2005 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? Fats and oils, together with carbohydrates and proteins, are important renewable resources compared to fossil and mineral raw materials, whose supply is finite. With the past availability of inexpensive petroleum feedstocks, the consumption of oils and fats as oleochemicals declined for most applications. The use of these renewable resources, however, has often been shown to have advantages when compared with petrochemical resources and as such fats and oils can be regarded as ideal raw materials for production of industrial products. Despite their ready availability and competitive price the domestic non-food use of fats and oils, which were once the primary sources of aliphatic carbon compounds used by industry, has remained stagnant in most applications. Of the approximately 101 million tons of fats and oils produced worldwide in 2004 only 14 million tons were utilized for oleochemicals. The United States is the world's largest producer of soybean oil and both edible and inedible tallow. Oils and fats are important US agricultural products, with an annual market size of roughly $12 billion, with soy oil and tallow dominating. Results from oleochemical research have show that the use of vegetable oils and fats allows the development of competitive products, which are both consumer and environmentally friendly. Despite their price advantage over other raw materials, total use of domestic fats and oils for such products as soaps, fatty acids and lubricants has remained stagnant over the past decade. The availability of domestic oils and fats, however, exceeds their consumption and export is relied upon for their disposition. Competition in the fats and oils market, however, poses serious challenges to US exports. In fact, palm oil now dominates world fats and oils trade with about 15 million tons being traded annually, versus 2 million for animal fats and 7 million for soybean oil. About 25% of the total fats and oils output of this country is destined for export. Traditionally, half the US production of tallow has been targeted for export to Asian markets. Increased competition from the palm oil-producing countries Malaysia and Indonesia, however, has put pressure on the pricing and demands of tallow in global markets. For example, US exports of tallow, both edible and inedible declined by 10% in 2003 continuing a trend seen in past years. Palm oil's continued growth in world fats and oils markets along with the incidences of mad cow (BSE) and foot and mouth outbreaks in Europe and the United States are cited as key factors responsible for this declining trend in US tallow exports. Continued decline in US export of tallow to the EU and Japan is forecasted with their decision to ban all animal-derived products in animal feeds. For the three-year period 1999-2001, US soybean oil production reached record annual levels averaging 8 MMT (consuming 7 MMT domestically and exporting 1 MMT). Despite the increased worldwide demand for soy oil, there are increased concerns over US soy oil usage because of the increased competition in soy oil domestic and export markets. These concerns arise from increased production of soybean oil in South America, notably Brazil, and China, and rapeseed and sunflower oils in Europe. With a projected decrease in exports, there will be a need to increase the demand for soy oil in domestic food and non-food uses. Another area of concern is the complete reliance on imported oxygenated oils, particularly castor oil, which is highly prized by US industries. One set of goals identified within this project is to devise new processes for expanding current uses, develop new higher-valued oleochemicals, and identify new food applications for fats and oils. Developing new processes and new outlets for these commodities is important to ensure the continued economic well being of the fats and oils production, processing, and industrial sectors of US agriculture. The project also develops methods for producing biodiesel using alternative technologies with low-cost feedstocks, and for producing oxygenated oils to replace imported oils. The project seeks to identify non-food uses targeted for creating new outlets and expanded markets for domestic fats and oils evaluating the application of biocatalysis and biomimicry (chemical reactions that mimic enzyme reactions) to fats and oils. The proposed research fosters the expanded use of domestic agricultural fats and oils in the production of new value-added products for food and non-food uses. Creation of new food outlets and non food biobased products from these renewable commodities would increase the economic return to the American farmer, address consumers' dietary concerns, increase