyl soy glycerides

David L. Compton1, david.compton@ars.usda.gov, John R. Goodell2, Steven Grall3,4, Kervin Evans1. (1) NCAUR-ARS-USDA, Peoria, Illinois, United States (2) iActive Naturals/Biotechnology Research and Development Corporation, Peoria, Illinois, United States (3) under contract with BRDC, Grall Co., Lemont, Illinois, United States (4) currently Food Plant Designs, LLC, Cincinnati, Ohio, United States

Soybean oil was transesterified with ethyl ferulate at 60 °C using the immobilized lipase Candida antartica lipase B (Novozym 435) to produce a mixture of feruloylated monoacylglycerols and feruloylated diacylglycerols homologues, collectively referred to as Feruloylated Soy Glycerides (FSG), a novel cosmeceutical ingredient used in the skin and hair care industries. A 1-tonne (1,000 kg)/yr, continuous, packed-bed bioreactor was designed and implemented for production of FSG. The substrate was fed to a series of four 10-L, packed bed columns containing Novozym 435 for over five continuous months. The FSG yield was ~ 65% over the course of the 450 kg production run with no loss of enzyme activity. FSG physicochemical characteristics from the most recent production run were compared to FSG produced five years ago to assess FSG stability. Antioxidant capacity (DPPH assay), oxidative stability (iodine number), and susceptibility to hydrolysis (Karl Fischer titration, total acid number, Lovibond color) were compared.

AGFD 166

Synthesis and lead discovery of pyrazolcyclohexanol derivatives

Guan Aiying1, guanaiying@sinochem.com, Changling Liu1, Yong Xie1, Zhi Huang1, Junfeng Wang1, Xiuli Wang1, Qin Sun1, Xufeng Sun1, jichun Yang1, yamei Wu2. (1) Shenyang Research Institute of Chemical Industry, Shenyang, China (2) Shenyang Sciencreat Chemicals Co., Ltd., Shenyang, China

It is well known that lead discovery plays an important role in R&D of novel agrochemicals. However, the discovery process is significantly longer while the success rate for commercial development is declining. Moreover, along with increasing resistance of plants to currently available fungicides, farmers around of world have been eager for novel crop protection products to increase yield. This promotes researchers in agrochemical field to continue searching for novel leads with innovated chemical

structure. Substituted pyrazole ring derivatives exhibit broad-spectrum of biological activities including herbicidal, antimicrobial and antitumor activities. The cyclohexanol moiety also has been used widely in discoveries of some filaricides, insecticides/acaricides and fungicides. A few pyrazolcyclohexanol compounds have been disclosed with probably insecticidal or herbicidal activity in a few prior arts. In this study, in order to find new lead compounds with fungicidal activity to lay foundation for further optimization, a highly efficient and feasible approach for discovering and developing novel agrochemicals, Intermediate Derivatization Methods (IDM) was adopted to direct and synthesize ten pyrazolcyclohexanol derivatives, Their structures were confirmed by 1HNMR, elemental analyses and mass spectral. Fungicidal bioassays were carried out to test all of the target compounds against five pathogens, cucumber downy mildew (CDM), wheat powdery mildew (WPM), southern corn rust (CSR), rice blast (RB) and cucumber grey mold (CGM). The results indicated that seven compounds with more than 80% fungicidal control are all worthy of being good lead compounds for further optimization to find candidates with promising activity.

AGFD 167

Design, synthesis, and bioactivity of novel aryloxypyridine pyrazole amide derivatives

Fan Yang, Chang-Ling Liu, liuchangling@sinochem.com, Guan Aiying, Wei Chen, jichun Yang, zhinian Li, Yuquan Song. Shenyang Research Institute of Chemical Industry, Shenyang, China

As an important member of heterocyclic compounds, the pyrazole amides has been deeply favored by agrochemical researchers continuously for many years. Particularly, since the successful development of the new broad-spectrum insecticide tolfenpyrad by Mitsubishi Chemical Corporation in 1988, pyrazole-5-carboxamide compounds attracted much attention of the agricultural chemical companies around the world. In order to develop novel agrochemicals with higher efficiency and lower toxicity from this kind of compounds, Intermediate Derivatization Method(IDM) was applied by replacing amino parts with aryloxypyridine moieties which possess reactive group. A series of tolfenpyrad analogues with novel structures were synthesized and bioassayed aiming at studying the structure-activity relationship comprehensively. The results of bioassays revealed that many of the compounds have been found with excellent fungicidal and/or insecticidal activity especially compound 5407, which showed 100% control of Blumeria graminis at 6.25mg/L, superior to many known high active compounds with similar structures at the same doses, the further field trials are in progress. These novel aryloxypyridine pyrazole amide derivatives with high activity are worthy to be further researched and developed as lead compounds.

AGFD 168

Design, synthesis, and fungicidal activity of pyrazolecarboxamide derivatives

Lizeng Wang, Chang-Ling Liu, liuchangling@sinochem.com, Yong Xie, zhinian Li, Jie Lan, Xufeng Sun, Guan Aiying. Shenyang Research Institute of Chemical Industry, Shenyang, China

Tolfenpyrad, a commercialized pyrazolecarboxamide compound, shows good insecticidal/acaricidal activity against Lepidoptera, Hemiptera, Thysanoptera and Eriophyidae mites in vegetables, ornamentals and fruit, also active against the fungus Sphaerotheca fuliginea. Chlorantraniliprole, discovered by Du pont, is a broad-spectrum insecticide with novel structure and new mechanism acting on RyR distributed in insects. Intermediate Derivatization Methods (IDM) was adopted by using 4-hydroxybenzonitrile as the starting material to design and synthesize twelve pyrazolecarboxamide derivatives combining some moieties of Tolfenpyrad and Chlorantraniliprole. Their structures were confirmed by 1HNMR, elemental analyses and mass spectral. Fungicidal bioassays were carried out to test all of the target compounds against five pathogens, cucumber downy mildew (CDM), wheat powdery mildew (WPM), southern corn rust (CSR), rice blast (RB) and cucumber grey mold (CGM). The test results of biological activity showed that most of the compounds have a broad spectrum fungicidal activity, especially to cucumber downy mildew at low doses. The present work demonstrates that pyrazolecarboxamide derivatives can be used as lead compounds for further optimization to find candidates with promising activity.

AGFD 169

Synthesis and lead discovery of phenylhydrazine derivatives

Huichao Li, Chang-Ling Liu, liuchangling@sinochem.com, Guan Aiying, zhinian Li, Sen Ma, Xiuli Wang, yamei Wu. Shenyang Research Institute of Chemical Industry, Shenyang, China

2,6-dichlorotoluene has a proven application as an important intermediate in several agrochemicals including bistrifluron (insecticide), methoxyfenozide (insecticide) and tembotrione (herbicide). With a simple chemical structure, 2,6-dichlorotoluene can be readily functionalized to generate many new intermediates. Phenylhydrazine derivatives have been reported with good fungicidal, insecticidal, acaricidal and/or herbicidal activities. With the aim of finding new lead compounds with potent bioactivity, eight novel phenylhydrazine derivatives were designed and synthesized from the starting raw material 2,6-dichlorotoluene by the approach of Intermediate Derivatization Methods (IDM), which has been recently reported and proved to be useful and efficient. The synthesized compounds were tested against five pathogens, cucumber downy mildew (CDM), wheat powdery mildew (WPM), southern corn rust (CSR), rice blast (RB) and cucumber grey mold (CGM). The results indicated that some compounds exhibited moderate activity against CDM. To improve the fungicidal activity, another ten compounds were prepared by further modification of the substituents of the phenyl ring. As a result, two lead compounds showing improved activity against CDM were found, and the activity profile were improved as well. The results of this study suggest that

further design and structural modification of the phenylhydrazine derivatives as fungicides is worthwhile.

AGFD 170

Design, synthesis, and biological activities of new strobilurin derivatives containing trifluoromethyl In the side chain

Miao Li2, Huiwei Chi2, jichun Yang2, zhinian Li2, Chang-Ling Liu1, liuchangling@sinochem.com. (1) Shenyang Research Inst of Chemical Industrial, Shenyang Liaoning, China (2) Shenyang Research Institute of Chemical Indus, Shenyang, China

The strobilurins are known as one of the most important classes of agricultural fungicides with broad fungicidal spectrum, lower toxicity towards mammalian cells and environmentally benign characteristics. Additionally, the strobilurin derivatives containing meta-trifluoromethyl substituted phenyl, pyridine and pyrimidine in the side chain display excellent fungicidal and acaricidal activity, such as fungicides trifloxystrobin, picoxystrobin and acaricide fluacrypyrim. To discover new strobilurin analogues with high activity, a series of new strobilurin derivatives containing trifluoromethyl in the side chain were synthesized and bioassayed. The compounds were identified by 1H and 19F nuclear magnetic resonance (NMR), IR, MS and elemental analysis. Preliminary bioassays indicated that some title compounds exhibited excellent fungicidal activities against Erysiphe graminis protecting wheat at 1.56 mg L-1 and 2-chloro-4-CF3-phenol analogues 2a showed a moderately high acaricidal activity against Tetranychus cinnabarinus at 10 mg L-1. The relationship between structure and biological activity is discussed in terms of effects of the substituent of the phenyl and pyridiny ring. The present work demonstrates that strobilurin analogues with substituted trifluoromethyl phenol or pyridinol side chains can be used as possible lead compounds for further developing novel fungicides and acaricides.

AGFD 171

Design, synthesis, and insecticidal evaluation of novel insecticidal aryloxy dihaloropropene derivatives

jichun Yang, Chang-Ling Liu, Miao Li, Xiuhui Chang, Guan Aiying, guanaiying@sinochem.com, Qiao Wu, Yuquan Song. Shenyang Research Institute of Chemical Industry, Shenyang, China

Controlling the development of insecticide resistance in pests and limiting the usage of synthetic chemicals for maintaining an eco-friendly environment necessitates the identification of alternative molecules with new structural features and action modes. Pyridalyl as a novel class of insecticides has unknown mode of action, and has very good control efficacy against populations of Heliothis virescens (F) and Plutella

xylostella (L) which are resistant to various currently used insecticides. It is well-known that the aryloxyphenyl derivatives have been widely used in agriculture. For example, pyriproxyfen, fenoxycarb, flufenoxuron and chlorfluazuron have been used as commercial insecticides, diclofop-methyl, haloxyfop-methyl and fluazifop-butyl as herbicides, famoxadone as fungicide for many years. So the terminal group replacement method which was key type of Intermediate derivatization methods (IDM) was applied by introducing aryloxyphenyl to pyridalyl to replace pyridyl. A series of new dihalopropene analogues with the substituted aryloxyphenyl were synthesized and bioassayed. The results of bioassays indicated that most of the compounds showed moderate to high activities at the tested concentration, especially some compounds displayed more than 75% insecticidal activity against P. xylostella at 6.25 mg/L, while pyridalyl showed 50% insecticidal activity at the same concentration. Two field trials showed the insecticidal activities of compound 10e(pyridyloxyphenyl) as a 10% emulsifiable concentrate (EC) was effective in the control of P. xylostella at 75–150 g a.i./hm2, and the mortality against P. xylostella of Compound 10e at 75 g a.i./hm2 was equivalent to reference samples of pyridalyl at 105 g a.i./hm2. So compound 10e can be a promising candidate for further development.

AGFD 172

Synthesis, characterization, and fungicidal activity of some new N-phenyl benzothiazolamine derivatives

Guan Aiying, guanaiying@sinochem.com, Chang-Ling Liu, Huichao Li, Qin Sun, Xufeng Sun, Junfeng Wang, Yong Xie, Fan Yang, jichun Yang. Shenyang Research Institute of Chemical Industry, Shenyang, China

It is well known that it is an emergency to quickly discover new structures to fight against the increasing resistance of plants to currently available fungicides. 2,6-Dichlorotoluene has been applied widely as the raw material for pesticides, such as herbicides dichlobenil, tembotrione and florasulame, insecticides methoxyfenozide and diflubenzuron, and the fungicide fluopicolide. In this study, 2,6-dichlorotoluene was selected as the key starting material due to its suitable functionality and its advantages of relatively low cost and wide commercial availability. By employing an effective new agrochemical discovery approach named ‘Intermediate Derivatization Methods’( IDM), the key intermediate 2,6-dichloro-3,5-dinitrotoluene was prepared from 2,6-dichlorotoluene, then reacted with substituted 2-aminobenzothiazoles to afford a series of N-phenyl benzothiazolamine compounds. Bioassays demonstrated that most of the synthesized compounds exhibited excellent fungicidal activities against cucumber downy mildew (CDM), rice blast (RB), cucumber grey mold (CGM) and rice blast (RB), particularly, the compound 3f exhibited 80% control at 0.03 mg/L concentration against RB, a little higher than commercial fluazinam, delivering a promising candidate for further development.

AGFD 173

Synthesis and herbicidal activity of novel substituted 3-(pyridin-2-yl)benzenesulfonamide derivatives

Yong Xie, Huiwei Chi, Guan Aiying, guanaiying@sinochem.com, Changling Liu, Hongjuan Ma. Shenyang Research Institute of Chemical Industry, Shenyang, China

Substituted 2-phenylpyridines which could be used as intermediates for drugs and agrochemicals showed good herbicidal activity. For instance, some 2-phenylpyridine derivatives could efficiently control velvet leaf (abutilon thephrasti), redroot pigweed (amaranthus retroflexus) and morningglory (ipomoea subspecies). In order to search for novel compounds which can be used for the targeted control of unwanted plants, a series of novel substituted 3-(pyridin-2-yl)benzenesulfonamide derivatives were designed and synthesized using 2-phenylpridines as the lead compound by Intermediate Derivatization Methods in an attempt to obtain novel compound candidates for weed control. The herbicidal activity assay in glasshouse tests showed several compounds could efficiently control velvet leaf, youth-and-old age, barnyard grass and foxtail. Especially, the activity of 708 and 801 were proved roughly equivalent to saflufenacil and better than sulcotrione at the same concentration. The result of the herbicidal activity assay in field tests demonstrated that 708 and 801 at 60 g/ha active substance could give the same effect as Bentazon at 1440 g/ha active substance to control dayflower and nightshade, meanwhile 708 showed better activity than Oxyfluorfen to control arrowhead. The present work indicates that novel substituted 3-(pyridin-2-yl)benzenesulfonamide derivatives could be used as candidates for potential herbicide.

AGFD 174

Synthesis and insecticidal activity study of 2-(2,6-dichlorobenzamido)-4-thiazolecarboxamide derivatives

Yong Xie, Shiying Xu, Guan Aiying, guanaiying@sinochem.com, Lizeng Wang, Changling Liu. Shenyang Research Institute of Chemical Industry, Shenyang, China

Thiazole which could be reactive group with excellent biological activity has already been widely used in pesticide molecular design. Many pesticide containing thiazole were reported in recent years, such as thiamethoxan, fluensulfone and ethaboxam. In order to search new highly active compound containing thiazole, 15 novel compounds of 2-(2,6-dichlorobenzamido)-4-thiazolecarboxamide derivatives were designed and synthesized from the starting material of Acetyl bromide ethyl formate by the Intermediate Derivatization Methods in an attempt to obtain novel compound candidates for insect and fungi control. The structures of all compounds were confirmed by 1H NMR, 19F NMR and elemental analyses. And all compounds were bio-assayed against insect and fungi. The preliminary assays showed that some compounds exhibited significant insecticidal and fungicidal activity. Especially, compounds D3 could excellently control Pyricularia oryza and Tomato late blight at the concentration of 400 mg/L, and compound D12 could effectively control Culex pippens pallens, Leucania

separate, Plutella xylostella and Myzus persicae at the concentration of 600 mg/L. This study will be useful for the design of new insecticides and fungicides.

AGFD 175

Adulteration and its detection of black raspberry products

Jungmin Lee, jungmin.lee@ars.usda.gov. USDA-ARS-HCRU worksite, Parma, Idaho, United States

We have continually researched improvements for commercially available cultivars of black raspberry (Rubus occidentalis L.; blackcap). During the past decade, we have analyzed fruit from over 1,000 black raspberry genotypes and cultivars, and found that the anthocyanin content to ranged from 39 to 996 mg of cyanidin-glucoside/100 mL (a 25-fold range). We documented ‘ORUS4143-rep 1,’ which has a rare anthocyanin profile deviation from the industry-standard ‘Munger’. Our records on black raspberry fruit and plants allowed our creation of an anthocyanin profile database that can be used for verifying authenticity of black raspberry products. Available black raspberry products in the US marketplace were each assessed for anthocyanin profile and concentration for quality evaluations. The results were alarming and indicate there are authenticity issues in US sourced black raspberry dietary supplements, and in Korean originated black raspberry products as well. Seven out of 19 black raspberry products (sold as dietary supplements) contained no fruit anthocyanins. Korean black raspberry products, many labeled to contain Rubus coreanus, actually comprised of R. occidentalis (American black raspberry), black carrot anthocyanins, or were too low in pigment concentration to classify. We are hopeful this research will aid in future black raspberry ingredient authenticity standards. This work was partially funded by a Specialty Crop Research Initiative (SCRI) grant number 2011-51181-30676 from USDA-National Institute of Food and Agriculture (NIFA).

AGFD 176

Effect of fresh and commercially processed orange juice on the oxidative status in healthy humans

Jacqueline Q. Silveira2, jacqueiroz@gmail.com, Thais B. Cesar2, Ana Lucia M. Nasser2, John A. Manthey1, Baldwin Elizabeth1. (1) Horticultural Research Laboratory, USDA-ARS , Ft. Pierce, Florida, United States (2) Food and Nutrition, Sao Paulo State University, Araraquara, SP, Brazil

Orange juice is a natural source of vitamins and flavonoids and regular consumption has been suggested to increase blood antioxidant activity. The objective of this study was to investigate the effect of ingestion of a single oral dose of orange juice on the markers of oxidative stress comparing two types of extraction: fresh squeezed (FOJ) and commercially pasteurized (POJ), analyzed by ABTS and TBARS. Twenty-four subjects participated in this study with a mean age of 27 ± 6 years, and 68.3 ± 11.7 kg

body weight. FOJ was extracted with a commercial juicer, and POJ, ready-to-drink, was pasteurized at 95ºC/15 s. Subjects ingested a dose of 11.5 mL/ body weight of FOJ, and after a 30 d washout, they ingested the same quantity of POJ. A fasting blood sample was collected initially, and following the ingestion of orange juice, blood samples were withdrawn at 4, 8, and 24 h. After intake of FOJ, the blood serum malondialdehyde (MDA) decreased significantly at 4h, while with POJ, MDA decreased after 8h. There was an increase in antioxidant capacity after consumption of FOJ and POJ, but this change was faster with FOJ (4h), while it delayed until 8h with POJ. In conclusion, a single dose of either FOJ or POJ improved the antioxidant status in healthy subjects, but this effect was faster with FOJ than with POJ.

AGFD 177

Luteolin-mediated apoptosis in leukemia cells involves PTTG1 oncoprotein and differential responses

Hsin Jung Tien1, tiencelia@gmail.com, Pei-Yi Chen2, Ji-Hshiung Chen1, Ming-Jiuan Wu3, Jui-Hung Yen1,4. (1) Department of Molecular Biology and Human Genetics, Tzu Chi University, Hualien, Taiwan (2) Center of Medical Genetics, Buddhist Tzu Chi General Hospital, Hualien, Taiwan (3) Department of Biotechnology, Chia-Nan University of Pharmacy and Science, Tainan, Taiwan (4) Institute of Medical Sciences, Tzu Chi University, Hualien, Taiwan

Overexpression of oncogenic proteins is involved in the proliferation promotion and the myeloid differentiation inhibition in acute or chronic myeloid leukemia. Luteolin (3’,4’,5,7-tetrahydroxyflavone), a flavonoid found in several vegetables and fruits, has been reported to possess anti-cancer properties in myeloid leukemia cells, however, the underlying mechanism remains unclear. In the present study, we first investigated the anti-cancer effect of luteolin on the undifferentiated or differentiated leukemia cells. We found that luteolin significantly induced cell death of HL60, THP-1 and K562 cells at concentration of 25-150 μM. However, when these cells induced differentiation by phorbol 12-myristate 13-acetate (PMA) or retinoic acid (RA), the cytotoxic effect of luteolin was markedly decreased. These findings suggested that luteolin induced cell death was more efficient in undifferentiated or immature leukemia cells. In our previous study, we demonstrated that pituitary tumor transforming gene 1(PTTG1), an oncoprotein overexpressed in leukemia cells, was significantly down-regulated during myeloid cell differentiation. Thus, we further investigate whether the level of PTTG1 expression is involved in the luteolin-mediated cell death. We found that the cytotoxic effects of luteolin in leukemia cells require PTTG1. Down-regulation of PTTG1 by shRNA impaired the luteolin-mediated cell death. In addition, PTTG1 knockdown cells reduced expression of apoptotic molecules in contrast to their control cells upon luteolin treatment. Taken together, our findings provide evidence that overexpression of oncogenic PTTG1 plays an important role in the mechanisms underlying luteolin-induced apoptosis in un-differentiated or early stage of leukemia cells. Thus, we suggest that the expression levels of PTTG1 may be a critical factor for application in anti-cancer therapy with luteolin in myeloid leukemia.

AGFD 178

Development of lecithin emulsion gel to enhance the oral bioaccessibility of nobiletin

Yuwen Ting1,2, pam.yt.ting@gmail.com, Yijun Pan1, Qingrong Huang2. (1) School of Food Science and Technology, Wuhan Polytechnic University, Wuhan, China (2) Food Science, Rutgers University, New Brunswick, New Jersey, United States

Nobiletin is one of major the polymethoxyflavones found in peel of citrus fruits and had been extensively studied for its potent biological efficacy in reducing risk of chronic diseases. However, oral consumption of nobiletin is greatly limited by its poor bioavailability as a result from its high melting temperature and highly crystalline nature. To overcome this problem, a novel emulsion gel system formed through lecithin molecule self-assembling interaction was developed to encapsulated nobiletin and to promote better bioaccessibility in the gastrointestinal track. Through varying the lecithin content, the gel structure and rheological property of the emulsion gel system was greatly affected. To realize the effect of different physical characteristics on the bioaccessibility of nobiletin, in vitro lipolysis model was used to study the intestinal digestion kinetic. The result from the lipolysis study indicated that lecithin emulsion gel with higher viscoelastic property can significantly delay the digestion process and could be used as a controlled release system for nobiletin. Moreover, the bioaccessibility of nobiletin was significantly improved when higher level of lecithin was used in the emulsion gel system.

AGFD 179

Updated exposure assessment for 4-methylimidazole (4-MEI) for the U.S. population based on quantitative data from foods

Daniel E. Folmer3, daniel.folmer@fda.hhs.gov, Diana L. Doell1, Hyoung S. Lee3, Gregory O. Noonan2, Susan E. Carberry3. (1) FDA, Alexandria, Virginia, United States (2) Food and Drug Admin, College Park, Maryland, United States (3) Office of Food Additive Safety, US Food and Drug Administration, College Park, Maryland, United States

4-Methylimidazole (4-MEI) is an impurity found in caramel colors produced using ammonium compounds (Class III and IV). 4-MEI can also form in food through Maillard reactions between reducing-sugars and amino acids that occur when a food is cooked, roasted, or heated. Caramel colors are the most widely used color additives in food. Results of toxicological testing in rats and mice conducted by the National Toxicology Program (NTP) on 4-MEI have raised safety concerns. The Food and Drug Administration (FDA) previously conducted a robust exposure assessment of 4-MEI from the use of Class III and IV caramel colors in foods, including beverages, based on the analysis of approximately 400 representative food products. In

the current study, our exposure estimate has been updated by including quantitative 4-MEI data from an additional 200 food products, focusing on foods other than beverages that contain caramel color, as well as select foods in which 4-MEI forms naturally as a result of the Maillard reaction. The 4-MEI level in all food samples collected was quantified using liquid chromatography-tandem mass spectrometry (LC-MS/MS). We have updated the dietary exposure estimate for 4-MEI for the U.S. population aged 2 years or more, as well as several subpopulations, using food consumption data from the combined 2007-2010 National Health and Nutrition Examination Survey (NHANES), as well as 14-day food consumption data, which are likely to be more representative of actual eating patterns in the U.S. This study summarizes the results of our updated dietary exposure assessment for 4-MEI.

AGFD 180

In-vitro digestion properties of Pickering emulsions stabilized by starch nanocrystals

rong liang, rongliang@jiangnan.edu.cn, Yanwei Jiang, Cheng Yang. Key Laboratory of Food Colloids and Biotechnology, Ministry of Education, School of Chemical and Material Engineering, Jiangnan University, Wuxi, Jiangsu, China

With the high storage stability, Pickering emulsions stabilized by particles are becoming more and more popular in different areas including food, cosmetic, medicine and chemical lately. However, with the lack of research about the in-vivo digestion and benefit of food-grade Pickering emulsions, the real application of these emulsions in food systems are still limited. The aim of this research was to investigate and reveal the digestion properties and mechanism of Pickering emulsions prepared by starch nanocrystals (SNCs) and three different type of oils, short chain triglycerides (SCT), medium chain triglycerides (MCT) and long chain triglycerides (LCT) by using an in-vitro digestion model. After digestion process, the percentage of starch digestion in three emulsions was decreased in the order of LCT > MCT > SCT, while the rate of oil digestion was increased in the order of LCT < MCT < SCT. Combined with the fluorescence photographs of particles during digestion, several reasons were analyzed to be related to these results: as for LCT emulsions, with the lowest percentage of oil digestion, more SNC would be combine at the oil-water interface which would be more benefit to increase the area of SNC to amylopsin. With respect to SCT emulsions, with the high rate of lipolysis, more bile salt would be incorporated into the micelles formed together with free fatty acids. So the precipitate formed by bile salt and calcium would be decrease and more calcium would be available in the digesta to induce the aggregation of SNC, further to inhibit the access of amylopsin to the glycosidic bond of SNC which corresponding to the lowest rate of starch digestion. As for the MCT emulsions, with the moderate lipolysis rate, the extent of SNC digestion was also between LCT emulsion and SCT emulsion. Overall, the digestion fate of Pickering emulsions were determined by the combination of different properties, including the type of oil and particles, the location of particles, calcium ions and bile salts. And this result

may also provide useful information for developing Pickering emulsions with different digeation and release profiles by adjusting the componets in the emulsions for functional compounds in food and pharmaceuticals.

AGFD 181

Tanshinone IIA modulates cell-surface LDLR level and LDL uptake via suppression of PCSK9 gene expression in HepG2 cells

Hung-Chen Chen1, janice16882@gmail.com, Ming-Jiuan Wu2, Pei-Yi Chen3, Yi-Chaun Chen4, Mi-Hsueh Tai1, Jui-Hung Yen1,4. (1) Department of Molecular Biology and Human Genetics, Tzu Chi University, Hualien, Taiwan (2) Department of Biotechnology, Chia Nan University of Pharmacy and Science, Tainan, Taiwan (3) Center of Medical Genetics, Buddhist Tzu Chi Hospital, Hualien, Taiwan (4) Institute of Medical Science, Tzu Chi University, Hualien, Taiwan

The secreted proprotein convertase subtilisin/kexin type 9 (PCSK9) has been demonstrated as a natural inducer of low-density lipoprotein receptor (LDLR) degradation and shown to regulate the level of LDLR protein, making it a critical modulator of cholesterol homeostasis. Inhibition of PCSK9 gene expression is becoming a novel strategy for lowing plasma cholesterol. Tanshinone IIA is one of the most bioactive constituents isolated from the rhizome of Salvia miltiorrhiza Bunge (Danshen), has been reported to prevent or slow progression of numerous diseases including cancer, hepatic fibrosis, neurodegenerative disorders and cardiovascular diseases. In the present study, we aim to investigate the hypocholesterolemic effect and molecular mechanism of tanshinone IIA. Our results showed that tanshinone IIA increased LDLR level on the cell surface, as well as LDLR activity; however, LDLR mRNA transcription was not affected in HepG2 cells. Furthermore, we found that tanshinone IIA significantly reduced PCSK9 mRNA and protein expression via suppression of PCSK9 promoter activity, leading to an increase in cell-surface LDLR and LDL uptake in HepG2 cells. The tanshinone IIA-responsive element of the PCSK9 promoter and the transcriptional modulators for tanshinone IIA-mediated PCSK9 gene regulation will be further investigated. Our current findings indicate that tanshinone IIA down-regulation of PCSK9 gene expression is associated with increases in cell-surface LDLR and LDLR activity in hepatic cells. Tanshinone IIA exhibits hypolipidemic activity and may serve as a useful phytochemical for prevention or treatment of hypercholesterolemia.

AGFD 182

Novel SERS-based approach to detect hydrogen peroxide scavenging activity

Weiping Qian, wqian@seu.edu.cn. Southeast University, Nanjing, China

The importance of evaluating the antioxidant activity has been widely reported in the past two decades not only for the extensive application of antioxidants in food industry

and food nutriology, but also because of the discoveries of the strong correlations between reactive oxygen species (ROS) and many pathogenesis of human diseases, e.g. nervous system injuries, atherosclerosis and cancer. Hydrogen peroxide (H2O2), a key part of ROS, is generated in vivo under physiological conditions by peroxisomes (a kind of oxidative enzyme including glucose oxidase and D-amino acid oxidase) and superoxide dismutase which catalyze the dismutation of superoxide radicals. Along with superoxide anion radical, H2O2 can damage many cellular components and further convert into more ROS such as hydroxyl radical. Thus, the evaluation of H2O2 scavenging activity is an important part for assessing the ROS scavenging activity. Generally, the evaluation of H2O2 scavenging activity is peroxidase based or enzyme-free but biomarker dependent. Many methodologies based on different mechanisms have been developed for detecting H2O2 and/or H2O2 scavenging activity, e.g. chemiluminescence, fluorometry, colorimetry, cyclic voltammetry, electro spin resonance, etc. Surface-enhanced Raman Scattering (SERS) is a new powerful analytical technique with ultrahigh sensitivity. In addition to the ‘‘whole-finger-prints’’ information and the low water background, SERS has high signal enhancement and fluorescence quenching effects, which are extremely desirable for the biological sample analysis. Because of its unique attributes, SERS has been used in a wide range of fields, including homeland security, food safety, clinical diagnosis, trace detection, chemical and biochemical monitoring, etc., since it was discovered in the late 1970s. In this work, we have developed a novel SERS-based approach to detect H2O2 scavenging activity by using gold nanoshell precursor nanocomposites (SiO2 /GNPs) as nanoprobes. Compared with conventional methods, the novel SERS-based method presented here with high sensitivity, rapid determination, relative ease of measurement may serve as an additional means for estimating the activities of multifunctional antioxidants.

AGFD 183

Simultaneous analysis of unregistered pesticides in Korea by liquid chromatography-tandem mass spectrometry

Sang-Hyeob Lee1, qqwe234@naver.com, Jeong-In Hwang1, Sang-Oh Jeon1, Jang-Eok Kim1, Young D. Lee3, Hee-Jung Kim2, Hyeon-Suk Lee2, Moon-Ik Jang2, Gyu-Seek Lee2. (1) School of Applied Biosciences, Kyungpook National University, Daegu, Korea (the Republic of) (2) National Institute of Food and Drug Safety Evaluation, Osong, Korea (the Republic of) (3) Division of Life and Environmental Science, Daegu University, KyungSan, Korea (the Republic of)

For introducing Positive List System (PLS) into Korea, it is required to develop rapid and sensitive methods that can simultaneously analyze unregistered pesticides in imported agricultural products using liquid chromatography-tandem mass spectrometry. Classical

multi-residue analysis method was established by Ministry Food and Drugs Safety (MFDS) in Korea, however, it did not have unregistered pesticides for simultaneous analysis. Therefore, we analyzed 47 unregistered pesticides using brown rice and orange as fatty and non-fatty representative samples with Korean Pesticide Residue Analysis (KOPRA) method developed in this study. The residual pesticides in samples were performed by extracting with acetonitrile, and a clean-up step using dispersive solid phase extraction with primary secondary amine and magnesium sulfate. Limits of quantification were 0.01 mg/kg for 46 target analytes, and 0.025 mg/kg for D-trans-allethrin. Recoveries in the fortification range of 0.1−0.5 mg/kg were within 60−130% with associated relative standard deviations below 30% for 47 pesticides. This method could be used as the official method for monitoring pesticides applicable to the PLS of imported agricultural products in Korea.

AGFD 184

Combination of pre-column nitro-reduction and ultraperformance liquid chromatography with fluorescence detection for the sensitive quantification of 1-nitronaphthalene, 2-nitrofluorene, and 1-nitropyrene in meat products

Kailin Deng2, kdengaa@connect.ust.hk, Wan Chan1,2. (1) Chemistry, The Hong Kong University of Science and Technology, Hong Kong, Hong Kong (2) Environmental Science Programs, The Hong Kong University of Science and Technology, Hong Kong, Hong Kong

Carcinogenic nitropolycyclic aromatic hydrocarbons (nitro-PAHs) are ubiquitous in the ambient environment. They are formed predominantly during incomplete combustion processes and by reaction of polycyclic aromatic hydrocarbons (PAHs) with nitrogen oxides and nitric acid. The emerging evidence that nitro-PAHs are uptaken by plants and bioaccumulated in the food chain aroused world-wide concerns. Therefore, analytical methods of high sensitivity are extremely important for assessing the risk of human exposure to nitro-PAHs. In this study, a simple and robust ultra-performance liquid chromatography coupled fluorescence detector (UPLC–FLD) method was established for the sensitive determination of nitro-PAHs in meat products. The method entails extraction with dichloromethane, sample clean-up by SPE, followed by pre-column reduction (Figure.1.), and quantification by UPLC−FLD analysis. The developed method was validated for extraction efficiency, accuracy, precision and detection limit; and has been successfully applied in quantifying 1-nitronaphthalene (1-NN), 2-nitrofluorene (2-NF), and 1-nitropyrene (1-NP) in fresh and cured meat products. The results showed that 1-NN was the most commonly identified nitro-PAH in the collected meat products. And for the first time, our study revealed high temperature food processing has a pronounced, but differentiated effect on nitro-PAHs in fresh and cured meat.

Figure. 1. Fe/H+-induced nitro-reduction of non-fluorescing nitro-PAHs forms fluorescent amino-PAHs for UPLC–FLD analysis.

AGFD 185

Updated exposure estimate for FD&C color additives for the U.S. population

Diana L. Doell, diana.doell@fda.hhs.gov, Daniel E. Folmer, Hyoung S. Lee, Kyla M. Butts, Susan E. Carberry. FDA, College Park, Maryland, United States

In the U.S., there are 7 synthetic organic color additives that are permitted for general use in food: FD&C Blue No. 1, FD&C Blue No. 2, FD&C Green No. 3, FD&C Red No. 3, FD&C Red No. 40, FD&C Yellow No. 5, and FD&C Yellow No. 6. These color additives are subject to batch certification by the Food and Drug Administration (FDA) and are permitted for use in foods at levels consistent with good manufacturing practice. In 2011, FDA convened its Food Advisory Committee (FAC) to consider data on the possible association between children’s consumption of these color additives in food and hyperactivity or other behavioral effects. The FAC concluded that a causal link between children’s consumption of synthetic color additives and behavioral effects had not been established based on the available data. However, the FAC recommended further research, including a comprehensive exposure assessment for these color additives, especially for children. Based on this recommendation, FDA previously

conducted a robust exposure assessment of the FD&C colors using data from the analysis of approximately 600 representative food products and 2-day food consumption data from the 2007-2010 National Health and Nutrition Examination Survey (NHANES). However, the use of 2-day food consumption data to assess chronic exposure can lead to an overestimation of exposure, especially for foods that are not commonly consumed. For this reason, data collected from a 14-day survey are considered to be more representative of actual consumption patterns in the U.S. Using 14-day food consumption data, the dietary exposure to FD&C color additives has been updated. Dietary exposures from the use of each FD&C color in food were estimated for the U.S. population aged 2 years or more, and several subpopulations, with a focus on children. This study summarizes the results of this exposure assessment.

