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? Commodity farming in the U.S. has been plagued by overproduction and maintained only by government subsidies. Even with heavy subsidies farm failures are still at an all time high in most regions of the U.S. Even more costly is the loss of diversity on the farm which has resulted in resistant pests and increased use of manufactured chemicals to maintain control. Of particular note is the emergence of corn root worm damage in first year corn. Utilization efforts for commodity raw materials has lent little relief to farmers who have seen commodity prices fall to record lows yet input costs for production are rising. New crops in rotation with commodity crops will provide an unsubsidized crop that will provide real farm income, introduction of new raw materials and provide ecological benefits through farm diversification. The challenges to new crop development are twofold. One, the agronomics of the crop must be favorable. Two, the raw materials produced must be novel (little or no domestic source) and have superior performance to their closest competitor. The development of new and alternative crops, enhancement of product quality during harvest storage and transport, development/improvement of methods of extraction, purification and manufacture, identification of new sources of natural products as nutriceutical and biopesticides and development of non-food products and undervalued by-products of agriculture are relevant to program components of National Program 306, Quality and Utilization of Agricultural Products. 2.List the milestones (indicators of progress) from your Project Plan. FY 2005 (Year 1): Complete the impact of cuphea rotation on corn root-worm in first year corn. Develop a partial seed analysis method for determining the fatty acid profiles of lesquerella which will direct germplasm selections. Develop Ulta Violet(UV) anthraquinone method and NIR method for oil, protein, and carbohydrate to guide sicklepod germplasm selections. Complete oil extraction studies for pilot scale processing of cuphea seed. Complete oil extraction and refining studies for pilot scale processing of lesquerella. Complete laboratory dehulling and fractionation of sicklepod seed. Complete oil extraction study of milkweed seed. Complete estolide number study for oleic monoestolides and the effect of capping chain length on pour points. Provide technical assistance for the commercial production of oleic estolides. Produce pilot scale quantities of lesquerella estolides. FY 2006 (Year 2): Provide oversight of 2,000 acres of cuphea production. Support germplasm development by pulsed Nuclear Magnetic Resonance (NMR), solid state NMR and partial seed Gas Chromatography (GC) analysis for selection of improved lesquerolic acid content and natural estolides. Transfer UV anthraquinone analysis method to breeders for sicklepod germplasm development. Complete cuphea oil refining studies. Crush 80,000 lbs. of lesquerella seed for oil which will supply initial market testing. Develop pilot scale fractionation methods to separate protein, oil and starch in sicklepod. Complete milkweed nematocide characterization. Study the effects of chainlength of capping group on pour points of diestolides. Provide oversight as needed for oleic estolide commercial production. Synthesize and explore physical properties of lesquerella estolides Begin the synthesis of viscosity modifier based on vegetable oils. FY 2007-2008 (Years 3 and 4): Provide oversight for the full commercial production of capric cuphea. Support germplasm development by pulsed NMR, solid state NMR and partial seed GC analysis for selection of improved lesquerolic acid content and natural estolides. Oversee commercial lesquerella extraction and processing. Oversee commercial cuphea extraction and processing. Develop method for the pilot scale isolation of anthraquinones from sicklepod. Complete the synthesis of milkweed nematocidal derivatives. Determine the effect of estolide linkage position on physical properties. Evaluate the physical properties of lesquerolic acid estolides. Complete the synthesis of viscosity modifiers and begin their performance evaluation. Provide oversight as needed for oleic estolide commercial production. FY 2009 (Year 5): Germplasm development of lauric cuphea. Support germplasm development by pulsed NMR, solid state NMR and partial seed GC analysis for selection of improved lesquerolic acid content and natural estolides. Oversee commercial lesquerella extraction and processing. Oversee commercial cuphea extraction and processing. Oversee commercial production of sicklepod gums. Complete the nematocide field trials. Support germplasm development by pulsed NMR, solid state NMR and partial seed GC analysis for selection of improved lesquerolic acid content and natural estolides. Produce pilot scale quantities of lesquerolic acid estolides. Complete evaluation of viscosity modifiers. Provide oversight as needed for oleic estolide commercial production. Evaluate physical properties of linkage position estolides. 