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? There is an ever-increasing need for trace element food composition data. Much of the trace element data in the latest National Nutrient Database for Standard Reference was generated many years ago and needs to be updated because foods have changed. Review of the data suggests that in some cases, unsophisticated analytical methodology produced less than accurate results due to problems with contamination, matrix interference, effects or instrumental background correction strategies. Graphite furnace-atomic absorption spectrometry (GF-AAS), inductively coupled plasma-atomic emission spectrometry (ICP-AES) and inductively coupled plasma-mass spectrometry (ICP-MS) are newer instrumental techniques that are readily available to commercial labs. All provide better detection limits for many elements as compared to flame atomic absorption spectrometry and the latter two offer the advantage of multielement determinations, allowing for the generation of accurate data for as many as 15-20 elements simultaneously. This project is organized into four objectives:. 1)development of robust multielement methods suitable of industry;. 2)development of sensitive, ultratrace methods for Co, Cr, Ni, and V;. 3)developement of a method for cobalamin species; and. 4)development of methods for the determination of Fe species. 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, in general, from an improved nutrition database for trace metals and increased knowledge regarding the concentrations and species of trace metals in foods. The field of analytical chemistry will benefit from improved methods for. 1)the simultaneous determination of Mn, Zn, Fe, Cu, Mg, Ca, and P;. 2)the determination of Co, Cr, Ni, and V;. 3)the determination of individual cobalamin species; and. 4)the determination of Fe as a porphyrin complex (heme Fe). In addition, the field of analytical chemistry will benefit from improved reference materials. 2.List the milestones (indicators of progress) from your Project Plan. Year 1 (FY 2004) Robust Multielement Methods *Develop and circulate a questionnaire for food industry analytical labs on methods for low levels of Mn, Zn, and Cu in foods. *Evaluate responses to questionnaire. Ultratrace Methods *Develop GF-AAS and ICP methods for Co and Cr. Cobalamin Speciation *Develop method for cobalamin species using capillary electrophoresis and ICP-MS. *Start development of sample preparation method for cobalamins focusing on solid phase extraction. Fe Speciation *Acquire standards for Fe species. *Initiate development of an High Performance Liquid Chromatography (HPLC) separation method for Fe species extracted from meat. Year 2 (FY 2005) Robust Multielement Methods *Complete evaluation of questionnaire. *Develop in-house food control materials for evaluation by food industry labs. Ultratrace Methods *Complete development of GF-AAS and ICP methods for Co and Cr. *Initiate development of GF-AAS and ICP methods for Ni, and V. Cobalamin Speciation *Complete development of sample preparation method for cobalamins. Fe Speciation *Continue development of HPLC method for Fe species extracted from meat. *Determine effect of cooking and acidification on heme Fe. *Set up in vitro digestion system to study heme Fe. *Initiate study of bioavailability of heme Fe. Year 3 (FY 2006) Robust Multielement Methods *Initiate development of ICP methods for food materials. Ultratrace Methods *Complete development of methods for Ni, and V. *Initiate development of in-house control materials for Co, Cr, Ni, and V. Cobalamin Speciation *Initiate determination of cobalamins in foods and supplements. *Develop a fresh frozen quality control materials. Fe Speciation *Continue study with in vitro digestion system for heme Fe. *Continue study of bioavailability of heme Fe. *Initiate quantification of total Fe and heme Fe in foods for database. Year 4 (FY 2007) Robust Multielement Methods *Develop Association Of Analytical Chemists (AOAC) method(s) for determination of food materials using ICP. Ultratrace Methods *Complete in-house control materials for Co, Cr, Ni, and V. *Initiate determination of Co, Cr, Ni, and V in food materials. Cobalamin Speciation *Initiate development of a dietary supplement quality control material for cobalamins. Fe Speciation *Finish study with in vitro digestion system for heme Fe. *Finish study of bioavailability of heme Fe. *Determine total Fe and heme Fe in foods for database. Year 5 (FY 2008) Robust Multielement Methods *Complete AOAC method(s) for determination of food materials using ICP. Ultratrace Methods *Complete determination of Co, Cr, Ni, and V in food materials Cobalamin Speciation *Complete development of dietary supplement quality control material for cobalamins. Fe Speciation *Determine total Fe and heme Fe in foods for database. 4a.What was the single most significant accomplishment this past year? Developed a method for the determination of heme Fe in meats using an acidified acetone extraction with HPLC-diode array detection. Heme Fe is more accessible in foods but a definitive method is lacking and there is no database for heme Fe. This method allows for the conclusive identification of heme Fe using the HPLC retention time and the molecular spectra and mass balance is achieved by comparing the Fe in the sample with the sum of the Fe in the extract and the remaining solids. This method will allow us to establishe a heme Fe database based on a nation wide sampling program implemented by the Nutrient Data Lab. 4c.List any significant activities that support special target populations. None. 4d.Progress report. Determined the effect of cooking on the integrity of myogolbin Fe and heme Fe in meats. This research addresses the question of what form of Fe reaches the small intestine. Initial results suggest that 100% of the myoglobine Fe is denatured and 30% of the heme Fe is destroyed. Collaborated with the Plant, Soil, and Nutrition Lab in Ithaca, NY, on evaluation of cooking on the bioavailability. Bile juices are critical to solubilizing heme Fe after denaturation of the myoglobin after cooking. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. Participated in the characterization of two new international food quality control materials providing trace element data to colleagues in Poland for a corn flower and a soya bean flour reference material. The materials both were fairly homogeneous and QC data all confirm the accuracy of the determinations. The only element which led to some problems was K at unusually low levels (in corn starch). Developed and circulated a questionnaire to colleagues from the National Food Processor's Analytical Chemists Subcommittee regarding low level trace element methods. Information gathered includes: methods used for trace element determinations, quality control strategies, interest in fresh food QC materials, and interest in participation in a study to improve low level trace element determinations for Mn, Zn, Fe and Cu. All of the laboratories contacted are commercial analytical labs. Many are associated with food producing companies and some are contract labs but all of them generate food composition data. The expected outcome of the study is development and technology transfer of robust methods suitable for industrial labs for the production of multielement food composition data. Another product is the production of much needed in-house food QC materials. Participated in an interlaboratory collaborative study determining nine trace elements (Ca. Cu, Fe, Na, K, P, Mg, Zn, and Mn) in a nutritional liquid (similar to Ensure) and a cereal (similar to Cheerios). We were 1 of 22 laboratories who participated and our data were in good agreement with the mean values from the study. Developed method for heme Fe in meats that employed the classic Hornsey extraction (acidified acetone) and determined the extracted Fe using atomic absorption spectrometry. Demonstrated that the use of spectrophotometry for the determination of Fe in the Hornsey extract gave results that are biased high by 15% to 40%. Initiated development of HPLC method to separate components extracted by the Hornsey method to verify that all the Fe was heme FE. 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. (NOTE: List your peer reviewed publications below). None. Review Publications Yanes Santos, E.G., Miller-Ihli, N.J. 2005. Parallel path nebulizer: critical parameters for use with microseparation techniques combined with inductively coupled plasma mass spectrometry. Spectrochimica Acta. 60:555-561.