AGFD 186

Preparation and rheological characterization of food grade gellan hydrogels with calcium ions as gelling agent

Emmanuel Neba N. Ambebila, ambebila@yahoo.com. Department of Chemical Engineering , University of Barcelona, Barcelona, Spain

Hydrogels are networks of hydrophilic polymer chains that are highly valued for their functionality and ability to tune physical properties in many industrial applications. Biopolymer-based hydrogels are relatively safe for applications in variety of industries for rheology control, encapsulation and targeted drug delivery due to their biocompatibility and nontoxic nature. Gellan is a bacteria exopolysaccharide capable of imbibing high volumes of water and swelling to form hydrogels with varying properties depending on the production method. In this work, gellan hydrogels were prepared via ionotropic gelation using calcium chloride as gelling agent and characterized by determining their rheological properties. The hydrogels were produced by ionotropic gelation of gellan solution using calcium ions as gelling agent under high shear rate (with an UltraTurrax T.25 basic IKA-WERKE). The rheological properties were characterised by performing amplitude sweeps, oscillatory frequency sweeps, yield point testing and shear rate sweeps with a modular advanced rheometer system (HAAKE-MARS) at 25oC. The gelation point was determined and the rheological properties showed a non-Newtonian behaviour with increase in gel strength as the quantity of calcium ions increased at constant gellan concentration but beyond a certain maximum Ca2+/gellan ratio, there was no further increase in gel strength. The hydrogels showed higher rheological properties (storage modulus and Yield point values) with an increase in the shear rate of the unltraturax. The gelation of gellan occurs at remarkably low concentrations compared to other polysaccharides making the system very favourable for use in food and cosmetic applications. Key words: gellan, hydrogels, gelling agent, rheology

AGFD 187

Saponins quantification in pigmented chickpeas cultivars

ada k. milan, keilamilan@hotmail.com, Sergio R. Serna Saldivar, Janet Gutierrez. Tecnologico de Monterrey, Monterrey, NL, Mexico

Chickpea (Cicer arietium L) is an important legume crop in the world. Legumes are the main source of dietary saponins, these bioactive compounds are a group of secondary metabolites with a structure constituting of steroid or triterpenoid aglycones and sugar moieties. For years, saponins had been considered anti-nutritional factors; nowadays, they have been attributed with multiple health promoting effects such as diminishing metabolic syndrome symptoms. In this study, the saponins concentration in 80% methanolic extracts of ten pigmented chickpeas cultivars (cv) were analyzed. The chickpeas studied had green (1 cv), black (3 cv), red (3 cv), brown (1 cv) and cream (2 cv) colored seed coat. The saponins were identified by their corresponding mass spectra obtained by HPLC-IT-MS and quantified by HPLC-ELSD using soyasaponin I as external standard. Three predominant saponins were found. Soyasaponin βg (m/z 1068) was identified as the most abundant saponin representing more than 90% of total content except by green cultivar (≈ 70%), and the content ranged from 866-164 μg/g in chickpea samples. These results agree with previous reports of saponins content in chickpea seeds. Moreover, Soyasaponin Bb (m/z 942) was found in all samples in less than 10% except for green cultivar (≈30%). Soyasaponin αg (m/z 1084) had been previously identified in chickpea but not quantified. However, the chickpeas of this study presented low (≈4%) amounts of this saponin. For the black and cream cultivars, there was an inverse association to seed size and saponins content. In this study, pigmented chickpeas showed soyasaponin Bb or αg in their profile, in contrast with other studies that not identified or quantified their presence in raw chickpea seeds. Therefore these chickpeas cultivars are a potential source of saponins for promoting health effects.

AGFD 188

Development and validation of analytical method of furan in seven different types of food matrices using SPME-GC/MS

YUN-JEONG SEOK, yunjung0314@naver.com, SOO YOUNG JEONG, Jae-Young Her, Kwang G. Lee. Dongguk University, Seoul, Korea (the Republic of)

Furan is a volatile compound formed during the maillard reaction and was recently classified as a possible human carcinogen (group 2B) by the International Agency for Research on Cancer. It has been reported to occur in various foods that undergo heat treatment. In this study, development and validation of analytical method of furan were carried out using solid phase microextraction-gaschromatography/mass spectrometry (SPME-GC/MS). To minimize the matrix effect, seven different types of food matrix such as water, peanut butter (semi-solid-fat), ham (solid-fat), corn oil (liquid-fat), rice soup (solid-non-fat), apple juice (liquid non-fat) and whole milk (liquid-lipoprotein) were selected for validation. The linearity values were obtained higher than 0.99 and recovery efficiencies were ranged from 46.0% to 113.9%. The limit of detection (LOD) was

ranged from 0.18 to 0.46 ng/g and the limit of quantification (LOQ) ranged from 0.54 to 1.38 ng/g. The inter-day precision of furan ranged from 5.1 to 17.6% and the intra-day precision of furan ranged from 3.0 to 15.0%. This research was supported by a grant (13162MFDS049) from Ministry of Food and Drug Safety in 2013 & 2014.

AGFD 189

Analytical advances in food-technology by establishment of a 14-C food –technology lab and kitchen

Matthias Kotthoff, matthias.kotthoff@ime.fraunhofer.de, Mark Bücking. Environmental and Food Analysis, Fraunhofer Institute for Molecular Biology and Applied Ecology, IME, Schmallenberg, Germany

Analyzing the fate and metabolism of ingredients or environmental and technical contaminations in complex matrices, such as food, is a major challenge in modern food processing. By using radioactively labelled substances any remains, metabolites and fragments can easily be monitored along the food processing chain. One major interest is the formation of flavour compounds and their fate during processing. Fraunhofer IME has set up a fully equipped laboratory kitchen and laboratory-scale food-processing facilities to be run under radioactive conditions. The core technical units are assembled around the four fields: Domestic / restaurant kitchen Fruit technology, beverages, wine, oil and respective microbiology (presses, separators, mills, fermenters and pasteurizers) Cereals, milling, baking technology (diverse mills, flexible extruder, separators, fermenters) Meat and meat technology (masticator, cutter, kettle, injector, piston filler) This approach allows us to monitor the fate and generation of flavouring compounds and the optimization of processing strategies. With these facilities on the other hand, we aim to identify precursors of desired and undesired cues and to optimize food processes to yield tastier, healthier and more sustainable food products.

AGFD 190

Carotenoid composition analysis in fruit of rose hip (Rosa glauca) by HPLC-DAD-APCI+-MS

Lijie Zhong, qingtianzlj@gmail.com, Karl-Erik Gustavsson, Marie Olsson. Plant Breeding, Swedish University of Agricultural Sciences, Alnarp, Skåne, Sweden

Rose hip fruit has traditionally been considered as healthy food, and used for e.g. tea, soup, and jam. It contains large amount of carotenoids as well as a high amount of vitamin C. The carotenoids from rose hip fruit were extracted, separated, identified and

quantified with a reverse-phase HPLC-DAD-APCI+-MS system. By comparing the chromatograms of saponified and untreated (unsaponified) samples, large amounts of xanthophylls were found to exist as the esterified form in rose hip fruit, including violaxanthin, 5,6-epoxy-lutein, zeaxanthin, rubixanthin, and β-cryptoxanthin. The carotenoid content in the investigated rose hip species was 1.68±0.19 mg/g dry weight. The carotenoid esters, constituting 37% of the total carotenoid content in the rose hip fruit was present in the largest amount, followed by lycopene 25%, β-carotene 17% and other carotenes 19%. Only small amounts of rubixanthin and β-cryptoxanthin were detected as free xanthophylls. The study of carotenoid composition revealed the potential for rose hip fruit to be developed as a natural carotenoid source for food products or nutraceuticals.

AGFD 191

Convenient, inexpensive measurement of free and copper-complexed hydrogen sulfide in wine

yi chen1, chenyi15@ncu.edu.cn, Jillian A. Jastrzembski2, Imelda Ryona3, Gavin L. Sacks4. (1) State key lab of food science and tehchnology, Nanchang University, Nanchang, China (2) Department of Food Science, Cornell University, Ithaca, New York, United States

Recent work suggests that copper-thiol complexes may serve as a latent source of free H2S and other malodorous volatile thiols during wine storage. However, current approaches to measurement of these complexes require the use of analytical tools like gas chromatography – mass spectrometry (GC-MS) that are unavailable to most wineries. To facilitate further studies, an inexpensive and convenient method for measurement of free and copper-complexed H2S in wine was developed using commercially available colorimetric gas detection tubes (GDTs). Calibration curves for free H2S in synthetic wine demonstrated good linearity (r2=0.9985), acceptable detection limits (0.34 μg/L), and good reproducibility (coefficient of variation = 3.9%). Interferences from sulfite or sulfate were not observed. Addition of CuSO4 resulted in near-instantaneous binding of free H2S, as expected. This complexed H2S could be measured by GDTs following disruption of the copper sulfide complexes, either by i) 10-fold dilution in concentrated NaCl brine or ii) addition of a strong Cu(I) chelator, neocuproine. These new approaches should be appropriate in both winery and research laboratory settings for studies of the role of copper-thiol complexes in causing sulfurous off-aromas during wine storage.

AGFD 192

Interaction between caseinophosphopeptides and theaflavin-3,3’-digallate and its impact on the antioxidant activity of theaflavin-3,3'-digallate

Yike Jiang2, yike9090@gmail.com, Yuwen Ting2, Jessica Li2, Qingrong Huang1. (1) Food Science, Rutgers Univ, New Brunswick, New Jersey, United States (2) Food Science, Rutgers University, New Brunswick, New Jersey, United States

Being found ubiquitously in food systems, the interactions between proteins and polyphenols during digestion can affect their bioavailability and bioefficacy. Although the interactions between protein/peptides and polyphenols have been extensively studied, the impact of these interactions on the bio-efficacy of polyphenols are still controversial. We selected the characteristic polyphenol theaflavin-3,3’-digallate (TF-3) from black tea and polypeptides caseinophosphopeptides (CPPs) from milk as the models to study their interactions at physiological condition. Further, the influence of their interactions on the antioxidant activity of TF-3 was evaluated. TF-3 is an important polyphenol in black tea, contributing greatly to its color and mouth feel. It has been documented to have multiple bio-efficacies including anti-oxidative, anti-inflammatory, cardiovascular protective and anti-carcinogenic effects. CPPs are a group of bioactive peptides hydrolyzed from caseins, the major proteins in milk. CPPs were detected in human stomach and small intestine, indicating that they are stable in human gastrointestinal tract. The interactions between CPPs and TF-3 were confirmed by fluorescence quenching study, and hydrogen bonding and van der Waals interaction were identified as the major interactions. Circular dichroism (CD) spectra in far-UV (190 – 260 nm) region revealed that association between CPPs and TF-3 did not change the secondary structures of CPPs. Near-UV (260 – 320 nm) CD spectra showed that the tertiary structures of CPPs were altered after interacting with TF-3. The conformation change resulted from the interactions between CPPs and TF-3 was further confirmed by Small Angle X-Ray Scattering (SAXS). Cellular Antioxidant Activity (CAA) assay revealed that the association between CPPs and TF-3 did not impair the antioxidant activity of TF-3.

AGFD 193

Micronanopores in diatomite fabricated by high energy electron beam and hydrothermal treatment to control the loss of pesticide

Xin Zhang1, xinzhang@ahau.edu.cn, Zhengyan Wu2, zhengyanwu@gmail.com. (1) School of Life Sciences, Anhui Agricultural University, Hefei, China (2) Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei, China

High energy electron beam (HEEB) irradiation and hydrothermal treatment (HT) were used to remove the impurities and enlarge the pore size of diatomite, resulting in plenty of micro-nanopores in diatomite. HEEB irradiation could make the impurities in the pores of diatomite loose and remove some of them, and HT could remove rest of them from the pores. A loss-control pesticide (LCP) by adding such modified diatomite to traditional pesticide. The micro-nanopores in diatomite are beneficial for the pesticide molecules to access and be adsorbed. This pesticide-diatomite complex tended to be retained by the rough surface of crop leaves, displaying a high adhesion performance

onto the leaves, so that the pesticide loss reduced, sufficient pesticide for crops was supplied, and the pollution risk of the pesticide could be lowered.

AGFD 194

Determination of protein-bound metabolites of nitrofurans by combining on-line precolumn derivatization and high performance liquid chromatography with fluorescence detection

WANG Yinan, yinan_wang2012@163.com, Wan Chan. Chmistry, Hong Kong University of Science and Technology, Hong Kong, Hong Kong

Nitrofurans (NFs) are broad-spectrum antibiotics that have been used extensively as food additives for food producing animals and as veterinary drugs treating protozoan and bacterial infections until NFs were observed to be human mutagenic and carcinogenic. However, illegal use or misuse of NFs exists. Therefore, simple and sensitive analytical methods are needed for identifying NFs contaminated food. In this work, we describe the development and validation of a novel and robust high performance liquid chromatography coupled fluorescence detector (HPLC–FLD) method for the sensitive determination of the protein-binding metabolites of NFs in fish muscle. After acid-hydrolysis releasing the protein-bound metabolites, the hydrolysates containing the NFs metabolites were protein precipitated and analyzed directly by HPLC–FLD analysis. The HPLC–FLD analysis combines on-line pre-column derivatization with fluorescence detection for NFs determination. Upon derivatizing with fluorenylmethyloxycarbonyl chloride (FMOC) harboring a fluorescent fluorenylmethyloxycarbonyl moiety, NFs metabolites forms FMOC conjugated that strongly fluoresce enabled their detection with fluorescence detection （Fig.1）. Our results showed that the HPLC-FLD analysis allows for sensitive quantification of NFs metabolites 1-aminohydantoin (AHD) and semicarbazide (SEM) at the detection limits of 0.84 and 0.28μg/kg, respectively, which are both below the minimum required performance limit (MRPL) set at 1 μg/kg by the EU. Using this new method, we can also evaluate the elimination of NFs residues in fish muscles after a contamination experiment. After 28 days of decontamination, a concentration of 143.9 μg/kg of AHD and a concentration of 9.8 μg/kg of SEM still remained in fish muscles.

AGFD 195

HPLC-MS of hesperidin metabolites in rat urine

Danielle Gonçalves2, Danielle.Goncalves@ars.usda.gov, Marilia Rodriques2, Thais Cesar2, John A. Manthey1. (1) USDA ARS-USHRL, Fort Pierce, Florida, United States (2) Food and Nutrition Department, Sao Paulo State University-UNESP, Araraquara, Brazil

The flavanone glycoside, hesperidin, in orange juice is heavily metabolized to flavanone glucuronides and mixed glucuronide/sulfates as well as a number of flavanone ring-fission products. These latter compounds occurred as the major metabolites of hesperidin in the urine of rats. Hesperidin was administered to adult male Wistar rats (200 mg/day) in the early evening and urine collected overnight. Studies of the metabolites in the urine samples were conducted by HPLC coupled with negative electrospray ionization-mass spectrometry. One main flavanone glucuronide was detected along with two other minor hesperetin-containing compounds. The main ring-fission metabolites were detected at 245, 243, 273, and 275 m/z, with fragment ions at 165, 163, 193, and 195 m/z. Each of these represented neutral losses of 80 amu, consistent with losses of sulfate groups. It is proposed that the fragment ions of 163 and 193 m/z are due to coumaric and ferulic acid groups, and that the 165 and 195 m/z fragment ions originate from substituted phenylpropionic acid conjugates. These compounds are being isolated for chemical structure determinations, for use as standards in pharmacokinetic studies, and analyses of biological actions in mammalian cells.

AGFD 196

Spectrofluorimetric study of the interaction of the mycotoxin citrinin with gold nanoparticles

Michael Appell2, michael.appell@ars.usda.gov, Wayne Bosma1, bosma@bradley.edu. (1) Dept of Chemistry and Biochemistry, Bradley University, Peoria, Illinois, United States (2) USDA-ARS, Peoria, Illinois, United States

Citrinin is a nephrotoxic secondary metabolite produced by certain fungal species from the Aspergillus, Penicillium and Monascus genera. Citrinin producing species occasionally contaminate grains, and may pose food safety risks. Analysis of this mycotoxin is frequently carried out by liquid chromatography-based fluorescence methods. Gold nanoparticles (AuNPs) offer a means to manipulate fluorescence detection and the materials provide a convenient platform for synthetic modification to tune emission signal. The influence of gold nanoparticles on the emission behavior of citrinin was investigated by steady-state fluorescence and ultraviolet absorption

spectroscopy in polar and aprotic solvents. Fluorescence quenching of citrinin by gold nanoparticles was observed in neat solvent. However, the fluorescence quenching generally associated with gold nanoparticle interactions was completely overcome under strongly acidic conditions. This behavior provides a cost-effective alternative to tune citrinin detection.

AGFD 197

Inhibitory effects of edible berry extracts on the formation of advanced glycation endproducts

Hang Ma1, mahangees@gmail.com, Weixi Liu2, Joel A. Dain2, Navindra P. Seeram1. (1) Bioactive Botanical Research Laboratory, Biomedical and Pharmaceutical Sciences, College of Pharmacy, University of Rhode Island, Kingston, Rhode Island, United States (2) Department of Chemistry, University of Rhode Island, Kingston, Rhode Island, United States

Glycation is a spontaneous process between reducing sugars and proteins that leads to the formation of advanced glycation endproducts (AGEs). The formation and in vivo accumulation of AGEs have been associated with several chronic human diseases including diabetes, inflammation, and neurodegenerative diseases. Emerging data suggest that phenolic-rich fruit, including berries, show great promise as natural anti-AGE agents. Here, we evaluated seven commonly consumed berries, including blackcurrant (Ribes nigrum), black chokeberry (Aronia melanocarpa), jamun berry (Syzygium cumini syn. Eugenia jambolana), lowbush blueberry (Vaccinium angustifolium), highbush blueberry (Vaccinium corymbosum), red raspberry (Rubus idaeus), and cranberry (Vaccinium macrocarpon) for their inhibitory effects on AGE formation. The berries were extracted with methanol and their crude extracts were further purified by adsorption chromatography (XAD-16 Amberlite Resin) to generate anthocyanins-free (ACF) and anthocyanins-rich (ACR) fractions. The crude berry extracts, along with their respective ACF and ACR fractions, were evaluated for anti-AGE effects by an intrinsic fluorescent assay using bovine serum albumin (BSA; as the model protein) and D-fructose (as the glycating agent). Aminoguanidine, a synthetic anti-AGE agent (at 100 µg/mL) was used as the positive control and showed 56.6% inhibitory effects against AGE formation. At a similar concentration (100 µg/mL), all of the extracts showed anti-AGE effects as follows: blackcurrant, black chokeberry, jamun berry, lowbush blueberry, highbush blueberry, red raspberry, and cranberry of 74.4, 70.7, 51.9, 43.6, 55.4, 72.0 and 78.7%, respectively. Also, at equivalent dried weight concentrations, the ACR fractions of each berry had a stronger inhibitory effect on AGE formation compared to their respective ACF fractions. This data suggests that anthocyanins are the major contributors to the anti-AGE activities of these berries.

AGFD 198

Pomegranate polyphenols inhibit the formation of advanced glycation endproducts and aggregation of beta amyloid

Hang Ma1, mahangees@gmail.com, Weixi Liu2, Daniel B. Niesen1, Joel A. Dain2, Navindra P. Seeram1. (1) Bioactive Botanical Research Laboratory, Biomedical and Pharmaceutical Sciences, College of Pharmacy, University of Rhode Island, Kingston, Rhode Island, United States (2) Department of Chemistry, University of Rhode Island, Kingston, Rhode Island, United States

Published studies from our group and others support the protective effects of pomegranate polyphenols against Alzheimer's disease (AD) but the mechanisms of actions are still unclear. The deposition of beta amyloid protein (βAP) plays a central role in the pathology of AD and the aggregation of βAP can be accelerated by several factors such as glycation, a non-enzymatic process which leads to the formation of advanced glycation end products (AGEs). Both AGE-modified βAP, and AGE itself, induce the aggregation of soluble βAP. Thus, the development of AGE inhibitors for the prevention of AD has attracted significant research attention. Here, we investigated the anti-AGE effects of pomegranate fruit polyphenols (namely, punicalagin, ellagic acid, gallic acid) and their human gut microbial derived metabolites (namely, urolithins A and B and their respective methylated derivatives). AGE levels were evaluated using MALDI-TOF, intrinsic fluorescence, and HPLC and βAP aggregation was evaluated by the Thioflavin T and Congo red assays. The compounds showed potent inhibitory effects on AGE formation, at all stages of glycation, and scavenged methylglyoxal, one of the most reactive among glycating agents and an AGE precursor. In addition, the compounds reduced the aggregation of βAP treated with methylglyoxal. This study suggests that the anti-AGE effects of pomegranate polyphenols and their gut derived microbial metabolites may play a role in its preventive effects against AD.

AGFD 199

Standardized food grade maple syrup extract (MSX) imparts lipid lowering and anti-inflammatory effects in mature differentiated mouse and human adipocytes

Pragati Nahar, pragaitnahar@gmail.com, Angela L. Slitt, Navindra P. Seeram. Bioactive Botanical Research Laboratory, Biomedical and Pharmaceutical Sciences, College of Pharmacy, University of Rhode Island, Kingston, Rhode Island, United States

Despite the availability of a wide variety of sweeteners, there is increasing consumer demand for natural sweeteners, among which pure maple syrup is highly regarded. Our laboratory has conducted extensive chemical characterization studies on maple syrup which led to the development of a standardized food grade maple syrup extract (named, MSX), which is safe and well tolerated (up to 1000 mg/kg per day in Sprague-Dawley rats). In this project, we evaluated the anti-lipogenic effects of MSX treatment in mature differentiated 3T3-L1 murine adipocytes and human visceral adipocytes. MSX treatment of mature adipocytes decreased lipid accumulation compared to control in both murine and human adipocytes. In 3T3-L1 adipocytes, this effect was associated with downregulation of adipo/lipogenic protein expression (e.g. PPARγ, Srebp1c). We also observed reduced mRNA expression of the pro-inflammatory mediators, namely IL-6, and TNF-α. Lastly, MSX downregulated the secretion of adipocytokines in

lipopolysaccharide-induced 3T3-L1 adipocytes. The current study adds to the growing body of in vitro and in vivo data supporting the biological effects and potential health benefits of the natural sweetener, maple syrup.

AGFD 200

Natural product derived brain absorbable RAGE inhibitors for Alzheimer’s disease: The case of the urolithins

Daniel B. Niesen, dan_niesen@my.uri.edu, Nishan Shah, Hang Ma, Navindra P. Seeram. Bioactive Botanical Research Laboratory, Biomedical and Pharmaceutical Sciences, College of Pharmacy, University of Rhode Island, Kingston, Rhode Island, United States

Alzheimer’s disease (AD) is a devastating neurodegenerative disease which is linked with an inflammatory component. The receptor for advanced glycation endproducts (RAGE), whose ligands include advanced glycation endproduct (AGE)-modified proteins and amyloid-β, which are elevated in AD brains, has been linked with increased neuroinflammation. This has led to our group’s search for anti-inflammatory natural product derived RAGE inhibitors (nRIs) as new pharmacological treatments of AD. The pomegranate (Punica granatum) fruit, a rich source of ellagitannins, has been reported by our group, and others, to exert anti-AD effects in animal models but the brain absorbable compounds are not known. Interestingly, while ellagitannins are not bioavailable in humans, they are biotransformed by gut microbiota into microbial metabolites, known as urolithins (dibenzopyranone derivatives) which achieve physiologically relevant concentrations and show anti-inflammatory effects in vivo. Therefore, given the above, we hypothesized that urolithins are the brain absorbable nRIs responsible for pomegranate’s protective effects against AD pathology. This hypothesis was supported by in silico (ACD/Labs to predict blood brain barrier penetration) and molecular docking (Autodock Vina to evaluate ligand-RAGE binding affinity) data. Using common motifs shared by known synthetic RIs, and functional group optimization, a library of urolithin derivatives was developed. Our group is currently pursuing ADMET, PK/PD, and further preclinical studies of these nRIs, and their related lead compounds, to advance their development as new anti-AD therapeutics. Also, this study underscores the importance of mining gut microbial metabolites, formed from host microflora metabolism of dietary natural products, for new therapeutic agents for chronic human illnesses including AD.

AGFD 201

Beyond L-DOPA: Bioactives in Mucuna pruriens for the treatment of Parkinson’s disease

Craig Hessler, craighessler@gmail.com, Daniel B. Niesen, Hang Ma, Navindra P. Seeram. Bioactive Botanical Research Laboratory, Biomedical and Pharmaceutical

Sciences, College of Pharmacy, University of Rhode Island, Kingston, Rhode Island, United States

Mucuna pruriens (commonly known as velvet bean) is a legume used in several traditional systems of medicines to treat a variety of ailments. The seeds of M. pruriens, which contains L-DOPA (3,4-dihydroxyphenalanine), has been used to treat Parkinson’s disease (PD) in Ayurvedic medicine. Notably, the anti-PD effects of M. pruriens seeds are supported by published animal and human clinical studies. Moreover, it has been reported that the anti-PD effects of this natural product are greater than when compared to synthetic L-DOPA alone. Whether there are other constituent/s, beyond L-DOPA, contributing to the anti-PD effects of this natural product is not known. Therefore, the current study was designed to comprehensively isolate and identify the phytochemical constituents in M. pruriens seeds. The isolates will be evaluated singly, and also combined as a whole M. pruriens seed extract (to evaluate additive and/or synergistic effects), in enzyme and cell based bioassays, to compare their anti-PD activities to L-DOPA alone. This project will contribute to the growing body of data supporting the anti-PD effects of this natural product.

AGFD 202

Methylglyoxal induced cell cytotoxicity inhibitory and scavenging properties of a standardized food grade maple syrup extract (MSX)

Weixi Liu2, wliu@chm.uri.edu, Zhengzi Wei1, Hang Ma1, Joel A. Dain2, Zahir Shaikh1, Navindra P. Seeram1. (1) Bioactive Botanical Research Laboratory, Biomedical and Pharmaceutical Sciences, College of Pharmacy, University of Rhode Island, Kingston, Rhode Island, United States (2) Department of Chemistry, University of Rhode Island, Kingston, Rhode Island, United States

Methylglyoxal (MGO) is a potent glycating agent involved in the formation of advanced glycation endproducts (AGEs). The in vivo accumulation of AGEs plays an important role in the pathology of several chronic illnesses including type-2 diabetes and Alzheimer’s disease. Emerging data suggest that phenolic-rich fruit, vegetables, and beverages show great promise as natural anti-AGE agents. While numerous anti-AGE studies have been conducted on these foods, there is lack of similar data on phenolic-containing natural sweeteners such as maple syrup. Our group has conducted extensive previous research on the identification and structure elucidation of a large number of bioactive polyphenols present in maple syrup. More recently, we reported the development of a nutraceutical food grade maple syrup extract (named, MSX) which has been chemically characterized and studied for safety and tolerability (in Sprague-Dawley rats at doses of up to 1000 mg/kg per day). Here, we show that MSX scavenges MGO (from biochemical assays and circular dichroism studies) similar to the positive control, aminoguanidine. Also, MSX protected normal human colon (CCD-18Co) cells from MGO-induced cell cytotoxicity and stimulated extracellular signal-regulated kinase (ERK) phosphorylation. Overall, this study supports the anti-AGE effects of phenolic-rich plant foods including the natural sweetener, maple syrup.

AGFD 203

3'-Hydroxypterostilbene simultaneously induces apoptosis and autophagy in human prostate cancer cells

Hui-Yun Tsai3, rhytsai@gmail.com , Tzou-Chi Huang1,4, Chi-Tang Ho2, Yu-Kuo Chen1, chenyk@mail.npust.edu.tw. (1) Department of Food Science, National Pingtung University of Science and Technology, Pingtung, Taiwan (2) Food Science, Rutgers University, New Brunswick, New Jersey, United States (3) Department of Food Science, Rutgers University, New Brunswick, New Jersey, United States (4) Department of Biological Science and Technology, National Pingtung University of Science and Technology, Pingtung, Taiwan

Prostate cancer is the second leading cause of cancer-related death among men in the United States. Therefore, more efforts should be dedicated to the development of preventive strategies to reduce prostate cancer prevalence and impact. Pterostilbene is a natural compound predominantly found in blueberries and grapes. 3'-Hydroxypterostilbene (OHPt), one of metabolites of pterostilbene, can also be isolated from whole specimens of the herb Sphaerophysa salsula. The objective of this study was to investigate the growth inhibitory effects of OHPt on PC-3 human prostate cancer cells. The results showed that OHPt treatments significantly decreased cell viability of PC-3 cells. OHPt also disrupted the mitochondrial membrane integrity and promoted the activities of caspase-3, -8 and -9. Moreover, OHPt increased the amount of proapototic protein Bax and decreased the anti-apoptotic protein Bcl-xL. These findings suggest that OHPt has strong growth inhibitory effect on PC-3 cells through induction of apoptosis pathway. Besides, OHPt also induced other cell death pathway-autophagy by increased the expression of Beclin-1, LC3B II proteins and autophagosome formation. Taken together, OHPt shows great potential as a novel and useful agent for the treatment and prevention of human prostate cancer cells. Further work is still required to clarify the molecular mechanism of apoptosis and autophagy induced by OHPt in vivo.

AGFD 204

Evaluating Raman spectroscopic data by using principal component analysis to determine the freshness of fish samples

Havva Tumay Temiz1, tumaytemiz89@gmail.com, Hasan M. Velioglu2, Ismail H. Boyaci1,3. (1) Faculty of Engineering, Department of Food Engineering, Hacettepe University, Ankara, Turkey (2) Faculty of Agriculture, Department of Agricultural Biotechnology, Namik Kemal University, Tekirdağ, Turkey (3) Food Research Center, Hacettepe University, Ankara, Turkey

Low content of saturated fats and cholesterol and high content of polyunsaturated fatty acids (PUFA) makes fish an indispensable element of a healthy diet. On the other hand, its high moisture content and weak muscle tissue does not allow the long-term storage. Freezing is the most commonly applied preservation method used for extending the

shelf life of fish. Improper practices of freezing and thawing, results with undesirable textural and functional changes, which mostly cannot be realized by consumers. Enzymatic and chemical changes as well as dielectric properties of fish samples were monitored in many studies in order to differentiate fresh and frozen-thawed fish samples. Fourier-transform infrared spectroscopy, impedance spectroscopy and near infrared spectroscopy are frequently used novel technologies with the advantages like rapidness and on-site usability. As one of the vibrational methods, which gives structural information and requires no sample preparation Raman spectroscopy is used in the present study in order to differentiate fresh and frozen-thawed fish samples. Unlike the reported studies, relatively short freezing (24h) and thawing (12h) periods were used in the present study. By using the advantage of its high sensitivity, very little changes in the lipid structure, which occurred during the very short storage periods, were monitored using Raman spectroscopy. Collected Raman data was analyzed by using Principal Component Analysis (PCA). Developed PCA models allowed us to obtain a successful differentiation between fresh, once frozen-thawed and twice frozen-thawed fish samples. Gas chromatography (GC) was used as reference method and fatty acid composition of fish samples were determined by GC analysis. A correlation was established between the results of GC and Raman spectroscopy in terms of changes in the lipid structure.

AGFD 205

Raman spectroscopic method for determination of erucic acid in canola oils

Elif Ercioglu1, elifercioglu@hacettepe.edu.tr, SERAP D. VELIOGLU2, Havva Tumay Temiz1, Hasan M. Velioglu2, Ismail H. Boyaci1,3. (1) Faculty of Engineering, Department of Food Engineering, Hacettepe University, Ankara, Turkey (2) Faculty of Agriculture, Department of Agricultural Biotechnology, Namik Kemal University, Tekirdağ, Turkey (3) Food Research Center, Hacettepe University, Ankara, Turkey

Erucic acid is the principal fatty acid in the rapeseed, which is commonly used for edible oil production. However, in vivo experiments reported that consuming erucic acid containing oils could lead to serious health problems. Due to these health concerns, a new strain of rapeseed known as canola which contains low erucic acid was developed using selective breeding. Today canola is often used with other vegetable oils for the production of edible oils, margarine or shortenings. However, studies related to the toxicity of erucic acid are still being conducted. Regarding the safety of vegetable oils, European Council require that canola oil must contain erucic acid less than 5 %, while US legislations had specified a maximum level as 2% in total fatty acids. Hence, establishing a rapid, simple and reliable method for determination of erucic acid is of great importance in terms of consumer safety and food legislations. Chromatographic methods namely, GC, GC-MS, and HPLC are commonly used for erucic acid determination. However, these methods mostly require time and chemical consuming pre-treatment procedures. To eliminate the disadvantages of traditional methods, vibrational methods have been widely used in food analysis. The aim of this study was

to evaluate the potential of Raman spectroscopy for determination of erucic acid percentage in total fatty acids of the canola oil. The erucic acid content of canola oils, between 0% and 33.56% (w/w) were determined by means of partial least squares (PLS) analysis. High coefficient of determination values were obtained for both calibration and validation graphs, 0.990 and 0.982 respectively. This study presents a rapid (45 s), non-destructive and accurate method for determination of erucic acid content in canola oil.

AGFD 206

Effects of KCl substitution on textural properties of Queso Fresco

Michael H. Tunick, Michael.Tunick@ars.usda.gov. USDA ARS, Wyndmoor, Pennsylvania, United States

Partial substitution of KCl for NaCl has been attempted in some common cheese varieties because of restrictions on sodium in the diets of some consumers. The changes in texture of Queso Fresco, a popular Hispanic cheese, were monitored during refrigerated storage after replacing some of the NaCl with KCl during the salting step. Queso Fresco is a high-moisture (49-58% water) non-melting variety containing up to 3% NaCl; in this study the NaCl/KCl percentages were 2.0/0, 0.75/0.75, 1.0/0.5, 1.0/1.0, 1.0/1.3, and 1.0/1.5. Texture profile analysis (TPA) revealed that hardness increased by approximately 3 N for every 1% increase in salt-to-moisture ratio (S/M). The pH of Queso Fresco is 6.0-6.4, as opposed to 5.2-5.6 for more common varieties, and this high value leads to a less cohesive and less springy cheese. The TPA cohesiveness and springiness did not change with S/M, indicating that these parameters are unaffected by type and amount of salt when the cheese pH is above 6.0. Small amplitude oscillatory shear analyses were also unaffected by KCl content because the curd is milled prior to packaging, which disrupts the casein matrix and eliminates long-range molecular interactions. Other than TPA hardness, the texture of Queso Fresco is not altered by replacing some of the NaCl with KCl.

AGFD 207

Biological activities of diterpeniods from Hyptis verticillata

Roy B. Porter, roy.porter@uwimona.edu.jm. Dept of Chem, Kingston Mona, Jamaica

Seven abietane diterpenoids were isolated from the roots of Hyptis verticillata (Labiatae). These are 7-acetoxy-16-benzoxyabieta-8,12-diene-11,14-dione (1), 11,14 dihydroxy-12-methoxyabieta-8,11,13-triene-7-one (2), 11,14-dihydroxy-12-methoxy-18(4→3βH) abeo-abieta-4(19),8,11,13-tetraene-7-one (3) 7-acetoxy-12-methoxyabieta-8,12-diene-11,14-dione (4), royleanone (5), 7,6-dehydroroyleanone (6) and 7-acetoxyhorminone (7). The results of their evaluations against the sweet potato weevil, Cylas formicarius elegantulus (Summers), their antioxidant activity using 1,1-diphenyl-2-

picrylhydrazyl (DPPH) assay and their antimicrobial potential using the disc diffusion assay against six human pathogenic microorganisms will be presented.