the opportunities for expanded rural development, decrease our dependence on imported petroleum, and address the environmental concerns affiliated with US reliance on fossil fuels. The project is also closely associated with CWU 1935-41000-067-00D, the approaches of which are fermentative. A second area of research that project scientists are addressing is the production of a renewable fuel, biodiesel, from agriculturally derived lipids. Biodiesel is an agricultural based diesel fuel replacement whose use is currently growing in commercial fuels markets worldwide. US production of biodiesel last year was approximately 30 million gallons with refined soybean oil being the primary feedstock for most of this production. Although soybean oil is the least expensive of the domestic vegetable oils, as well as the most abundant, biodiesel produced from it is still more expensive than petroleum-based diesel fuel. Currently, biodiesel production when using refined fats and oils as feedstocks is not cost-competitive with petroleum diesel. This price differential is the single greatest barrier to the more widespread adoption of biodiesel as a fuel. One focus of the research is aimed at lowering the economic barrier to biodiesel production. The approaches are three-fold: (a) the development of methods to allow the use of less expensive sources of vegetable oils and animal fats as biodiesel precursors; (b) the development of new processing schemes that reduce the cost of biodiesel production from conventional feedstocks such as soy oil; and (c) the construction of economic models for the processes developed. Another obstacle to the use of biodiesel is that, relative to fossil-based diesel fuel, it causes an elevation in the engine emission of oxides of nitrogen (NOx). Project scientists aim to reduce the production of this regulated air pollutant through the identification of compounds that could be added to fuel and interrupt the chemistry of NOx formation. Quality parameters of biodiesel from these feedstocks including residual glycerol and sulfur content in the fuel also are addressed. Other work focuses on the utilization of glycerol, a co-product from biodiesel production, and the production of biodegradable materials for use is industrial applications such as polymers, coatings, adhesives,and lubricants. The overall objectives of the project are highly relevant to National Programs #306 (Quality and Utilization of Agricultural Products), to which the project is officially coded, and #307 (Bioenergy and Energy Alternatives). Research conducted within the project associated with National Program #306 is closely aligned with the National Program's Component II on New Processes, New Uses, and Value-Added Foods and Biobased Products, and Component I, Quality Characterization, Preservation, and Enhancement. Work under National Program #307 is aligned with Component II on biodiesel fuels. 2.List the milestones (indicators of progress) from your Project Plan. FY2005, Milestone chart, Project 1935-41000-066-00D, 15 months (6-18-04-9-30-05): Objective 1, Structured lipids: Develop methods for isolating bioactive isomers of CLA using food-approved reagents. Produce a series of structured lipids containing short, medium and long-chain fatty acids for use as low-calorie fats and/or nutritional supplements. Optimize immobilization techniques for each biocatalyst/catalyst studied. Objective 2a&2b, Alkyl- and/or alkoxy-branched fatty acids: Conduct bench-scale experiments to identify new catalytic approaches to branched-chain FA's for prospective use as lubricants, emollients, and hydraulic fluids. Objective 3, Functionalized lipids: Identify "new" epoxidation catalysts that use the "green" oxidants hydrogen peroxide and oxygen, and epoxidize common unsaturated FA's/esters and compare their activity to established epoxidation catalysts. Objective 4, Biofuels and additives: Investigate water removal to decrease alcohol requirements in the in situ transesterification of intact oilseeds. Identify alternative feedstocks for in situ biodiesel production. Produce biodiesel from animal products and determine the fuel properties. Screen lipases for best activity in removing free fatty acids in greases prior to conversion into biodiesel. Determine the ability of fuel additives to reduce NOx emissions from biodiesel combustion. Develop analytical methods for determining blend level of biodiesel in petrodiesel. Objective 5, Glycerol utilization: Identify methods for the preparation of glycerol-dibasic acid polyester prepolymers using selected enzymes and or catalysts. FY2006, Milestone chart, Project 1935-41000-066-00D, 27 months (10-01-05-9-30-06): Objective 1, Structured lipids: Optimize reaction conditions to maximize