4a.What was the single most significant accomplishment this past year? Commercial production of cuphea began in 2005 with initial acreages grown in the upper midwest. Seeds harvested from this grow-out will serve two purposes. First, to supply a high value product from cuphea oil. Secondly, to provide sufficient quantities of seed for increased production in 2006. 4b.List other significant accomplishments, if any. Examined the lab synthesis and optimization of coco-canola and canola estolides which had excellent pour points in the -36 degree C range. These new estolides are comparable with the simple oleic and coco-oleic estolides in terms of their oxidative stability. Completed the synthesis of estolides from oleic fatty acid, lesquerella and castor oils. The estolides will be evaluated as a potential cutting fluid and have its rheological values measured by the Cereal Products Research Unit at NCAUR. Physical properties of lesquerella oil estolides completed. Properties of castor and lesquerella estolides were comparable. Best pour points of -36 degree C were found for hexanoic capped lesquerella estolides. Bad oxidative stability was observed for these materials unless they were hydrogenated. After hydrogenation, these estolides gave very good RBOTs (58 min) without antioxidant. Completed the synthesis of a very cost effective lard-oleic estolide with cold temperature properties comparable with oleic estolides. These new animal-plant estolides have good oxidative stability properties (RBOT) at a relatively low cost. Completed the synthesis and physical properties of estolides from lesquerella/castor fatty acids. Best estolides in term of physical properties were capped with 2-ethylhexanoic acid (pour points in the -60 degree C range). Large scale canola fatty acid synthesis for potential license. Removal of the tin 2-ethylhexanoate catalyst used in the lesquerella (oil and fatty acid) estolide synthesis with the use of a carbon filter. Scaled-up of coco-oleic estolides in pilot plant. Cuphea was successfully grown and harvested on sandy irrigated soils, Havana IL (3 acres). Cuphea grown in the greenhouse did not host the corn root-worm. About 150 gallons of crude cuphea oil was pressed from 7,000 lbs of seeds, largest supply of cuphea oil in the world. Started to synthesize new branching functionality in fats. Directed toward lubricants application. Started the isolation of the 22:2 F.A.’s from meadowfoam. Directed toward viscosity modifiers and lower viscosity lubricants. Natural estolides from lesquerella auriculata isolated and characterized by NMR. Contains 95% estolide in the crude oil as a mixture of estolides. Completed esterification rate study of petroselenic acid and resolution study with a series of normal and branched chain alcohols. Method with isoamyl alcohol gave the best esterification rates and excellent separation of oleic from petroselenic. Examined lesquerella estolides and lesquerella oil as a potential viscosity modifier in oils. Started the synthesis of oleic high EN polyestolides with EN ~3. Evaluated seeds and press cake of cuphea, lesquerella and milkweed for proteins. The dominant protein classes in cuphea seeds were prolamins and glutelins. In lesquerella seeds, the dominant protein classes were globulins, prolamins, and albumins. Milkweed seed proteins were primarily albumins and globulins. Sicklepod seed endosperm processing (kilogram) scale-up is in progress for the separation and isolation of gums, protein and anthraquinones. Milkweed seed meal nematicide isolation and identification is in the final bioassay stage for confirmation and the spectroscopy phase is underway. Analysis of density-graded cuphea seeds. Seed fractions weighing over 2.5 g/1,000 seeds contained up to 35% oil. The oil had as much as 70% capric acid. Lighter seeds (less than 1.5 g/1,000 seeds) had 50% less oil, 13% less capric acid and higher amounts of palmitic and linoleic acids. There was little difference in protein contents between heavier (21-22%) and lighter (17-20%) seed fractions. 4c.List any significant activities that support special target populations. New crops research offers economic opportunities to small and limited resource farmers through the development of cuphea, milkweed, and lesquerella crops. 