AGFD 208

Study of the encapsulation of aroma compounds from starch emulsions by reversed flow gas chromatography (RFGC)

JOHN KAPOLOS1, jkapolos@teikal.gr, ATHANASIA KOLIADIMA2, GEORGE KARAISKAKIS2. (1) FOOD TECHNOLOGY, TECHNOLOGICAL INSTITUTE OF PELOPONNESE, Kalamata, Greece (2) CHEMISTRY, UNIVERSITY OF PATRAS, Patras, Greece

Starch consists of two biopolymers, amylose (linear polymer of 1000 glucose units) and amylopectin (branched polymer of 4000 glucose units). Glucose units linked by α-(1-4) glycosidic bonds. Usually, amylose is about 25% while amylopectin is about 75% of the total mass. These data, however, can vary depending on the origin of the starch. So, amylopectin vary from 95% in types of starch with low amylose content to a few percentages in types of starch with high amylose content. As it is well known, amylose in neutral aqueous solutions, forms helices in which encapsulated low molecular weight aroma compounds. However, amylopectin does not form such helices. In addition, most foods are watery therefore it is of particular importance to study the behaviour of different types of starch versus aroma compounds in the presence of water. In this study Reversed Flow Gas Chromatography (RFGC) technique was used for studying the encapsulation of aroma compounds in starch emulsions. Slurries of starch granules from different origin (rice, potato and corn) were produced and their concentrations where determined. Experiments at different temperatures (303.15 K, 313.15 K, 323.15 K and 333.15 K) in order to investigate the interaction between aroma compounds (d-limonene, diacetyl, 1-hexanol) and starch emulsions were carried out. Following appropriate mathematical analysis the calculation of diffusion coefficients of aroma compounds in starch emulsion and the coherence coefficients between the aroma compounds and starch emulsion were extracted. Additionally, information on the effect of amylose content of the capture and release of aroma compounds was investigated. References [1] Katsanos N.A.; Kapolos J. (1989). Diffusion coefficients of gases in liquids and partition coefficients in gas liquid interphases by reversed-flow gas chromatography Anal. Chem., 61, 2231-2237. [2] N.A. Katsanos and G. Karaiskakis. (2004). Inverse Gas Chromatography and its applications. New York: HNB Publising. Acknowledgment This research has been co-financed by the European Union (European Social Fund – ESF) and Greek national funds through the Operational Program "Education and

Lifelong Learning" of the National Strategic Reference Framework (NSRF) - Research Funding Program: ARCHIMEDES III.

AGFD 209

Stereochemical determination of methamidophos and ruelene, organophosphorus compounds

Melinda C. Chiu, melinda.chiu@jjay.cuny.edu, Kristi Tami, Cristina Kinahan, Anita Ng, Gloria Proni. Science Department, John Jay College - CUNY, New York, New York, United States

Many commercially sold insecticides include a racemic mixture of chiral organophosphorus (OP) compounds. The compounds’ two enantiomers may have different ways in which they either degrade or accumulate in the environment. This becomes an issue when realizing that the enantiomers may have certain toxicities toward several species of animals and insects. In this presentation the HPLC methodology to collect enantiomerically pure OP compounds of methamidophos, and ruelene will be presented. In addition, the absolute configuration of the pure enantiomers was established by electronic circular dichroism (CD) and optical rotatory dispersion (ORD).

AGFD 210

Analysis of carcinogenic 4(5)-methylimidazole in various commercially available foods and beverages

Sumin Lee, Jae-Young Her, jyher83@gmail.com, Min-chul Jung, Kwang G. Lee. Dongguk University, Seoul, Korea (the Republic of)

4(5)-methylimidazole (4(5)-MI) is a nitrogen-heterocyclic compound and byproducts produced during the manufacture of caramel color used to darken food products such as soy sauces. 4(5)-MI is classified by the international agency for research on cancer (IARC) as group 2B. The objective of this study was analysis of carcinogenic 4(5)-MI in various foods and beverages. A gas chromatography-mass spectrometry (GC-MS) was used for the quantification of 4(5)-MI. The ion-pair extraction was carried out using bis-2-ethylhexylphosphtate. 4(5)-MI was derivatized with isobutylchloroformate. Quantification was conducted by internal standard method with 2-ethylimidazole (2-EI). The linearity value (r2) of a standard curve for quantitative analysis was 0.9999. The limit of detection (LOD) was 11.06 ug/kg and the limit of quantitation (LOQ) was 36.87 ug/kg. 4(5)-methylimidazole was detected in various commercially available foods and beverages containing caramel colorant. Amount of 4(5)-MI in 144 commercially available foods and beverages such as soy sauces, coffee, caramel syrup, red ginseng juice, plum juice ranged from 63.31 μg/L ± 9.30 to 1925.48 ± 73.92 μg /L.

AGFD 211

Metaboilic exploration about blueberry, raspberry, and blackberry

Woosoek Kim1, Jeongjoo Pyo2, Jae-Young Her1, jyher83@gmail.com, Kwang G. Lee1. (1) Dongguk University, Seoul, Korea (the Republic of) (2) Korea University, Seoul, Korea (the Republic of)

The various berries which was used to health functional foods, beverages, yogurts, and jams have antioxidant, antimicrobial, anticancer. And they were consumed by many peoples. In this study, we identified components of blueberry, raspberry and blackberry using gas chromatography-mass spectrometer (GC-MS) and mass profiler professional (MPP). Mass spectrometry based metabolite profiling in combination with multivariate data analysis was introduced to quantity changes in metabolic patterns. The data of analyzed berries were processed by principle component analysis (PCA) as multivariate analysis to determine whether separate classes of extraction solvents. PCA score plot were identified through retention time and m/z data pair. At the results, PCA score plot obtained from hexane and ethyl ether extracts was the good discrimination between berries. And then, loading plot analysis performed to increase accuracy of data processing was collected 139 peaks. Among them, 28 peaks of main component were categorized in the significant level using ANOVA test (p<0.001). Although we have not yet identified the component of berries, tridecanal, methyl eugenol, 2-nonadecanol, 1-methylbutyl palmitate, 2-heptacosanone and 2-undecanone were searched using library. Methyl eugenol is known as antiallergic reaction compound in blueberry, raspberry, and blackberry.

AGFD 212

Formation and reduction of furan in soy sauce (ganjnag) according to the time of addition of food additives

Min Yeop Kim, Jae-Young Her, jyher83@gmail.com, Jun Young Lee, Kwang G. Lee. Dongguk University, Seoul, Korea (the Republic of)

Soy sauce is a traditional Korean food ingredient, widely used for many Asian cusines. Previous work reported the relatively high level of furan found in soy sauce, which may have been derived from the Maillard reaction in natural soy sauce making process. The objective of this study was to understand the formation of furan in soy sauce according to sterilization and fermentation temperatures. The effect of various food additives on the reduction of furan in soy sauce model system was further investigated. Soy sauce was manufactured using a standard manufacturing procedure using Koji (Aspergillus oryzae) fermentation. A level of furan was determinated by Headspace-Solid phase microextration(SPME) followed by Gas chromatography-Mass Spectrometry(GC-MS), every 5 days up to 30 days. Five food additives (magnesium sulfate, calcium sulfate, sodium sulfite, dibutyl hydroxyl toluene (BHT), and butylated hydroxyanisole (BHA)) were added to the soy sauce model system under two different conditions (simultaneous fermentation and with food additives prior to sterilization) in order to determine the effect of food additives on

reduction of furan. The addition of magnesium sulfate, calcium sulfate, BHT and BHA to soy sauce showed a significant reduction of furan level both two conditions, compared to control sample (p<0.05). However, sodium sulfite under simultaneous fermentation showed effective reduction ratio of furan levels compared to when processing this additive prior to sterilization. (Reduction of simultaneous fermentation: 94.6%~95.2%, sterilization with additive: 15.0%~17.0%, respectively) To exploit this analytical method, the present study will provide guidance to the food companies and help to reduce furan studies.

AGFD 213

Validation of an analytical method for quantification of Benzo(a)pyrene in two different types of food matrices using GC/MS

shinwoong Park1, Jun-Hyun Jeong2, Jae-Young Her1, jyher83@gmail.com, Kwang G. Lee1. (1) Dongguk University, Seoul, Korea (the Republic of) (2) Dongguuk University, Seoul, Korea (the Republic of)

Benzo(a)pyrene is hydrocarbons—organic compound containing only carbon and hydrogen formed during the imperfect combustion and was recently classified as group 1 by the International Agency for Research on Cancer. Benzo(a)pyrene has been widely used as a marker of exposure to total carcinogenic polycyclic aromatic hydrocarbons (PAH). They have been reported to occur in various foods that undergo heat treatment. In this study, development and validation of analytical method of Benzo(a)pyrene was carried out using gaschromatography/mass spectrometry (GC/MS). To minimize the matrix effect, two different types of food matrices such as apple juice (non-fatty group) and sausage (fatty group) were selected for validation. The linearity value was obtained higher than 0.99 and recovery efficiencies were ranged from 94.4% to 110.2%. The limit of detection (LOD) was 0.01 ng/g and the limit of quantification (LOQ) was 0.05 ng/g. The inter-day precision of PAHs ranged from 5.56 to 7.77% and the intra-day precision of PAHs ranged from 1.20 to 14.48%. This research was supported by a grant (14162예방안072) from Ministry of Food and Drug Safety in 2013.

AGFD 214

Formation and reduction of ethyl carbamate in soybean paste (Doenjang) model system

Su jeong Lee, Ha Yan Song, Jae-Young Her, jyher83@gmail.com, Kwang G. Lee. Dongguk University, Seoul, Korea (the Republic of)

Ethyl carbamate (Urethane, EC) occurs naturally in many alcoholic beverages and fermented foods. Due to its carcinogenic nature, many researches had been conducted to develop analysis method of EC in many matrices (solid, semi-solid, liquid) of fermented food such as soybean paste, soy sauce and vinegar. The aim of this study was to determine the effect of urease in the formation of EC, in the presence citrulline,

ethanol and urea using Doenjang model system. Doenjang was made in the laboratory using a standardized manufacturing method. Previously reported precursors (citrulline, ethanol, urea) were added to Doenjang with and without urease, and the level of EC in Doenjang model was determined by Gas Chromatography-Mass Spectrometry / Selective Ion Monitoring mode (GC-MS/SIM) for 30 days. The level of EC increased after 30 days, and the degree of increase was significantly higher in Doenjang with added precursors. Within the precursor, the more precursor were added, the more EC were detected. Regardless of precursor type, addition of urease decreased the level of EC in Doenjang. Addition of urease seems effective method for reduction of EC in Doenjang model. Future work is needed to develop alternative method for EC reduction in Doenjang model.

AGFD 215

Development of an analytical method for quantification of biogenic amines in fermented soybean paste (Doenjang)

Yong Gun Kim, Jun Young Lee, Jae-Young Her, jyher83@gmail.com, Kwang G. Lee. Dongguk University, Seoul, Korea (the Republic of)

Biogenic amines (BAs) are toxic nitrogenous compounds and formed by the microbial decarboxylation of amino acids during fermentation. BAs can cause several toxicological problems such as nausea, respiratory distress and hypo- or hypertension. Consuming high amount of fermented foods high in BAs, can pose these adverse health effects. Development of analysis method to enhance the sensitivity of detecting BAs in fermented food matrix is crucial. In this study, development of analytical method to quantify the BAs was presented in fermented soybean paste (Doenjang) model using High Performance Liquid Chromatography (HPLC). Seven Doenjang samples were selected for validation of this method, and each was manufactured in a traditional method. The matrix effect was considered for validation. The linearity values from current method were higher than 0.99, and recovery efficiencies ranged from 97.0% to 107.2%, respectively. The method detection limit (MDL) ranged from 0.37 to 2.50 ug/g and the limit of quantification (LOQ) ranged from 1.23 to 8.35 ug/g. The inter-day precision of BAs ranged from 0.84 to 8.91% and the intra-day precision of BAs ranged from 0.43 to 8.98%, respectively. The mean values of biogenic amines including tryptamine, β-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine found in commercial Doenjang samples were 55.0, 49.0, 263.2, 9.0, 229.0. 72.5, 29.5, and 10.6 ug/g, respectively. The analytical method presented by current study can give valuable guidance to the food industries by providing the direct applicable method for BAs in various fermented food ingredients.

AGFD 216

Oil lipolysis process controlled by formation of Pickering emulsion

Weiping Jin1,2, jinweiping1988@163.com, Yike Jiang2, Bin Li1, Qingrong Huang2. (1) College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China (2) Food Science, Rutgers University, New Brunswick, New Jersey, United States

Pickering emulsion, formed by solid nanoparticles absorbed at fluid interfaces, has been renewed in recent 10 years, especially for health and cosmetics application where the use of surfactants is undesirable. Up to now, food-grade particles for stabilizing Pickering emulsion, such as zein, whey protein and modified starch particles, are largely emboldened. In my work, a kind of food-grade nanoparticle was fabricated by gelatin and polyphenol self-assembly, and it displayed good interfacial property and anti-oxidative capacity. Four kinds of Pickering emulsions were stabilized by gelatin/polyphenol nanoparticles with different oil phases (medium chain triglyceride (MCT), canola oil, corn oil and olive oil respectively). Due to various oil polarities, emulsion droplet sizes showed differences from 80 to 120μm. Their lipid digestion kinetics evaluated using an in vitro lipolysis analyses, including mimic fasted state and fed state respectively. The kinetics of fatty acid release was calculated. Lipid hydrolysis in canola oil and corn oil emulsions was faster and the plateau values of total percentage of fatty acid release were much higher than pure oil. However, Pickering emulsion suppressed the process of olive oil lipolysis. The influence of emulsions kinds on lipid hydrolysis could be attributed to several mechanisms: (i) smaller oil droplet size increases contacted surface area of bile salts and lipase, (ii) strong adsorption of stabilized nanoparticles at the interface possesses steric hindrance and competition between lipase, (iii) network formation by Pickering emulsion droplet bridge may reduce lipid hydrolysis kinetics. Those results inspired us that Pickering emulsion could use for controlling oil lipolysis and regulating hydrophilic nutrients delivery.

AGFD 217

Use of fat compost from dairy industry wastewater as a new organic amendment for pepper (Capsicum annuum L.) crop

Maria Laura Fiasconaro, Maria Eugenia Lovato, Carlos Martin, cmartin@santafe-conicet.gov.ar. Instituto de Desarrollo Tecnologico para la Industria Quimica, Santa Fe, Argentina

Fat compost was obtained by aerobic composting from dairy industry wastewaters mixed with wood chips and green grass. The aim of this study was to analyze the impact of different doses of compost obtained from fats waste of dairy industry mixed with commercial substrate and with soil on pepper plants development. Furthermore, this investigation attempted to determine the effects of these mixtures on germination and composition of pepper fruits. FC extract resulted free of phytotoxicity. Different growing media were prepared by mixing 0, 10, 20 and 40% of FC with commercial substrate (CS) or soil. Results showed that higher doses of FC improved yield and several characteristics of fruit as dry matter, diameter and concentrations of some carotenoids. The mineral composition of pepper fruit was also improved by application

of different doses of FC and the incorporation of this residue did not increase the heavy metals concentration of pepper fruit.

AGFD 218

Anti-inflammatory effect of resveratrol metabolite, δ-viniferin, on LPS-stimulated murine macrophage

Pei-Hsuan Hsieh3, hsiehelen@gmail.com, Min-Hsiung Pan1, Chi-Tang Ho2. (1) National Taiwan University, Taipei, Taiwan (2) Food Science, Rutgers University, New Brunswick, New Jersey, United States (3) Food Science, Rutgers the State University of New Jersey, New Brunswick, New Jersey, United States

Inflammation is a physiological and pathological process in response to stimulus, such as tissue injury, chemical irritation and microbial pathogen infection. Acute inflammation is a rapid response and self-limiting. In general, a controlled inflammatory response is a beneficial defense system, which provides protection against infection. However, it can become detrimental if prolonged and dysregulated called chronic inflammation. Over the past several years, chronic inflammation has been recognized may be the root of various human diseases, including neurological disorders, metabolic disorder disorders, obesity, cardiovascular diseases and even cancers. Resveratrol is a well-known natural anti-oxidative and anti-inflammatory compound from grapes. In the previous study, the anti-oxidative properties of resveratrol metabolites have been suggested to be more active than resveratrol. δ-Viniferin is a main resveratrol dehydrodimer and has been identified in grape cell cultures and wines. It possesses two resorcinol skeletons, which are determinants of radical scavenging and anti-oxidative potential. Hence, we researched in the anti-inflammatory effect of δ-viniferin. We prepared this compound in vitro by the oxidative dimerization of resveratrol with horseradish peroxidase in the presence of H2O2. We found that δ-viniferin suppressed the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), as well as the down-stream product, nitric oxide, in murine RAW 264.7 cells induced with lipopolysaccharide (LPS). Moreover, the inhibitory effects of δ-viniferin on inflammation associated signaling pathways, such as the blockade of LPS-induced IκBα phosphorylation and degradation, the decrease of Akt and PI3K phosphorylation, and the reduction of LPS-induced transcriptional activity of NF-κB. According to the results, δ-viniferin may have potential to be developed into an effective anti-inflammatory agent.

AGFD 219

Density functional theory study of the formation mechanism of acrylamide with glyoxal and asparagine as precursors

Fu M. Tao1, ftao@fullerton.edu, Jinhong Wu1,2, zhengwu wang2. (1) California State University, Fullerton, California, United States (2) Shanghai JiaoTong University, Shanghai, China

The theoretical mechanism for acrylamide formation with glyoxal and asparagine precursors was studied by density functional theory methods. Transition state theory with intrinsic reaction coordinate analysis was used to explore the minimum energy reaction pathways of the potential energy surface thus elucidating the thermodynamic and kinetic properties along those paths. Our B3LYP/aug-cc-pVDZ calculations reveal two distinct pathways as feasible in the formation of acrylamide. Path I involves N-glycosylation, dehydration, decarboxylaion and directly decomposition of decarboxylated Schiff base to form acylamide, while Path II goes on the first three same steps as the path I and then proceeds to form acrylamide via the formation of 3-Aminopropionamide and elimination of ammonia. Analysis of thermodynamic and kinetic parameters for these pathways suggest the rate limiting step in path I being tautomerization of an azomethine ylide and in path II, elimination of ammonia. Here, path I is found to be more favorable in comparison to path II. In addition, participation of explicit water molecules is found to decrease of activation energy barrier for the majority of the elementary kinetic steps in this reaction, thus increasing the overall rate constant.

AGFD 220

Characterization and quantification of flavonoids and organic acids throughout fruit development in American cranberry (Vaccinium macrocarpon) using HPLC and APCI-MS/MS

Yifei Wang2, yifei@scarletmail.rutgers.edu, Jennifer Johnson-Cicalese3, Ajay P. Singh2, Nicholi Vorsa1. (1) Rutgers Univ, Chatsworth, New Jersey, United States (2) Plant Biology&Pathology, Rutgers University, New Brunswick, New Jersey, United States (3) Department of Plant Biology & Pathology, Rutgers University, Philip E. Marucci Center for Bluberry & Cranberry Research & Extension, Chatsworth, New Jersey, United States

Cranberry fruit constituents, particularly the flavonoids, have been identified as beneficial for human health. Individual flavonoids, including flavonols and flavan-3-ols, and organic acids were characterized and quantified in cranberry fruit from fruit set to ripening in eight cranberry varieties. Among the eight flavonol glycosides characterized by APCI-MS/MS, no general pattern of accumulation was observed. However, the major flavonol, quercetin-3-galactoside (representing 36% of total flavonols) initially decreased, followed by an increase during fruit ripening; while myricetin-3-galactoside (24%) and quercetin-3-glucoside (3%) increased slightly over the season, and quercetin-3-arabinofuranoside (11%) decreased. Varietal variation was evident for total flavonols on four out of eight sampling dates. Flavan-3-ols, including epicatechin monomer and polymers with degree-of-polymerization (DP) up to DP-11, were quantified. Concentrations of individual flavan-3-ols were highest in early fruit development stages and declined rapidly through early fruit maturation, but increased slightly during fruit ripening. Significant varietal variation was observed among the eight varieties tested for individual DP and total proanthocyanidins, ranging almost two-fold. Organic acids including quinic, citric, malic and benzoic acids were identified and quantified by reverse-phase HPLC. Concentration of quinic acid declined to approximately half the level from early fruit development to final ripening, while malic

acid increased slightly. Benzoic acid was detected in low concentrations from first sampling date to late July, and accumulated substantially during fruit ripening. Harvest date and cranberry variety would appear to significantly influence the levels of various health related constituents in resulting cranberry products.

AGFD 221

Flavor chemical analysis of shrimp from near-shore Louisiana Gulf Coast estuaries

Kevin H. Driggers, kdrigg1@tigers.lsu.edu. School of Nutrition and Food Science, Louisiana State University, Baton Rouge, Louisiana, United States

Shrimp can have discernable flavor characteristics that correlate to conditions of species, diet, and environment at the time and location of harvest. Identification of the flavors of shrimp could be used to optimize shrimp farming and harvesting methods. Identification of shrimp from a location that offers superior flavor offers a new business opportunity marketing premium quality shrimp. Low concentrations of flavor compounds in shrimp can cause difficulties for measurement. A method for concentrating volatile flavor compounds to a detectible level is needed for accurate and meaningful analysis. Shrimp harvesting in the Gulf of Mexico region of the US is a $400 million USD industry. This study is focused on populations of shrimp harvested in Louisiana coastal areas. Variables will include: flavor compound composition, geography, time of year, water composition, and harvest depth. Spectroscopic measurements of each shrimp sample will be compared to identify variances in flavor compound composition. Information from these comparisons will be used to guide a sensory evaluation, to determine the impact of shrimp flavor compound composition to shrimp flavor. Preliminary experiments included using solid-phase micro-extraction (SPME) and simultaneous distillation extraction (SDE) to concentrate the volatile flavor compounds. SDE was chosen as the better method. Shrimp samples from estuaries with diverse ecologies were collected. Raw shrimp were homogenized, combined in aqueous solution with a reference tri-methyl pyrazine (TMP), and an antifoaming agent dimethyl sodium methoxide (DMSM). The volatile compounds of the shrimp were extracted using simultaneous distillation extraction (SDE). Analysis was performed using Gas Chromatography-Mass Spectroscopy (GC-MS). Fractions of shrimp samples will also be analyzed using Liquid Chromatography-Mass Spectroscopy (LC-MS) to measure and identify non-volatile flavor compounds.

AGFD 222

Time-resolved determination of physicochemical quantities for physically adsorbed or chemisorbed aroma compounds on starch granules, by inverse gas chromatography

ATHANASIA KOLIADIMA2, akoliadima@chemistry.upatras.gr, JOHN KAPOLOS1, GEORGE KARAISKAKIS2. (1) TECHNOLOGICAL INSTITUTE OF KALAMATA, Kalamata, Greece (2) CHEMISTRY, UNIVERSITY OF PATRAS, Patras, Greece

The main factor influencing the fixation of aroma compounds on food are the nature of the solid. These interactions are dominated by three mechanisms: a) the distribution of aroma compounds between different phases of food, b) the diffusion of aroma compounds through the bulk of food and c) the linking of aroma compounds with food components. Starch is one of the food components which have the possibility to interact with aroma compounds because of its ability to bind these compounds in two different ways: a) by simple absorption and b) by physical encapsulation in the bulk. The mechanism of this interaction depends on the type of compound the morphological and energetic properties of the surface and of the composition of the starch molecules. Surface characterization of starch granules is of great importance in order to investigate this interaction. So physicochemical parameters for adsorption of aroma compounds at starch granules are measured as a function of time. Local isotherms θ against adsorption energy ε, fractional changes of adsorption sites f(ε) /c*max against ε, θ against f(ε)/c*max, and distribution functions θ f(ε)/c*max over adsorption energy values ε, were calculated. The method uses only chromatographic experimental data obtained by the inverse gas chromatography technique known as reversed-flow gas chromatography. It was applied to study the adsorption of d-limonene and diacetyl onto particles of starch from wheat, rice, potato and corn at different temperatures. Also from the results the time separation of experimental surface energy together with the time-independent rate constants for adsorption and desorption of aroma compounds on starch granules is described. The geometrical mean of the London parts of the total surface free energy (γL

1γL2)1/2 of the

adsorbed aroma compounds and the solid surface, accompanied by the relevant probability density functions over time are also calculated and the results can be used as a good measure for surface characterization. References N. Katsanos, G. Karaiskakis. (2004). Time-Resolved Inverse gas chromatography and its applications. New York: HNB Publishing. Acknowledgment This research has been co-financed by the European Union (European Social Fund–ESF) and Greek national funds through the Operational Program "Education and Lifelong Learning" of the National Strategic Reference Framework (NSRF)-Research Funding Program: ARCHIMEDES III. Investing in knowledge society through the European Social Fund

AGFD 223

Early detection of milk spoilage via volatile organic compound analysis using multidimensional gas chromatograph/mass spectrometry

Kevin Rochford, rochford.kevin1@gmail.com. W. T. Woodson High School, Fairfax, Virginia, United States

Early detection of milk spoilage using trace volatile organic compounds (VOCs) as indicators poses a challenge because other VOCs from milk could form complex backgrounds. Separation using one gas chromatograph (GC) column is frequently insufficient to obtain the desired detection sensitivity with a mass selective spectrometer (MSD). Here, a two-dimensional GC separation method with a heart cut technique is applied to detect trace VOCs generated from milk and detect their changes in concentration during a slow spoilage process. VOC samples of milk samples were collected via headspace sampling using a 50/30 μm Divinylbenzene/Carboxen SPME. The two-dimensional separation was achieved by using a SGE BPX-5 column (280°C, 56 m × 530 µm × 0.5 µm) for the first dimensional separation based on boiling points and a SGE SolGel WAX column (280°C, 30 m × 530 µm × 0.5 µm) for the second dimensional separation based on polarities. Samples extracted from a 1-minute heart cut window were further separated in the second column before being analyzed using a 5975C MSD. Using this method, a variety of VOCs were identified that commonly exist at the early stage of milk spoilage caused by different bacteria (B. subtilis, Lactobacillus sp., and Staph. aureus), including 2-nonanone, benzaldehyde, hexanoic acid, octanoic acid, decanoic acid, 3-methyl butanoic acid, and phenylethyl alcohol. Furthermore, the concentrations of these VOCs were found to be correlated with pH changes during the milk spoilage. The method development and the dependence of VOC detection sensitivity on sampling conditions and column separation parameters will be discussed. This work lays a foundation for developing a non-invasive and generic method for early detection of milk spoilage caused by different bacteria.

AGFD 224

Structural properties of B-type procyanidin oligomers and their ability to scavenge free radicals: A DFT study

Ana M. Mendoza-Wilson, mwilson@ciad.mx, Sergio I. Castro-Arredondo, Rene Renato R. Balandran-Quintana. Coordinacion de Tecnologia de Alimentos de Origen Vegetal, Centro de Investigacion en Alimentacion y Desarrollo, Hermosillo, Sonora, Mexico

Procyanidins (PCs) are one of the most abundant class of phenolic compounds produced by plant tissues and have proven to be more effective free radical scavengers than vitamins C and E, so they have beneficial effects on the fruits and vegetable quality, in addition to the health of consumers. There is growing evidence indicating a strong relationship between the antioxidant ability of PCs and structural factors such as degree of polymerization and conformation, but there are also many controversies about this issue. Currently exists a special interest to know the differences in the ability to scavenge free radicals among monomers, dimers and trimers, as these represent the most bioavailable forms of PCs. However this is an arduos task since by their polyhydroxylated structure these compounds generate many intermediates with distinct conformations and very short half-life, which do not allow their isolation through experimental methods, so it is necessary to use computational methodologies. In this work the structural properties and ability to scavenge free radical of monomers, dimers

and trimers of PCs was studied through the three basic mechanisms: hydrogen atom transfer (HAT), sequential proton-loss electron-transfer (SPLET) and single electron transfer followed by proton transfer (SET-PT), employing the Density Functional Theory (DFT) computational method. In summary it was found that the ability of PCs to scavenge free radicals by the HAT and SET-PT mechanisms tended to increase with the degree of polymerization (monomer < dimer < trimer) and was totally dependent on the conformation as it was higher in the compact and compact-compact conformations of dimers and trimers, respectively. In the SPLET mechanism the relationship of the structural properties with the ability of PCs to scavenge free radicals did not show a well-defined trend as in the HAT and SET-PT mechanisms.

AGFD 225

Determination of the antiradical and chelating potential of a phenolic extract and a procyanidin-rich fraction of apple peel by experimental and computational methods

Ana M. Mendoza-Wilson1, mwilson@ciad.mx, Angelica Espinosa-Plascencia2, Refugio Robles-Burgueño2, Rene Renato R. Balandran-Quintana1, Maria d. Bermudez-Almada2. (1) Coordinacion de Tecnologia de Alimentos de Origen Vegetal, Centro de Investigacion en Alimentacion y Desarrollo, A.C., Hermosillo, Sonora, Mexico (2) Coordinación de Ciencia de los Alimentos, Centro de Investigacion en Alimentacion y Desarrollo, A.C., Hermosillo, Sonora, Mexico

The apple peel is an important source of phenolic compounds among which the procyanidins are the most abundant. Both compounds have a variety of chemical and biological properties, such as the abilities to scavenge free radical (antiradical) and to chelate metals that give them potential uses as antioxidants in the areas of foods and health, depending of their composition and structure. In this study were characterized the phenolic compounds and procyanidins extracted from the apple peel of a Red Delicious variety, and their antiradical and quelating potential was determined employing experimental and computational methods. It was found that the phenolic extract consisted of 45% of flavan-3-ols, 42.55% of flavonols, 11.51% of dihydrochalcones, 0.22% of anthocyanidins and 0.7% of phenolic acids. The procyanidin-rich fraction consisted of 71.78% of procyanidin oligomers in the range of dimers to heptamers, 27.5% of monomers of flavan-3-ols and 0.72% of a gallated dimer. The mean degree of polymerization of the procyanidin-rich fraction was 4.12. The procyanidin oligomers were constituted of 97% of (-)-epicatechin as extension and terminal units and 3% of (+)-catechin as terminal unit. The antiradical potential determined through the inhibition of lipid peroxidation of linoleic acid and the ABTS·+

radical, was significantly higher in the procyanidin-rich fraction than in the phenolic extract. On the other hand, the chelating ability measured by the formation of complexes with Cu(II), was slightly greater for the procyanidin-rich fraction than for the phenolic extract. The thermochemical data obtained computationally by the PM6 semiempirical method, corroborated that procyanidins have higher antiradical potential than the major phenolic compounds of apple peel.

AGFD 226

New active packaging film from natural resources

Ana Machado, avm@dep.uminho.pt. University of Minho, Guimaraes, Portugal

A new antioxidant active packaging material for food has been developed, consisting of a co-extrusion film of two layers polylactic acid (PLA) containing a natural antioxidant (NAO). The latter was extracted from aromatic herbs from Portuguese land fields with a mixture of hexane/ethanol, 4:3 (v/v). The antioxidant was previously incorporated in the polymers matrices using a twin-screw extruder and a double layer film was produced by co-extrusion. The mechanical tests demonstrate that the mechanical properties were not affected by the incorporation of NAO. Organoleptic properties and color, texture, and physical characteristics of fat meet packaged with the new active film were measured to evaluate the shelf life of the packaged product. Results showed that, compared to film without NAO, the active film efficiently enhanced the stability against oxidation processes, thus being a promising way to extend the products shelf life.

AGFD 227

Development of a novel biomagnetic separation method for rapid detection of Escherichia coli by phage display technique

ZIYUAN WANG, ziyuanw@foodsci.umass.edu. Food Science, University of Massachusetts Amherst, Sunderland, Massachusetts, United States

Detection of food-borne pathogens is of great importance to reduce the health risk for customers. Thus, rapid and sensitive assay techniques are needed. Conventional culture methods are time consuming, while other biochemical analysis also requires pre-enrichment step resulting in a total assay time of at least 24 h. This study developed a rapid detection and separation method for target bacterial cells by combining biomagnetic separation (BMS) with bacteriophage assay. In this method, bacteriophage T7 was genetic modified to display a peptide and phage infection cycle was utilized for the detection of Escherichia coli (E.coli). The surface of beads coated with streptavidin ligands can bind with biotinylated bacteriophage T7 via the strong binding between streptavidin and biotin molecular, this complex is then utilized for the specific capture of target bacterial. Several factors, including the ratio of magnetic particles with phage particles, reaction time and washing steps were optimized. Then this phage-based biomagnetic separation was evaluated for detecting E.coli from artificially contaminated broth. The concentration of E.coli cells was determined indirectly by detecting progeny phage after infection. Detection limit of this assay in broth is less than 104 CFU/ml, and the assay can be performed in 6 to 7 hours. The results of this study demonstrate that the biomagnetic assay is a rapid, simple, and sensitive technique for the detection of E.coli which can be applied to detect food samples without pre-enrichment.

AGFD 228

Microheterogeneity and microrheological properties of high-viscosity oat β-glucan solutions

Jingyuan Xu1, james.xu@ars.usda.gov, George Inglett1, Yiider Tseng2, Denis Wirtz3. (1) NCAUR/ARS/USDA, Peoria, Illinois, United States (2) Uniersity of Florida, Gainsville, Florida, United States (3) The Johns Hopkins University, Baltimore, Maryland, United States

Soluble fibre β-glucan is one of the key dietary materials in the healthy food products known for reducing serum cholesterol levels. The micro-structural heterogeneity and micro-rheology of high-viscosity oat β-glucan solutions were investigated by monitoring the thermally driven displacements of well-dispersed microspheres via video fluorescence microscopy. By comparing the distribution of the timedependent mean-square displacement (MSD) and ensemble-averaged MSD of polystyrene microspheres imbedded in four concentrations of β-glucan solutions, we found that the solutions exhibited perfectly homogeneous behaviour at 61%, but showed a certain degree of heterogeneity at 2%. Micro-rheology investigation revealed that β-glucan solutions displayed nearly perfect viscous behaviour at 61%, but the property changed into viscoelastic at 2%. Both micro-structural heterogeneity and micro-rheological property shifts occurred over a small concentration range, between 1% and 2%, of β-glucan.

AGFD 229

Synthesis of quinolactacide, penicinoline, penicintoam, and their analogs as potential insecticides

Sivappa Rasapalli, Ronald Mastrolia, rmastrolia@umassd.edu. University of Massachusetts Dartmouth, North Dartmouth, Massachusetts, United States

Endophytes, especially endophytic fungi, have been proved to be rich sources of biologically active secondary metabolites. Bioassay-guided fractionation of the extracts of endophytic fungi of mangrove plants have resulted in the isolation of a new and unusual pyrrolyl 4-quinolinone alkaloids. Among these, structurally related metabolites such as penicinoline and its methyl ester, quinolactacide and penicintoam are the important metabolites with insecticidal acitivities. All these share quinolone phramacophore and have expressed impressive and other useful biological activities. We have initiated a synthesis program to tap the biomedicinal potential of this family based on the isatoic-anhydride chemistry. Focused library of analogues of quinolones have been generated and are being evaluated for insecticidal activities. The key steps of the total synthesis were oxonitrile genmreation and their condensation with various isatoic anhydrides in DMF in presence of NaOMe to produce quinolones. Cyclizations of nitriles/esters under basic conditions to provide tetracyclic lactams that are also potential topoisomaerase inhibitors (image 2). Details will be presented.

AGFD 230

Inhibitory activity of Enterococcus faecalis PL9003 on oxidation and melanogenesis

Yeonhee Lee, yhlee@knrrc.or.kr, Hakmi Lee. Seoul Women's University, Seoul, Korea (the Republic of)

Aims: Characterization of inhibitory activity of lactic acid-producing Enterococcus faecalis PL9003 on oxidation and melanin production. Materials and Methods: Anti oxidation activity including SOD production, resistance to superoxide anions, eliminating ability of hydroxyl radical, superoxide dismutase activities, total antioxidant activities were assayed. The heat stability of antioxidant activities were tested at various temperatures for various time intervals. Melanogenesis inhibition activity was assayed by measuring the L-tyrosinase inhibition activity with in vitro assay and inhibition of melanin contents with in situ assay. Results: Anti oxidant activity and inhibition activity on melanogenesis of PL9003 were very stable at high temperature. It turned out that PL9003 had two different kinds of SOD. Conclusions: PL9003 can be used as a starter for fermentation of natural plants in cosmetic industry or directly used for dietary consumption.