individual CLA isomers and incorporate into triacylglycerols. Determine physical properties of low-calorie structured lipids and, with collaborators, evaluate their use in selected applications. Objective 2, Branched fatty acids: Synthesize mid-chain branched-fatty acids/esters and conduct preliminary testing in lubricant applications. Develop analytical procedures for characterizing branched-chain fatty acids. Objective 3, Functionalized lipids: Subject fatty acid epoxides to mild hydrolysis and optimize polyol FA yields and Immobilize oxidation catalysts for production of diols and polyols from unsaturated FAs. Objective 4, Biofuels and additives: Produce sufficient spent soy flakes to determine nutritional quality of soy flakes after in situ transesterification. Optimize bench scale in situ transesterification of distillers' dried grains with solubles (DDGS). Complete economic analysis of bean-to-biodiesel model. Optimize conditions for lipase removal of free fatty acids greases and use as pretreatment step in biodiesel production. Determine if there is a relationship between fuel structure and NOx emissions. Develop a simplified field test for determining biodiesel blend levels. Objective 5, Glycerol utilization: Optimize polyester prepolymer synthesis. Initiate work on the development of methods for producing glycerol-based prepolymers and for their conversion to water-soluble hyper-branched polymers for use in bioremediation. FY2007, Milestone chart, Project 1935-41000-066-00D, 39 months (10-01-06-9-30-07): Objective 1, Structured lipids: Use low-temperature transesterification for the enrichment of selected CLA isomers. Develop options for the removal or elimination of trans-fatty acid isomers from fat and oil products. Objective 2, Branched fatty acids: Optimize conditions for scale up synthesis of targeted branched-chain FA's and devise purification schemes. Evaluate branched-chain oxygenated fatty acids and esters as lubricant additives or base (carrier) fluids. Objective 3a, Functionalized lipids: Complete polyol studies and prepare sufficient quantities to determine potential performance as additives in metalworking fluids and/or as lubricant additives. Objective 3b, Phospholipid applications: Evaluate selected phopholipid (PL)-derived components as lubricant additives. Objective 4, Biofuels and additives: Develop continuous in situ transesterification of soy flakes, DDGS, meat & bone meal (MBM). Develop pressure-catalyzed soapstock saponification method for producing biodiesel. Initiate work to improve the low-temperature properties of animal fat-derived biodiesel fuels. Identify best additized or iodine value-adjusted biodiesel candidate for reducing NOx emissions. Conduct engine testing of biodiesel fuels. Objective 5, Glycerol utilization: Create a series of glycerol-based polyols and polyesters and convert them into a series of linear and /or dendrimeric polymers. FY2008, Milestone chart, Project 1935-41000-066-00D, 51 months (10-01-07-9-30-08): Objective 1, structured lipids: Identify and overcome bottlenecks that reduce isomer separation or final purity of the targeted nutraceuticals or tailored lipid products. Objective 2, Branched fatty acids: Formulate selected branched-chain fatty acid products targeted for specific applications. Objective 3, Functionalized lipids: Evaluate selected polyols for use as metalworking or fuel additives, and /or polydispersants. Complete preparation of polyol materials with different structures and correlate structure-function properties to enhance performance. Identify best-modified PL structures or combined PLs. Objective 4, Biofuels and additives: Process sufficient DDGS to conduct feeding trials of spent meals from in situ transesterification. Optimize in situ transesterification of canola seed in batch reactions. Conduct continuous pressure-catalyzed saponification of soapstock for conversion into biodiesel. Objective 5, Glycerol utilization: Undertake collaborative testing studies of glycerol-based polymers; optimize production parameters, and patent technology for licensing. 