4d.Progress report. A pilot plant scale seed cleaning and oil crushing facility was completely finished at NCAUR. This pilot facility addressed a crucial need in U.S. new crop development by creating a location where small scale crushing of new crops seed could be accomplished. Three drums of crude cuphea oil have been produced in 2005 from pressing about 7,000 lbs of cuphea seed. The oil was used to obtain a potential CRADA/end-user for cuphea oil in cosmetics industry as well as to support future milestones. Also, 60,000 lbs of lesquerella seed were cleaned and are ready for pressing early this next report year 2006. The oil will be used to further develop lubricant work at NCAUR and Terresolve Technologies Ltd., as well as provide research samples for further market development in cosmetics and coatings. In addition, 5,000 lbs of milkweed seeds were cleaned and pressed at NCAUR to yield about 200 lbs of crude oil. The oil was sent to Natural Fibers Corp for oil recovery and market analysis evaluation. Cuphea grow-outs went to commercialization in 2005. NCAUR and the North Central Soil Conservation Research Lab in Morris, MN, transferred all growing conditions and procedures to Technology Crops Inc. NCAUR directly oversaw and managed 5 farms of cuphea (~20 acres) in the Peoria area. NCAUR personnel were responsible for field cultivation, planting, scheduling spraying, row cultivation, and harvest, as well as post-harvest management - seed drying and transporting seed back to NCAUR while utilizing existing corn and soybean farm production equipment. Seeds were processed in NCAUR crushing facility and oil used for development work. Synthesis and physical property data on estolides, lesquerella-oleic estolide 2-ethylhexyl esters, from lesquerella fatty acids had pour points in the -60 degree C range. These pour points exceed other derivatives and will meet the pour point requirements of some of the most demanding environmental applications (i.e. military). Lesquerella triglyceride estolides were synthesized using an optimized tin 2-ethylhexanoate catalyst (0.1 wt%) which reduced reaction temperatures from 200 degree C to 130 degree C. This temperature reduction in the synthesis of estolides allowed for scale-up of these materials in our pilot plant. Physical properties of castor and lesquerella estolides were comparable. The best pour point -36 degree C for hexnoic capped lesquerella oil estolides. These lesquerella oil estolides usually had bad oxidative stability unless hydrogenated where they then gave very good RBOT and do not require any antioxidant to their 58 minute RBOT. Gallon size reactions (conducted in the pilot plant) of lesquerella oil with myristic acid have yielded C-14 capped lesquerella oil estolides in high yield. In addition, this synthesis was further improved by the ability of being able to remove the tin 2-ethylhexanoate catalyst from the product stream. When manufactures examine the performance of the engine, the presence of tin is used as an indicator that there is unnecessary wear taking place in the engine. By removing the tin we can now incorporate this lubricant additive into engines that must pass extensive tests. An estolides structure study was initiated in 2005, where the estolide size was limited to examine the effect of chain length on pour point when (Estolide Number) EN = 1 is held constant. The estolide synthesis by nature forms a wide range of estolides sizes when conducted under the straightforward published conditions. Through a rigorous synthesis, estolides have been synthesized with EN = ~1 but the physical properties still need to be collected and analyzed. Also initiated a study on lesquerella estolides and lesquerella oil as a potential viscosity modifier in oils with no success in 2005. Sicklepod seed endosperm processing (kilogram) scale-up is in progress for the separation and isolation of gums, protein and anthraquinones. Natural estolides from lesquerella auriculata isolated and characterized by NMR. NMR showed crude oil contains 95% estolide as a mixture of estolides. A partial seed method was developed for determining elite hydroxy breeding lines using GC (Gas Chromatography). Esterification of petroselenic acid was competed with a rate study of esterification and resolution study with normal and branched chain alcohols. An esterification method was developed with isoamyl alcohol which yielded good rates and excellent separation of oleic from petroselenic. Evaluated seeds and press cake of cuphea, lesquerella and milkweed for proteins. