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? The essential nature of genetically coded amino acids containing sulfur (methionine and cysteine) and selenium (selenocysteine) is well established. However, the health promoting effects of non-genetically coded sulfur and selenium amino acids (i.e., amino acid secondary metabolites) is unproven. The lack of conclusive proof with respect to these effects is largely due to a lack of data on the levels of these compounds in commonly consumed plant materials. A method, or series of methods, is needed to determine all S and Se amino acids and amino acid secondary metabolites in plant materials. More complete data is needed for methionine and cysteine in the National Nutrient Database for Standard Reference (NNDSR) and a provisional database is needed for selenocysteine, selenomethionine, and selected non-protein, amino acid secondary metabolites. The research to be undertaken falls under National Program 107 - Human Nutrition and addresses performance goal 3.1.2.B as described in the National Program Action Plan. Goal 3.1.2 is "Food Composition and Consumption: Develop techniques for determining food composition, maintain national food composition databases, monitor the food and nutrient consumption of the U.S. population, and develop and transfer effective nutrition intervention strategies." This project directly addresses Priority Objective B. Develop Analytical Methodology for Nutrients. The field of nutrition and public health will benefit from a better understanding of the distribution of S and Se in proteins, free amino acids, and amino acid secondary metabolites in foods. New data in the National Nutrient Database will provide the public with accessible information on the S and Se content of foods. There will be improved knowledge of the exchange of S and Se in biological systems. The field of analytical chemistry will have more accurate and comprehensive methods for the determination fo S and Se compounds in foods. 2.List the milestones (indicators of progress) from your Project Plan. Year 1 (FY 2004) * Initiate development of in-house S and Se standards. * Initiate development of a total sulfur method. * Initiate development of methods for free S and Se amino acids. * Initiate evaluation of the CNBr method for S and Se amino acids. Year 2 (FY 2005) * Continue development of in-house S and Se standards. * Complete development of a total sulfur method. * Continue development of methods for free S and Se amino acids. * Continue evaluation of the CNBr method for S and Se amino acids. Year 3 (FY 2006) * Complete development of in-house S and Se standards. * Complete development of methods for free S and Se amino acids. * Complete evaluation of the CNBr method for S and Se amino acids. * Initiate development of method for determination of S and Se in proteins. * Initiate determination of S and Se compounds in foods for database. Year 4 (FY 2007) * Complete development of method for determination of S and Se in proteins. * Continue determination of S and Se compounds in foods for database. Year 5 (FY 2008) * Continue analyzing S and Se compounds in foods for database. 3.Milestones: A. List the milestones (from the list in Question #2) 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. Year 1 (FY 2004) * Initiate development of in-house S and Se standards. * Initiate development of a total sulfur method. * Initiate development of methods for free S and Se amino acids. * Initiate evaluation of the CNBr method for S and Se amino acids. All the milestones scheduled to be completed in Year 1 (FY 2004) were accomplished. B. List the milestones (from the list in Question #2) that you expect to address over the next 3 years (FY 2005, 2006, & 2007). What do you expect to accomplish, year by year, over the next 3 years under each milestone? Year 2 (FY 2005) * Continue development of in-house S and Se standards. * Complete development of a total sulfur method. * Continue development of methods for free S and Se amino acids. * Continue evaluation of the CNBr method for S and Se amino acids. Year 3 (FY 2006) * Complete development of in-house S and Se standards. * Complete development of methods for free S and Se amino acids. * Complete evaluation of the CNBr method for S and Se amino acids. * Initiate development of method for determination of S and Se in proteins. * Initiate determination of S and Se compounds in foods for database. Year 4 (FY 2007) * Complete development of method for determination of S and Se in proteins. * Continue determination of S and Se compounds in foods for database. The milestones scheduled for FY 2005 through FY 2007 are still valid. There is a strong need for characterizing the S and Se distribution in proteins and amino acid secondary metabolites. Evaluation of the health-promoting capability of these compounds will depend on accurate analytical methods and databases. 4.What were the most significant accomplishments this past year? A. Single most significant accomplishment during FY 2004: None. B. Other significant accomplishments: None. C. Significant accomplishments/activities that support special target population: None. D. Progress Report: The CNBr method was applied to the determination of selenomethionine in variety of wheat flours (wheat gluten, Durum wheat, hard red spring wheat, and soft winter wheat). Despite total Se concentrations differing by as much as a factor of 45, the selenomethionine content was a constant fraction, approximately 46 microgram/gram. Initiated collaboration with the National Research Council of Canada on the characterization of the selenomethionine content of a Se enriched yeast reference material. Initiated development of a method for the routine determination of S and Se amino acids in foods, using ethylchloroformate derivatization and gas chromatography. Identified elution times for methionine, cysteine, selenomethionine, and selenocysteine. Separated alliin (the active S component in garlic) from alanine using an altered temperature program. Initial determination of S amino acids in selenized yeast and broccoli. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. This is the first year of a new CRIS. Accomplishments same as Question 4A. 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? None. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. None. Review Publications Rybak, M.E., Calver, E.M., Harnly, J.M. 2004. Quantitative determination of allicin in garlic: supercritical fluid extraction and standard addition of alliin. Journal of Agricultural and Food Chemistry. 52:682-687. Harnly, J.M. 2003. Organosulfur and organoselenium compounds: occurrence, importance, and analysis. [abstract]. Paper No. PO40. Harnly, J.M., Rodriguez, L.E., Wolf, W.R. 2004. Sulfur and selenium: nutrients, bioactive elements, and analytical challenges. [abstract]. October: Paper No. P4. Wolf, W.R., Goldschmidt, R.J. 2004. Selenomethionine contents of nist wheat reference materials. Analytical and Bioanalytical Chemistry. 378(5):1175-1181.