4a.What was the single most significant accomplishment this past year? Measuring blends of biodiesel in petrodiesel: Biodiesel (oils or fats converted into simple alkyl esters) is most commonly utilized as a blend of 20 vol % in petrodiesel (B20); other blend levels such as B5 and B2 also are employed. As the marketing of biodiesel blends grows, facile methods are needed for determining or verifying the blend level of the marketed fuel. A high performance liquid chromatographic method for quantifying blends of biodiesel in petrodiesel was developed; it employs a short column with a simple isocratic mobile phase and is completed within 20 minutes. Quantitation can be made using either an evaporative light scattering detector (ELSD) or UV detector. The method is quantitative over a broad range blends (1 to 30 v/v % biodiesel). The method also can be used to quantitate similar levels of unconverted oils or fats in petrodiesel. The results of this work have the potential of becoming a standard test method, which currently does not exist, for determining the actual percentage of biodiesel in a blend. (NP307, Biodiesel Component; Milestone: Objective 4) 4b.List other significant accomplishments, if any. Economic modeling of biodiesel processes. CWU researchers, in collaboration with on-site engineers, initiated the development of capital/operating cost models for industrial scale production of biodiesel (soy methyl esters) from soy oil using conventional technology (oil extraction followed by alkali-catalyzed esterification) and by direct in situ esterification of the oil in intact soy flakes. The models are being developed to identify cost–saving steps in the production of biodiesel so as to improve its competitiveness with petrodiesel. Comparison of the models will allow for an economic assessment of the relative cost of the in situ method for biodiesel production compared to the conventional technology used in biodiesel production. (NP307, Biodiesel Component; Milestone: Objective. 4) Improved fatty acid functionality: The limited functionality of domestic fats and oils severely restricts there usage in many industrial applications. To overcome this deficiency, specialty oils such as castor oil are imported or domestic oils are oxygenated, often using hazardous chemical processes. CWU researchers have addressed this problem using an enzyme termed peroxygenase, which is found in oat seeds. When purified or partially purified, this enzyme can use hydrogen peroxide as an oxidant for oxygenating unsaturated fatty acids. With impure peroxygenase commercial organic hydroperoxides are used. Project researchers have concentrated their efforts on using ground oat seeds as a source of impure peroxygenase because of the low cost of using ground oat seeds. In this way they have been able to prepare good yields of epoxides from all tested unsaturated fatty acids and fatty amides. The peroxygenase shows unique reaction selectivity. It produces cis epoxides from cis double bonds, but is only weakly active on trans double bonds. Furthermore, peroxygenase is most active on isolated double bonds. If two or more double bonds are present and are separated from each other by a single methylene group, then only one double bond is epoxidized rapidly; the adjacent double bond is relatively unreactive. This feature of peroxygenase catalysis determines the structure of polyols prepared after hydrolysis of the epoxides. The ready availability of these highly functionalized fatty materials should expand there usage in applications such as metalworking fluids, where domestic fatty acids have previously had limited usage. Because of considerable industrial interest in this relatively simple method of producing fatty polyols, the researchers have applied for patent protection. (NP306, Component II; Milestone: Objective 3) Biodiesel fuels having improved emissions and stability: Fatty acid methyl esters (FAME) of lard, beef tallow, and chicken fat were prepared for use as biodiesel fuels. The research was undertaken to address concerns of elevated NOx emissions and storage stability of vegetable oil-derived biodiesel fuels. In-house nitrogen oxide (NOx) emission tests were conducted with the animal fat-derived esters and the results were compared to those from soy oil biodiesel as 20 vol% blends in petroleum diesel. The data indicated that all three animal fat-based biodiesel fuels had lower NOx emission levels than did the soy-based biodiesel. In fact, NOx emission levels for tallow-based biodiesel gave the same values as those from petroleum diesel. Selected cold-flow properties, lubricity, and oxidative stability of the fat-derived fuels also were measured. In general, the cold temperature properties of the fat-based biodiesel fuels were poorer than those of soy-based biodiesel (methyl soyate), but the lubricity and oxidative stability of fat-based esters were superior to the soy esters. These results should further the use of these renewable fats as biodiesel feedstocks. (NP307, Biodiesel Component; Milestone: Objective 4) 4c.List any significant activities that support special target populations. None. 4d.Progress report. Though a highly desirable renewable fuel, biodiesel does increase engine emissions of oxides of nitrogen (NOx), which are a federally regulated air pollutant. In a research diesel engine at this facility, using a 20% blend level biodiesel in petrodiesel resulted in a 