2004 Annual Report 1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? Soybean oil is the major food oil of the U.S. and the rest of the world. In order to provide functional properties and resistance to thermal and oxidative breakdown, soybean oil must be hydrogenated which in turn produces trans fatty acid isomers. The presence of trans isomers in food oils represents a major problem for the industry since trans has been implicated as serum cholesterol elevating agents in humans. On July 11, 2003, the U.S. Food and Drug Administration (FDA) finalized food labeling regulations. By Jan. 1, 2006, food manufacturers will be required to list trans acids on labels. The trans issue has prompted the industry to seek to formulate food oils including interesterification, plant breeding, and genetic modification. The objectives of this project include: (1) develop basic information permitting the expansion of structurally modified oilseed crops into functional foods, (2) development of basic information on triglyceride functionality obtained by hydrogenation, biocatalytic, chemical, and genetic modification, (3) development of new technologies to produce food oils low in trans acid by catalytic hydrogenation through control of selectivity by pressure, temperature, substrate, and catalyst concentration and use of binary gas systems, and (4) investigate the use of starch lipid complexes as emulsifiers for food oil systems. Approaches to solving these problems involve using highly sophisticated tools such as mass spectrometry, high pressure liquid chromatography, and pulsed nuclear magnetic resonance spectroscopy, to characterize and assess the functional role of triglycerides in food oil systems. Information on fatty acid and triglyceride composition will be developed in order to guide plant breeders and geneticists in designing more functional seed oils. The presence of trans fatty acids in hydrogenated food oils is a major concern of the edible oil industry because they have been linked to elevated serum cholesterol levels in humans. As a result the industry is faced with labeling requirements and possibly seeking the processing technologies to replace hydrogenation. Among those are interesterification and/or genetic modifications. Basic technical information on the fatty acid composition and triglyceride structures of genetically modified oils required for specific end uses are lacking. Such information is of great importance to plant geneticists, breeders, and edible oil processors attempting to design more functional, nutritional, and healthful food oils and products. This project contributes to National Program 306 on "Quality and Utilization of Agricultural Products" under Component 1 for "Quality Characterization, Preservation, and Enhancement," Problem Area 1a., "Definition and Basis for Quality". This research contributes to developing new knowledge to understand the roles of product composition, molecular structure, and physical state for optimum end use performance of edible vegetable oils. Technologies are developed to produce new food oil products with increased nutrition and other quality traits consistent with consumer demands. The research also contributes to meeting the ARS goal for genetic improvement of oilseeds for quality attributes for good quality products. 2.List the milestones (indicators of progress) from your Project Plan. Year 1: Completion of structured triglyceride synthesis and characterization of structural, thermal, dilatometric, and crystallization behavior. Year 2: Completion of chilling/crystallization studies on interesterified and structurally modified oils. Year 3: Completion of starch-lipid complex emulsifier studies. Year 4: Completion of low trans oils by pressure controlled hydrogenation in binary gas mixtures. Year 5: Completion of all studies with appropriate documentation. 3.Milestones: A. List milestones that were scheduled to be addressed in FY 2004. How many milestones did you fully or substantially meet in FY 2004 and indicate which ones were not fully or substantially met, briefly explain why not, and your plans to do so. Work on the use of binary gas mixtures for trans acid suppression (CO2/H2, N2/H2) was deferred to FY05 because the equipment necessary to conduct hydrogenation in the pilot plant has been dismantled and would require a complete reassembly which was not possible because of personnel limitations within the CRIS and collaborators (New Crops and Processing Technology Research), other milestones including synthesis of elaidic acid containing triglycerides for characterization of functional properties in food oil systems was successfully completed along with development of methodology for separating and purifying them. Milestone #5 allowed for completion of studies and preparation of manuscripts. Currently a number of manuscripts describing the solubility and phase behavior of a number of functional triglycerides are being prepared. In addition, a number of commercially prepared specialty lipids have been characterized for composition, structure and physical properties. A considerable amount of effort has been expended in cooperative efforts with other groups. R.O. Adlof has offered expertise in lipid synthesis and characterization to S. Peterson (USDA-ARS-MWA-NCAUR-Fermentation Biotechnology Research, Peoria, IL) and R. Dunn (USDA-ARS-MWA-NCAUR-Food and Industrial Oil Research, Peoria, IL). B. List the milestones that you expect to address over the next 3 years. In January 2004, a new project entitled "Improved Functional Food Oils via Novel Processing Technologies" was submitted to the Office of Scientific Quality Review for inclusion in the National Program 306, Quality and Utilization of Agricultural Products Panel Review. In March 2004, the project was reviewed with a score of 7.0 (No Revision) and approved in May 2004. This new project (3620-41000-112-00D) will continue research to develop technologies to reduce the trans and saturated acid contents of food oils. The milestones and expected results are as follows: FY 2005: Identify and establish parameters necessary to suppress trans acid formation during the catalytic hydrogenation of substrate and completion of work on binary gas mixtures and novel substrates. FY 2006/2007: Investigate computer modeling to prepare candidate for food applications. Prepare and evaluate food products for acceptance, performance, and sensory texture attributes. 