AGFD 231

Studies on the discovery of agriculturally active compounds from marine endophytic fungi

Hao-fen Sun, fyqfyx@163.com, Chao-xi Wang, Qian Song, Yi-zhen Tang, Yan Xia. Qingdao Technological University, Qingdao, China

Marine fungi metabolites are new and important resources for the discovery of novel structures and environment-friendly agriculture antimicrobial substances. From the coastlines near Qingdao, China, we obtained nearly 100 fungi strains from marine organisms. Then, using HPLC and TLC methods, we screened them against 12 strains of plant pathogenic fungi by the flat confrontation experiment and 4 strains of bacteria by coating extracts on filter papers. We discovered marine fungus CC-1 with unique morphology, isolated from marine red alga Chondria Crassicaulis, and HR-1, isolated from marine shellfish abalone, Haliotis Rubra, and found rich metabolic and significant in vitro farm antimicrobial activities. The dominant alga, Chondria Crassicaulis contained a large number of secondary metabolites with strong biological activities and its endophytic fungi and active metabolites have potential biological control abilities for plant diseases. This report describes our work on the bioactivity-guided isolation, structural elucidation of secondary metabolites, investigation of structure-activity relationship, and lead compounds for the discovery of green agriculture fungicides and bactericides.

AGFD 232

Study of vitro digestion on desiccated coconut

Hengyuan Wu1, Jian Xiong1, 940383467@qq.com, Jun Ye2. (1) School of Light Chemistry and Food Science, South China Uni. of Techn., Guangzhou, Guangdong,

China (2) State Key Laboratory of Pulp and Paper Engineering, South China Uni. of Techn., Guangzhou, Guangdong, China

Desiccated coconut, the edible solid portion inside coconut, contains cellulose and coconut oil. In this study, vitro simulation experiment was conducted on defatted and fat desiccated coconuts under the conditions of 37 ℃ and different pH values (1,2,3,4). The digestion of desiccated coconut and the pH changes of the acids were investigated, which would provide reference for applications of desiccated coconut as hyperacidity alleviator and dietary fiber additive. The result was showed that in 30s, the pH values of all the samples rose, and the rise varied with initial pHs, in which the samples (pH=2 and 3）of desiccated coconut had the largest increases that both were 1.3; and in 6h, the pH values of the samples (PH≥3) stabilized at 4.5±0.2, while the pH values of samples (pH=1) decreased compared to those in 30s. The residues of two kinds of desiccated coconut showed different change rules. The residues of defatted desiccated coconut decreased with the increase of pH, and then stabilized at pH=3, while the residues of fat desiccated coconut increased after decrease, because the oil decomposed more with the increase of pH. Acknowledgement: Supported by the National 973 project under Grant No. 2010CB732201, the National Natural Foundation of China under Grant No. 31270617.

AGFD 233

Quantitation of chiral heterocyclic key aroma compounds in cooked Alliaceae varieties using a stable isotope dilution assay

Mario Flaig1, mario.flaig@gmx.de, Michael Granvogl2, Peter H. Schieberle3,1. (1) Deutsche Forschungsanstalt für Lebensmittelchemie, Freising, Germany (2) Tech University of Munich, Freising Bavaria, Germany (3) Technical Univ of Munich, Freising, Germany

Onions, leek, shallots and other Alliaceae-varieties are important and widespread culinary ingredients as vegetables and spices in numerous processed foods all over the world. Among various sulfur compounds mainly characterizing the overall aroma of raw and processed onions (Allium cepa L.), several chiral, heterocyclic alkyl-substituted sulfur compounds, cis-/trans-3,5-diethyl-1,2,4-trithiolan, cis-/trans-3,6-diethyl-1,2,4,5-tetrathian and cis-/trans-4,6-diethyl-1,2,3,5-tetrathian, were identified as characteristic and important aroma compounds in cooked onions and were also reported in other Alliaceae-varieties. Due to the well-known aroma properties of sulfur compounds, they often hold a special position as potent aroma compounds with characteristic odor qualities. Furthermore, the stereochemistry is a well-known aspect influencing their sensory properties. Up to now, no comprehensive qualitative and quantitative data are available on the amounts of cis-/trans-3,5-diethyl-1,2,4-trithiolan, cis-/trans-3,6-diethyl-1,2,4,5-tetrathian

and cis-/trans-4,6-diethyl-1,2,3,5-tetrathian. Hence, the concentrations of the above mentioned odorants were investigated after cooking of commercial white and red onions, leek, shallots, chives and spring onions. In connection with method development for enantiomer separation, all target compounds were identified in comparison to authentic reference compounds combining gas chromatography with mass spectrometry and sensory techniques. Afterwards, a stable isotope dilution assay (SIDA) was developed for quantitation purposes applying multidimensional GC-MS techniques. Finally the target compounds were synthesized and their sensory properties were evaluated with special emphasis on odor-thresholds of the isomers in air and odor qualities.

AGFD 234

Peri-receptor modulation of the human salivary proteome by taste stimuli

Theresa Stolle, theresa-stolle@gmx.de, Matthias Bader, Thomas Hofmann. Chair of Food Chemistry and Molecular Sensory Science, Technische Universität München, Freising, Germany

To measure and predict a food’s taste signature, it is fundamental to understand peri-receptor events taking place in the oral cavity and controlling the effective concentration of taste and trigeminal stimuli to be available for chemoreceptor activation. In this context, salivary proteins are believed to interact with key food taste molecules and, by doing so, seem to impact taste receptor activation. In order to gain some more detailed insight into the impact of proteins on stimulus perception and, vice versa, the influence of a stimuli-induced modulation of salivary proteins, healthy volunteers were challenged with a series of taste-active compounds and saliva flow was measured time-dependent. Among the compounds tested, citric acid (sour), 6-gingerol (pungent), and α/β-hydroxy sanshools (tingling) showed the highest saliva-stimulating activity. Time-dependent changes in the salivary proteome were further analysed by tryptic in-gel digestion and nano-HPLC-MS/MS to identify the proteins modulated upon tastant challenge. Surprisingly, a stimuli-dependent regulation of the overall protein content and the concentration of individual salivary proteins were observed instantaneously upon stimulation, thus suggesting the release of proteins from preformed vesicles and not from de novo synthesis. Some of the proteins affected are supposed to interact with flavour molecules and hence, modulating the concentration of the free flavour compound in the oral cavity. Other proteins were found to exhibit antimicrobial and anti-inflammatory activities, thus triggering innate protective mechanisms in mucosal immunity and in non-immune mucosal defense.

AGFD 235

Modeling the leachability of pH-dependent ionizable organic contaminants from municipal sewage sludge

Arjun Venkatesan, avenka21@asu.edu, Rolf U. Halden. Center for Environmental Security, Biodesign Institute, Arizona State University, Tempe, Arizona, United States

Of the 6.5 million metric tonnes of municipal sewage sludge (MSS) or biosolids produced annually in the U.S., >50% are applied on land as a soil amendment. An increasing concern regarding this practice is the presence in MSS of chemical and biological agents posing environmental and human health concerns. Hazardous agents leachable from MSS have the potential to pollute surface and groundwater or be subject to uptake by feed and food crops. A meta-analysis of contaminants detected in nationally representative samples showed the presence of 312 organic contaminants in U.S. MSS, of which 23% (n = 71) were identified as ionizable organic contaminants (IOCs) contributing 82% of the established total contaminant mass loading. Among these were 58 pharmaceuticals and personal care products (PPCPs), 12 perfluoroalkyl substances (PFCs) and the surfactant 4-nonylphenol (NP). The load of these IOCs released to U.S. soils via land-application of MSS was calculated to equal approximately 2,000 tonnes. A partitioning model developed to assess the role of pH in the potential mobility of contaminants revealed that between 36 and 85% of the mass of individual classes of IOCs potentially could desorb from land-applied MSS due to ionization forced by pH changes in the environment. Model output further suggested that up to 42 metric tonnes per annum of IOCs potentially could be mobilized from land-applied MSS. This model may serve to identify the fraction of leachable contaminants in MSS at different pH regimes and to estimate the potential contamination of surface/ground water near MSS-applied agricultural fields.

AGFD 236

Alkaloid profiles of hairy root cultures of Catharanthus roseus differ when generated by different strains of Agrobacterium rhizogenes

John de la Parra1, delaparra.j@husky.neu.edu, Noreen Rizvi2, Roger A. Kautz1, Poguang Wang3, Roger Giese3, Carolyn W. Lee-Parsons1,2. (1) Chemistry and Chemical Biology, Northeastern University, Cambridge, Massachusetts, United States (2) Chemical Engineering, Northeastern University, Boston, Massachusetts, United States (3) Pharmaceutical Sciences, Northeastern University, Boston, Massachusetts, United States

The Madagascar periwinkle plant (Catharanthus roseus) produces the valuable anti-cancer compounds vincristine and vinblastine at very low yields. One approach to increasing the production of these unique alkaloids is the genetic engineering of the plant's biosynthetic pathway using the plant pathogen, Agrobacterium rhizogenes (the related bacteria, Agrobacterium tumefaciens, causes the crown gall tumor in plants). The alkaloid profile differs between C. roseus hairy root cultures generated by different strains of A. rhizogenes. Their alkaloid profiles, including a previously uncharacterized metabolite, are compared and characterized by MS/MS and NMR.

AGFD 237

Ensuring coffee freshness in portioned coffee system

Luigi Poisson1, Sylvie Legrand1, Yves Wyser1, Frederic Mestdagh1, Frederic.Mestdagh@rdor.nestle.com, Britta Folmer2, Josef Kerler1. (1) Nestle, Orbe, Switzerland (2) Nespresso, Lausanne, Switzerland

Many portioned coffee systems are offered today to consumers. Main advantages of single sealed coffee portions for consumers are the convenience of preparation, and the variety and choice. During storage of roast and ground coffee (R&G) various degenerative alterations can take place such as oxidation and/or intrinsic reactions of aroma compounds leading to a fading of the fresh coffee aroma and/or the development of off-notes[1]. In order to ensure that the consumer can enjoy the desirable fresh coffee aroma during the whole shelf life the coffee must be stored in absence of oxygen and light. Various packaging materials are used to pack portioned coffee, ranging from aluminium to various plastics. In some cases, the barrier is provided by a secondary pouch made of high barrier laminate. A recent study on the role of the packaging material on the shelf-life of roasted coffee showed that aluminium preserves best the freshness[2]. The aim of present study was to link the alteration in the aroma composition to the perceived sensory impact from a consumer perspective. Roasted coffee was stored under different conditions and using different packing material (whole beans, R&G and portioned) during one year shelf life. The quantitative evaluation of freshness and oxidation markers implied the use of stable isotope labelled molecules as internal standards, and analysis by solid phase micro extraction (SPME) combined with gas chromatography/mass spectrometry (GC/MS). Sensory analysis was performed as a Two-Alternative Forced Choice test (2-AFC) to detect changes in coffee quality, such as loss of aroma intensity and the appearance of off-notes. A causal link was found between the alteration of evaluated aroma compounds and the point in time where a change in the sensory profile was significantly perceived. In addition, a modelling study was conducted to assess the impact of oxygen presence during processing of the coffee, the amount of oxygen consumed by the coffee during its shelf life and packaging permeability on the residual oxygen and on freshness loss. It is concluded that processing coffee in the absence of oxygen is key for highest initial coffee quality and that storage in hermetically sealed capsules with very high barrier properties allow to retain freshness over the entire shelf life. 1 Poisson et al. (2006). Proceedings of 21st ASIC conference. 2 Glöss et al. (2014). Chimia, 68, Nr. 3.

AGFD 238

Red shortening: Characterization and utilization in formulating novel functional biscuits

Hany Abou Gharbia, samehmawad@gmail.com. Food Science, Alexandria University, Alexandria, Egypt

Functional biscuits were manufactured by replacing white shortening (WSh) with red shortening (RSh) by about 80%. White shortening was found to be comparable to RSh in terms of moisture content, refractive index, iodine value, saponification value and unsaponifiable matter. On the other hand, WSh was different from RSh in cloud point, color, slip melting point, acid and anisidine values as well as free fatty acids. The fatty acid composition of WSh and RSh was different. The total saturated fatty acids in WSh and RSh was 57.4 and 50.8%, respectively, whereas, the total unsaturated fatty acids for WSh and RSh amounted for 42.4 and 49.0%, respectively. It was obvious that RSh possessed higher content of antioxidants than WSh, the total α-tocopherol and tocotrienols were 654 and 530 ppm, respectively. Moreover RSh contains 465 ppm of carotenes as compared to only 2 ppm present in WSh. In conclusion, biscuit samples formulated in the present study could be stored up to 10 months at room temperature without any significant deterioration in terms of oxidative stability, peroxide ansidine and acid values.

AGFD 239

Analysis of lipid transfer proteins in Arabidopsis thaliana by means of epitope tags to decipher the role of LTP4's lipid in plant senescence

Johanna Bautista, joba1025@gmail.com. Biochemistry, California State University, Los Angeles, Los Angeles, California, United States

Through microarray assays Dr. Vellanoweth's lab found two specific lipid transfer proteins that are upregulated during the bolting stage in Arabidopsis thaliana. RNAi knock down of these genes produce a mutant plant that continues to flower after having already completed senescence and apparent death. We hypothesize that LTP4 carries a lipid signal that mediates meristem death. The objective of this work is to insert an epitope tagged lipid transfer protein 4 (LTP4) gene into the Arabidopsis thaliana plant which will allow us to use a reagent to specifically pull out this lipid carrying protein for analysis. We expect that this approach will help us uncover the identity of the LTP4-bound lipid. Also, using the epitope tags on the LTP4 protein, we will track the process in which the LTP4 protein carries the unknown lipid through the plant in senescence. Currently, the entry clone with the LTP4 gene of interest has been constructed. Through the lytic reaction of the entry clone and various destination vectors we expect to obtain our expression vectors. The expression vectors will then be used to transform AGL1 Agrobacterium tumefaciens that will introduce the genes into the plant via the floral dip method. This will give us our final transgenic plants that will have an epitope tagged LTP4 gene.

AGFD 240

Identifying T-DNA insertion site in Arabidopsis thaliana LTP-4 and LTP-3 mutants by thermal asymmetric interlaced PCR (TAIL-PCR)

Israel Santana, santanaisrael83@gmail.com, Jessica Ortiz, robert l. vellanoweth. chemistry and biochemistry, california state university los angeles, Los Angeles, California, United States

Significant phenotypic differences such as extended life span, multiple floral meristems and greater vegetative growth have been observed in transgenic LTP-4 RNA-i Arabidopsis thaliana plants. We attempt to confirm these phenotypes are a result of short interfering RNA-i (siRNA) knockdown and not the result of the transgenic DNA (T-DNA) insertion disrupting normal gene function. Thermal asymmetric interlaced PCR (TAIL-PCR) will be used to identify the location of the transgenic DNA insert within the LTP 4 and LTP3 Arabidopsis genomic lines. We aim to amplify a sequence that contains both known vector sequence and adjacent unknown genomic sequence. TAIL-PCR utilizes three specific forward primers (T1, T2, and T3) in consecutive reactions together with short arbitrary degenerate (AD) primers. The primers are designed to have varying annealing temperatures so that the alternating high and low temperature cycles yield higher concentrations of the specific product over nonspecific products. Preliminary results show products amplified by TAIL-PCR for LTP-3 are still within vector sequence and do not amplify the adjacent genomic region. LTP-4 T-DNA insert was found inserted within a Leucine rich repeat-containing protein gene nested in chromosome 5.

AGFD 241

Capillary electrophoresis coupled with inductively-coupled plasma mass spectrometry as an analytical tool for arsenic speciation in rice

Haiou Qu, haiou.qu@fda.hhs.gov, Thilak Mudalige, Sean Linder. Office of Regulatory Affairs/Arkansas Regional Lab, U.S. Food And Drug Administration, Little Rock, Arkansas, United States

In recent years there is an increasing concern about the present of arsenic as a contaminant in foods. Because of unique farming practices, rice has the potential to accumulate more arsenic from the soil when compared to other crops. Dimethylarsinic acid (DMA), monomethylarsonic acid (MMA), arsenite [As(III)], and arsenate [As(V)] are four arsenic species that are commonly found in rice and rice products. Due to species dependent toxicity, it is necessary to quantify the concentration of each species in order to accurately determine the risk associated with eating rice and rice products. Currently, liquid chromatography coupled with inductively coupled plasma mass spectrometry

(ICPMS) been demonstrated to be a feasible method for arsenic speciation in rice. A common problem of this method is that the column can deteriorate after many injections, which compromises subsequent measurements. Here we have developed an alternative methodology using capillary electrophoresis, coupled with ICPMS to separate and quantify common arsenic species in rice and rice cereal. An additional arsenic containing compound, o-arsanilic acid, which is not inherently present in natural sample, was added as an internal standard to adjust for run-to-run variation generated during the injection step. After an enzyme-assisted microwave extraction, DMA, MMA, As (III) and As (V) in rice and rice cereal were successfully separated under basic buffer condition (pH=11) and a applied voltage of 20 kV. The peak area was found to have a linear relationship with the concentration of each arsenic species. Two rice standard reference materials (SRM 1568 b and CRM 7503-a) were analyzed to validate the method, and the determined concentration of DMA, MMA and inorganic arsenic were within 5% difference of the certified values of the reference materials.

AGFD 242

Prevention and treatment of Staphylococcus aureus biofilm formation using Russian Propolis ethanol extracts

Julia Bryan, jbryan@mail.saintpeters.edu, christian Traba, Michael J. Castaldi. Chemistry, St. Peters University, Lyndhurst, New Jersey, United States

In this study, the exposure of three Russian Propolis ethanol extracts from different regions in Russia were used in: 1) the prevention and 2) treatment of Staphylococcus aureus biofilms on various surfaces. It was found that despite similar activities against biofilm prevention (planktonic bacteria), drastic differences were observed in biofilm treatments. The antibacterial and anti-biofilm results indicated that: 1) antibacterial and 2) anti-biofilm activity of Russian propolis ethanol extracts may be governed by different types of chemical compounds found in the propolis ethanol extracts. Further analysis indicated that the mechanism of action of propolis ethanol extracts appear to be complex and involves the degradation of the extracellular polymeric matrix, exposing the bacteria within the biofilm to the antibacterial agents found in the propolis extracts. Kinetic studies conducted under optimal experimental conditions, revealed that bacterial cells in Staphylococcus aureus biofilms were killed (>99.9%) by propolis after 12 hours of treatment. The sterilization ability of propolis for the prevention and treatment of biofilm related contaminations on various materials were confirmed and information gathered from this study provides insights into the anti-biofilm mechanism of propolis.

AGFD 243

Orange juice reduces oxidative stress and inflammatory markers in patients with chronic hepatitis C

Danielle R. Gonçalves1, danirgoncalves.nutri@gmail.com, Cláudia G. Lima1, Paula S. Ferreira1, Paulo I. Costa2, Thais B. Cesar1. (1) Food and Nutrition, Sao Paulo State

University, Araraquara, Brazil (2) Clinical Analysis, Sao Paulo State University, Araraquara, Sao Paulo, Brazil

Chronic infection by HCV causes hepatic inflammation and alterations in liver metabolism, resulting in biochemical and oxidative stress disorders. Supplementation with antioxidants might minimize the deleterious effects of the disease and contribute to liver protection. Orange juice includes nutrients and bioactive compounds which exhibit antioxidant and anti-inflammatory properties. The present study assessed whether orange juice may improve the inflammation markers in patients with chronic hepatitis C. Liver enzymes such as ALT, AST, FAL and γ-GT; oxidative stress (ABTS, TBARS) and CRP were measured before and after eight consecutive weeks of supplementation with 500 mL of 100% commercial pasteurized orange juice (Pera Rio variety) in 66 chronic hepatitis C carriers (orange juice group, n = 46 and control group, n = 20), who were being treated with antiviral therapy. Following regular intake of orange juice as a dietary supplement, CRP decreased in the patients’ juice group; furthermore, oxidative stress was reduced and the levels of the liver enzyme AST decreased in those patients who had high levels before the intervention. We suggest that orange juice intake should be encouraged in patients with chronic hepatitis, once the orange juice has minimized some of the metabolic alterations caused by HCV, possibly due to its bioactive components. This recommendation is based on the positive effects of this food on inflammation markers and oxidative stress.

AGFD 244

Characterization of constituents from cranberry non-dialyzable material that inhibit bacterial co-aggregation and adhesion

Kelsey Penndorf1, kpenndorf@umassd.edu, Catherine C. Neto1, cneto@umassd.edu, Mark Feldman2, Shiri Meron-Sudai4, Zichria Rones2, Doron Steinberg2, Micha Fridman3, Isaac Ginsburg2, Itzhak Ofek4, Ervin Weiss5. (1) Chemistry and Biochemistry, University of Massachusetts Dartmouth, North Dartmouth, Massachusetts, United States (2) Institute of Dental Sciences, Hebrew University-Hadassah, Jerusalem, Israel (3) Schl of Chemistry, Tel Aviv University, Tel Aviv, Israel (4) Clinical Microbiology and Immunology, Tel Aviv University, Tel Aviv, Israel (5) Golschlager School of Dental Medicine, Tel Aviv University, Tel Aviv, Israel

Previous studies have shown that a high molecular weight nondialyzable material (NDM) from cranberry juice concentrate exhibits potent anti-adhesion activity of a number of microbial species including oral bacteria, uropathogenic Escherichia coli, Helicobacter pylori, and influenza virus. However, the compounds responsible for such activity were not identified. In the present study, fractionation of cranberry NDM on Sephadex LH-20 yielded two major fractions: a 75% acetone soluble fraction (NDMac) that inhibited bacterial co-aggregation and exhibited binding affinity to bacterial surfaces, and a 50% ethanol soluble fraction (NDMet) that showed little activity in these assays. NDMac inhibited co-aggregation of oral bacteria Streptococcus sanguinis with Fusobacterium nucleatum and Fusobacterium nucleatum with Porphyrymonas gingivalis

at MIC of 0.47 mg/mL and 0.94 mg/mL respectively. MALDI-TOF MS analysis of the NDMac fraction showed ion m/z consistent with the presence of proanthocyanidins (PACs), primarily of 3-6 degrees of polymerization with an A-type linkage, and composed mostly of (epi)catechin units, with some (epi)gallocatechin and anthocyanin units also present. Ions detected in the NDMet fraction had m/z values consistent with xyloglucans rather than PACs, and its 1H NMR spectrum showed higher signal intensities in the polysaccharide region. Further fractionation of NDMac on MCI CHP-20P with water/methanol elution revealed that the anti-adhesion activity determined by coaggregation of oral bacteria resides in less polar fractions, eluting with 70% or higher methanol. Characterization of the active constituents will enhance our understanding of the full potential of cranberry NDM as an anti-adhesion agent.

AGFD 245

Inhibition of colon cancer growth and inflammation in cellular and mouse models by cranberry extracts (Vaccinium macrocarpon)

Sarah Frade1, sfrade@umassd.edu, Anne Liberty1, Anuradha Tata1, Xian Wu2, Mingyue Song2, Xiaokun Cai2, Hang Xiao2, Catherine C. Neto1. (1) Chemistry and Biochemistry, Cranberry Health Research Center, University of Massachusetts Dartmouth, North Dartmouth, Massachusetts, United States (2) Food Science, University of Massachusetts Amherst, Amherst, Massachusetts, United States

The ability of cranberry fruit extracts to inhibit colon carcinogenesis is under investigation using a combination of in vitro and in vivo approaches. Extracts and compounds from cranberry fruit (Vaccinium macrocarpon) were observed to decrease the proliferation of HCT116 and HT-29 colon tumor cells, while growth of CCD-18 normal colon cells was impacted to a lesser extent. A cranberry feeding study was conducted using AOM/DSS mice, a model of colitis-associated colon carcinogenesis. Mice were treated with azoxymethane and dextran sodium sulfate to induce inflammation-driven colon carcinogenesis, while receiving AIN-93 diet containing whole cranberry powder (WCP), a polyphenol-rich polar extract, a lipid-rich nonpolar extract, or no cranberry. After 20 weeks on WCP diet, the number of tumors and tumor volume per mouse were significantly decreased, by more than 50% compared to control, accompanied by a reduction in tissue markers of inflammation IL-1 and IL-6. Significant reductions in tumors and inflammation were also observed with the polar and nonpolar extracts. The identities of cranberry constituents and metabolites responsible is under investigation. The effects of the cranberry extracts and some of their major constituents, including proanthocyanidins and ursolic acid, on expression of IL-6 were determined in the colon cell lines using ELISA. Metabolic profiling using HPLC-DAD and LC-MS is also underway to determine changes in the composition of mouse colonic metabolites resulting from cranberry diets. These studies demonstrate potential for dietary cranberry to inhibit colon carcinogenesis through decreased inflammation.

AGFD 246

NMR-based metabolomic analysis and quantification of phytochemical constituents in North American cranberry fruit (Vaccinium macrocarpon)

Andrew Milstead1, amilstead@umassd.edu, Liang Xue1, lxue1@umassd.edu, Kimberly L. Colson2, Catherine C. Neto1. (1) Chemistry and Biochemistry, Cranberry Health Research Center, University of Massachusetts Dartmouth, North Dartmouth, Massachusetts, United States (2) Bruker BioSpin Corporation, Billerica, Massachusetts, United States

Cranberries have been a focus of natural products research for the past two decades, as cranberry extracts have many reported bioactivities including reduction of bacterial adhesion, inflammation and tumor growth. The compounds responsible for these activities are under investigation. Content of phytochemicals in the fruit may vary based on cultivar, season and growing region. Targeted and non-targeted NMR methods were used to determine variation in constituents, including those that are difficult to quantify by other methods. Cranberry fruit of eight different cultivars was harvested in Massachusetts, New Jersey, Oregon, and Wisconsin in 2011, and extracted in ethanol at room temperature to yield crude mixtures containing a broad range of constituents. Analysis was carried out using 400 MHz 1D 1H NOESY spectra and Assure-RMS software (Bruker Biospin). Methods were developed to quantify ursolic acid, oleanolic acid, malic acid, and benzoic acid; these averaged 18.7, 15.0, 127.9, and 1.6 mg/g fruit extract, respectively. Methods for hyperoside and procyanidin A2 are also being developed. Ursolic acid, an anti-inflammatory and antiproliferative constituent, ranged from 9.3-31.8 mg/g extract, and one-way ANOVA found WI samples higher in ursolic acid than MA or OR samples. Principal component analysis (PCA) of NMR data provided non-targeted analysis of sample-to-sample variation, revealing differences between cultivars and regions. Variance between Demoranville and Early Black cultivars evident in the PCA scores plot was associated with a bucket in the loadings plot at 1.08 ppm, attributable to ursolic acid. ANOVA confirmed that ursolic acid content differed significantly between these cultivars. NMR methods can provide efficient, repeatable and environmentally friendly analyses of phytochemical extracts.

AGFD 247

Dietary exposure of nonphthalate-based plasticizers from use in food contact material

LaShonda T. Cureton, clashon@vt.edu, Allan B. Bailey. US Food and Drug Administration, College Park, Maryland, United States

Plasticizers have long been added in the industrial manufacture of polymers to facilitate processing and increase flexibility and toughness by acting as an “internal lubricant” between the polymer chains. Plasticizers help balance performance and costs—materials can be processed using a variety of techniques at a lower cost with mass versatility in the global market segment. China is the single-largest plasticizer market in the world, accounting for nearly 38% of world consumption in 2012. The US

consumption of plasticizers was approximately 0.8 million metric tons, of which approximately 60% were phthalates, in 2012. About 80-90% of the plasticizer market is dedicated to polyvinyl chloride (PVC), which is a highly versatile thermoplastic that can be transformed into both rigid and flexible products having a combination of engineering properties. While materials produced from rigid PVC do not contain any plasticizer, articles manufactured from flexible PVC are traditionally compounded with plasticizers to achieve ductility, while maintaining strength in construction. The most commonly used plasticizers are phthalates, accounting for over 78% of the world consumption of plasticizers in 2012. Given the significant use of flexible PVC in many commodity products, the FDA has an interest in determining the dietary exposure and toxicological effects of phthalate-based plasticizers and potential “drop in” substitutes, like epoxidized soybean oil (ESBO), adipates and terephthalates. This technical review summarizes the identity, regulatory status and studies on levels in food and food simulants, and previous estimates of cumulative exposure, as available, to a specific class of non-phthalate plasticizers, namely adipate-based and ESBO-based plasticizers. Review of the literature and agency files shows that di-2-ethylhexyl adipate (DEHA) is the most widely used adipate-based plasticizer and the cumulative dietary concentration (CDC) is currently 2400 ppb (primarily from PVC cling film). ESBO is typically used in flexible PVC for film wrap and container gaskets and literature results from a host of migration and food concentration studies indicate that migration of ESBO into fatty foods in highest. Comparatively, both plasticizer types result in higher migration with higher plasticizer levels, a larger food contact surface area, at higher temperature during food contact and with the amount time the food contact material has been in contact with food.

AGFD 248

Preparation of metal chelating active food packaging materials by laminated photografting

Jason Z. Lin3, linzhuangsheng@gmail.com, Maxine Roman3, Fang Tian3, Eric A. Decker1, Julie M. Goddard2. (1) Univ of Massachusetts, Amherst, Massachusetts, United States (2) Food Science, University of Massachusetts Amherst, Amherst, Massachusetts, United States (3) Food Science, University of Massachusetts-Amherst, Amherst, Massachusetts, United States

Transitional metals, especially iron and copper, can induce lipid oxidation in bulk and emulsified oil systems such as mayonnaise and salad dressings; the synthetic metal chelator EDTA is, therefore, commonly added as an antioxidant. To address the increasing consumer demand for ‘clean label’ food products, non-migratory metal chelating active packaging materials were developed to control transition metal induced lipid oxidation. Metal chelating polypropylene films were made by photografting of acrylate monomers to produce materials with surface immobilized metal chelating ligands of carboxylic acids (PP-g-PAA) and hydroxamic acids (PP-g-PHA). Both PP-g-

PAA and PP-g-PHA chelating films inhibited lipid oxidation in oil-in-water emulsion by delaying the formation of lipid hydroperoxides and hexanal. Previously, the photografting of the metal chelating films were conducted in oxygen-free environment with nitrogen inerting, which hinders possible industrial scale-up. In this project, a laminated photografting technique was developed to enable atmospheric photografting (ie: eliminate the nitrogen inerting step). The PP-g-PAA film was made by photografting a laminated layer of acrylic acid and benzophenone (BP) mixture underneath an UV-ozone functionalized polypropylene film. The PP-g-PHA film was made by photografting a laminated layer of methyl acrylate and BP mixture, followed by a post graft modification to add hydroxamic acids. Surface modification was confirmed using ATR-FTIR, SEM and contact angle analysis. The PP-g-PAA film had ferric ion chelating activity of 182 ± 29 nmol/cm2 and cupric ion chelating activity of 84 ± 4 nmol/cm2 at pH 5. The PP-g-PHA film had ferric ion chelating activity of 89 ± 10 nmol/cm2 and cupric ion chelating activity of 262 ± 16 nmol/cm2. The reported laminated photografting technique may be adapted for roll-to-roll manufacture of non-migratory metal chelating active packaging materials on an industrial scale that may enable removal of EDTA from food and consumer products.

AGFD 249

Rapid detection of Salmonella using a redox cycling-based electrochemical method

Danhui Wang2, danhuiwang@foodsci.umass.edu, Amanda Kinchla2, Sam R. Nugen1. (1) 246 Chenoweth Laboratory, University of Massachusetts, Amherst, Massachusetts, United States (2) Food Science, University of Massachusetts Amherst, Amherst, Massachusetts, United States

An electrochemical method based on redox cycling combined with immunomagnetic separation and preconcentration was developed for the rapid and sensitive detection of Salmonella. Both approaches were used to provide a rapid detection scheme with a low limit of detection. Magnetic beads modified with anti-Salmonella antibodies were used for separation and preconcentration of Salmonella from phosphate buffered saline (PBS) and agricultural water. Then anti-Salmonella antibodies conjugated with alkaline phosphatase (ALP) were used for labelling the Salmonella captured by magnetic beads. Alkaline phosphatase catalyzed the substrate L-ascorbic acid 2-phosphate (AAP) to electroactive species L-ascorbic acid (AA) while tris(2-carboxyethyl)phosphine (TCEP) facilitated the regeneration of AA on the gold electrode to form redox cycling resulting in a continued and amplified signal. Under the optimal conditions, the Salmonella in PBS buffer as well as agricultural water from two different sources were detected. The limit of detection of this approach was approximately 7.6 × 102 CFU/mL and 6.0 × 102 CFU/mL in PBS buffer and agricultural water, respectively, without preenrichment. The entire assay time was less than three hours. If the agricultural water was pre-enriched for four hours, the limit of detection was approximately 10 CFU/mL.

Schematic representation of the immunomagnetic preconcentration and electrochemical detection based on redox cycling. The detection procedure contained three main steps. The first step was immunomagnetic separation and preconcentration of Salmonella from sample matrix. The second step was immunological reaction with anti-Salmonella antibodies conjugated with ALP. The third step was enzyme reaction and electrochemical detection.

AGFD 250

Metal oxide gas sensor array combined with a miniaturized gas chromatographic system for fast detection of volatile quality indicators

Matthias Kotthoff1, matthias.kotthoff@ime.fraunhofer.de, Mark Bücking1, mark.bücking@ime.fraunhofer.de, Jörg Bruckert1, Marie-Luise Bauersfeld2, Jürgen Wöllenstein2. (1) Environmental and Food Analysis, Fraunhofer Institute for Molecular Biology and Applied Ecology, IME, Schmallenberg, Germany (2) Fraunhofer Institute for Physical Measurement Techniques IPM, Freiburg, Germany

Food production involves complex processes that need to be controlled to ensure food quality. This involves time-consuming and expensive steps and generates financial burdens. So-called on-line methods, integrated into the production process, save time and money, but are restricted to measuring the simplest chemical / physical parameters. More complex systems are under development, but are not reliable enough for commercial application. In cooperation with the Fraunhofer IPM a robust and cost effective gas sensor array for rapid detection of volatile compounds was developed. The gas sensor array is intended for application in various areas of food industry, also directly on the farm-side (i.e. for coffee, cocoa, tea). We used commercially available, inexpensive metal oxide sensors, which are already used e.g. in fire detectors and combined them into a system that follows gas chromatographic principles, i.e. this system uses a chromatographic separation column which splits the gas mixture and feeds the sensors with the individual substances [1]. Working scheme: First, selected food samples undergo a gas chromatographic evaluation. E.g. green coffee is an interesting candidate, because of the limited number of volatile compounds, which is of advantage for a fast detection approach. After this GC/MS analysis, characteristic standard substances are tested with the portable low-cost gas chromatographic measurement system including the gas sensor array. In a third step the final industrial application is developed. First results showed that the reproducibility of the sensor approach was comparable to the reference analysis, but the sensitivity was up to ten-fold higher.

AGFD 251

Metabolomics application for rapid screening and authentication of Asian palm civet coffee (Kopi Luwak)

Udi Jumhawan1, udi_jumhawan@bio.eng.osaka-u.ac.jp, Sastia Prama Putri1, Yusianto Yusianto2, Takeshi Bamba1, Ei-Ichiro Fukusaki1. (1) Department of Biotechnology, Osaka University, Suita Osaka, Japan (2) Indonesian Coffee and Cocoa Research Institute, Jember, East Java, Indonesia

In our previous report, selection of discriminant markers for authenticity evaluation of Asian palm civet coffee, world’s priciest ‘premium’ coffee, was done through application of metabolite profiling employing gas chromatography/mass spectrometry (GC/MS). However, relatively expensive instrument and maintenance has become one of the major obstacles for pilot application in industries. Furthermore, compound identification, an essential step in metabolite profiling approach, has been infamously considered as tedious and challenging. Therefore, there is a need to develop rapid and reliable method for quality assessment of civet coffee. An alternative analysis utilizing gas chromatography coupled with a universal and cost-effective detector, flame ion detector (GC/FID), via metabolite fingerprinting has been established for screening of 37 commercial and non-commercial coffee beans. A similar chromatographic profile with higher sensitivity of reported discriminant markers from GC/MS analysis and important coffee metabolites, such as citric acid malic acid, had been obtained by using GC/FID. Principal component analysis (PCA) and orthogonal projection to latent structure-discriminant analysis (OPLS-DA) were then applied for the GC/FID data. The predictive model exhibited good fit and reproducibility in comparison to the GC/MS data, for differentiation of civet coffee, regular coffee and coffee blend without characterization of metabolite identities [figure 1 and 2]. Taken together, the results suggested that the GC/FID system offered rapid coffee authenticity screening and could be beneficial for industrial routine.