5% elevation in NOx production. These results are in line with those reported by others in certified diesel engines. In work previously reported here, the ability of antioxidants, added at a concentration of 1000 ppm, to decrease NOx emissions was tested. Butylated hydroxyanisole (BHA) was the most promising of the compounds examined, causing a 4.4 percentage point decrease in the NOx emissions of an 80/20 blend of petrodiesel/soy biodiesel. This represents elimination of nearly 75% of the NOx increase caused by biodiesel. During the current period, various BHA blend levels were investigated to determine if the additive was effective at lower levels, or if the observed effect was magnified at higher levels. Tested additive levels were 500, 750, 1000 and 2000 ppm in B20. Although some effect was observed at 750 ppm and 2000 ppm, maximum NOx reduction was observed at 1000 ppm BHA. As noted above, 1000 ppm BHA in B20 eliminated nearly completely the NOx elevation seen with antioxidant-free biodiesel. Engine studies demonstrated that the combustion of soybean oil cyclohexyl esters at a 20% blend level in petrodiesel increased NOx emissions. The increase was greater than that seen with soybean oil methyl ester biodiesel: 11% vs. 5% elevation relative to the NOx output of petrodiesel. It was confirmed that the substitution of the methyl group with a cyclohexyl group actually increased the bulk modulus rather than decreasing it. Thus the results obtained here are consistent with previous conclusions that increasing bulk modulus increases NOx output. The work demonstrates the difficulty of predicting the effect of chemical structure on bulk modulus. The general applicability of the recently developed in situ method for biodiesel production was applied to the production of biodiesel from distillers dried grains with solubles, the predominant co-product of ethanol production. Production of biodiesel using low-cost alternative feedstocks should lower the cost of biodiesel. Progress in this area sould not only lower biodiesel cost but also expand biodiesel supplies as well as improve the economics of fermentative ethanol production. Commercial preparations of conjugated linoleic acid isomers (CLA) are composed almost entirely of equal amounts of the two bioactive CLA isomers. For use in formulated foods or as dietary supplements it is best that the pure isomer be used. Accordingly, project researchers have shown that the individual isomers can be enriched in a process comprising selective esterification with alcohols and the lipase (Candida rugosa). Selective esterification of CLA isomers was studied using a linear series of alcohols with increasing hydrophobic character (carbon chain-length of C8-18). The reaction mixture was then fractionated into an ester fraction and free fatty acid fraction using column chromatography. In this way it was found that the selectivity of esterification for one isomer increased with increased chain-length of the alcohol donor. This result led to the use of higher molecular weight food-approved alcohols for the separation of the two bioactive CLA isomers. Branched-chain fatty acids have important applications in selected areas since they have better oxidative stability than their linear unsaturated counterparts yet lower melting points than saturated linear chain fatty acids. Because of these important properties, therefore, they are ideal for use as biodegradable lubricants. Accordingly, work in this potentially important area of fatty acid chemistry has been initiated during this reporting period. Initial results indicate that fatty acids such as oleic acid can be converted to their branched-chain isomers in high conversions. Self-metathesis of unsaturated fatty acids presents an important and efficient pathway for the synthesis of long-chain, symmetrically unsaturated alpha-omega-dicarboxylic acids (C18, C20, C22, C26), which serve as important intermediates for the synthesis of biodegradable polyesters and polyamides. Project researchers have found that second generation Grubbs catalyst can effectively catalyze the solvent-free self-metathesis of monounsaturated fatty acids of varying purity (from 90% to 99%) to afford two important classes of products: unsaturated alpha-omega-dicarboxylic acids and unsaturated hydrocarbons in high conversions (up to 87%). This solvent-less self-metathesis reaction also works for monounsaturated fatty acids containing additional functionalities. The reactions also work well with catalyst loading as low as 0.005 mol % and a turnover number as high as 10,800 has been achieved. Cross metathesis (ethenolysis) of methyl oleate in supercritical ethylene in the presence of Grubbs catalysts also was examined. Ideally, such an environmentally benign ethenolysis reaction can lead to the production of methyl-9-decenoate and 1-decene, which serve as important intermediates to value-added products such as detergents. Initial results show that ethenolysis of methyl oleate in supercritical ethylene in the presence of Grubbs catalysts gave methyl-9-decenoate and 1-decene with 99% selectivity and conversion up to 28%. The researchers will continue to optimize the experimental conditions in order to maximize the yields of methyl-9-decenoate and 1-decene. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. This project is in its first year and represents a reorientation of work from its predecessor CRIS project (1935-41000-060). Briefly, selected examples of the underlying principles from that project that form the foundation of the current project include the following: Identification and characterization of lipases; their use in the development of enzymatic processes for the production of nutraceutical lipids; the lipase-mediated enrichment of industrially important fatty acids; and the production of low-calorie structured lipids. The combining of physical fractionation and enzymatic fractionation processes to produce tailored lipids from natural fats and oils. The reaction of unsaturated fatty acids with lipoxygenase enzymes converted them to their hydroperoxy and epoxy derivatives. The latter oxygenated fatty acids were subsequently converted to hydroxy-epoxy, polyol, and dicarboxylic acid derivatives. The immobilization of enzymes for use as biocatalysts in the lipase-catalyzed production of simple alkyl esters of fats, oils, and greases for use as biodiesel and the oxygenation of unsaturated fatty acids. Designing processes for the conversion of soapstock to 'high-acid acid oils' and to simple esters for use as biodiesel. Construction of quantitative computer models to estimate the cost of biodiesel production from refined vegetable oil, from vegetable oil soapstock, and from whole soybeans. 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? CWU scientists presented an overview of project work on enzymatic processes for the production and characterization of functional fatty acids to the staff and students at the University of Missouri, Columbia (UMC). As a result of this visit, a pre-proposal was prepared in collaboration with Dr. Galen Suppes, UMC, and Mr. Jim Martin, United Soybean Board and Omni Tech International, and submitted to the Biomass Research and Development Initiative of the U.S. Department of Energy in partnership with the U.S. Department of Agriculture for funding. The pre-proposal, however, was not selected for full proposal submission. The following procedures have been transferred to the general scientific community through CWU publications and presentations at various scientific conferences: Procedures for making epoxides with peroxygenase in microsomes, immobilized peroxygenase technology, the technology for making fatty epoxides with ground oat seeds, and procedures for hydrolyzing fatty epoxides to produce polyols. In addition project researchers have used for the first time various HPLC/MS analytical procedures for the analysis of fatty epoxides and fatty polyols. A patent application was filed on the selective epoxidation of non-proximate cis carbon-carbon internal double bonds in a compound containing at least two cis carbon-carbon internal double bonds that are mono-methylene-interrupted. The process involves the novel concept of contacting a polyunsaturated fatty acid a with a suspension of ground oat seeds and continuing the contacting for a period of time such that only one of the carbon-carbon double bonds is epoxidized. A collaborating partner is evaluating products prepared with the technology described in this patent. A confidentiality agreement was entered into with a major producer of fatty esters for both the food and chemicals industry. The interest of the company resides in CWU research on the development of new uses for glycerol. Discussions were had on the potential of blending various ester feedstocks for improving the low-temperature and combustion properties of biodiesel. Strong interest has been expressed in the biodiesel economic modeling work FOAC researchers have undertaken. The first model was developed to estimate the costs of plant construction and the process costs for the production of biodiesel from soy oil. This is a predecessor of the modeling work presently underway, described above. In the past year project researchers have distributed approximately a dozen copies of the first model, in advance of formal scientific publication, to interested parties upon request. These inquires came from individuals and firms interested in building biodiesel production facilities. Scientific