4.What were the most significant accomplishments this past year? A. Single most significant accomplishment during FY 2004: Characterization of functional triglycerides found in hydrogenated soybean oils by High Pressure Liquid Chromatography/Atmospheric Pressure Chemical Ionization Analysis (HPLC/APCI) and synthesis of model compounds. Hydrogenated soybean oil (IV 65) was shown by analytical/preparative HPLC and atmospheric pressure chemical ionization in mass spectrometry APCI to be a complex mixture of triglycerides containing oleic, elaidic, palmitic, and stearic acids with trace of linoleic acid present. Nine triacylglycerols were identified with the majority consisting of mixed oleic/elaidic isomers (where O = oleic, E = elaidic), and POO, OOS, and POS (where P = palmitic, S = stearic) and 92% of the trans fatty acid are accounted for by OOO/EEE isomers, POO, OOS and POS. The physical properties of elaidic containing triglycerides have not been well characterized and reported in the literature. ESE, EES, EEO, EEL and PPE were synthesized in high purity and high melting point, solid fat, solubility, and heats of fusion are being determined to shed light on their functional roles in food oil systems. The synthesis and subsequent purification of these triglycerides has been found to be a difficult and considerable analytical methodology development to obtain high yields and purity. B. Other significant accomplishment(s), if any: None. C. Significant activities that support special target populations: None. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. New technologies were developed for synthesis, analysis and solubility and phase behavior of structured and naturally occurring triacyglycerols. This information will be useful to edible oil processors, plant breeders, geneticists and food technologists. Technologies developed to produce foods with zero or low trans acid content are of much interest to consumers and the edible oil industry because of health/nutritional issues and food labeling regulations effective Jan. 1, 2006. Studies on the formulation of margarines/spreads from hi-stearic acid soybean oils have shown that products formulated from them have suitable physical and sensory properties, thus providing an outlet for these crops estimated to have a market value in excess of $130,000,000. Pressured controlled hydrogenation and hydrogenation in binary gas systems (CO2 and H2) have shown great promise in reducing and suppressing trans acid formation. In contrast to commercially prepared basestocks containing 40% trans, hydrogenation at moderately high pressures produces products containing about 17% trans. When blended with additional liquid oil, soft spreads with about 5% trans result. Compared to commercial products containing about 10% trans, this represents about a 50% reduction. Initial studies made on hydrogenation of high oleic/soybean blends have shown promise in reducing the trans acid contents. Preliminary work on the stability of emulsions made from Fantesk (c) in which lecithin is the lipid portion has shown great promise as an emulsifier system for low fat spreads. 6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? A confidentiality agreement has been signed with Beatrice Foods, Indianapolis, IN, to allow formulation of low fat spreads with Fantesk as an emulsifier. The work with Beatrice Foods is expected to commence soon. A draft Cooperative Research and Development Agreement (CRADA) has been sent to A.C. Humko, Memphis, TN, to develop low trans hydrogenation techniques. The CRADA with A.C. Humko is dependent on funding from the industrial partner. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. List, G.R. 2004. Decreasing trans and saturated acid content of food oils. Food Technology. 58:23-31. Adlof, R.O. Winner, Dutton Award for 2004, Analytical Division of AOCS, for significant contributions to the analysis of fats and oils - contributions reflected in current analytical methodologies. Adlof, R.O. 2004. Adlof Shares 30 Years of Fats and Oils Research; Dutton Award speech. Inform. 15(7):489-490. Adlof, R.O., Copes, L.C., Walter, E.L. 2003. Characterization of triacylglycerol positional isomers in structured lipids by Ag-HPLC [abstract]. 25th World Congress and Exhibition of International Society for Fats Research, Bordeaux, France. p. 6. Adlof, R.O. 2004.Characterization of Structured Triacylglycerols by Silver-ion HPLC [CD-ROM]. ACS Annual Meeting, Anaheim, CA. ANAL 145. Adlof, R.O. 2004. Synthesis, Physical Properties and Solubilities of Some Palmitic Acid Containing Functional TAG in Soybean Oil [abstract]. AOCS Annual Meeting, Cincinnati, OH. p. 22. Review Publications List, G.R. 2004. Processing and reformulation strategies to decrease trans and saturated acids in food oils. Food Technology. 58:23-31. Farr, W.E., List, G.R. 2004. Chapter 17: Hydrogenation techniques. In: Dunford, N.T. editor. Book Chapter for Nutritionally Enhanced Processing. Champaign, IL:AOCS Press. p. 261-218. List, G.R. 2004. Giants from the past: Ralph H. Potts (1900-1981). Inform. 15:168-169. Adlof, R.O., List, G.R. 2004. Analysis of triglyceride isomers by silver-ion HPLC: effect of column temperature on retention times. Journal of Chromatography A. 1046:109-113. List, G.R. 2003. Trans fats: quality, safety and industrial trends. Proceedings of the International Conference & Exhibition of Nutraceuticals & Functional Foods. p. 119-123. List, G.R. 2003. Giants of the past: Michel Eugene Chevruel. Inform. 14:564-565. List, G.R. 2003. Giants from the past: David Wesson. Inform. 14:638. Emken, E.A., Adlof, R.O., Duval, S.M., Shane, J.M., Walker, P.M., Becker, C. 2004. Effect of triacylglycerol structure on absorption and metabolism of isotope labeled palmitic and linoleic acid by humans. Lipids. 39:1-8. Chu, F.E., Lund, E.D., Harvey, E., Adlof, R.O. 2004. Arachidonic acid synthetic pathways in the oyster protozoan parasite Perkinsus marinus: Evidence of usage of a delta-8 pathway. Molecular and Biochemical Parasitology. 133:45-51. Adlof, R.O. 2004. Internet resources for the lipid chemist. Lipid Technology. 16:29-34. Zhao, C., Adlof, R.O., Lofstedt, C. 2004. Sex pheromone biosynthesis in the pine caterpillar moth, Dendrolimus punctatus (lepidoptera: lasiocampidae). Journal of Insect Biochemistry and Molecular Biology. 34:261-271. List, G.R., Steidley, K.R., Neff, W.E., Snowder, G.D. 2003. Physical properties of mutton tallow. Grasas Y Aceites Monograph. 54:81-83. List, G.R. 2004. Processing and reformulation for nutrition labeling of trans fatty acids. Lipid Technology. 16(8):175-179.