[figure 1] OPLS-DA models of GC/FID and GC/MS data

[figure 2] PCA score plot of GC/FID

AGFD 252

Designing the rolling circle amplification based surface-enhanced Raman spectroscopy method for 35S promoter maize gene detection

BURCU GUVEN1, burcuguven@hacettepe.edu.tr, Ismail H. Boyaci2,3, Ugur Tamer4, Esra Acar Soykut3. (1) HACETTEPE UNIVERSITY, Ankara, Turkey (2) Faculty of Engineering, Department of Food Engineering, Hacettepe University, Ankara, Turkey (3) Food Research Center, Hacettepe University, Ankara, Turkey (4) Faculty of Pharmacy, Department of Analytical Chemistry, Gazi University, Ankara, Turkey

In this study, we developed the genetically modified organism detection method by using the combination of rolling circle amplification (RCA) and surface-enhanced Raman spectroscopy (SERS) and compared the developed RCA-SERS assay with traditional-SERS assay. As part of the study, gold surfaces and gold nanorods were used to form sandwich structure. Gold nanorods were labeled with 5,5’-Ditiobis(2- Nitrobenzoik asit) (DTNB) to enhance SERS intensity. Quantification of the target concentration was performed via SERS spectra of DTNB on the nanorods. SERS spectra of target molecules were enhanced through the RCA reaction and the detection limit was found to be 6.3 fM. The sensitivity of developed RCA-SERS assay was compared with traditional SERS assay and the detection limit was found to be 0.1 pM. For the developed RCA-SERS assay, the specificity tests were performed using the 35S promoter of Bt-176 maize gene and nonsense sequence. It was found out that the developed RCA-SERS sandwich assay method is quite sensitive, selective and specific for target sequences in model and real systems.

AGFD 253

Modeling the human colon: An automated multistage fermentation approach

Laurel A. Doherty1, laurel.a.doherty.civ@mail.mil, Steven Arcidiacono1, Kenneth Racicot1,2, Jason W. Soares1. (1) U.S. Army Natick Soldier Research Development & Engineering Center, Natick, Massachusetts, United States (2) Tufts University, Medford, Massachusetts, United States

Growing evidence shows the importance of the gut microbiome to human health; however, characterization of the effects of gut bacteria is constrained by experimental limitations inherent in in vivo studies. In vitro fermentation models afford comparatively greater flexibility, facilitating broader and more varied investigation of native gut community dynamics such as metabolism and function. Several in vitro gut models have been developed with varying degrees of control and accuracy and are often the product of ad hoc construction. Here, we present our efforts toward a fully automated, event-driven, multiple-stage gut model using a DASGIP parallel bioreactor station to provide a physiologically relevant system mimicking domain-dependent metabolic behavior of the gut microbiome. Initial efforts focused on establishment and validation of single-stage fermentations mimicking the distal colon. Inocula were prepared from fresh fecal donations of three individuals within 2 hrs of collection by processing in reduced phosphate buffer supplemented with glycerol and immediately storing at -20oC. Automated batch fermentations were performed utilizing 2% (w/v) pooled fecal inocula, to normalize individuals’ microbiota, in a nutrient-rich anaerobic media at 37oC under controlled oxygen-free atmosphere. Fed-batch fermentations under distal colon conditions were also performed under various rates of nutrient delivery; results indicated enhanced biomass production and elongated growth phase compared to batch distal cultures. Characterization of fermentation byproducts, as well as the influence of dietary inputs on gut microbiota population and function, have been conducted. More recent efforts encompassing continuous fermentation of the three domains of the colon, separately and in sequence, as well as equilibration studies to establish a stable

microbial community, are ongoing. Use of the DASGIP parallel bioreactor platform to develop a multiple-stage human gut model will facilitate investigation of the effect of Army-relevant stressors on gut health, and insight gleaned from the model will inform nutritional strategies to restore Warfighter gut homeostasis.

AGFD 254

Antioxidant activity and inhibition of amylase by Washington navel oranges

Swathi Kommein, kommeins@yahoo.com, Bhimu Patil. Vegetable and Fruit Improvement Center, Horticultural Sciences , Texas A&M University , College Station, Texas, United States

Free radicals are formed naturally in our body and alter many normal cellular processes including all major components such as DNA, proteins, lipids, and cell membranes. The alteration of cellular process may play a role in the development of cancer and terminal illnesses. This can be prevented by the consumption of natural antioxidants such as b-carotene, lycopene, vitamins A, C and E from fruits and vegetables. It is well known that citrus fruits are rich in vitamin C, carotenoids and other phenolic compounds. Among the citrus fruits, Washington Navel oranges are one of the sweetest seedless oranges present in supermarkets. In the present study, antioxidant activity and amylase inhibition was investigated for Washington Navel oranges. Lyophilized Washington Navel fruit samples were extracted sequentially with hexane, chloroform, acetone, methanol and methanol:water (8:2) using a Soxhlet apparatus for 8 hours each separately. All extracts were concentrated, freeze dried and used for Invitro bioassays. The total phenolic content of the extracts was determined by Folin–Ciocalteu method and antioxidant activity of the extracts was assayed using in vitro models such as antioxidant capacity by radical scavenging activity using α,α-diphenyl-β-picrylhydrazyl (DPPH), 2,2'-azino-bis 3-ethylbenzothiazoline-6-sulphonic acid (ABTS), and phosphomolybdenum method. The total phenolic contents of the fruit extracts were expressed as catechin equivalents. Antioxidant capacity of five extracts were expressed as equivalent to ascorbic acid µmol/g of extract, which was in the order of chloroform> methanol: water> acetone > methanol > hexane, by phosphomolybdenum method. Recent studies have shown that plant-based foods containing high total polyphenolic compounds and flavonoids were linked to α-amylase inhibitory activities. All tested five extracts showed very moderate to good amylase inhibition activity. The results indicated that, the Washington Navel extracts found to possess very good antioxidant and amylase inhibition activity, thus these fruits can be further explored for understanding the health promoting properties. This research is based upon work supported by USDA-NIFA No.2010-34402-20875, “Designing Foods for Health,” through the Vegetable & Fruit Improvement Center.

AGFD 255

Determination of microbial volatile organic compounds patterns from virulent and hypovirulent Cryphonectria parasitica isolates by headspace-SPME-GC-MS

Jinyan She1, shejinyan@hotmail.com, Mary King1, Beth Stokes3, Yi Jiang4, Richard Baird3, Todd E. Mlsna2. (1) Chemistry, Mississippi State University, Mississippi State, Mississippi, United States (3) Department of Biochemistry, Molecular Biology, Entomology and Plant Pathology, Mississippi State University, Mississippi State, Mississippi, United States (4) Civil and Environmental Engineering Department, Mississippi State University, Mississippi State, Mississippi, United States

American chestnut tree comprised over 25% of Appalachian Mountain forests plays an important role in local ecological system. It was virtually eliminated by chestnut blights in 1940s. Natural control involves usage of debilitated virus-infected strains of pathogen hypo-virulent (HV) isolates to convert the lethal virulent (V) forms into HV’s. Traditional determination method of HV strains is labor intensive and time consuming. Studies of volatile organic compounds (VOCs) metabolites from fungi isolates using Solid Phase Micro Extraction (SPME) have been accomplished by many research institutions. However, no study has been reported on Microbial Volatile Organic Compounds (MVOCs) produced by virulent and hypo-virulent Cryphonectria parasitica isolates. Herein, we report the determination of MVOCs patterns by headspace solid phase microextraction (HS-SPME) with an 85 µm Carboxen/Polydimethylsiloxane (CAR/PDMS) fiber coupled with gas chromatography- mass spectrometry (GC-MS). From the study, the volatile organic compounds patterns of five virulent C.parasitica isolates and three hypo-virulent C.parasitica isolates were determined. Future works will focus on multivariate analysis for data interpretation, model validation for MVOCs pattern recognition, and pattern determination of Cryphonectria hypovirus (CHV) isolates.

AGFD 256

Kinetic stability of bean and pea proteins: Effect on protein digestibility and bean resistance to environmental conditions

Ke Xia1, ke.xia@hotmail.com, Jennifer Wilcox2, Sandy Pittelli2, Wilfredo Colon2,1. (1) Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York, United States (2) Dept of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, New York, United States

Kinetically stable proteins (KSPs) are hyperstable, and therefore, are highly resistant to degradation. KSPs appear to arise in nature as a mechanism for organismal adaptation and survival against harsh conditions, and are often found in extremophiles. We have demonstrated a correlation between kinetic stability and a protein’s resistance to the denaturing detergent SDS, and developed a powerful diagonal two-dimensional (D2D) SDS-PAGE assay that in combination with proteomics analysis allows the identification of KSPs in different cells or organisms. We have applied this method to a group of 12 legumes that are important to the agricultural and food industries, and identified the KSPs present in each legume. Our results show that legumes with higher content of KSPs are more difficult to digest whereas legumes with low amounts of KSPs are highly digestible. In addition, we found that several proteins linked to resistance against biotic

stress are kinetically stable, including all three major bio-defensive lectins. Overall, our results have implications for our fundamental understanding of the role of KSPs in legume adaptation to weather and biotic stress. Furthermore, this study has implications for understanding the nutritional efficiency of legumes (e.g. food intolerance), and the role of KSPs in legume bio-defense, with potential applications for enhancing legume nutrition and the development of green non-toxic pest control strategies.

AGFD 257

Antioxidative compounds from Garcinia buchananii stem bark

Mathias Salger, mathias.salger@tum.de, Timo D. Stark, Junichiro Wakamatsu, Thomas Hofmann. Chair of Food Chemistry and Molecular Sensory Science, Freising, Germany

Garcinia buchananii (Clusiaceae) is a plant native to Sub-Saharan Africa and its crude extracts are traditionally used by local inhabitants to treat diarrhea, dysentery, abdominal pain, and various infectious diseases. Thus, a broad variety of biological activities was assumed, effected by compounds of various extracts. By means of activity-guided fractionation using in vitro antioxidative assays of an aqueous ethanolic stem bark extract we isolated three previously unknown compounds, isomanniflavanone, an ent-eriodictyol-(3α→6)-dihydroquercetin-linked biflavanone, 1,5-dimethoxyajacareubin and the depsidone garcinisidone-G. Furthermore, six known compounds, (2’’R,3’’R)-preussianon, euxanthone, 2-isoprenyl-1,3,5,6-tetrahydroxyxanthone, jacareubin, isogarcinol, and garcinol were found for the first time in Garcinia buchananii. MPLC and HPLC separation techniques enabled us to isolate these compounds and corresponding structures were elucidated by means of UPLC-TOF-MSe, 1D- and 2D-NMR as well as ECD spectroscopy, and polarimetry. Antioxidative activities of all compounds were determined by H2O2 scavenging, H-/L-TEAC, and H-/L-ORAC assays and revealed high potency stronger or comparable to strong antioxidants like L-ascorbic acid, epicatechin, quercetin and BHT.

AGFD 258

Innovative microwave-assisted procedure for the extraction and purification of policosanols from beeswax

Virginia Brighenti1, virgivalle@alice.it, Arianna Chiossi3,4, Alberto Venturelli4, Federica Pellati2. (1) Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy (2) University of Modena and Reggio Emilia, Modena MO, Italy (3) University of Bologna, Bologna, Italy (4) Tydock Pharma s.r.l, Modena, Italy

Policosanols are a mixture of long chain aliphatic primary alcohols that have hypocholesterolemic properties, thanks to which there is an increasing interest in their extraction process. To meet this rising interest the research focused on the extraction

and purification of policosanols from beeswax taking advantage of microwave technology, which hitherto has never been applied to this mixture. The developed process comprises three main steps: microwave-assisted trans-esterification; microwave-assisted hydrolysis; fast purification with preparative LC. The final step is responsible for the increased purity of policosanols, thanks to the removal of undesired non-polar compounds. Predominant alcohols investigated in this work are tetracosanol, hexacosanol, octacosanol, triacontanol and dotriacontanol. Compound identification was performed using GC-EI-MS, while GC-FID analysis was chosen for the quantification of the main fatty alcohols present in the process products. This new method represents a useful tool for the production of PCs from beeswax to be used in nutraceutical products.

AGFD 259

Determination of hydrogen bonding acidity values and distribution doefficients for flavonoids with multiple hydroxyl substituents.

Cody E. Earp1, earp@gotarleton.edu, William L. Whaley1, Michael H. Abraham2. (1) Chemistry, Geo Sciences Environ. Sci, Tarleton State Univ, Stephenville, Texas, United States (2) Chemistry, University College of London, London, United Kingdom

The flavonoids are a large class of natural products, and many of these compounds are now considered lead structures for drug development. The logarithm of the distribution coefficient, LogDOct, and Abraham hydrogen bonding acidity, A, are important parameters for predicting the absorption, distribution, metabolism and excretion (ADME) properties of potential drugs. The values of A for mono-hydroxy flavones and mono-hydroxyisoflavones were previously determined by a method based on NMR spectroscopy. These parameters were used to estimate LogDOct and A values for flavonoids bearing multiple hydroxyl substituents. Key intramolecular hydrogen bonding interactions were identified that are important for accurate prediction of these parameters by the computer program ACD Absolve. The intramolecular hydrogen bond between the position 5 hydroxyl group and the carbonyl group at position 4 of both the flavone and isoflavone ring systems is particularly strong. This confers a value of A that is near zero for the position 5 hydroxyl group. The hydroxyl group at position 7 has an A-value of 0.68; however, when a second hydroxyl group is present at position 5, there is a slight enhancement of the value of A (for the 7-OH group of both flavones and isoflavones). The position 4 hydroxyl group has a value of A equal to 0.69 for both flavones and isoflavones. The values of LogDOct (at pH = 7.4) were measured for a series of catechol flavones and then used to estimate values of A for these compounds. A complementary approach using experimentally determined parameters along with calculations by ACD Absolve was a productive approach for the characterization of complex flavonoids. (This research was financially supported by the Welch Foundation Departmental Research Grant Program and the Tarleton Organized Research Grant Program.)

AGFD 260

Characterization and phytoremediation of a crude oil contaminated wetland

Eucharia O. Nwaichi1, nodullm@yahoo.com, Lasbrey Ikechukwu Opara2, Paul Aleruchi Nwoha1. (1) Biochemistry, University of Port Harcourt, Port Harcourt, Nigeria (2) HSE, Macphed Ventures, Port Harcourt, Nigeria

Levels of metal and hydrocarbon contaminants were determined in water from a wetland around an abandoned oil-well in Mgbuoba Community in the Nigerian Niger Delta. The performance of Hevea brasilensis at clean – up of resulting effluent was also investigated. H. Brasilensis was grown hydroponically in the effluent for forty-three (43) days in the presence of white light and a salted variant while growth indices were measured. Deionized water was used as control and all experiment was done in triplicates. Levels of Phenol and some heavy metals were determined at 0d and 43d to ascertain performance. Observed levels of Fe, Pb, Cd, Phenol and total hydrocarbons were statistically significant at p≤0.05 and reduced by 99.8%, 99.9%, 99%, 99.9% and 80.01% respectively. Cyanide, mercury and chromium were not detected for all samples. Salted regimes showed markedly higher uptake of heavy metals. Also, produced biomass compared favourably with control, and is indicative of remarkable tolerance to induced phytotoxicity. Generally results indicate excellent phytoextraction of studied contaminants, leaving relatively low values in the effluent, and could be explored in the management of all important wetland in the functioning of ecosystem vis a vis agriculture.

AGFD 261

Stabilization of whey protein isolate (WPI) by sugar beet pectin (SBP) through a Maillard-type reaction in solution

Phoebe X. Qi1, Phoebe.Qi@ars.usda.gov, Yingping Xiao1,2. (1) Dairy and Functional Foods, USDA-ARS-ERRC, Wyndmoor, Pennsylvania, United States (2) Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences, Hangzhou, Zhejiang, China

Proteins are often used by the food industries as emulsion forming and stabilizing agents, whereas polysaccharides are mainly used as thickening and water-holding agents. In particular, formulations containing mixtures of milk proteins and polysaccharides can be found among the ingredients of a wide range of colloidal food systems such as salad dressing, mayonnaise, beverages and ice cream. Although it has been established that the emulsification properties of proteins can be improved by forming conjugates with pectin through a Maillard-type reaction in dry-state, an in-depth molecular understanding on the structural changes in proteins as a result of the reaction is still lacking. In this work, we investigate the physical, chemical and structural changes of whey protein isolate (WPI), a by-product from the cheese making process, that occurred upon reacting with sugar beet pectin (SBP) in solution at near neutral pH. Conjugates were prepared by heating the mixtures at a fixed WPI concentration (5%, w/w) but varying ratios of SBP (%, w/w) at 1, 1.7 and 5 at 60oC for 48 hours. Chemical

changes of the conjugates including free sulfhydryl (SH) and primary (NH2) and secondary (NH) amines contents were determined by DTNB and ninhydrin assays respectively, and showed noticeable decreases at all levels of SBP compared to the WPI control. SDS-PAGE showed lower levels of SBP (1.0% and 1.7%) not only significantly prevented the formation of intermolecular disulfide among different whey proteins but also mitigated changes in the secondary structure of WPI caused by heating, especially b-sheet structure, as studied by far-UV circular dichroism (CD) and FTIR spectroscopies. The effect of conjugation on the tertiary structure of WPI was characterized by Trp intrinsic fluorescence, and demonstrated disruptions in the tryptophan residue environment compared to the WPI control. The thermal stability of the conjugates was also studied by CD and fluorescence, and showed an increased level of heat resistance over WPI. This work established a molecular basis for an optimal weight ratio for forming conjugates between WPI and SBP in solution, which bears implications on devising approaches for the purpose of improving functional properties of WPI through interactions with SBP.

AGFD 262

Adsorption of clay microparticles at the interface of PEG/dextran aqueous biphasic systems: Formation of clay-stabilized aqueous-aqueous emulsion droplets

Fatma Pir-Cakmak, fup110@psu.edu, Christine D. Keating. Penn State University, University Park, Pennsylvania, United States

Figure 1. Microscope images showing clay particles in PEG/dextran ATPS. Left images show overlay of fluorescence channels and right images show transmitted light for samples containing: (A) Kaolinite (KGa-2), (B) Na-rich montmorillonite (Swy-2), (C) Ca-rich montmorillonite (STx-1b) and (D) NX-illite in 20%/20% w/w PEG-Dext ATPS. Fluorescent-labeled dextran (red) and PEG (green) has been included to aid visualization of the dextran-rich phase droplets and PEG-rich continuous phase. Particle assembly at liquid/liquid interfaces is well known for oil/water systems and has been observed in aqueous biphasic systems.1 We are exploring the assembly of clay mineral particles in poly (ethylene glycol) (PEG)/dextran aqueous two-phase systems (ATPS). Clay minerals are of particular interest because they have been hypothesized as a catalyst for formation of nucleobases on the early earth.2 We compared the three different main types of clay: kaolinite, montmorillonite and illite, which differ in their particle size, crystal structures and layering as well as their swelling properties. In order to understand clay particle assembly in the ATPS, confocal optical microscopy, zeta potential and interfacial tension measurements were performed. Understanding how these particle types interact with the ATPS will lead to improved understanding of Pickering type emulsions in all-aqueous systems and may have relevance to compartmentalization of catalytic activity for protocell. 1-) J. H. Jiang, Y. Zhu, Z. G. Cui, and B. P. Binks, Angew Chem Int Edit 52 (47), 12373

(2013). 2-) J. P. Ferris and G. Ertem, Science 257 (5075), 1387 (1992).

AGFD 263

Rechargeable antimicrobial N-halamine coatings for food contact surfaces

Luis J. Bastarrachea1, lbastarr@foodsci.umass.edu, Julie M. Goddard2. (1) Food Science, University of Massachusetts, Amherst, Massachusetts, United States (2) Food Science, University of Massachusetts Amherst, Amherst, Massachusetts, United States

Every year in The United States of America about 48,000,000 people get sick from foodborne diseases. From that number, about 128,000 are hospitalized, 3000 of whom die. According to the Centers for Disease Control and Prevention, contaminated surfaces in the food industry represent one of the main risk factors for foodborne outbreaks. Herein, the effectiveness of N-halamine antimicrobial coatings applied on common food contact surfaces is reported. N-halamines are a type of antimicrobial composed by a nitrogen atom bound to a halogen (normally chlorine). They are able to inactivate microorganisms by releasing the corresponding halogen atoms once they come into contact with microorganisms. Their most important feature is the ability to be rechlorinated continuously, which makes them effective for many cycles of disinfection. A nitrogenous coating composed by polyethyleneimine (PEI) and poly(acrylic acid) (PAA) was applied over the surface of stainless steel and polyethylene (PE) through layer-by-layer deposition. This coating was able to inactivate Listeria monocytogenes in more than 5 logarithmic cycles upon N-halamine activation. However, due to the hydrolytic effect of the diluted bleach solution used to activate the N-halamines, poor stability was observed towards multiple rechlorination cycles. After modifying the coating preparation method, it was possible to produce a robust coating able to resist at least 100 rechlorination cycles and at least 300 washing cycles under different levels of pH without changing significantly its N-halamine holding capacity. The mentioned PEI-PAA coating was substantially improved, but it still exhibited some disadvantages related to its preparation. An alternative method to prepare an antimicrobial coating for food contact materials is reported. Alternating layers of PEI and styrene maleic anhydride copolymer (SMA) were applied onto the surface of polypropylene (PP). The resulting coating had low surface energy and presented enhanced antimicrobial character due to the presence of both cationic and N-halamine forming moieties. In its cationic form, the coating inactivated Listeria monocytogenes by ~ 3 logarithmic cycles. In the form of N-halamines > 5 logarithmic cycles were reached. The modified surface was able to be rechlorinated for at least 10 cycles. Such rechargeable antimicrobial coatings may support improving food safety by reducing cross-contamination of microorganisms from food processing equipments.

AGFD 264

Sodium diffusion in potatoes

Janam k. Pandya, kpandya@umass.edu, Amanda Kinchla. Food Science, University of Massachusetts, Amherst, Amherst, Massachusetts, United States

In recent days, people of the United States have become concerned about sodium intake in their diet. Research studies show that the average American takes 3,400 to 4,500 mg/day of sodium in their diet against dietary recommendation of 1,500 to 2,300 mg/day. Studies also show that 70-75% sodium in the diet comes from the processed foods. Excess sodium in the diet can potentially cause health issues such as hypertension, heart attack, kidney failure and bone problems. With a view of that our study focuses on quantifying the diffusion of sodium in potatoes as a model food and to change process variables to determine the possibilities of sodium diffusion reduction. The study was designed on potatoes to quantify sodium diffusion at different cooking temperatures in different concentration of salt solutions. Sodium diffusion was also examined with presence of different anions larger than sodium chloride, by cooking potatoes in different sodium salt solutions. Different temperature combinations of 56oC, 73oC and 100oC in combination with 0.86M and 0.086M Na+ strength solutions were evaluated. Sodium salts other than sodium chloride such as phosphate, sulfate, tripolyphosphate, metabislufite and alginate were incorporated in the study to measure anionic response in sodium diffusion. Sodium analysis was performed with Sodium Ion Selective Electrode. Sodium adsorption was quantified using linear correlation equations, derived from simplifications of Fick’s second law. Sodium mass transfer coefficient (ks) was calculated from linear regression analysis. Among different parameters incorporated in the test, changes in temperature ranging above and below the starch gelatinization temperature resulted in the largest effect on sodium diffusion (36% reduction after cooking for 40 minutes in 0.86M Na+ solution), followed by introduction of anion species larger than chloride (from 31% reduction in case of sodium tripolyphosphate to 5% reduction with sodium alginate after cooking for 40 minutes at 73oC in 0.86M Na+ strength). The future extension of this study can be conducted on different complex food systems like meats which are major source of sodium in the diet.

AGFD 265

Determination of furan levels in commercial orange juice products and its correlation to the sensory and physiochemical characteristics

Mina Kim, minakim@dongguk.edu, Min Yeop Kim, Kwang G. Lee. Dept Food Science and Biotechnology, Dongguk University, Seoul, Korea (the Republic of)

Orange Juice (OJ) is well-accepted fruit juice and consumed widely from children to adults due to its unique flavor and high vitamin C contents. While most of commercial OJs undergoes the thermal processing step, limited work has been conducted to investigate the furan levels in the commercial OJs in Korea. The objective of current study was to determine the furan levels in commercial OJ products and correlate them to the sensory and physiochemical characteristics of OJs. Further, the factors among sensory and physiochemical attributes that showed high correlation to the furan in OJs were identified. Furan was quantitatively and qualitatively analysed by Solid Phase

Micro Extraction (SPME) followed by Gas Chromatography-Mass Spectrometry (GC-MS). A descriptive analysis was conducted using a highly trained panel (n=5) to evaluate 17 OJ samples in triplicates. Physiochemical characteristics including pH, color (L*, a*, b*), °Brix, and Vitamin C content (total ascorbic acid, dehydroascorbic acid, and ascorbic acid content) were analysed using standard method of analysis. Univariate and multivariate statistical analyses were conducted for data analysis. Freshly-squeezed type OJs (n=4) had significantly lower furan levels than other OJs treated with heat processing before packaging (p<0.05). Sensory characteristics of 17 commercially available OJs were significantly different and were grouped by their distinct flavor characteristics. Vitamin C content, specifically, dehydroascorbic acid, as well as medicinal sensory flavor attribute showed higher correlation to the furan level in OJs, and these can be used as indicators for the furan level in OJs.

AGFD 266

Optimization of essential oils properties by enzymatic modification of their chemical composition: Toward sustainable processes for the production of new fragrant ingredients

Sylvain Antoniotti, antoniot@unice.fr. CNRS - University of Nice, ICN UMR 7272, NIce cedex 2, France

Essential oils are mixtures of dozens of organic compounds, mainly terpenes, terpenoids and phenyl propanoids. They present a great diversity of chemical compositions, and as a consequence, of properties. When essential oils are used in perfumery, if the most relevant property is the olfactory character, other aspects such as toxicity and allergenicity have to be considered. In many cases, a given property is due to a single compound, or a small group of compounds, in the mixture. For example, chemicals such as methyleugenol, safrole, atranol, or estragole are under surveillance because of thier toxicity, and their use is fragrance and cosmetic products is regulated. On an other side, high impact odorants such as α- and β-santalol in sandalwood, or β-damascenone and rose oxide in rose are known to contribute the most significantly to the organoleptic quality of the material. If a modification of the composition is necessary, either to decrease or increase the concentration of a given compound, very few viable technological solutions exist. The main technique industrially used in this regards is distillation, with a certain lack of selectivity, a risk of degradation of sensitive materials and a high energy cost. In this context, we have developped biocatalysis-based strategies to modifiy the chemical composition of rose, palmarosa, and sandalwood essential oils. With the use of enzymes, highly specific and/or selective reactions could be carried out in mild conditions to accurately tune the composition of essential oils. Moreover, the resulting products remain natural products, a highly valuable grade of ingredients with strong marketing advantages. We will present our results in a study on the removal of eugenol from rose essential by a peroxidase-based strategy, as well as a lipase-based treatement of palmarosa essential oil to tune its olfactory notes from floral to fruitty.

Bouhlel, C.; Dolhem, G.A.; Fernandez, X.; Antoniotti, S. J. Agric. Food Chem. 2012, 60, 1052-1058. Ramilijaona, J.; Raynaud, E.; Bouhlel, C.; Sarrazin, E.; Fernandez, X.; Antoniotti., S. Chem. Biodiv. 2013, 10, 2291–2301. Antoniotti.S. Molecules, 2014, 19, 9203-9214.

AGFD 267

Analytical chemistry, formation, reduction, chemoprevention, and in vivo exposure of acrylamide

Yu Zhang, yuzhangzju@163.com. Department of Food Science and Nutrition, Zhejiang University, Hangzhou, Zhejiang, China

The analytical chemistry, formation profile, kinetic elucidation, quantitative structure-activity relationship (QSAR), chemoprevention and in vivo exposure of acrylamide have been investigated since 2004. The main outcomes include: (i) The simultaneous UHPLC-MS/MS analysis of Maillard reaction products of acrylamide and mercapturie acid metabolites have been established. (ii) The inhibitory effect of flavonoids on the formation of acrylamide and its association with antioxidant properties has been demonstrated. (iii) The impact of flavonoids on the kinetics of acrylamide has been revealed. (iv) Factors contributing to the inhibitory effect of flavonoids, including number and position of aromatic hydroxyls, presence of C-glycosides and spatial topological structure, have been indicated using QSAR modelling. (v) The internal exposure of acrylamide and glycidamide in rats and humans has been investigated. In summary, systematic outcomes on the presence of acrylamide in food have been achieved via our 10-year investigation, which points out the direction of food safety chemistry researches.

AGFD 268

Synthesis and development of a new respiratory inhibition-type fungicide

Won Hyung Lee, genius1982@nate.com, Joo-Kyung Kim, Ho-Cheol Shin, In-Cheon Hwang. Central research institute, Kyung Nong Co, Gyeong-jusi, Korea (the Republic of)

The strobilurins, first isolated in 1977 from fermentations of Stroblurus tenacellus, are one of the most important classes of agricultural fungicides. These possess a wide range of fungicidal activities as a consequence of their ability to inhibit electron transfer between mitochondrial cytochrome b and cytochrome c1. The purpose of this study is to synthesize and develop new strobilurin compounds. A series of methoxyiminoacetamide derivatives possessing 2,2-dichlorovinyl side chains have been synthesized and their biological activities against various plant fungal pathogens have been evaluated. A compound having the strongest activity is selected and commercialization is in progress with other tests, including toxicity and formulation.

AGFD 269

Differentiation of red Port wines categories according to their volatile carbonyl compounds

Nathalie Moreira2, Isabel Vasconcelos2, Frank Rogerson3, Paula Guedes de Pinho1, pguedes@ff.up.pt. (1) Toxicology, faculty Pharmacy Porto University, Porto, Portugal (2) Universidade do Porto, aUCIBIO@REQUIMTE/Laboratório de Toxicologia, Departamento de Ciências Biológicas, Faculdade de Farmácia, Rua Jorge Viterbo Ferreira, 228, 4050-313, Porto, Portugal (3) Travessa Barão de Forrester 86, cSymington Vinhos, S.A., Apartado 26, 4431-901, V.N.Gaia, Portugal

The aim of the present study was to determine carbonyl compounds in different categories of port wines. For this purpose, a central composite design (CCD) (1, 2) was applied in order to investigate the influence of the principal parameters affecting the HS-SPME extraction of volatile carbonyl compounds associated to port wine aroma. Thirty two Port wines from different categories were analyzed: Vintage (n=5), Late-Bottled Vintage (LBV) (n=8), Colheita wines (aged between 1935 and 2002) (n=8), Tawny (10, 20, 30 and 40 years) (n=6) and Ruby (n=5). Forty five volatile compounds including aldehydes (saturated and unsaturated), dialdehydes (glyoxal, methylglyoxal, diacetyl) and ketones (b-damascenone, b-ionone, 2-butanone) gave optimal extraction using the following procedure: 7 min incubation of 2 mL of wine with 2000 mg/L of PFBHA, followed by HS-SPME extraction for more 45 min at 53 ºC. The method was validated with regard to the linearity, repeatability, inter and intra-day precision and accuracy. Results showed that LBV wines presented the highest levels of 2-methyl-1-propanal, 2-methyl-1-butanal and 3-methyl-1-butanal. Ruby wines and some of the LBV wines analyzed presented a similar volatile profile. Vintage wines presented the highest levels of b-damascenone and the lowest levels of 2-methyl-1-butanal and 3-methyl-1-butanal, when compared to the other categories. Colheita and Tawny wines were characterized by high levels of 2-phenylacetaldehyde and benzaldehyde. (1) Barros, E.P., Moreira, N., Pereira, G.E., Leite, S.G., Rezende, C.M.R., Guedes de Pinho, P. 2012. Development and validation of automatic HS-SPME with a gas chromatography-ion trap/mass spectrometry method for analysis of volatiles in wines. Talanta, 101:177–186. (2) Moreira, N., Meireles, S., Brandão, T., Guedes de Pinho, P. 2013. Optimization of HS-SPME-GC-IT/MS method using a central composite design for volatile carbonyl compounds determination in beers. Talanta, 117:523-531.

AGFD 270

Influence of plant-based protein diet on orange-spotted grouper (Epinephelus coioides) white muscle proteome profile

Yuan-Tih Ko1, irisko@ntou.edu.tw, Chyng-Hwa Liou2, chliou@ntou.edu.tw, Fu-Hsiung Huang1, Bartosz Kazlowski1, Sh-Chien Shu1, Yi-Teng Tan1, Yi-Siyuan Luo1, Ing-Feng Sie1. (1) Department of Food Science, Biotechnology Division,School of Life Sciences, National Taiwan Ocean University, Keelung, Taiwan (2) Department of Aquaculture,

School of Life Sciences and Center of Excellence for, National Taiwan Ocean University, Keelung, Taiwan

There is a growing demand for alternatives to fish meal to reduce production costs in many aquaculture enterprises. Plant-based protein supplements were reported as potential replacements for fish meal. In this study, we conducted two feeding trials to determine the effects of low protein diet (LP diet) and plant-based protein diet (wheat and corn gluten, PP diet) on white muscle proteome profile of grouper (Epinephelus coioides). Partial replacement of proteins from brown fish meal by starch (LP diet) resulted in significant down-regulation of specific structural proteins (αMHC, tropomyosin 1 and actin muscle) and creatinine kinase (CK) enzymes. This observation was further supported by impaired weight gain and feed efficiency in fish fed LP diet. Our results suggest that LP diet may have negative effect on grouper growth and metabolism. Fish fed PP diet showed significant increase in tropomyosin 1, and stable expression of other structural proteins similar to control diet. In addition, PP diet caused overexpression of soluble adenylyl cyclase and chromosome undetermined SCAF7390. Our findings demonstrate that proteomic changes induced by plant-based protein diet are similar to fish meal diet indicating its potential use as a replacement for fish meal protein in fish feed.

AGFD 271

Food’s combinatorial odor codes – new knowledge on how nature recruits volatiles to make our foods smell so good

Andreas Dunkel4, andreas.dunkel@tum.de, Martin Steinhaus1, Matthias Kotthoff2, Bettina Nowak1, Dietmar Krautwurst3, Peter Schieberle1, Thomas Hofmann4. (1) DFA Lebensmittelchemie, Freising, Germany (2) Environmental and Food Analysis, Fraunhofer Institute for Molecular Biology and Applied Ecology, IME, Schmallenberg, Germany (3) German Research Center for Food Chemistry, Freising, Germany (4) Chair of Food Chemistry and molecular sensory, Technical University of Munich, Freising, Germany

Advances in chemical trace analysis and post-genomic progress revealed odor qualities of nature’s chemosensory entities to be defined by odorant-induced olfactory receptor activity patterns. Beyond traditional views, a recent meta-analysis combined with multivariate analytics now demonstrates that the typical food odors are combinatorially constructed by 3 to 40 genuine key food odorants (KFOs) per food item, from a toolbox of ~230 KFOs in total out of ~10,000 food volatiles. These odorants are detected by ~270 OR out of our ~400 odorant receptor genes in total, and are translated as unitary constructed olfactory objects in the brain. Moreover, abundance analysis revealed a group of 16 wide-spread generalist odorants, which are detected in more than 25% of the analyzed food items and are primarily released from ubiquitous food bulk components (carbohydrates, amino acids, fatty acids). In comparison, ~150 specialist odorants were found to be present only in less than 5% of all food items and are mainly generated from distinctive precursor molecules comprising polyphenols, isoprenoids

and other largely unknown biosynthetic metabolites. This presentation gives insight into nature’s chemical signatures of smell and beyond addresses various economic implications and, furthermore, will outline how the key findings of this meta-analysis can guide future odorant receptor research, target selection in the biotechnological odorant production, as well as fundamental studies decoding how these chemical signatures are translated into hedonic preference/aversion of food flavors.