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. The development of saturated capped estolides has increased the functional property range of vegetable based lubricants with pour points that now reach -60 degree C. Because of the decrease in the iodine values of this material the oxidative stability has been greatly improved with RBOT times that now exceed most common synthetic oil based lubricants. The lubricity and biodegradability of the oils was maintained. Lastly, the cost of producing these materials will make them very competitive against synthetic oils currently in the market. An end-user has demonstrated that this material lowers emissions when used as an additive to vegetable oil based lubricants. Large scale synthesis of these estolides has been conducted and is working to transfer this process to and industrial partner. Natural estolides were found by solid state NMR in whole lesquerella auriculata seed. Natural estolide may provide a lower cost route to highly functionalized lubricants from vegetable oils. A milkweed nematicidic fraction was identified from meal fractions. This isolate possessed higher activity than the original meal and characterization of this fraction is in progress. Cuphea development has continued to proceed at a rapid pace during this project from not being a commercial crop to having commercial acreages in 2005. Many agronomic problems still exist in the crop but initial farming practices have been established and a market for both the capric cuphea (300,000 acres required) and the lauric cuphea (2 million acres required) have been identified as well as industrial partners. Lesquerella development has continued with 40 acre field plots harvested in 2005. Products from lesquerella oil have been developed to aid in the commercialization of this new crop. Lesquerella oil estolides and lesquerella fatty acid estolides were synthesized and physical properties were determined. Lesquerella estolides could be useful in lubricant applications and may provide an avenue for large quantities of oil to be consumed thus providing a key driving force to lesquerella commercialization. We have discovered that several new crop seed meals are effective against agricultural pests. One example is that milkweed seeds (after their oil is recovered for another product) and ground pods kill a pest (nematodes) that causes a great deal of damage on potatoes in the Northwestern U.S. We have ongoing crop development groups for cuphea and lesquerella with everyone involved from growers to end-users to make sure the best information is available to everyone. 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? New Crops and Processing Technology personnel gave presentations to farm cooperatives in Illinois to demonstrate the potential of new crops in the Midwest, participated in a cuphea and lesquerella development groups that includes other ARS scientists, university and industrial cooperators. Canola and coco-canola estolides have be synthesized and transferred to potential licensees. The potential cooperator seeks uses in lubricants, and full scale commercial production should begin in FY 2006. Cuphea grow-outs went to commercialization in 2005. NCAUR and the North Central Soil Conservation Research Lab in Morris, MN, have transferred all growing conditions and procedures to a private company. 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). Making great lubricants with estolides, The Bulletin, Oct-Dec 2004. Alternative harvest – IWU Ag experts are studying various crops such as milkweed, okra and sunflowers and will present findings at field day, Peoria Journal Star, July 20, 2004. Flowering plant could be cash crop of future, Pekin Daily Times, February 5 & 6, 2005. NCGA’S Glass attends Opening of Value-Added Pilot Plant, National Corn Growers Association, http://www.ncga.com/news/2004/September/090804.htm. Review Publications Cermak, S.C., Isbell, T. 2004. Estolides - The next bio-based functional fluid. Inform. 15(8):515-517. Mohamed, A., Gordon, S.H., Harry O Kuru, R.E., Palmquist, D.E. 2005. Phospholipids and wheat gluten blends: interaction and kinetics. Journal of Cereal Science. 41:259-265. Cermak, S.C., Isbell, T.A., Isbell, J.E., Akerman, G.G., Lowery, B.A., Deppe, A.B. 2005. Batch drying of cuphea seeds. Industrial Crops and Products. 21(3):353-359. Gesch, R.W., Cermak, S.C., Isbell, T., Forcella, F. 2005. Seed yield and oil content of cuphea as affected by harvest date. Agronomy Journal. 97:817-822. Harry-O'Kuru, R.E., Wu, Y.V., Evangelista, R., Vaughn, S.F., Rayford, W., Wilson, R.F. 2005. Sicklepod (Senna obtusifolia) seed processing and potential utilization. Journal of Agricultural and Food Chemistry. 53:4784-4787. Harry-O'Kuru, R.E., Gordon, S.H., Biswas, A. 2005. A facile synthesis of aminohydroxy triglycerides from new crop oils. Journal of the American Oil Chemists' Society. 82:207-212. Wu, Y., Abbott, T.P. 2005. Gum and protein enrichment from sicklepod (Cassia obtusifolia) seed by fine grinding and sieving. Industrial Crops and Products. 21:387-390. Wu, Y.V., Nichols, N.N. 2005. Fine grinding and air classification of field pea. Cereal Chemistry. 82(3):341-344. Wu, Y., Hojilla-Evangelista, M.P. 2005. Lesquerella fendleri protein fractionation and characterization. Journal of the American Oil Chemists' Society. 82:53-56.