publication is scheduled for fall, 2005. Approximately two dozen inquiries have been fielded from existing and potential biodiesel producers regarding the in situ transesterification method for biodiesel production developed in this CRIS project. Many of these parties have been sent copies of the project's scientific publications describing this work. The largest constraints to adoption are the absence of a full economic analysis of the technology and the lack of data addressing the suitability as an animal feed of the spent seed meal exiting the process. Project researchers are presently working to address both of these bottlenecks, the latter in collaboration with animal nutrition laboratories.. A collaborative arrangement was entered into with a state-funded research organization to transfer the technology for in situ transesterification and to assemble a consortium of collaborators to further investigate the suitability of the meal co-product as an animal feed. A formal collaboration was established with Philadelphia Fry-o-Diesel, a start-up program of the Philadelphia Energy Cooperative. The role of the CWU in this local energy initiative is to provide technical collaboration to assist them in their establishment of biodiesel production from low-value lipids. CWU researchers are assisting a major biodiesel producer to help that company identify the causes of undesirable precipitation in their final biodiesel product. This work is part of CWU efforts in identifying and potentially rectifying the cause of the low-temperature problems associated with biodiesel fuels. Multiple inquiries have been received regarding CWU-patented technology describing the production of biodiesel from low cost feedstocks such as soapstock by sequential hydrolysis-esterification of this lipid feedstock. This technology is ready for licensing at this time. Constraints to adoption are technical matters, and seem to be catalyst costs and wastewater disposal costs, items that are scheduled for study later in the life of this CWU. Transfer of CWU technology to the industry was made in a series of meetings held with a consultant to the fats and oils industry and the owner of a small firm specializing in oleochemical lubricants. These interactions resulted in submission of a collaborative SBIR developmental grant to investigate current and past technologies for the production of polyol and branched-chain fatty esters as biodegradable lubricants and/or metal working fluids. CWU researchers presented an overview of their work on nutraceutical-type lipids and low-calorie lipids and on the dry fractionation of natural fats and oils to produce fractions enriched in mono- and polyunsaturated triacylglycerols to representative of a major oleochemical company. The company, which is a major supplier of methyl esters to the industry, has interest in the project's work on biodiesel from alternative feedstocks. Presentations made by CWU researchers also covered their newly developed methods for biodiesel analysis. Considerable interest was expressed on polyol fatty acid and ester production using peroxygenase enzyme present in oat seeds. A Confidentiality Agreement for exploring the potential use of the enzymatic method of polyol production was implemented. In continuation of collaborative efforts with a major supplier of lubricants and lubricant additives to the metal working industry, material transfer agreements were entered into with the company for their evaluation of selected polyesters prepared by CWU scientists. Other products identified at this meeting were triacylglycerol fractions obtained by dry fractionation of selected animal fats for testing as lubricant-type base fluids. Polyol samples also were given to two other companies for evaluation in applications other than metal working fluids. A confidentiality agreement was entered into with a major producer of fatty esters for both the food and chemicals industry. The interest of the company resides in the use the development of new uses for glycerol. Toward this goal CWU scientists collaborated with Rohm and Haas Co. (Spring House, PA) on a green chemistry proposal entitled New Sustainable Chemistry for Adhesives, Elastomers and Foams submitted to the DOE and USDA Joint Biomass Research and Development Initiative Program. The proposal was awarded funding for a two-year period to conduct research in this area. The role of the CWU research is to identify and collaborate on studies directed to the use of selected fats and oils and crude biodiesel glycerol co-product for the production of biobased adhesives. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). Agricultural Research magazine, Vol. 53, no. 4, April, 2005, p. 13, contained a description of the in situ method of biodiesel production. This was subsequently excerpted to independent but similar articles in Renewable Fuel News (Vol. XVI, No. 15, April 18, 2005, p. 1, 8); Biodiesel Magazine (Vol. 2, No. 5, June 2005, p. 48-50); Feedstuffs (Vol. 77, no. 27, July 4, 2005, p. 15); Biocycle (Vol. 46, no. 5, May 2005, p. 70); and Linkages (Vol. 1, Issue 4, Feb. 2005). These articles led to the substantial public sector interest in CWU accomplishments, noted above. By invitation, CWU researchers presented a description of the in situ method for biodiesel production to the Governors of the National Biodiesel Board at their meeting in Ft. Lauderdale, FL, in February 2005. An overview of the data on in situ transesterification, on the effects of antioxidant addition on biodiesel NOx output, and the use of various fuel feedstocks was given at invitation at a workshop designed to determine biodiesel research opportunities and challenges, January 2005, in Chicago. The same information also was presented at the National Biodiesel Board meeting in Fort Lauderdale, FL. An overview of CWU research on biobased products and biofuels was presented at the USB/USDA joint meeting in Rosemont, IL, October 2004. Approximately 60 attendees from government, industry, and university were present. CWU work on the production of biobased lubricants and lubricant additives was made before 80-100 participants at the Lubricants and Fluids Technical Advisory Panel Meeting, United Soybean Board, Chicago, 2004. A CWU scientist served as an invited instructor for Short Courses on Biodiesel Production, sponsored by the American Oil Chemists Society in Salt Lake City, UT, May 1, 2005, and at the 26th World Congress and Exhibition of the ISF - Modern Aspects of Fats and Oils - A Fascinating Source of Knowledge, Prague, Czech Republic, Sept. 25-28, 2005. A CWU scientist presented an overview of project work on enzymatic processes for the production and characterization of functional fatty acids to the staff and students at the University of Missouri, Columbia (UMC). As a result of this visit, a pre-proposal was prepared in collaboration with Dr. Galen Suppes, UMC, and Mr. Jim Martin, United Soybean Board and Omni Tech International, and submitted to the Biomass Research and Development Initiative of the U.S. Department of Energy in partnership with the U.S. Department of Agriculture for funding. A provisional patent application entitled Methods of selectively epoxidizing carbon-carbon double bonds has been filed. The patent describes a process for the enzymatic oxygenation of polyunsaturated fatty acids to epoxidized fatty acids and their subsequent conversion into poly fatty acids. This technology is available for commercial development and adoption. Review Publications Foglia, T.A., Jones, K.C., Nunez, A., Phillips, J.G., Mittelbach, M. 2004. Comparison of chromatographic methods for the determination of bound glycerol in biodiesel. Chromatographia. 60:305-311. Hsu, A., Jones, K.C., Foglia, T.A., Marmer, W.N. 2004. Continuous production of ethyl esters of grease using an immobilized lipase. Journal of the American Oil Chemists' Society. 81(8):749-752. Piazza, G.J., Foglia, T.A. 2004. Synthesis of a fatty tetrahydroxyamide using peroxygenase from oat seeds. Journal of the American Oil Chemists' Society. 81(10):933-937. Foglia, T.A., Haas, M.J., Mcaloon, A.J., Marmer, W.N. 2004. Cost modeling of biodiesel production: effect of feedstock and processing. Proceedings of the United States-Japan Cooperative Program in Natural Resources, Protein Resources Panel Annual Meeting. p. 50-54. Haas, M.J., Foglia, T.A. 2005. Alternate feedstocks and technologies for biodiesel production. In: Knothe, G., Krahl, J., and Van Gerpen, J., editors. The Biodiesel Handbook. Champaign, IL: AOCS Press. p. 42-61. Nunez, A., Moreau, R.A., Foglia, T.A. Liquid chromatography/mass spectrometry for the analysis of biosurfant glycolipids secreted by microorganisms. Book Chapter. W. C. Byrdwell (ed) in Modern Methods for Lipids Analysis by Liquid Chromatography/Mass Spectrometry and Related Techniques, pp.447-471, 2005. Lee, K., Foglia, T.A., Lee, J. 2005. Low calorie fat substitutes: synthesis and analysis. In: Hou, C.T., editor. Handbook of Industrial Biocatalysis. Boca Raton, FL:CRC Press. p. 16-1:16-19. Hess, M.A., Haas, M.J., Foglia, T.A., Marmer, W.N. 2005. The effect of antioxidant addition on nox emissions from biodiesel. Energy and Fuels. 19(4):1749-1754. Haas, M.J. 2005. Animal fats. In:Shahidi, F., editor. Bailey's Industrial Oil and Fats Products. Hoboken, NJ:John Wiley & Sons, Inc. p. 161-212. Bloomer, S., Haas, M.J. 2004. How to write a scientific paper. Inform. 15(12):761-763. Piazza, G.J., Foglia, T.A. 2004. 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