AGFD 272

Application of sensomics to commercial flavour analyses

Megan Thornton1, megant@deakin.edu.au, Monica Bueno-Fernandez2, James Addison3, Kathleen Chu3, Lewis Jones3. (1) Centre for Advanced Sensory Science, Deakin University, Melbourne, Victoria, Australia (2) Laboratory for Flavour Analysis and Enology, University of Zaragoza, Zaragoza, Spain (3) Mars Petcare, WALTHAM Centre for Pet Nutrition, Waltham-on-the-Wolds, Leicestershire, United Kingdom

Delivery of a consistent odor is important for consumer acceptability of food. Many commercial companies use the “Sensomics” approach developed by the Technical University of Munich to identify and quantify key odorants in their products [1]. This approach combines bio-activity guided detection of key odorants using gas chromatography-olfactometry (GC-O), accurate odorant quantification, followed by aroma reconstitution and sensory omission experiments. There are over 100 peer-reviewed publications using this approach, identifying key odorants in over 200 food products. Food companies tend to use Sensomics differently to academic researchers and its application to two food products, as well as some modifications to speed the analysis, are described here. This paper used the GC-O step in the Sensomics approach to study the changes in odorant compounds in a thermally processed product (roasted macadamia nut) and a stored/ matured product (fragrant rice). The results successfully identified the key changes in odorants between unroasted and roasted macadamia nuts and the batch to batch variation in fragrant rice samples Two GC-O methodologies were compared; Aroma Extract Dilution Analysis (AEDA), as often employed in Sensomics publications, where odorant priority is assigned by calculation of the Flavour Dilution (FD) factor; and Modified Frequency (MF), a more time-efficient methodology, where odorant priority is assigned by the frequency of odor detection and odor intensity. A statistical comparison of the data from AEDA and MF showed no significant differences between the assigned priorities of odorants identified in each method. However, MF was able to identify more GC-O odor peaks in a shorter time scale. In summary, Sensomics is a useful tool for commercial flavor researchers and, in the examples given, the GC-O stage can be accelerated by using MF. 1. Dunkel et al., Nature’s Chemical Signatures in Human Olfaction: A Foodborne Perspective for Future Biotechnology. Angewandte Chemie International Edition, 2014. 53: 7124-7143.

AGFD 273

Insights into the chemical composition of processed food products

Andrew J. Taylor, flavometrix@btconnect.com, Nadine Wollmann, James W. Marshall. Mars Petcare, Melton Mowbray, United Kingdom

Many food products are formulated from mixtures of biological materials, so the chemical composition of the food typically comprises thousands of different compounds. Heat processing causes further changes, which increase the chemical complexity through reactions like oxidation, hydrolysis and Maillard. Analysis of the many different chemical classes in raw and manufactured foods requires multiple extraction and separation techniques followed by identification of the compounds present. This type of analysis is very time consuming and, usually, analysis is focused on some key, known compounds which provides limited information on the chemistry taking place. A new approach will be described, which seeks to analyse as wide a selection of compounds as possible using a simple solvent extraction and advanced mass spectroscopy. Novel data analysis tools have been developed to help visualise and deconvolute the chemical data and provide new insights into the chemical composition of foods, the chemistry occuring during processing and how processing conditions affect the composition of the finished product.

AGFD 274

Needle in a haystack: Flavor analysis in complex food systems

James W. Marshall, james.marshall@effem.com, Taylor Andy, Amanda Obee. Waltham Centre for Pet Nutrition, Mars Petcare UK, Melton Mowbray, United Kingdom

The understanding of foods and their flavour characteristics has been advancing at a rapid pace for decades. Analytical chemistry developments have played a crucial role in the field of human research and many of the important human flavour compounds, nutrients and other functional chemicals in foods are well understood[1]. In the food industry, grand challenges are often more complicated than simply understanding the known flavour chemicals present in samples and this is particularly true when the consumer is not human. The published literature suggests that felines respond differently to some flavour cues compared to humans but also are deeply in tune with other factors[2] that affect food preference over single and multiple exposures. The same situation is likely to be true for humans, which means only multi-disciplinary research will result in a full understanding of the effect of food chemistry (including flavour) on consumers and in determining their preference choices in the long term. This level of multi-disciplinary research requires analytical tools which look beyond the flavour chemicals and nutrients we already know are important. To go beyond the known, researchers must be able to search the complex chemical landscape of foods for the key differences between samples, and connect the analytical data, and interpretation of that data, to the consumer. This talk outlines an industrial approach to

finding the needle in the haystack – investigating the key differences between real food products. 1. Dunkel, A., et al., Nature’s Chemical Signatures in Human Olfaction: A Foodborne Perspective for Future Biotechnology. Angewandte Chemie International Edition, 2014. 53(28): p. 7124-7143. 2. Hewson-Hughes, A.K., et al., Consistent proportional macronutrient intake selected by adult domestic cats (Felis catus) despite variations in macronutrient and moisture content of foods offered. Journal of Comparative Physiology B, 2013. 183(4): p. 525-536.

AGFD 275

From model food to real food systems: Advances and challenges in relating sensory measurements to in vivo flavor release

Jean-Luc Le Quere, jllequere@dijon.inra.fr, Elisabeth Guichard, Pascal Schlich. Center for Taste and Feeding Behavior, INRA, Dijon, France

The relationships existing between aroma perception and in vivo aroma release have been the subject of a number of studies mainly conducted on model foods. It has been acknowledged for a long time that flavor perception is a dynamic process during which the intensity and the quality of the perception vary with time. Moreover, during food consumption intrinsic intra- and inter-individual variability has been evidenced. However, the dynamics of perception have been considered only scarcely in comparison to aroma release over time measured by mass spectrometry techniques (nosespace analyses). Among available dynamic sensory methods, Time-Intensity (TI) is not able to capture multidimensionality of perception, whereas Temporal Dominance of Sensations (TDS) can (1). Simultaneous data recordings have been conducted only in a few studies essentially on flavored model foodstuffs and rarely on real foods. Although some temporal links have been evidenced in these studies, no clear and complete relationships between the two sets of data could be robustly established. Illustrated by various results obtained in our group with model and real foods, the present presentation will focus on various aspects of the relationships existing between aroma perception and in vivo aroma release obtained with nosespace analyses. Physiological parameters important for food oral processing, such as mastication behavior and food bolus formation with incorporated saliva, which could explain inter-individual perception variability will be emphasized. The influence of texture on aroma release and its relation with sensory analyses of aroma and texture descriptors will be discussed. By combining nosespace and TDS sensory analyses various temporal and descriptive links that could be proposed between sensory and release data will be discussed. A new way of analysing conjointly these two sets of data at individual level, as proposed recently (2), will be presented and discussed from data obtained with flavored model candies and commercial cheeses. Finally, some challenges in terms of sensitivity of available techniques and in terms of understanding the complexity of flavor perception of real foods from in vivo flavor release data will be presented and discussed.

1. Pineau, N. et al., Food Quality and Preference 2009, 20, (6), 450-455. 2. Schlich, P. et al., In The 14th Weurman Flavour Research Symposium, Cambridge, UK, 2014.

AGFD 276

Mechanism of flavor release from a pet food: Learnings and challenges

Punyatoya Mohapatra, pmohapatra@afbinternational.com, Cassandra Cox, Bridgette Bennett. AFB International, Saint Charles, Missouri, United States

The pet food industry applies flavors or palatants on kibbles, wet diets, treats, and frozen foods to enhance palatability. The palatants add appealing taste to the pet food and drive the animals towards the bowl. Hence an in-depth knowledge of the aroma and taste components from the surface of the kibble is important to understand the drivers of palatability. Several analytical methods are in place to identify and understand a pet food flavor profile. However, most of these techniques require sample preparation (grinding, heating, cooling, addition of solvents) which is often inconsistent with real world animal experience. This study aims to understand the mechanism of flavor release from the surface of a whole kibble in order to support use of analytical methods that better mimic what the animal experiences. An experiment was designed to understand the effect of kibble type (whole and ground), medium (water and synthetic saliva), temperature (35oC and 45oC), and extraction time (1 minute and 5 minute) on flavor profile using electronic nose and electronic tongue respectively. Specific ions (71 and 84) showed peak intensities reflecting differences in the volatile profile of ground versus whole kibble. The instrument showed strong coorelation between kibble type, medium, and temperature.The ion response from electronic nose showed temperature as a significant factor by itself.The electronic tongue consisted of seven sensors, of which three sensors, X, Y, and Z, showed strong interaction between kibble type and medium. Sensor D showed interaction between all three factors, kibble type, medium, and extraction time. A combination of electronic nose and tongue provided useful information about factors affecting the flavor release from the surface of a kibble. It was conluded that flavor profile of a whole kibble is different than a ground kibble and in-line with the real world animal experience.

AGFD 277

Using in vivo analytical techniques to screen for masking and enhancing solution and to understand flavor interactions on compound level in a complex food flavor system

Catrienus de Jong1, catrienus.dejong@nizo.com, Angelique Villiere2,3, Laurent Lethuaut2,3, Carole Prost2,3. (1) Flavour, Food and Beverages, NIZO food research, Ede, Netherlands (2) Food Science and Engineering, ONIRIS, Nantes, France (3) Food Aroma Research Group, CNRS UMR 6144 GEPEA, Nantes, France

A better understanding of the flavor interaction within a complex food flavor system is key in the preparation of more healthy and complex foods with low sugar and salt levels while maintaining an excellent flavor perception1. It will also facilitate enhancing or masking typical flavor characteristic and optimize flavor balance. We describe two in vivo approaches to study aroma-aroma interactions on compound level in food systems. The first approach called the Olfactoscan2 in which a GC and olfactometer are coupled to screen for typical compounds which are able to mask a typical off flavor in orange juice (potato) and meat (boar taint). The second approach is based on a new original device called GC-INNOSCENT, designed by ONIRIS, that permits to recover selected volatile extracts3. Furthermore, it allows to know the sensorial impact of selected volatile compounds on the global expression aroma on the basis of addition or omission approaches. The combination of these two techniques are both used to help industry to get a better understanding and solution to optimize their current products or used for new product developments. Where confidentiality permits our approaches are illustrated with results from actual cases. 1 Poinot P., Arvisenet G., Ledauphin J., Gaillard J.L., Prost C. (2012). How can aroma-related cross-modal interactions be analysed? A review of current methodologies. Food Quality and Preferences, 28(1), 304-316. 2 Burseg, K.; de Jong, C. (2009), Application of the Olfactoscan method to study the ability of saturated aldehydes in masking the odor of methional. Journal of Agricultural and Food Chemistry; 57(19):9086-90. 3 Villière A., Arvisenet G., Lethuaut L., Prost C., Serot T., (2012). Selection of a representative extraction method for the analysis of odourant volatile composition of french cider by GC-MS-O and GcxGC-TOF-MS, Food chemistry, 131, 1561-1568.

AGFD 278

Covalent polyphnol-protein intractions – challenges and research needs

Sascha ROHN, rohn@chemie.uni-hamburg.de. Hamburg School of Food Science, University of Hamburg, Hamburg, Germany

Plant phenolic compounds are hypothesized to possess a lot of different physiological properties such as antioxidativity, immunmodulation, antimutagenicity, or even anticarcinogenicity. Unfortunately, these health promoting effects are mostly determined with pure substances. Unfortunately, such approaches do not consider any changes of the phenolic compounds, although alterations cannot be excluded during food processing. Depending on the conditions, phenolic compounds are able to react with other food ingredients such as proteins. It has been shown that this reaction leads to a change of nutritional quality [1,2]. Complexity and number of the reaction products are high, as the reactivity remains enabling the formation of protein crosslinking [3]. Many of these products are colored in various shades, but it is not clear to which extent these reactions contribute to color of processed food. Therefore, characterization of the reaction products with their specific binding sites is

quite challenging. So far, the derivatives formed have been characterized in terms of changes in their physicochemical and structural properties. Methods applied included measurement of free amino and thiol groups, as well as the determination of the tryptophan content, protein solubility, estimation of the hydrophilic/hydrophobic character, and in the case of enzymes, measurement of their activity against standard substrates. However, those approaches were only a kind of indirect characterization of the covalent interaction. With regard to modern analytical methods, mass spectrometry, NMR spectroscopy, and fluorescence quenching methods have been developed for the analysis of polyphenol-protein interactions [1,2]. As an alternative tool, even hyphenated HPTLC can be used [5,6]. [1] Xiao (2012) Crit. Rev. Food Sci. Nutr., 52: 85-101. [2] Le Bourvellec (2012) Crit. Rev. Food Sci. Nutr. 52: 213-248. [3] Rohn (2006) Mol. Nutr. Food Res. 50: 696-704. [4] Rawel (2010) Phytochem Rev. 9: 93-109. [5] Tscherch (2013) Phytochem Anal. 24:436-445. [6] Rohn (2014) Food Res. Int. 65: 13-19.

Oxidation of a catechol moiety as a pre-requiste for the formation of reactions products.

AGFD 279

Do dietary Maillard reaction products play a role in the progression of noncommunicable diseases?

Veronika Somoza, veronika.somoza@univie.ac.at, Ann-Katrin Holik. Nutritional and Physiological Chemistry, University of Vienna, Vienna, Austria

Noncommunicable diseases (NCDs), also known as chronic diseases, are not passed from person to person. They are of long duration and generally slow progression. The four main types of NDCs are cardiovascular diseases (like heart attacks and stroke), cancers, chronic respiratory diseases (such as chronic obstructed pulmonary disease and asthma), and diabetes mellitus Type 2 (T2DM). A Western diet containing high amounts of processed, Maillard reaction product (MRP)-rich foods and low amounts of vegetables has been associated with a greater risk of insulin resistance and risk factors of metabolic syndrome, cardiovascular complications, and obesity in T2DM patients (1) and in healthy subjects (2, 3). The current question is whether the intake of compounds generated upon heat treatment, especially MRPs, contribute to mechanisms regulating food intake, body weight maintenance, as well as lipid and glucose homeostasis. Recent studies have shown that intake of severely heat-treated foods postprandially decrease leptin and adiponektin, both involved in the regulation of energy homeostatsis and food intake, in T2DM patients (4). In one of our own studies, we could demonstrate that dietary MRPs from bread crust impair insulin sensitivity, affect leptin plasma concentrations and leptin receptor expression in the hypothalamus, and increase total food intake (5). Customized DNA-microarray analysis in brain cells and adipocytes revealed changes in the expression of genes relevant for insulin sensitivity, satiety and craving after treatment with high concentrations of Nε-carboxymethyllysine, one of the MRPs formed in bread crust and other heat-treated foods. These results were confirmed by functional assays in adipocytes and enterocytes. Thus, chronic intake of high-temperature processed foods might pose a risk for obesity, and two of the NDCs, cardiovascular diseases and T2DM, in compromised subjects. Heat-processing of foods may need to be optimized to mitigate concentrations of harmful compounds while increasing the beneficial ones. Esmaillzadeh A et al., Am J Clin Nutr (2007) 85:910-18. Vlassara H et al., Proc Natl Acad Sci (2002) 99:15596–601. Birlouez-Aragon I et al., Am J Clin Nutr (2010) 91:1220-6. Stirban A et al., Ann N Y Acad Sci. (2008) 1126:276-9. Sebeková K et al., Physiol Behav (2012) 105:693-701.

AGFD 280

Kinetic modeling of acrylamide formation during the finish-frying of french fries with variable sugar content

Dimitrios P. Balagiannis1, d.balagiannis@reading.ac.uk, Jane K. Parker1, Jeremy Higley2, Tony Henson2, Gordon Smith3, Bronislaw L. Wedzicha4, Donald S. Mottram1. (1) University of Reading, Reading, United Kingdom (2) ConAgra Foods, Omaha, Nebraska, United States (3) Kansas State University, Manhattan, Kansas, United States (4) University of Leeds, Leeds, United Kingdom

Acrylamide is formed from the reaction of reducing sugars and asparagine in the course of the Maillard reaction. Potatoes contain considerable amounts of these precursors, so deep frying of potato strips results in the formation of acrylamide. However, the rate of the formation of the latter is a function of multiple factors, with precursor concentration, moisture content and frying temperature being the most important ones. Furthermore, temperature and moisture gradients, as well as precursor and acrylamide concentration gradients are generated along the body of the French fry, due to the shape of the potato strips and the limited frying time. Taking all the above factors into account, the kinetics of the formation of acrylamide from the Maillard reaction during deep frying of French fries were studied. Multiresponse Kinetic Modelling was implemented in order to elucidate the kinetically important chemical pathways that lead to acrylamide formation. Initially, a kinetic model was built based on fried potato strips with varied initial concentration levels of glucose and fructose, the reducing sugars which naturally occur in potatoes. Then, the kinetic model was extended and revised to allow for variations in the maltose content. Even though maltose does not occur naturally in white potatoes, it was considered as an important carbohydrate to manipulate sugar levels. In this context, potato was used as a test matrix to understand the kinetics of acrylamide formation from maltose in relation to glucose and fructose, and to investigate the potential for acrylamide formation in maltose-containing potato products.

AGFD 281

Producing low acrylamide risk potatoes: A three-year public/private sector collaborative project focused on genetics, agronomy, and storage

Nigel Halford, nigel.halford@rothamsted.ac.uk. Rothamsted Research, Harpenden, United Kingdom

Acrylamide is a processing contaminant that was discovered in a range of popular foods in 2002, with fried, baked and roasted potato and cereal products, and coffee being the main contributors to dietary intake. It forms from free asparagine and reducing sugars via similar pathways to the flavors and aromas associated with browning, but is classified as ‘probably carcinogenic to humans’. Indicator levels for its presence in foods were set by the European Commission in 2011 and revised in 2013. Results will be presented from a three-year project called ‘Producing Low Acrylamide Risk Potatoes’, which was undertaken in the United Kingdom and involved a consortium of public sector research teams (from Rothamsted Research, James Hutton Institute and University of Reading) and private sector organisations. The study showed that varietal selection could be a powerful tool in ensuring regulatory compliance of potato

products for acrylamide formation. It highlighted the effect of long-term storage on acrylamide risk, the complex relationship between precursor concentration and acrylamide formation in potato products, and the link between acrylamide formation, browning and flavor. It also investigated the effects of fertilisation regimes and water availability during cultivation on acrylamide-forming potential. Genetic studies included the analysis of a potato breeding population that showed segregation of acrylamide-forming potential, in order to identify quantitative trait loci that control the trait. It is important that potato breeders engage on the acrylamide issue to enable compliance with future regulatory scenarios. Improving the storage characteristics of potatoes, reducing the concentrations of free asparagine and reducing sugars, and making the concentrations of these metabolites less responsive to crop management and environmental factors, should all be breeding targets. However, reducing acrylamide formation while not affecting the qualities that define products and are demanded by consumers is likely to be difficult, and breeders must take this into account.

AGFD 282

Flavor and Acrylamide Formation

John S. Elmore, j.s.elmore@reading.ac.uk. University of Reading, Reading, United Kingdom

Flavor formation in potato chips is dependent to a large degree on the Maillard reaction. However, the Maillard reaction is also responsible for the formation of acrylamide, a potential carcinogen. Since the presence of acrylamide in cooked foods was discovered in 2002, food manufacturers have tried to reduce acrylamide formation while maintaining desirable flavor. This talk will describe the relationship between acrylamide and flavor formation and examine how potato variety and tuber storage conditions can influence chip quality.

AGFD 283

Reducing the acrylamide-forming potential of wheat and rye

Tanya Curtis, Tanya.curtis@rothamsted.ac.uk, Jennifer Postles, Nigel Halford. Rothamsted Research, Harpenden, United Kingdom

Acrylamide is a processing contaminant that was discovered in a range of popular foods in 2002. It forms from free asparagine and reducing sugars via similar pathways to browning colors, aromas and flavors, during high-temperature cooking and processing, but is classified as ‘probably carcinogenic’ to humans. Cereal products account for 30 – 45 % of dietary acrylamide intake. Projects have been initiated on reducing the acrylamide-forming potential of wheat and

rye, involving groups from Rothamsted Research, John Innes Centre and the University of Reading, and a consortium of private sector organisations. The studies have shown significant differences between varieties of wheat and rye with respect to acrylamide-forming potential. In these crops, free asparagine concentration is the limiting factor for acrylamide formation, and this is the parameter on which varietal selection should be based. The genetic control of asparagine accumulation in the grain is being investigated. Environmental factors (E) also have significant effects, on their own and in combination with varietal differences (G × E), and crop management is important: Sulfur deficiency, for example, causes a massive accumulation of free asparagine in wheat grain, although it does not affect free asparagine concentration in rye under field conditions. Nitrogen fertilization, however, increases free asparagine concentration in both wheat and rye. Plant breeders must engage on the acrylamide issue or risk losing market share to those who do, but the problem is made more difficult by the link between browning, flavor and acrylamide formation. It must also be remembered that cereals are hugely important to global food security and significant health benefits are associated with eating wholegrain cereal products.

AGFD 284

Impact of water deficit on volatile composition of grapes and wine

Michael C. Qian1, michael.qian@oregonstate.edu, Krista Shellie2. (1) Oregon State Univ, Corvallis, Oregon, United States (2) Horticultural Crops Research Laboratory, USDA-ARS, Parma, Idaho, United States

Availability of water has become an increased concern for agriculture, including wine and wine grape production. Water status will directly affect plant nutrient uptake, and change vine canopy size. The canopy size may alter the microclimate within the vine canopy and change vine physiology. The change in vine and grape physiology will impact the secondary metabolism, including the volatile composition of the grapes. The influence of vine water status during berry development on the volatile and precursor composition of Merlot grapes was studied in semi-arid, southwestern Idaho over multiple growing seasons. Vines were provided with different percentages of their estimated evapotranspiration water demand (ETc) throughout berry development. The results demonstrated that the severity of vine water deficit during berry development affected grape volatile composition despite vintage differences in volatile composition. Free C6 compounds, which give herbaceous aroma, decreased with increasing severity of vine water stress. The severity of vine water deficit affected the free and glycosidically bound terpenols. Moreover, vine water deficit severity significantly increased free and bound β-damascenone as well as bound 3-hydroxy-β-damascenone, TDN, vitispirane. Wine produced from deficit-irrigated vines had an increased amount of branch-chained fatty acid esters and β-damascenone relative to wine produced from well-watered vines.

AGFD 285

Influence of sunlight exposure on Pinot noir grape and wine volatile composition

Michael C. Qian1, Fang Yuan2, fang.yuan@oregonstate.edu. (1) Oregon State Univ, Corvallis, Oregon, United States (2) Food Science and Technology, Oregon State University, Corvallis, Oregon, United States

Canopy manipulation practices can change canopy microclimate of grapevines to affect plant temperature, photosynthesis, metabolic activity, and disease resistance. Cluster-zone leaf removal can increase fruit sunlight exposure, and change the microclimate of the vine. It has been shown that cluster-zone leaf removal for Pinot noir at West Oregon had little influence on vine growth, or grape maturity in terms of total soluble solids (TSS), pH or titratable acidity (TA) at harvest. However, leaf removal at fruit zone affected both volatile and nonvolatile composition of the grape fruits. Multiple year study showed that fruit zone leaf removal resulted in higher concentrations of quercetin glycoside, petunidin- and malvidin-3-monoglucoside anthocyanins in grapes. In addition, in addition, leaf removal resulted in higher concentrations of β-damascenone, and some bound-form terpenoids in both grapes and wines. The increases in β-damascenone were positively correlated to the increased sunlight exposure. The effect of sunlight exposure could be associated with UV light. A separate study demonstrated that UV exposure significantly increased the Brix and pH in the grape juice, and increased wine color density, anthocyanins, total pigment, total phenolics and tannin content. UV exposure resulted in increase of nerol, geraniol and citronellol but not linalool in the wine.

AGFD 286

Not your ordinary terroir - the role of pathogenesis related proteins (PRPs) in limiting tannin extraction across winegrape varieties and regions

Lindsay F. Springer2, Gavin L. Sacks1, gls9@cornell.edu. (1) Food Science, Cornell University, Ithaca, New York, United States (2) Food Science and Technology, Cornell University , Dundee, New York, United States

Condensed tannins (CT) impart important organoleptic qualities, particularly astringency, to red wines. Unlike many other grape-derived flavor compounds, the quantity of CT in fruit is not well correlated with its concentration in wine. We have previously reported differences in CT binding to insoluble grape-derived materials from a variety of V. vinifera and interspecific (V.vinifera x Vitis spp.) cultivars, and identified several tannin-binding pathogenesis related proteins (PRPs) in wines by NanoLC-MS/MS. In the current work, proteins from 33 wines were quantified by SDS-PAGE. Wines produced from wild grapes had the highest quantity of protein (Vitis riparia, 667 ± 119 mg/L), 20-fold higher than concentrations in V. vinifera wines (25.2 ± 22 mg/L). Intermediate PRP quantities were observed in interspecific hybrid (V.vinifera x V.vitis

spp.) cultivars (105 ± 49 mg/L). We observed a significant correlation between PRPs and CT loss from wines following CT addition (p<0.05), suggesting that grape-derived PRP concentration is a key factor controlling tannin extraction or retention. Because PRP expression is induced by certain environmental factors (cold, disease pressure), this work suggests a unique way for how the sum of local conditions (terroir) can affect wine qualities.

AGFD 287

Accumulation of exogenous volatiles in Vitis vinifera fruit and leaves as nonvolatile glycoconjugates

Kerry Wilkinson1, kerry.wilkinson@adelaide.edu.au, Renata Ristic1, Julie Culbert1, Lieke van der Hulst1, Ana Pardo-Garcia2, Gonzalo Alonso2, Rosario Salinas2, Natoiya Lloyd3, Yoji Hayasaka3. (1) School of Agriculture, Food and Wine, The University of Adelaide, Glen Osmond, South Australia, Australia (2) Universidad de Castilla-La Mancha, Albacete, Spain (3) The Australian Wine Research Institute, Adelaide, South Australia, Australia

Grapevines are capable of absorbing volatile compounds present in the vineyard during the growing season and these volatiles can subsequently influence the composition and sensory properties of grapes and wine. For example, 1,8-cineole (eucalyptol) has been identified in fruit harvested from grapevines grown in close proximity to eucalyptus trees1; while camphor was identified in wine, following grapevine exposure to lavandin hydrolat, an essential oil byproduct2. In some cases, exogenous volatiles accumulate in the fruit and/or leaves of grapevines in non-volatile glycoconjugate precursor forms; i.e. with one or more sugar moieties attached. For example, several recent studies have demonstrated the presence of glucosylglucosides, glucosides, diglycosides and rutinosides of smoke derived volatile phenols in grapes and wine, following grapevine exposure to smoke3,4. It has been suggested that glycosylation may increase the solubility of volatile compounds, facilitating their transportation and/or storage, as well as limiting any potential toxicity5. This paper concerns the application of quantitative liquid chromatography-tandem mass spectrometry methods in order to understand the glycosylation of exogenous volatiles in grapevine fruit and/or leaves, following: (i) vineyard exposure to bushfire smoke; (ii) grapevine exposure to smoke under experimental conditions; and (iii) foliar applications of different volatile compounds to grapevines. Evidence for the accumulation of a glycoconjugate precursor to an exogenous volatile compound without a free hydroxyl functional group will also be presented. 1. Capone, D. L.; Jeffery, D. W.; Sefton, M. A. J. Agric. Food Chem. 2012, 60, 2281–2287. 2. Martínez-Gil, A. M.; Pardo-García, A. I.; Zalacain, A.; Alonso, G. L.; Salinas, M. R. Food Res. Int. 2013, 53, 391–402.

3. Wilkinson, K.L.; Ristic, R.; Pinchbeck, K.A.; Fudge, A.L.; Singh, D.P.; Pitt, K.M.; Downey, M.O.; Baldock, G.A.; Hayasaka, Y.; Parker, M.; Herderich, M.J. Aust. J. Grape Wine Res. 2011, 17,S22–S28. 4. Hayasaka, Y.; Parker, M.; Baldock, G. A.; Pardon, K. H.; Black, C. A.; Jeffery, D. W.; Herderich, M. J. J. Agric. Food Chem. 2013, 61, 25–33. 5. Winterhalter, P.; Skouroumounis, G. K. Adv. Biochem. Eng. Biotechnol. 1997, 55, 73–105.

AGFD 288

Changes in orange juice flavor volatile and non-volatile compounds in response to citrus greening or Huanglongbing (HLB) disease and disease management strategies

Baldwin Elizabeth5, Liz.Baldwin@ars.usda.gov, Anne Plotto5, Jinhe Bai3, John A. Manthey2, Smita Raithore4, Huqing Yang5, Sophie Deterre1, Sharon Dea5. (1) UDSA ARS, Fort Pierce, Florida, United States (2) USDA ARS, Fort Pierce, Florida, United States (3) USDA, ARS, HRL, Fort Pierce, Florida, United States (4) USDA-ARS, Fort Pierce, Florida, United States (5) Horticultural Research Laboratory, USDA-ARS , Ft. Pierce, Florida, United States

Citrus greening or Huanglongbing (HLB) disease has decimated citrus production around the world and is currently impacting the Florida citrus industry. At its height, Florida was producing around 250 million boxes of citrus fruit per year, but declined to 106 million boxes last year. In Florida, 90% of the fruit (mostly oranges and grapefruit) are juiced with about 10% of the industry being in fresh fruit. This large reduction in citrus production has been due to many factors, but mainly to HLB, and to a smaller extent, citrus canker disease. Further impacts of HLB are the increased costs of production to combat the disease, which has also resulted in lost acreage, as well as the closure of many packinghouses and several juice processing plants. In addition, HLB disease results in off-flavor of orange juice due to decreased sugars, sometimes increased acids and increased levels of bitter limonoids and astringent flavonoids. The aroma volatile profile of the juice and the essential oil by-product is also altered by HLB disease, favoring terpenoid compounds at the expense of aldehyde and ester top notes. Flavor management strategies for processors are being developed to predict quality of incoming fruit juice, such as use of electronic tongue and qPCR for determination of pathogen DNA, so processors can make informed decisions for blending purposes. Efforts are continuing to identify other off-flavor compounds and to develop methods to mitigate their off-flavor effects. The effects of foliar nutritional sprays, used by growers to mitigate disease symptoms on trees was investigated for effects on juice quality, although no consistent results were found.

AGFD 289

Postharvest practices to alleviate flavor loss of tomatoes under current marketing systems

Jinhe Bai1, jinhe.bai@ars.usda.gov, Baldwin Elizabeth1, Anne Plotto1, Libin Wang1,2. (1) USDA, ARS, HRL, Fort Pierce, Florida, United States (2) Nanjing Agricultural University, Nanjing, Jiansu, China

Flavor of tomatoes is best when the fruit are harvested full ripe and delivered to consumers at room temperature. However, under the current marketing system, fruit usually are harvested at the mature green (MG) stage and shipped at low temperature to slow ripening and prevent fruit losses due to bruising and decay, thereby extending storage life. The consequence is that tomatoes in today’s marketplace lack of flavor. We conducted a series of studies to alleviate the flavor loss by harvesting fruits at advanced maturity stages, and applying non-chemical (hot water) and chemical fumigation treatments including methyl salicylate (MS), and 1-methylcyclopropene (1-MCP) at harvest. Results showed that 1) harvesting at breaker (BR) stage eliminated immature green (IG) fruit (not capable of ripening with good quality) which are often difficult to distinguish from MG, and are responsible for much of the low flavor quality fruit. A combination of BR harvest and 1-MCP fumigation resulted in better flavor quality with adequate shelf life; 2) hot water treatment prior to chilled storage for MG tomatoes improved fruit sanitation, color development and mitigated chilling-induced flavor loss upon ripening. Fruit preheated before chilling had higher levels of 6-methyl-5-hepten-2-one, 2-methylbutanal and 2-phenylethanol than non-heated fruit and were perceived to have more tomato flavor; 3) MS treatment prior to chilling alleviated chilling-induced flavor loss. Chilling suppressed production of oxygen, nitrogen and sufur-containing heterocyclic compounds, ketones, alcohols, and aldehydes, including 13 important aroma components to tomato flavor, however, MS-treated fruit had higher concentrations of these important aroma contributors.

AGFD 290

Molecular assessment of metabolome changes in carrots (Daucus carota L.) induced by abiotic stress challenges

Corinna Dawid1, Corinna.Dawid@tum.de, Andreas Dunkel2, Thomas Nothnagel4, Detlef Ulrich3, Barbara Singldinger1, Daniela Günzkofer1, Thomas Hofmann1. (1) Chair of Food Chemistry and Molecular Sensory, Technical University of Munich, Freising, Germany (2) Chair of Food Chemistry and molecular sensory, Technical University of Munich, Freising, Germany (3) Institute for Ecological Chemistry, Plant Analysis and Stored Product Protection, Julius Kühn Institute (JKI), Federal Research Centre for Cultivated Plants, Quedlinburg, Germany (4) Institute for Breeding Research on Horticultural and Fruit Crops, Julius Kühn-Institut (JKI), Federal Research Centre for Cultivated Plants, Quedlinburg, Germany

With a current annual world production of more than 30 million tons and a total growing area of about 1.5 million hectares the cultivated carrot (Daucus carota L.) ranks among the top ten vegetable crops. Although, its typical sweet flavor is favored all around the world, the palatability of carrots and its products might be strongly decreased by a sporadic bitter taste which is often the reason for consumer complains. This off-flavour

is induced by abiotic and biotic stress factors during harvesting, transportation, storage and processing. Although recent application of a SENSOMICS approach gave first insight into individual bitter tasting phytochemicals in carrots, their up-regulation during abiotic stress still remains elusive. In order to gain a more comprehensive knowledge on the chemical mechanisms involved in quality changes of cultivated carrots in response to abiotic stress factors like water stress or post-harvest mechanical (transport-) stress, numerous non-volatile key metabolites of different stressed and native carrot genotypes were comparatively characterized by means of UPLC-TOF-MS metabolic profiling. Software-assisted marker molecule selection in stressed carrots, followed by preparative chromatographic purification revealed the chemical structures of bitter tasting phytochemicals by means of LC-MS, LC-MS/MS, and 1D/2D-NMR experiments. Accurate quantitation of these target molecules by means of UPLC-MS/MSMRM in carrots before and after stress challenge of the same genotypes revealed for the first time novel insights into the stress-induced metabolic response of distinct carrot genotypes. These results might help to navigate carrot breeding programs and to optimize post-harvest treatment of carrots from producer to consumer/processor towards the production of sensorially preferred carrot products.

AGFD 291

Flavour formation in skimmed milk powder in a low-moisture model system

Ashleigh Stewart1, cv010167@pgr.reading.ac.uk, Angela Ryan2, Alistair Grandison1, Jane K. Parker1. (1) Food and Nutritional Sciences, University of Reading, Reading, United Kingdom (2) Nestle PTC York, York, United Kingdom

The Maillard reaction is a cascading set of browning reactions that occur during thermal processing between reducing sugars and amino groups, leading to the formation of many compounds, some of which produce flavour. The thermal reactions of milk and milk powder have been studied in aqueous systems, either using a complete product[1, 2] or simple model systems[3, 4]. Preliminary work showed that the Maillard reaction of skimmed milk powder (SMP) is enhanced in non-aqueous conditions. A low-moisture system was achieved by reconstituting SMP in inert oil. Heating this system for 2 hours at 115 °C produced thermally-derived flavour compounds, such as maltol and furaneol, in significant amounts. After demonstrating the high flavour potential of SMP in an oil-based system, model systems were used to explore the key components for flavour formation. The Maillard reaction of milk takes place between lactose and free amino groups in milk proteins, primarily the e-amino group of lysine. However, there are other components in SMP that may play an important role in flavour formation but have not yet been studied. Combinations of lactose, casein, free amino acids, whey proteins and monosaccharides were studied, in addition to the effect of pH and salts. By heating these model mixtures, in concentrations equivalent to those in SMP, it was possible to elucidate the key

components that effect the Maillard reaction in SMP. Flavour formation was monitored after heating using SPME-GC/MS. References 1. Boekel, M.A.J.S.v. and H.E. Berg. Kinetics of the early Maillard reaction during heating of milk. in Proc. 5th Int. Symp. on the Maillard Reaction, Minnesota, 1993: Maillard reactions in chemistry, food, and health, T.P. Labuza et al. (eds.) (1995) 170-175. 1995. 2. Fenaille, F., et al., Modifications of milk constituents during processing: A preliminary benchmarking study. International dairy journal, 2006. 16(7): p. 728-739. 3. Van Boekel, M.A.J.S., Kinetic modelling of sugar reactions in heated milk-like systems. Neth. Milk Dairy J. 50 (1996) 245-266., 1996. 4. Brands, C.M.J. and M.A.J.S. van Boekel, Kinetic modelling of reactions in heated disaccharide–casein systems. Food Chemistry, 2003. 83(1): p. 13-26.

AGFD 292

Use of gas chromatography with quadrupole time-of-flight mass spectrometry (GC/Q-ToF) to compare odor formation in meat and model meat systems following the addition of precursor flavors

Lewis Jones1, Lewis.Jones@eu.effem.com, James Addison1, Nathan Hawkins2, Kathy Ridgway2. (1) Waltham Centre for Pet Nutrition, Near Melton Mowbray, United Kingdom (2) Anatune, Cambridge, Cambridgeshire, United Kingdom

Flavor precursors are added to food, before processing to generate the required desirable flavors in the finished product. Amino acids and reducing sugars are common flavor precursors used with thermally processed food, as they are the starting reagents of the Maillard reaction, which produces a wide range of odorous compounds. The choice of flavor precursors is often based on fundamental knowledge generated through the use of simplified model systems. However, real foods are chemically diverse and may present different physical environments. The Maillard reaction itself is a complex set of inter-related chemical pathways that produce hundreds of compounds. Therefore, applying the fundamental knowledge of flavor precursors to product is a challenge. This work investigates the difference in volatile compounds generated using process flavors added to a real food product (canned liver) and to model systems. This is achieved with both targeted and non-targeted analysis using gas chromatography coupled with quadrupole time-of-flight mass spectrometry (GC/Q-ToF) and single quadrupole mass spectrometry (GC/Q). The key odorants of canned and thermally processed liver were determined using a targeted “sensomics” approach. Using these data, three separate precursor flavors were formulated using the amino acids; isoleucine, methionine and cysteine to produce the key liver odorants; 3-methylbutanal, 3-(methylthio)propanal; and 2-acetylthiazole respectively. The precursor flavors were added before thermal processing to liver, a

model system (comprising the reducing sugars and free amino acids determined in liver) and water. The amount of key odorants formed from the precursor flavors was significantly lower when part of the liver system, as compared to the model system. Untargeted, chemometric analysis was used to investigate differences in all volatiles generated between the systems. Results show the effect of free amino acids, reducing sugars and the liver matrix itself on odorant and volatile formation. The advantage of using GC/Q-ToF as compared to GC/Q for both targeted and untargeted analysis will be discussed.

AGFD 293

Parameters impacting flavour profile and shelf-life of dairy ingredients

Martine Trotin, martine.trotin@mdlz.com, Andreas Czepa, Barbara Suess, Pierre R. Guilet, Jochen Pfeifer. Mondelez International, Reading, United Kingdom

Dairy ingredients are key to many snacking and confectionary products delivering flavour and texture functionalities. It is therefore important for the industry to be able to evaluate the dairy raw materials based on parameters which impact product quality. Both the actual flavour profile of dairy ingredients as well as the shelf-life will influence the selection of dairy ingredients for specific applications and therefore need to be characterised with relevant analytical and/or sensory measurements. It is well known that the drying technology which is used to transform the wet dairy ingredient into a dry system has an influence on the resulting flavour profile and shelf-life. Spray drying is the most widely used technology in the dairy industry due to its favourable processing costs. However, it can result in changes to the initial flavour profile and have detrimental effect on shelf-life if not carefully managed. Another important aspect is to understand and predict the final flavour profile and shelf-life of products containing dairy ingredients as well as whether the incorporation of dairy ingredients into products such as chocolate, caramel, and biscuit can delay undesirable reaction which could occur during the shelf-life of dairy ingredients. This presentation will provide a comprehensive review of existing knowledge including the impact of drying technology of typical dairy systems on loss of flavour compounds, generation of off-notes and generation of precursors for oxidation, the comparison between different technologies such as spray drying and freeze drying, hypotheses on potential mechanisms behind the differences, and relevant measurements to characterise the dairy flavour profile and shelf-life. The presentation will also provide initial results from a new shelf-life study of dairy ingredients as well as the resulting outcome from the application into products.

AGFD 294

Sensory and instrumental analysis of sweet potato fries

Jane K. Parker1, j.k.parker@reading.ac.uk, Stella Lignou1, J S. Elmore1, Dimitrios P. Balagiannis1, Jeremy Higley2, Gordon L. Smith3, Donald S. Mottram1. (1) Department of

Food and Nutritional Sciences, University of Reading, Reading, United Kingdom (2) ConAgra Foods, Omaha, Nebraska, United States (3) Department of Grain Science and Industry, Kansas State University, Manhattan, Kansas, United States

Sweet potato fries are increasingly popular in the US, yet their flavour is relatively unexplored, both from a sensory and an instrumental perspective. In this work, two varieties of sweet potato, which are commercially viable and phenotypically diverse, were prepared with and without a carbohydrate-based coating which is used to improve texture. Raw sweet potato strips were par-fried, frozen and then finish fried in canola oil at 175 °C for 140 s. These were served immediately to a sensory panel, who performed a quantitative descriptive analysis on the four samples. Although several of differences were texture driven, a caramel flavour was found to be significantly higher in the variety with the higher sugar content, a spicy aroma was correlated with the batter coated fries and four attributes (perfume/apple aroma and flavour as well as sweet potato flesh aroma and flavour) were correlated with the uncoated fries. These data will be correlated with GC-MS and GC-Olfactometry data obtained from SAFE extracts of the same batches of fries and discussed in relation to the precursors which were determined in the raw sweet potato. The non-homogeneity of the fries and the role of the batter both present a challenge in interpreting the underlying chemistry

AGFD 295

1-p-Menthen-8-thiol, the grapefruit character impact volatile, is a thermally generated artifact in citrus juices

Fatima Jabapurwala2,4, Jianming Lin2,1, Russell L. Rouseff2,3, rrouseff@ufl.edu. (1) Firmenich Inc, Plainsboro, New Jersey, United States (2) CREC, Univ of Florida, IFAS, Lake Alfred, Florida, United States (3) Citrus Research/Food Science, Southwest University, Chongqing, China (4) Independent Consultant, Delhi, India

In 1982, 1-p-menthen-8-thiol, 1-PM-8-T, was reported as a “powerful flavor impact compound (which) displays a genuine unmistakable aroma of fresh grapefruit juice”(1). Unfortunately the authors did not identify this potent aroma compound from fresh grapefruit, GF, but rather from 100 L of canned grapefruit juice, GFJ, after using the artifact inducing Likens-Nickerson extraction. Historically, most consumers have first experienced GFJ as a non-refrigerated, canned product. Canned GFJ is hot filled and undergoes flavor changes which produce an aroma profile very different from fresh squeezed juice. 1-PM-8-T was subsequently reported to be one of the most potent aroma compounds in “fresh, hand squeezed GFJ”(2). The “fresh” grapefruit came from a supermarket in Germany. However, fresh “from the tree” GFJ has no measurable 1-PM-8-T, but increasing concentrations can be detected in these juices after heating. Furthermore, using GC-sulfur selective pulsed flame photometry detection, PFPD, as well as GC-olfactometry we have identified 1-PM-8-T in thermally abused orange juice. Anaerobic conditions are required for optimal 1-p-menthen-8-thiol formation. In the presence of oxygen, the 2,8-epithiol is preferentially formed. To determine why 1-PM-8-T is found in supermarket fresh GF but not fresh from the tree GF, we measured

internal GF volatiles using a modified SPME procedure. The coating applied to supermarket GF to increase shelf life is a water (and oxygen) barrier. Under these conditions fruit respiration shifts to an anaerobic state and under these conditions 1-PM-8-T can be detected. References 1. Demole, E. and Enggist, P., Ohloff, G., 1-p-Menthene-8-thiol: A Powerful Flavor Impact Constituent of Grapefruit Juice (Citrus paradisi MACFAYDEN), Helvetica Chimica Acta, 1982, 65,1785. 2. Buettner, A. and Schieberle, P., Characterization of the Most Odor-Active Volatiles in Fresh, Hand-Squeezed Juice of Grapefruit (Citrus paradisi Macfayden), J. Agric. Food Chem. 1999, 47, 5189

AGFD 296

Chiral mono-terpene profile in Pinot Gris and Riesling wines determined by head phase-solid phase micro-extraction-multidimensional gas chromatography-mass spectrometry (HS-SPME-MDGC-MS)

Mei Song, Ying Xia, Elizabeth Tomasino, elizabeth.tomasino@oregonstate.edu. Oregon State University, Corvallis, Oregon, United States

Quantification of chiral mono-terpenes in white wine was achieved using head-space-solid phase micro-extraction-multidimensional gas chromatography-mass spectrometry (HS-SPME-MDGC-MS) with stable isotope dilution analysis. Fifteen chiral mono-terpenes were separated and quantified, including: (S)-(-)-limonene, (R)-(+)-limonene, (+)-(2R,4S)-cis-rose oxide, (-)-(2S,4R)-cis-rose oxide, (-)-(2R,4R)-trans-rose oxide, (+)-(2S,4S)-cis-rose oxide, furanoid (+)-trans-linalool oxide, furanoid (-)-cis-linalool oxide, furanoid (-)-trans-linalool oxide, furanoid (+)-cis-linalool oxide, (-)-linalool, (+)-linalool, (-)-α-terpineol, (+)-α-terpineol and (R)-(+)-ß-citronellol. It was found that sugar content of wine impacted measurement of these compounds, and different calibration curves were needed based on the target wines sugar content. The chiral mono-terpene profile was investigated in 10 Pinot Gris wines and 10 Riesling wines. Of the 15 chiral mono-terpene compounds, 12 were present in the wines. Results showed diverse chiral mono-terpene profiles in investigated wines, with significant differences on the concentration of each detected chiral compounds. It is possible that the differences in chiral mono-terpenes profile may be driving the varietal differences in white wines. Further study will be focused on if the enantiomeric ratios of each chiral compound and different residual sugar content will contribute to the varietal differences in more white wines.

AGFD 297

Understanding the effects of ethanol-flavor interactions on flavor perception in alcoholic beverages

Chelsea Ickes, homes2@illinois.edu, Keith R. Cadwallader. Dept. Food Sci. Human Nutr., University of Illinois, Urbana, Illinois, United States

Ethanol is the key component of alcoholic beverages. Aside from its obvious effects on the body, ethanol also plays an important role in the aromas of these beverages. Alcohol content varies greatly among the various types of alcoholic beverages. For example, the ABV (alcohol by volume) of beer and wine is around 3-12%, while distilled spirits contain around 40%. The interactions of ethanol with water as well as other matrix constituents such as sugars and dissolved solutes can have a significant effect on the perceived aromas of these beverages. Ethanol-water interactions have been well studied in the field of physical chemistry. A change in the alcohol concentration of a beverage has been shown to significantly affect the release of volatiles into the headspace of the product. This is common knowledge in the industry, as it is standard practice to dilute a distilled spirit to an ABV of 23% prior to evaluating its sensory (aroma) characteristics. Furthermore, the odor detection thresholds of aroma compounds are known to be affected by the alcohol content of the matrix. As alcohol strength changes some aroma attributes significantly increase in intensity while others undergo a significant decrease. Additionally, ethanol may create or disturb odor synergies or mask certain aroma attributes. Aside from impacting aroma release, ethanol at high concentrations can also cause alcohol burn to the oral and nasal cavities. This talk will present the current understanding of the effects of ethanol on flavor perception in alcoholic beverages and will discuss analytical approaches aimed at better understanding these effects.

AGFD 298

Enantiomeric analysis of volatile chiral compounds in ready-to-drink tea beverages during storage using multidimensional gas chromatography

Fei He3, fei.he@oregonstate.edu, YanPing L. Qian2, Michael C. Qian1. (1) Oregon State Univ, Corvallis, Oregon, United States (2) Crop and Soil, Oregon State University, Corvallis, Oregon, United States (3) Food Science and Technology, Oregon State University, Corvallis, Oregon, United States

Tea is one of the most widely consumed beverages in the world. Ready to drink (RTD) tea is a growing segment in the beverage market due to its improved flavor and health benefit. However, tea flavor is very complex due to tea variety and process conditions that affect both the flavor profiles and the shelf life. In this study, flavor and flavor stability were evaluated in two commercial RTD teas in an 8-week storage study at four different temperature conditions (room temperature, 35 °C, 40 °C, 45 °C). Solid-phase microextraction–gas chromatography-mass spectrometry (SPME-GC-MS) was performed to identify the aroma compounds and quantify their concentrations in fresh and aged tea samples, as well as to characterize the aroma changes during storage. The results showed geranial and neral decreased very quickly during storage, whereas citronellol and linalool decreased slowly. β-ionone had very little change during storage. The isomeric ratio changes of 6 pairs of volatile compounds (linalool, limonene, α-

terpineol, citral, citronellol, 4-terpineol) during 8 weeks storage were further investigated using multidimensional GC-MS (MDGC-MS). Storage temperature had a dramatic impact on the flavor stability, including the conversion of chiral isomers. For example, during 8 weeks storage, some of the (S)-limonene was converted to the (R)-limonene, a conversion of (R)-linalool to (S)-linalool was also observed during storage. Enantiomer purity ratios also varied significantly under different storage time and temperature for most of compounds. These results may aid in monitoring the quality and safety of tea beverages.

AGFD 299

Changes in the key aroma compounds of Shiitake and Oyster mushrooms induced by a thermal treatment

Philipp Cornelius Schmidberger, philipp.schmidberger@lrz.tum.de, Peter Schieberle. Chair of Food Chemistry, Technische Universität München, Freising, Germany

Due to their easy cultivation and their attractive aroma edible mushrooms are a popular food all over the world. However, knowledge on the modification of the overall aroma profile induced by thermal treatment is rather scarce. By application of the aroma extract dilution analysis (AEDA) on distillates prepared by extraction/SAFE-distillation from either unprocessed and processed Shiitake (Lentinula edodes) and Oyster mushrooms (Pleurotus ostreatus), respectively, the key aroma compounds could be characterized and differentiated into those biochemically formed and others generated thermally. Application of aroma extract dilution analysis (AEDA) on the volatile fraction isolated from pan-fried Shiitake mushrooms revealed 42 odor-active compounds in the flavor dilution (FD) factor range of 8-8192. Among them 3-hydroxy-4,5-dimethylfuran-2(5H)-on, 1,2,4,5-tetrathiane, 4-hydroxy-2,5-dimethylfuran-3(2H)-one, phenylacetic acid, 3-(methylthio)propanal and trans-4,5-epoxy-(E)-2-decenal showed the highest FD factors. In pan-fried Oyster mushrooms, 40 odor-active compounds were newly identified. In order to quantitate identified aroma compounds, stable isotope dilution analyses (SIDA) were performed. In the course of this, four sulfurous compounds were synthesized as isotopically labeled standards for the first time. Odor activity values (OAV) were calculated as ratio of their concentrations to their odor thresholds in water in order to identify key odor compounds. Quantitation experiments on the most important aroma-active compounds in raw and pan-fried Shiitake and Oyster mushrooms revealed considerable changes during thermal treatment.

AGFD 300

Molecular mechanisms of disease chemoprevention by polymethoxyflavones

Min-Hsiung Pan1, mhpan@ntu.edu.tw, Ching-Shu Lai1, Chih-Yu Lo3, Shiming Li2, Chi-Tang Ho2. (1) National Taiwan University, Taipei, Taiwan (2) Food Science, Rutgers

University, New Brunswick, New Jersey, United States (3) National Chiayi University, Chiayi, Taiwan

Chemoprevention been recognized as a promising strategy to prevent human disease by using natural compounds to prevent, block, inhibit, reverse, or retard the process of disease. Chemopreventive agents act through a variety of mechanisms to prevent and improve diseases, including anti-oxidative, anti-inflammation, anti-proliferation, anti-metastasis, anti- angiogenesis and induction of program cell death via modulation of signaling cascades, gene expressions and protein function. Polymethoxyflavone (PMFs) and hydroxyl PMFs (OH-PMFs) are a unique class of flavonoids that almost exclusively exists in citrus genus, especially in the peel. Moreover, OH-PMFs can be formed from their corresponding PMFs counterparts by hydrolysis during storage. Our studies have revealed both PMFs and OH-PMFs exhibit a broad spectrum of biological activity, such as inhibition of cancer cells growth, suppression of inflammatory mediators and skin carcinogenesis as well as colonic tumorigenesis, and inhibit adipogenesis in 3T3-L1 adipocytes and high fat diet-induced obesity. In addition, we found OH-PMFs exhibit epigenetics regulation properties. Understanding of the chemopreventive mechanisms of PMFs and OH-PMFs might promote further application for prevention and treatment of various human diseases.

AGFD 301

Lipid-lowering activity of citrus polymethoxylated flavones is mediated by down-regulation of lipogenic genes

Zhen-Yu Chen2, zhenyuchen@cuhk.edu.hk, Lin Lei1. (1) The Chinese University of HK, Hong Kong, Hong Kong (2) The Chinese University of Hong Kong, Hong Kong, China

We investigated the underlying mechanism of lipid-lowering activity of polymethoxylated flavones (PMF). Thirty-four male hamsters were divided into four groups and fed a non-cholesterol diet (NCD) or one of the three high cholesterol diets with addition of 0% PMF, 0.5% PMF (L-PMF), and 1.0% PMF (H-PMF), respectively, for 8 weeks. Results showed PMF could lower plasma triacylglycerols (TG) by 32-46% but it had no effect on plasma cholesterol in hamsters. This was accompanied by down-regulation of the mRNA of liver sterol regulatory element binding protein-1c (SREBP1c), fatty acid synthase (FAS), and peroxisome proliferator activated receptor alpha (PPARα) while up-regulation of liver lipoprotein lipase (LPL). In addition, PMF supplementation reduced the body weight gain and the relative weights of white adipose tissue pads by 23-39%, possibly by decreasing the gene expression of FAS and LPL in epididymal fat pad. It was concluded that TG-lowering activity of dietary PMF was mediated by down-regulation of genes involved in lipogenesis.

AGFD 302

Citrus polymethoxyflavones and monodemethylated polymethoxyflavones inhibit adipogenesis in 3T3-L1 adipocytes

Shu-Mei Lin2, Pei-Shiaun Chen2, Min-Hsiung Pan3, Shiming Li1, Chi-Tang Ho4, Chih Y. Lo2, chihyu27@hotmail.com. (1) Hubei Key Laboratory of Economic Forest Germplasm Improvement and Resources Comprehensive Utilization, Huanggang Normal University, Huanggang, Hubei, China (2) Food Science, National Chiayi University, Chiayi City, Taiwan (3) Institute of Food Science and Technology, National Taiwan University, Taipei, Taiwan (4) Food Science, Rutgers University, New Brunswick, New Jersey, United States

Obesity is considered as a low level chronic inflammation and its formation is found to increase the risk of metabolic syndrome. Polymethoxyflavones (PMFs) and monodemethylated polymethoxyflavones (OH-PMFs) exist exclusively in the citrus genus, particularly in citrus peels. Many studies showed that the anticarcinogenic activities of 5-OH-PMFs were greater than PMFs. Using 3T3-L1 adipocyte cell model, our study investigated the role of the PMFs and OH-PMFs on adipogenesis, differentiation-related transcription factor during differentiation and the secretion of adipokines. Based on the results, PMFs and OH-PMFs from citrus peels can be used as a natural resource for obesity prevention.

AGFD 303

Anti-adipogenesis effect of 5-demethylnobiletin and its acetylated derivative in 3T3-L1 preadipocyte model

YEN-CHEN TUNG3, emjine@gmail.com, Guor-Jien Wei4, Shiming Li5, Min-Hsiung Pan1, Chi-Tang Ho2. (1) National Taiwan University, Taipei, Taiwan (2) Food Science, Rutgers University, New Brunswick, New Jersey, United States (3) Food science, Rutgers University, Highland park, New Jersey, United States (4) Department of Nutrition and Health Sciences, Kainan University, Taoyuan, Taiwan (5) College of Life Sciences, Huanggang Normal University, Hubei, China

Obesity is an abnormal accumulation of fat as triacylglycerol in our body especially in adipose tissue. It is highly related with diabetes, cancers, cardiovascular diseases and inflammatory related diseases. Therefore, reducing the prevalence of obesity is an important issue. Nobiletin and 5-demethylnobiletin are polymethoxyflavonoid (PMF) and hydroxylated PMF, exclusively exist in the citrus peels. It has been demonstrated that nobiletin can suppress adipogenesis in vitro. Many recent studies showed that hydroxylated PMF had better biological activities than PMF. However, hydroxylated PMFs have poor aqueous solubility and low oral bioavailability. We used chemical modification to produce 5-acetylated nobiletin to improve its bioactive efficiency in vitro. In our researches, we demonstrated that 5-acetylated nobiletin had better anti-adipogenic activity than 5-demethylnobiletin via AMP-activated protein kinase (AMPK) pathway in 3T3-L1 preadipocyte model.

AGFD 304

Polymethoxyflavones from aged orange peels: Extraction, formulation, and bioefficacy

Qingrong Huang, qhuang@aesop.rutgers.edu. Food Science, Rutgers Univ, New Brunswick, New Jersey, United States

Aged orange peels (Chenpi) have long been used in traditional Chinese medicine to prevent inflammation related diseases. They are rich in essential oils and polymethoxyflavones (PMFs), the category of flavones containing two or more methoxy groups attached to the 15-carbon benzo- γ-pyrone skeleton structure with a carbonyl group on the C4 position. In this talk, PMFs were extracted by using either supercritical fluid or sub-critical fluid method. The profiles of PMFs were affected by both aging time and processing techniques. Due to the highly crystalline nature of PMFs, PMFs have poor solubility in water, which limits their broad applications in food. To address this issue, and also to improve the bioavailability of PMFs, PMFs from Chenpi extracts were formulated to form oil-in-water emulsions. Their digestion profiles and bioaccessibility were evaluated by using in vitro lipolysis. The prevention and treatment of obesity and Type II diabetes using Chenpi PMFs were also carried out by using the in vivo high-fat diet-induced obese and type II diabetic mice model.

AGFD 305

Molecular characterization of the interactions between polymethoxylflavones and kappa casein

Changchu Ma2, changchu@foodsci.umass.edu, Hang Xiao1, Lili He3. (1) Food Science, University of Massachusetts, Amherst, Massachusetts, United States (2) Food Science, University of Massachusetts,Amherst, Amherst, Massachusetts, United States (3) Food Science, University of Massachusetts, Amherst, Amherst, Massachusetts, United States

The molecular interactions between flavonoids and proteins play important roles in the biological fate and bioactivities of flavonoids. The objective of this study is to investigate molecular interactions between polymethoxyflavones (PMFs) and kappa casein. Two PMFs, nobiletin (NBT) and 5,3′,4′-tridemethylnobiletin (TDN), and two states of protein, native and thermal denatured, were used in this study. Surface enhanced Raman spectroscopy (SERS) and fluorescence spectroscopy were used to characterize and quantify the interactions, respectively. The SERS study shows that TDN has stronger binding to casein than NBT. SERS peak shifts assigned as OH bend in position C5 were observed when TDN interacting with casein. Additionaly, increased binding with thermal denatured casein(by heating at 95°C for 30 minutes) was observed for NBT. TDN, however, behaved in an opposite way, in which its binding affinity to thermal denatured casein decreased. Binding was also quantified by tryptophan quenching fluorescence spectroscopy, and quenching constant (Ksv) is calculated by the Stern–Volmer equation. The Ksv of NBT with casein before and after heating are 0.0478 and 0.0522 µM-1, while the Ksv of TDN with casein before and after heating are 0.1012 and 0.0592µM-1. Our results suggest that hydroxyl groups on PMFs played a key role in the

interactions between PMFs and casein. NBT with no hydroxyl group interacts with kappa casein through hydrophobic interaction. Therefore, after casein denatured, increased interaction was observed due to the increased hydrophobic interaction. TDN with three hydroxyl groups interacts with kappa casein through hydrogen bonding and hydrophobic interaction, and hydrogen binding is the driving force. While heating destroyed the hydrogen binding sites of proteins with TDN, its interaction with denatured protein decreased. This study demonstrates the significance of the flavonoid structure and protein status on the flanoids-protein interactions.

AGFD 306

Gastrointestinal biotransformation enhances biological effects of polymethoxyflavones

MInqi Wang, Mingyue Song, Xian Wu, Zili Gao, Fei Xu, Yong Cao, Hang Xiao, hangxiao@foodsci.umass.edu. Food Science, University of Massachusetts, Amherst, Massachusetts, United States

Accumulating evidence indicated the important role of biotransformation in influencing biological effects of food components. We investigated the gastrointestinal biotransformation pathways of major polymethoxyflavones in rodents. In general, the major sites of biotransformation for polymethoxyflavones are small intestine and colon. In the small intestine, polymethoxyflavones undergo extensive phase I metabolism to produce multiple demethylated metabolites. These phase I metabolites further undergo phase II metabolism to produce sulfate and/or glucuronide conjugates. Therefore, the majority of polymethoxyflavones exist as phase II conjugates in the small intestine. However, in the colon, the phase II conjugates are converted back to phase I metabolites by the action of gut microbiome, thus only phase I metabolites can be found in the colon. Most interestingly, our results demonstrated that phase I metabolites of polymethoxyflavones have potent biological effects such as anti-inflammation, anti-proliferation, pro-apoptosis, anti-angiogenesis, anti-oxidation, etc. and these effects are even stronger than those produced by their parent polymethoxyflavones. Overall, our findings indicated that GI biotransformation of polymethoxyflavones results in high levels of phase I metabolites in the colon that may elicit enhanced biological effects.

AGFD 307

Metabolite profiling of barley grain: Impact of induced drought stress

Karl-Heinz Engel1, K.H.Engel@wzw.tum.de, Alexandra Lanzinger1, Thomas Frank1, Gabriela Reichenberger2, Markus Herz2. (1) Technische Universitaet Muenchen, Freising, Germany (2) Bavarian State Research Center for Agriculture, Freising, Germany

Barley genotypes were field-grown in three consecutive seasons under normal weather conditions and under induced drought stress, using a Rain-Out-Shelter. Samples were

subjected to a gas chromatography-mass spectrometry based metabolite profiling, allowing the analysis of a broad spectrum of low molecular weight metabolites. The comparative assessment of the data by means of multivariate and univariate approaches demonstrated that there are grain constituents which are significantly influenced, either consistently increased or consistently decreased, by drought stress. In addition, the results indicate that free amino acids may serve as potential markers for barley genotypes differently adapted to drought stress.

AGFD 308

Effect of growing environment on the characteristics of soybeans for food uses

Sam K. Chang, schang@fsnhp.msstate.edu, Shi Meng. Dept. Food Science, Nutrition and Health Prom, Mississippi State University, Mississippi State, Mississippi, United States

Soybeans have been used for thousands of years for human food consumption in Asia, even though world production of soybean is mostly for oil and feed use. The protein meal after oil removal also is used for functional ingredients incorporated in numerous foods. Since the FDA’s health claim in 1999 that soy protein may reduce cardiovascular disease, global utilization of soybean and soy protein products has increased tremendously. Recent understanding of isoflavones and other bioactive compounds in soybean further strengthens the potential of soybean and soy-based products as a health food. Soybean cultivars, containing high protein content and desirable protein subunits have been developed for food uses. However, soybean protein and various subunits are affected by planting location and year, which has significant impact on chemical composition due to different climate and soil conditions. We have tested the effect of two locations (Stoneville, Mississippi, and Puerto Rico) on seed quality, seed size, and protein content and subunit composition of 22 accessions (plant introductions) of soybean from the USDA National Soybean Germplasm Collection Center. The results showed significant effects of location on the characteristics of individual soybeans in terms of seed quality, seed size, protein content and protein subunit compositions. In addition, our and other research also showed that isoflavones and phenolic composition and antioxidant capacity were affected by cropping year and location. Selection of soybean varieties that perform consistently in various locations is important for food application.

AGFD 309

Growing conditions affect flavonoid concentration and yield in American skullcap (Scutellaria lateriflora)

Dennis A. Shannon1, shannda@auburn.edu, Arsene Similien5, Agnes M. Rimando2, Edzard van Santen1, C. W. Wood3, Nirmal Joshee4, Barbara W. Kemppainen6. (1) Crop, Soil and Environmental Sciences, Auburn University, Auburn, Alabama, United States (2) USDA ARS, University, Mississippi, United States (3) West Florida Research and

Education Center, University of Florida, Jay, Florida, United States (4) Agricultural Research Station, Fort Valley State University, Fort Valley, Georgia, United States (5) Agricultural Division, SEED Ministries, Les Cayes, Haiti (6) Anatomy, Physiology and Pharmacology, Auburn University, Auburn, Alabama, United States

American skullcap (Scutellaria lateriflora L.) is a plant species native to the Eastern United States that has sedative properties associated with flavonoids. Information on how growing conditions affect flavonoid content was lacking. A 2x2x3 factorial experiment was conducted in a randomized complete block design (r = 4) with a split plot restriction on randomization to explore effects of light, irrigation and nutrient application on flavonoid concentration and yield in American skullcap. Treatment factors were shade (40% shade vs. no shade), irrigation (applied at 30 kPa vs. no irrigation) and nutrients (no fertilizer vs. fertilizer (100 kg N, 68 kg P, 42 kg K ha-1) and chicken litter (100 kg N, 50 kg P, 123 kg K ha-1). Shade formed the main plot units; irrigation and nutrients were randomized within subplots. Above-ground biomass was harvested twice in both 2007 and 2008. Dried, finely ground whole plant samples were extracted and analyzed for flavonoid concentration using HPLC. The flavonoid baicalin was present in highest concentration and yield, followed by baicalein; wogonin and chrysin were present in only small amounts. Flavonoid concentration was 26% higher in full sun than under shade, 20 % higher with irrigation than without, and 29% lower with added nutrients. Significant interactions of shade x irrigation and shade x nutrients were observed. Flavonoid yield was 26% higher under shade, 97% higher with irrigation and 44% higher with added nutrients. Significant interactions of shade x irrigation, shade x nutrients and irrigation x nutrients were also observed. Highest concentrations of baicalin and total flavonoids measured were obtained by growing skullcap in full sun with irrigation with or without manure. Highest flavonoid yields were obtained under shade with irrigation and manure or fertilizer. This research demonstrates the importance of growing conditions on the bioactivity of medicinal plants.

AGFD 310

Chemical characterization of pigments in three guava (Psidium guajava) Colombian varieties

Ivonne Alejandra González2, Antonio Melendez1, Francisco J. Heredia1, Coralia Osorio Roa2, cosorior@unal.edu.co. (1) Department of Nutrition & Food Science. Facultad de Farmacia, University of Seville, Sevilla, Spain (2) Departamento de Química, Universidad Nacional de Colombia, Bogota, Colombia

Guava (Psidium guajava L.) is a very nutritious and tasty tropical fruit. It has a high content of pectin, dietary fiber, minerals, essential aminoacids, and vitamin C. In Colombia, there are three commercial varieties commonly known as “Regional roja” (pink-fleshed pulp), “Regional blanca” (white-fleshed pulp), and “guayaba-pera” (Palmira ICA-1). The last one is a genetically improved variety that is technically produced in hot and humid climates. The fruit is pear-shaped and relatively large with a weight ranging from 100 to 130 g, with a deep pink flesh and low content of seeds. In contrast, the

other two varieties are round-shaped, with a 100 g average weight and stone-like seeds inside. They are produced in mountainous regions with average temperature of 24°C. Thus, the pigment composition (carotenoid and chlorophylls) of the three Colombian guava varieties, in unripe, turning and full ripe maturity stages was studied. Lycopene was identified as the major carotenoid of Regional roja and Palmira ICA-1 (18.25 and 28.07 mg kg-1 fruit, respectively in full ripe fruits) although a small amount of lutein was also found in the two varieties. b-Carotene was only found in Palmira ICA-1, whilst no detectable levels of carotenoids were found in the white-fleshed guava (Regional blanca). A good correlation between the total carotenoid content and the colour parameters of guava peel was found. Financial support: Ministerio de Agricultura y Desarrollo Rural de Colombia.

AGFD 311

Fresh ginger vs. dry ginger: The impact of temperature on the bioactive components in ginger

Shengmin Sang, ssang@ncat.edu. Center for Excellence in Post-Harvest Technol, North Carolina AT State University, Kannapolis, North Carolina, United States

Ginger (Zingiber officinale Rosc.) has received extensive attention as a botanical dietary supplement in the USA and Europe due to its antioxidative, anti-inflammatory, and anti-cancer activities. According to the practice of Traditional Chinese Medicine, fresh ginger and dried ginger have different beneficial functions. The secret is the major components in fresh ginger, gingerols, are unstable during thermo process to generate the dehydrated products, shogaols. We and others have reported that gingerols and shogaols are so different from each other in terms of biotransformation, bioavailability and biological functions. This presentation will cover the impact of temperature on the chemical changes of ginger and the differences between gingerols and shogaols in terms of their biotransformation and biological functions.

AGFD 312

Differentiating organic and conventional oregano using ultraperformance liquid chromatography mass spectrometry (UPLC-MS), headspace gas chromatography with flame ionization detection (headspace-GC-FID), and flow injection mass spectrum (FIMS) fingerprints combined with multivariate date analysis

Boyan Gao2,1, raphaelgao1985@gmail.com, Weiying Lu1, Liangli L. Yu2,1. (1) Department of Agriculture and Biotechnology, Shanghai Jiaotong University, Shanghai, China (2) Univ of Maryland, College Park, Maryland, United States

Chromatographic and mass spectrometry flow injection fingerprints were combined with multivariate data analysis to elucidate the chemical composition of both volatile compounds and nonvolatile compounds in oregano samples. The combination of these technologies is also applied to differentiate organic grown oregano from their

conventional counterparts. The results indicated that headspace-GC-MS could effectively identify and quantify the volatile compounds in oregano samples without any sample pre-treatment; UPLC-MS might provide the chemical profile of nonvolatile compounds in oregano; FIMS provided a fast test and could be potentially used for high-throughput examinations of oregano. These technologies can be used together as an effective analytical method in clarifying the composition of volatile and nonvolatile components in botanical samples, as well as differentiating plant samples with different chemical composition or concentration.

AGFD 313

Detection of Escherichia coli in drinking water using T7 bacteriophage-conjugated magnetic probe

Juhong Chen4, juhong@foodsci.umass.edu, Ziwen Jiang5, Samuel D. Alcaine2, Vincent M. Rotello1, Sam R. Nugen3. (1) Univ of Massachusetts, Amherst, Massachusetts, United States (2) Food Science, University of Massachusetts, Amherst, Massachusetts, United States (3) 246 Chenoweth Laboratory, University of Massachusetts, Amherst, Massachusetts, United States (4) Food Science, University of Massachusetts, Amherst, Amherst, Massachusetts, United States (5) Chemistry, University of Massachusetts, Amherst, Amherst, Massachusetts, United States

In this study, bacteriophage (phage) immobilized on magnetic beads was introduced to detect bacteria cells in drinking water. T7 phage is a lytic phage with a high specificity to E. coli. The T7 bacteriophage-conjugated magnetic beads were able to specifically bind to and separate E. coli BL21 from drinking water using external magnetic field. Meanwhile, the phage was able to infect E. coli resulting in lysis and the release of β-galactosidase (β-gal) from bacterial cells into a buffer solution. The released β-gal was able to initiate a colorimetric reaction, which was quantified using UV-Vis spectrophotometer. Using this strategy, we have been able to detect bacteria cells at concentration of 1 × 104 CFU/mL within 2.5 hours. Under such experimental condition, 10 CFU/mL bacteria cells in drinking water could be detected after 6 hours of pre-enrichment in Luria Bertani (LB) broth with isopropyl β-D-thiogalactopyranoside (IPTG). The colorimetric results can also be determined visually, allowing for the rapid detection of bacteria in resource limited settings.

Schematic representation of detection of E. coli in drinking water using T7 bacteriophage-conjugated magnetic probe.

(a) photographs of bacteria detection at different concentrations before and after magnetic separation. (b) photograph of colorimetric response and (c) absorbance intensity upon different bacteria concentrations.

AGFD 314

Electrospun water soluble nanofibers for dehydration and storage of bacteriophage for decontamination of agricultural water

Charmaine Koo2, ckoo@foodsci.umass.edu, Sam R. Nugen1. (1) 246 Chenoweth Laboratory, University of Massachusetts, Amherst, Massachusetts, United States (2) Food Science, University of Massachusetts, Amherst, Massachusetts, United States

Contaminated agricultural water due to farm run-off for irrigation is an ongoing health issue as polluted produce puts consumers at risk for foodborne illnesses. One potential method to decontaminate irrigation water is the addition of bacteriophage, which is

capable of infecting and lysing target bacterial cells, which effectively kills the pathogen. Most bacteriophage can be kept viable by lyophilization, but freeze drying is an expensive, high energy and time consuming process. Conversely, electrospinning is a dehydration process where a potential is used to draw nano-sized fibers from a liquid. This can rapidly dehydrate solutions and effectively preserve the bacteriophage. Thus, we have investigated electrospun water-soluble nanofibers as a more efficient, economical, rapid and scalable process for the dehydration of bacteriophage in solution. Polyvinylpyrrolidone, a water soluble polymer, was used for electrospinning because it is certified as “generally regarded as safe” (GRAS) which is required for use in agricultural water. A storage study showed the effective use of storage media buffers containing sugars and salts which act as protectants during spinning and storage. The loss of bacteriophage infectivity was determined immediately following electrospinning and during storage using agar overlay plating and plaque counting. Furthermore, the morphology and size of the nanofibers were observed under scanning electron microscopy. With the ability to maintain a considerable amount of viable bacteriophage after electrospinning relative to lyophilization, electrospinning could be a potential method for the preservation and storage of bacteriophage.

Figure 1. Scanning electron micrographs of dehydrated bacteriophage in water-soluble electrospun nanofibers spun with a) Polyvinylpyrrolidone in deoionzed water b) Polyvinylpyrrolidone in storage media buffer with sucrose.

AGFD 315

Pectin homogalacturonans: Nanostructural characterization of methylesterified domains

Randall G. Cameron2, randall.cameron@ars.usda.gov, Yang Kim3, Ashley Galant4, Gary Luzio2, Jason Tzen1. (1) National Chung-Hsing University, Taichung, Taiwan (2) US Dept of Agriculture, Fort Pierce, Florida, United States (3) Food Science & Technology and Carbohydrate Bioproduct Research Center, Sejong University, Seoul, Korea (the Republic of) (4) Busch Agrucultural Resources, LLC, Moorhead, Minnesota, United States

Functionality of pectic hydrocolloids is largely dependent on the two major domains commonly found in their homogalacturonan regions, i.e., methylester protected domains (MPDs) and non methylesterified domains (NMDs). MPDs can participate in hydrogen bonding and hydrophobic interactions but unlike the NMDs they are incapable of calcium mediated cross-linking. MPDs also are inaccessible to endo polygalacturonase attack while NMDs are susceptible. We isolated and characterized MPDs contained within homogalacturonans that had been demethylesterified either by plant pectin methylesterases or chemically by base saponification. Molecular weight (~ 9500 – 14,000 Da) and degree of methylesterification (DM; ~ 63% – 94%) of remnant MPDs were measured. Subsequently the MPDs were base saponified at 4 °C. The resulting demethylesterified oligomers were separated, visualized and quantified. Population parameters estimated were the average number of MPDs per pectin molecule, the sum of galacturonic acid units in MPDs per molecule, the average MPD size and the number of “average” size MPDs per molecule. Analysis of variance and Tukey’s Multiple Comparison Test indicated significant differences in MPDs based on DM and the method of demethylesterification. Pearson’s correlation coefficients were calculated for MPD parameters, previously published structural data from NMDs and rheological properties of the demethylesterified homogalacturonans from the enzyme treatments. Significant correlations were observed for DM, absolute degree of blockiness, G′, NMD and MPD parameters. When coupled with methods to characterize NMDs these techniques provide an opportunity to statistically describe both highly charged and more neutral domains in a population of pectin molecules.

AGFD 316

In-product anti-counterfeiting agrochemicals using phase change nanoparticles

Miao Wang, wang.mia@husky.neu.edu, Ming Su, m.su@neu.edu. Northeastern University, Boston, Massachusetts, United States

This research describes a new nanoparticles-based in-product barcode system, in which a panel of phase change nanoparticles with discrete and sharp melting peaks is added as a barcode into in a variety of agrochemicals. Environmentally degradable organic solids are used to make nanoparticles. The barcode can be readout by detecting melting peaks of nanoparticles using differential scanning calorimetry. This method has high labeling capacity due to small sizes of nanoparticles, sharp melting peaks, and large scan range of thermal analysis. The in-product barcode can be effectively used to protect agrochemical products from being counterfeited due to its large coding capacity, technical readiness, covertness, and robustness.

Phase change nanoparticles are used as thermal barcode for argochemicals.

AGFD 317

Functional diet ginger (Zingiber officinale Roscoe, Zingiberaceae)

Hui-Min Wang, davidw@kmu.edu.tw. Kaohsiung Medical University, Kaohsiung, Taiwan

Ginger, the rhizomes of the herb Zingiber officinale Roscoe (Zingiberaceae), is a perennial herb and is used widely as a spice throughout the world. There are many species of ginger, mainly cultivated in tropical countries of Asia. We published that biologically active compounds found in diet ginger has been shown to have a number of pharmacological activities including anti-oxidant, anti-pigmentation, anti-cancer, anti-bacterial and to reduce the metabolic syndrome. It has important economic value and a wide range of pharmacological effects. Keywords: Zingiber officinale Roscoe, anti-oxidant, anti-pigmentation, anti-cancer, anti-bacterial, metabolic syndrome

AGFD 318

Is our salad safe? Efficacy of disinfection techniques to decontaminate spinach leaves and reduce cross-contamination

Nichola Kinsinger1, nkins001@gmail.com, Sharon L. Walker2. (1) University of California Riverside, Riverside, California, United States (2) University of California, Riverside, Riverside, California, United States

Each year bacterial contamination impacts consumers, causing food related bacterial outbreaks endangering countless lives and often resulting in death. The Center for Disease Control estimates that 1 in 6 people become ill annually and 3,000 peopled die from foodborne diseases. Cross-contamination from 0.1% of produce accounts for 99% of illnesses due to ineffective disinfection rinses used post-harvest. There have been several recent outbreaks of Escherichia coli O157:H7in leafy green products such as prepackage salad and baby spinach. The attachment and detachment kinetics of E. coli from both whole baby spinach leaves and isolated epicuticle layers of the spinach is investigated using a parallel plate flow chamber. Specifically, mass transfer rate coefficients for both attachment and detachment from the leaves or leaf epicuticle layers are being determined as a function of a range of relevant environmental parameters. The impact of water chemistry and common disinfection rinses on the removal and inactivation of E. coli on spinach leaves is also investigated. The spinach leaf texture has been incorporated within a COMSOL model to evaluate disinfectant concentration gradients that may limit rinsing efficacy and result in dangerous foodborne outbreaks. Optimization of TiO2 photocatalyst is evaluated as a supplemental and alternative

technology to reduce cross-contamination during produce rinsing. This project is at the food safety-water nexus, where understanding the contribution of water chemistry will ensure proper food safety through irrigation, harvesting and processing.

AGFD 319

Authentic milk powder variance study and detection of melamine adulteration using Raman spectroscopy and chemometrics

Sanjeewa R. Karunathilaka1, SanjeewaRasikaK.RanasinghePathirajage@fda.hhs.gov, Samantha Farris1, Magdi Mossoba2, Betsy J. Yakes3. (1) U.S Food and Drug Administration, Center for Food Safety and Applied Nutrition, U.S Food and Drug Administration, Center for Food Safety and Applied Nutrition, Collage Park, Maryland, United States (2) U.S Food and Drug Administration, Center for Food Safety and Applied Nutrition, Collage Park, Maryland, United States

The urgent need to develop rapid targeted and/or non-targeted tools to screen milk powder for economically motivated adulteration was emphasized by the tragic 2008 melamine adulteration incident involving milk powders.1,2 The United States Pharmacopeia (USP) has led a collaborative team that is currently investigating Raman spectroscopy as one of the potential analytical methods for the detection of adulterants in milk powders with focus on melamine. An understanding of the physiochemical variability within authentic skim milk powder (SMP) and non-fat dry milk (NFDM) is key, prior to the development of targeted and/or non-targeted classification models. The observed spectral data were interpreted by principal component analysis (PCA). The possible sources of variance were explored by the trends in principal component (PC) scores and the spectral features in PC loading plots. PC calibration models were built with a set of 40 SMP samples for the variance study. Interpretation of the PC scores plots revealed no clear trends for various milk powder categories, including days of analysis, SMP vs NFDM, supplier, and condensing temperatures, except for four samples from a single manufacturer being separated from the rest. The elucidation of PC loadings revealed the chemical source of variance resulting from fat, lactose, and proteins. A PC calibration model was built with a set of 40 SMP samples for the development of a targeted classification model for melamine adulteration. Milk powder samples spiked with 0.1-2 % by weight of melamine were then projected into the calibration model. Based on the 95 % confidence limit for calibration Q residuals, samples spiked with melamine at 0.25 % or higher could be classified correctly. Extended work with wet-blended samples is underway. References: Botros, L. L.; Jablonski, J.; Chang, C.; Bergana, M.M.; Wehling, P.; Harnly, J. M.; Downey, G.; Harrington, P.; Potts, A. R.; Moore, J. C. Exploring authentic skim and nonfat dry milk powder variance for the development of nontargeted adulterant detection methods using near-infrared spectroscopy and chemometrics. J. Agric .Food Chem. 2013, 61, 9810-9818.

Scholl, P. F.; Farris. S. M.; Mossoba, M. M.; Rapid turbidimetric detection of milk powder adulteration with plant proteins. J. Agric .Food Chem. 2014, 62, 1498-1505.

AGFD 320

Thermal dependence of riboflavin photodegradation in amorphous sucrose matrices

Yan L. Wang, yylacm@gmail.com, Maria Corradini, Richard D. Ludescher. Food Science, Rutgers University, New Brunswick, New Jersey, United States

Our previous research identified the sensitivity of riboflavin phosphorescence towards temperature induced molecular mobility changes in amorphous sucrose films, which suggests the potential application of riboflavin as a GRAS optical probe for molecular mobility. Additionally, continuous monitoring of riboflavin phosphorescence during heating and cooling cycles revealed differences in the delayed luminescence emission spectra, likely due to a higher rate of irreversible photodegradation of riboflavin at high temperatures. If properly characterized, the thermal dependence of riboblavin photodegradation can potentially be operationalized in sensors for temperature abuse. The objective of this study was to characterize the photodegradation kinetics of riboflavin in solid sucrose films under different temperature profiles using by phosphorescence spectroscopy. The amorphous films were prepared so that a molar ratio of riboflavin to sucrose was equal to or smaller than 1:104 to avoid probe-probe interaction. Delayed fluorescence emission spectra were recorded and fitted with a log-normal function. The maximum emission intensity (Imax) was estimated from the model. Under a constant temperature, the physical properties of the matrix and the quantum yield of delayed fluorescence remained constant. Therefore, the progressive change in Imax due to photoexcitation can be assumed to be solely due to riboflavin degradation. The photodegradation rate at each temperature was calculated from the relationship between maximum emission peak intensity (Imax) and number of flashes. Imax decreased in an exponential manner as a function of the number of flashes. The estimated photodegradation rates were 1x10-4, 3x10-4, 4x10-4, 1x 10-3, and 2x10-3 flash-1 at 0, 20, 40, 60, and 80˚C, respectively. The temperature dependence of the photodegradation rate showed a biphasic pattern, increasing gradually from 0 to 40˚C and dramatically from 40 to 80˚C. Implications of these findings on the applicability of phosphorescence from riboflavin in solid matrices as a sensor for temperature abuse will be discussed.

AGFD 321

Investigating the potato's defensive shield: Metabolites profiling and solid-state NMR compositional analysis of suberin-enriched wound-healing tissues

Keyvan Dastmalchi1, drk1dast@yahoo.com, Linda R. Kallash1, Van C. Phan2, Wenlin Huang1, Olga Serra3, Ruth Stark1. (1) Department of Chemistry, The City College of New York, New York, New York, United States (2) Department of Natural Sciences, Hostos Community College, Bronx, New York, United States (3) Departament de Biologia, Facultat de Ciències, University of Girona, Girona, Spain

Potato (Solanum tuberosum L.) is a worldwide food staple, but substantial waste accompanies its cultivation due to mechanical damage and suboptimal healing conditions. Motivated by both economic and nutritional considerations, metabolite profiling of wound tissue extracts and quantitative 13C NMR of solid residues aims to improve understanding of wound periderm formation. In the current investigation, polar and nonpolar extracts of wound periderm tissues from four potato cultivars with contrasting skin russeting patterns (Atlantic, Chipeta, Norkotah Russet, and Yukon Gold) were profiled at day-3 and day-7 stages of wound-induced periderm development. The materials were assessed using LC-MS, TOF-MS, GC-MS and NMR spectroscopic methods. Multivariate analyses revealed that distinctions among the cultivars' metabolite profiles were less pronounced at day 7 compared to day 3 post wounding. Biomarkers for cultivar type and wound healing time point included polyphenolic amines, flavonoid glycosides, phenolic acids, glycoalkaloids, fatty acids, α,ω-fatty diacids, fatty alcohols, alkanes and glycerol esters. In the polar extracts, polyphenolic amines are markers at day 3 for the russeted Norkotah Russet and Atlantic cultivars, whereas this marker class appears only at later stages for the smoother Yukon Gold and Chipeta varieties. The abundant long-chain fatty acids in nonpolar extracts and solids from the smooth-skinned Yukon Gold cultivar suggested accelerated suberization; this hypothesis was supported by high proportions of arenes, alkenes, and carbonyl groups in the solid and among the polar biomarkers. The absence of many marker classes in nonpolar extracts and interfacial Atlantic solids suggested slower healing.

AGFD 322

Neuroprotective effect of heptamethoxyflavone in the mouse brain

Satoshi Okuyama1, sokuyama@cc.matsuyama-u.ac.jp, Yoshiaki Amakura2, Morio Yoshimura2, Takashi Yoshida2, Atsushi Sawamoto1, Mitsunari Nakajima1, Yoshiko Furukawa1. (1) Department of Pharmaceutical Pharmacology, College of Pharmaceutical Sciences, Matsuyama University, Matsuyama, Ehime, Japan (2) Department of Pharmacognosy, College of Pharmaceutical Sciences, Matsuyama University, Matsuyama, Ehime, Japan

We have recently revealed that 3,5,6,7,8,3',4'-heptamethoxyflavone (HMF), one of the citrus flavonoids, has neuroprotective abilities in the central nervous system (CNS). HMF exerts a protective effect against neuronal dysfunction in brain ischemia. To generate an ischemic animal model, mice were subjected to bilateral common carotid artery occlusion (2VO) for 12 min. To investigate the effect of HMF on brain function, we analyzed the hippocampus by immunohistochemical methods and Western blotting.

Hippocampus was chosen as the region of study because it deals with memory formation and is one of the regions vulnerable to global brain ischemia, resulting in cognitive dysfunction. HMF was continuously administered for 5 days before and for 3 days following the surgery, HMF: 1) induced the phosphorylation (activation) of extracellular signal-regulated kinases (ERK) and cAMP response element-binding protein (CREB), and 2) enhanced the expression of brain-derived neurotrophic factor (BDNF), a representative neurotrophic factor in the hippocampus. These results suggested that HMF has the ability to accelerate ischemia-induced BDNF production in the hippocampus after brain ischemia, and this may be mediated by activation of ERK and CREB. ERK is a component of the mitogen-activated protein kinase signaling cascade, and CREB is a downstream transcription factor of activated ERK; in addition, CREB phosphorylation appears to be a critical step in the signaling cascade that is involved in BDNF synthesis, synaptic plasticity and memory function in the CNS. When HMF was continuously administered for 3 days immediately after the 2VO surgery, HMF: 1) rescued neuronal cell death in the hippocampus, 2) increased the production of BDNF, and 3) suppressed the activation of microglia, whose activation has been shown to significantly aggravate neurological deficit scores for ischemic mice. In the Y-maze test, HMF showed protection against ischemia-induced short term memory dysfunction. These findings suggested that HMF has a neuroprotective effect following brain ischemia that is induced by BDNF production and anti-inflammatory effects. Although the detailed mechanisms of HMF still remain unclear, our findings present the possibility that the citrus compound HMF might be beneficial as a neuroprotective agent in neurological disorders.

AGFD 323

Citrus polymethoxyflavones preventing the development of Alzheimer’s disease by regulating Aβ metabolism

LIMIN GUO1,2, guolm_xj@163.com, Lei Wang1,3, Wensheng Zhang1,3, zws@bnu.edu.cn, Hang Li1,3, Shiming Li4. (1) State Key Laboratory of Earth Surface Processes and Resource Ecology, Beijing Normal University, Beijing, China (2) Institute of Agro-products Storage and Processing, Xinjiang Academy of Agricultural Sciences, Urumqi, Xinjiang, China (3) Beijing Area Major Laboratory of Protection and Utilization of Chinese Medicine Resources, Beijing Normal University, Beijing, China (4) Food Science, Rutgers University, New Brunswick, New Jersey, United States

Alzheimer`s Disease (AD) is a progressive degenerative dementia particularly in elder people. There are estimated 6.25% of population worldwide with the symptoms of AD and the number of AD patients keeps growing. Because of the increasing lifespan and the severe detrimental effects of AD on human health, it is currently one of the most important study areas to prevent AD development and to improve the living conditions of AD patients. The Aβ cascade hypothesis states that the AD starts with the loss of brain neurons caused by Aβ aggregation and plaque deposition in the brain. The accumulation of Aβ, resulting from its metabolic disorders, is the core pathogenesis of AD. Based on this mechanism, we have developed a neuron cell line which can

produce a stable overexpression of β-amyloid precursor protein (APP) and investigated the regulation of Aβ metabolism by citrus polymethoxyflavones. Our results have shown that comparing with EGCG from green tea and theaflavins from black tea , citrus polymethoxyflavones could regulate key metabolic enzyme activity more effectively, and also reduced the intracellular Aβ levels, indicating that citrus polymethoxyflavones have the potential to delay the onset and to slow the progression of AD.

AGFD 324

5-Demethylnobiletin synergistically enhances the anticancer activity of paclitaxel in non-small cell lung carcinoma (NSCLC)

Chi-Chen Lin1,2, lincc@dragon.nchu.edu.tw, Shiming Li3, Chi-Tang Ho4. (1) Institute of Biomedical Science and Rong Hsing Research Center for Translational Medicine, National Chung-Hsing University, Taichung, Taiwan (2) Department of Medical Research and Education, Taichung Veterans General Hospital, Taichung, Taiwan (3) College of Life Sciences, Huanggang Normal University, Huanggang, China (4) Food Science, Rutgers University, New Brunswick, New Jersey, United States

The discovery and development of novel antitumor drugs for non-small cell lung carcinoma (NSCLC) treatment is urgent in order to improve the effectiveness of lung cancer therapy and prognosis. In the current study, we investigated the antitumor activity of 5-demethylnobiletin and its combination with paclitaxel in human CL1-5 lung cancer cells and in vivo model of lung cancer. The combination index method was used to examine the interaction of 5-demethylnobiletin and paclitaxel. Data were correlated with alternation in cell apoptosis and cell cycle arrest. The effect of the drugs on microtubule tubulin polymerization and activation of caspases were also studied. Furthermore, CL1-5 xenograft-bearing mice were treated with either 5-demethylnobiletin, paclitaxel or a combination of both given concurrently to evaluate the antitumor efficacy of this combination. The results revealed that 5-demethylnobiletin and paclitaxel interact synergistically and caused cancer cell growth, apoptosis and G2/M cell cycle arrest in CL1-5 cells. In addition, the combination of 5-demethylnobiletin with paclitaxel triggered microtubule distortion and polymerization dramatically as compared to single drug treatment. In CL1-5 xenograft bearing treated mice, the combination is significantly superior to any single agent. Immunohistochemistry analysis of cacaspe-3 show increased apoptosis in mice treated with the combination relative to either drug alone. In conclusion, results of the current study demonstrates that the potency of 5-demethylnobiletin combining with concurrent low dose of paclitaxel for clinical NSCLC therapeutics.

AGFD 325

5-Acetyloxy-6,7,8,4'-tetramethoxyflavone, a tangeretin derivative, inhibits cell growth in human prostate cancer PC-3 cells

Yu-Kuo Chen2, ykchen@rci.rutgers.edu, Yin-Bo Chen2, Jia-Lin Guo2, Tzou-Chi Huang2,3, Shiming Li4,5, Chi-Tang Ho1. (1) Food Science, Rutgers University, New Brunswick, New Jersey, United States (2) Department of Food Science, National Pingtung University of Science and Technology, Pingtung, Taiwan (3) Department of Biological Science and Technology, National Pingtung University of Science and Technology, Pingtung, Taiwan (4) Department of Food Science, Rutgers University, New Brunswick, New Jersey, United States (5) Hubei Key Laboratory of Economic Forest Germplasm Improvement and Resources Comprehensive Utilization, Huanggang Normal University, Huanggang, Hubei, China

In the present study, we investigated the growth inhibitory effect of 5-acetyloxy-6,7,8,4'-tetramethoxyflavone (5-ATAN), an acetylated derivative of tangeretin (TAN), on human prostate cancer PC-3 cells. The results showed that 5-ATAN revealed stronger cytotoxic effects than that of TAN on cell viability of PC-3 cells. After 48 h treatment, the IC50 of 5-ATAN and TAN were determined as 5.2 and 25.9 µM, respectively. Moreover, treatment of 5-ATAN increased the sub-G1 phase population and induced G2/M phase arrest in the cell cycle of PC-3 cells. We also found that 5-ATAN triggered the activation of caspase-8, -9 and -3 and disrupted mitochondrial membrane potential in PC-3 cells, suggesting induction of apoptosis. In cell wound healing test, 5-ATAN decreased the cell migration dose-dependently. Overall, we infer the mechanism of cell growth inhibition of 5-ATAN in human prostate cancer PC-3 cells may due to induction of the cell cycle arrest and apoptosis that can be regarded as a potentially useful agent for the treatment and prevention of human prostate cancer.

AGFD 326

Distribution investigation of polymethoxyflavones in citrus peels

Tao Long1, long_tao@126.com, Lanying Xu1, Hui Zhao2, Chi-Tang Ho3, Shiming Li1. (1) Chemistry and Chemical Engineering, Huanggang Normal University, Huanggang, Hubei Province, China (2) Food Science, Tianjin University of Commerce, Tianjin, Tianjin City, China (3) Food Science, Rutgers University, New Brunswick, New Jersey, United States

Citrus peels provide rich and sole source of polymethoxyflavones (PMFs). It has been found in the past decades that PMFs have many health beneficial properties, such as antioxidant activity, preventive effects of obesity by influencing microbiota, functioning as anti-tumor and cancer chemopreventive agents, suppression of tumor growth by activating apoptosis or blocking cancer cell cycle progression. PMFs from citrus peel extract (CPE) also exhibit potent efficacious inhibition effects against inflammation in animal models. Much more research results regarding the biological activity of PMFs have been published and more explorations of health promoting properties of PMFs are undergoing at an accelerated pace. Furthermore, the analyses of citrus peel PMFs have been progressed well in recent years due to fruitful research in the field of biological activity and the need for identity identification of PMF compounds from citrus genus, particularly polymethoxyflavones. Recent research results and broad application of

PMFs in the areas of nutraceuticals, pharmaceuticals, and functional foods as well have prompted us to search rich content of PMFs among different sections of a particular citrus peel, such as aged tangerine peel, and a variety of citrus peels to have a direct comparison of PMF content among the samples analyzed. In this presentation, we will report our findings of the rich source of PMFs to serve as an efficient references for future PMF demand for potential functional food development from citrus peel PMFs.

AGFD 327

Chemistry and nutraceutical properties of polymethoxyflavones from citrus peels

Shiming Li1,3, shiming3702@yahoo.com, Chi-Tang Ho3, Min-Hsiung Pan2. (1) College of Life Sciences, Huanggang Normal University, Huanggang, China (2) National Taiwan University, Taipei, Taiwan (3) Food Science, Rutgers University, New Brunswick, New Jersey, United States

The peels of citrus genesis are almost the exclusive resource of a variety of polymethoxyflavones (PMFs), which are flavones with multiple methoxyl groups instead of hydroxyl groups in common flavonoids. For past decades, it has been found that PMFs possess a myriad of health promoting properties such as anticancer and anti-oxidant activity, attenuation of tumor growth, inhibition of inflammatory biomarkers, reduction of hyperglycemia and hyperlipidemia, prevention and reduction of Alzhemer's diseases among others. In this presentation, we will give an overview of the chemistry, bioavailability, metabolism and biological activities of polymethoxyflavones and demethylated polymethoxyflavones. The aim of this overview is to introduce the basic chemical and physical properties of PMFs and to welcome in-depth reports for the biological activities of PMFs from various research groups worldwide. The results from the bioactivity research of citrus peel PMFs will demonstrate the potential use of citrus peels as both functional foods and medicinal products.

AGFD 328

Biomimicking the stratum corneum to engineer edible oleogel

Tzu-Min Wang, littlekwei@gmail.com. Food Science, Rutgers University, Highland Park, New Jersey, United States

Stratum corneum (SC) containing free fatty acids, ceramides and cholesterols is mimicked to exploit the co-crystallization of these molecules into oleogels to make a hardstock fat replacers, which could be used as new forms of topical skin therapy for dermatoses. The structure and physicochemical properties of oleogels comprised of various ratios of stearic acid (SA), beta-sitosterol (beta-ss) and ceramide III (CerIII), as the molecular gelators, and canola oil as the primary liquid phase are investigated herein. The overall weight percentage of the structurant remained constant at 8 wt%, while the ratio of SA, beta-ss and CerIII were varied compiling a total of 21 different combinations. In this study, 8 of 21 samples formed elastic opaque oleogels; and of

those combinations capable of gelling, 7 had ‘fiber-like’ crystallites as their primary building blocks to generate a space filling three-dimensional crystal networks effectively entraining the canola oil. From the relationship between the ratio of the structurants and gel forming ability, SA and CerIII are the structurants that drove fiber formation. Their physical properties differed drastically, differential scanning calorimetry showed that varying the ratio of the three structurants, the onset of melting ranged between 27 - 40 °C and the onset of crystallization was between 19 – 29 °C. The oleogels examined herein had an elastic modulus (G') of 103-104 Pa, which was greater than the loss modulus (G'') (102-103 Pa). The remaining 13 ratios did not form oleogels and remained as opaque, viscous solutions. Although these ratios were incapable of forming oleogels they still had interesting microstructures that did not form a continuous 3D matrix capable of immobilizing the oil.

AGFD 329

Water alkalinity and hardness in beer brewing

Roger Barth, rbarth@wcupa.edu. Chemistry, West Chester University, West Chester, Pennsylvania, United States

Trace ions in water have a significant influence on the enzymatic reactions in the brewing process. In particular, alkalinity (bicarbonate) and hardness (calcium and magnesium ions) affect the pH, which is especially significant in the hydrolysis of starch during mashing. The optimal pH for mashing is in the range of 5.2-5.5. Bicarbonate raises mash pH and hardness lowers it. The net result is usually summarized in a parameter called the residual alkalinity, developed by Kolbach in 1941. The residual alkalinity is given by: RA = Alkalinity – {Ca2+}/3.5 – {Mg2+}/7, where all concentrations are customarily given in normality or CaCO3 equivalents, or the like. Kolbach’s equation, developed for beer wort prepared from Pilsner malt after boiling, needs to be revised to be applied to the mash and to darker grades of malt. Our measurements show that the effective alkalinity, as measured under laboratory mashing conditions, is much less sensitive to hardness ion concentration than predicted by the traditional formula. Our measurements include Pilsner, pale ale, and Munich malts, which are progressively darker and more acidic.

AGFD 331

Influence of molecular structure on interactions of dietary polyphenols and an immunodominant gluten peptide

Charlene Van Buiten2, charlene.vanbuiten@gmail.com, Carlos N. Pacheco1, Emmanuel Hatzakis1, Ryan Elias2. (1) Department of Chemistry, Penn State University, State College, Pennsylvania, United States (2) Food Science, The Pennsylvania State University, University Park, Pennsylvania, United States

Polyphenols have been shown to have anti-obesigenic, anti-inflammatory and antioxidative properties, all of which have been argued to exert a beneficial impact on human health when consumed in the diet. In addition to these direct interactions with the body and its metabolic functions, polyphenols have also been shown to have anti-nutritional properties by interacting with other dietary (e.g., minerals, proteins). Proteins rich in proline have been shown to be particularly susceptible to interaction with polyphenols, resulting in the formation of protein-polyphenol complexes and even precipitation. As proteins are often the basis for food allergies and intolerances, the anti-nutritional effects of polyphenols may actually be beneficial to some individuals through the binding and sequestration of these proteins. The aim of this study was to confirm and characterize the binding of dietary polyphenols to a proline-rich gluten peptide that has been previously implicated as immunodominant in Celiac disease pathogenesis. The dietary polyphenols chosen for this study – gallic acid, epicatechin, epigallocatechin, epigallocatechin gallate and theaflavin – vary in both structural characteristics and molecular weight. Saturation transfer difference nuclear magnetic resonance (STD-NMR) was used to identify the binding epitopes, or areas of interaction, of each polyphenolic ligand with the gluten peptide as well as protein-ligand dissociation constants.

AGFD 332

Sugar dialdehydes as glutaraldehyde analogs for cross-linked and immobilized chymotrypsin

Dana E. Wong, dewong@foodsci.umass.edu, Julie M. Goddard. Food Science, University of Massachusetts Amherst, Amherst, Massachusetts, United States

Enzyme immobilization can improve optimal working conditions, environmental tolerance, and aids in recovery and reuse after processing. Many traditional enzyme immobilization methods utilize the cross-linker glutaraldehyde to covalently attach enzymes by their terminal amine group amino acids. Although allowable for food processing, glutaraldehyde is not an ideal material due to its known toxicity, and has been associated with most activity loss in immobilized enzymes. This work aims to synthesize homobifunctional aldehydes produced by periodate oxidation of carbohydrates commonly found in food production waste streams (monosaccharides, disaccharides, long chain carbohydrates), that act as cross-linker analogues to replace the need for glutaraldehyde use in bioprocessing. Glucose, galactose, sucrose, lactose, starch, and dextran were oxidized to form multiple aldehyde groups. The aldehyde content and therefore cross-linking ability of the modified carbohydrates was determined. The cross-linker analogues were utilized in preparing carrier-free immobilized enzymes, or cross-linked enzyme aggregates (CLEA) of chymotrypsin. Chymotrypsin enables vast improvements in catalytic efficiency of a number of important food processing reactions including protein hydrosylate conversion for ingredient use as surfactants and foaming agents. Protein yield and chymotrypsin activity were measured to verify the effectiveness of the cross-linker analogues when compared to glutaraldehyde. CLEA produced from dextran aldehyde had a 64.4%

protein yield whereas CLEA produced from glutaraldehyde yielded <1% of original chymotrypsin solution. Chymotrypsin retained up to 65.8% activity as CLEA produced with the sucrose aldehyde compared to <1% prepared with glutaraldehyde calculated next to free enzyme. Homobifunctional cross-linkers prepared from food grade and waste stream carbohydrates represent an alternative to glutaraldehyde when used in enzyme immobilization that can maintain activity, and create a value-added process to the food and agriculture industries.

AGFD 333

DNA-comprising iron oxide/silica particles as tags against extra virgin olive oil adulteration

Michela Puddu2, michela.puddu@chem.ethz.ch, Daniela Paunescu2, Wendelin J. Stark3, Robert N. Grass1. (1) ICB, ETH Zurich, Zurich, Switzerland (2) ICB, ETHZ, Zurich, Switzerland (3) ICB ETH Zurich, Zurich, Switzerland

We developed a method to produce inexpensive, magnetic, high-performance, DNA comprising tags against oil adulteration.1 The nanoengineered tags consist of Fe2O3 cores, a silica shell, and dsDNA sequences layered in between. The iron oxide accounts for the magnetic properties of the obtained particles, while the outer silica matrix offers surface functionality and provides long-term DNA stability.2 DNA could be recovered unharmed upon particle dissolution in fluoride comprising buffers, and analyzed by qPCR and Sanger sequencing. The new material is an optimal tool for in-product labeling. The use of DNA-based barcodes ensures high versatility of coding. The particles are inert, resistant, and harmless, since iron oxide and amorphous silica are routinely used as food additives. The magnetic core of the particles facilitates handling/separation, and allows for sample pre-concentration prior to analysis. We decided to utilize the novel nanomaterial as label for extra virgin olive oil, since nowadays adulterated and fake olive oil are one the biggest food fraud. To this aim, the DNA including particles were functionalized with hydrocarbon chains to achieve dispersibility in hydrophobic liquids. The particles were dispersed in the oil and further retrieved by magnetic separation, followed by particles dissolution and DNA analysis by qPCR. We could successfully retrieve and detect the tags by qPCR in the oil and statistically discriminate 10 fold dilution steps of the oil suspensions, down to a concentration of 1 µg taggant per liter of oil (i.e. ppb levels). The procedure, which can be used for any oil, was additionally tested with gasoline and bergamot oil. Therefore, we developed a potent and low-cost platform for tracing/tagging oil items which enables to discriminate with reliability adulterated oil (e.g. diluted products) and determine oil origin/legitimacy. 1. Puddu, Paunescu, Stark, Grass, ACS Nano 2014. 2. Paunescu, Puddu, Soellner, Stoessel, Grass, Nat. Protoc. 2013.

AGFD 334

Biological soil quality indicators and conditioners in the phytoremediation of crude oil polluted agricultural soil

Eucharia O. Nwaichi1, nodullm@yahoo.com, Lasbery I. Opara2, Emmanuel O. Anosike1. (1) Biochemistry, University of Port Harcourt, Port Harcourt, Nigeria (2) HSE, Macphed Engineering Services, Port Harcourt, Nigeria

Owing to vital roles played by enzymes in maintaining soil ecosystem quality and functional diversity, the influence of organic manure on the resultant biological quality of a crude oil polluted agricultural soil from a 90d phytoremediation pot experiment was investigated. A 4 - factor phyto – assisted clean up of crude oil polluted agricultural soil was designed with options of manure to boost micobial activities. Profiles of β – glucosidases, proteases, dehydrogenases, phosphomonoesterases and respiration were investigated. Application of soil conditioner gave no marked dehydrogenase activity, which increased with depletion of available P. Marked increases in CO2 release and alkaline phosphatase activity with soil conditioning may implicate beneficial relationship with the abundance of microbial populations. Flooding of some soils correlated with β – glucosidases and respiratory acitivities. We found a direct relationship between cellulose breakdown, measurable with β – glucosidase activity, organic matter and CO2 release, measurable with respiratory activity within all soils in the present study.

AGFD 335

Ginkgo biloba: A new look at an old plant

Jack D. Williams1, jwilliams@mercyhurst.edu, Gregory R. Boyce2. (1) Chemistry and Biochemistry, Mercyhurst University, Erie, Pennsylvania, United States (2) Chemistry, Florida Gulf Coast University, Fort Myers, Florida, United States

The aroma profiles of raw and roasted seeds obtained from Ginkgo biloba berries were investigated using solid phase microextraction gas chromatography mass spectrometry (SPME-GC-MS). Changes in volatile composition were monitored with respect to seasonal variation in both roasted seeds and ripe berries. In addition GC-MS and LC-MS were used to identify the presence of 4'-O-methylpyridoxine (Ginkgotoxin) in the raw and roasted seeds.