Source: Agricultural Research Service submitted to
DETERMINATION OF BIO-ORGANIC NUTRIENTS IN FOODS
Sponsoring Institution
Agricultural Research Service/USDA
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
0405814
Grant No.
(N/A)
Project No.
1235-52000-043-00D
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Jul 3, 2002
Project End Date
Feb 21, 2004
Grant Year
(N/A)
Project Director
WOLF W R
Recipient Organization
Agricultural Research Service
BLDG 003 BARC W RM 331
BELTSVILLE,MD 20705-2351
Performing Department
(N/A)
Non Technical Summary
(N/A)
Animal Health Component
50%
Research Effort Categories
Basic
25%
Applied
50%
Developmental
25%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
70150102000100%
Knowledge Area
701 - Nutrient Composition of Food;

Subject Of Investigation
5010 - Food;

Field Of Science
2000 - Chemistry;
Goals / Objectives
To develop precise and robust methods for the analysis of folates, non-digestible oligosaccharides, resistant starches, and inositol phosphates. To test newly developed analytical procedures on a variety of foods for eventual technology transfer.
Project Methods
Non-digestible oligosaccharides will be extracted into various solvents, separated and quantified using high performance anion exchange chromatography (HPAEC). Compare the extent of starch hydrolysis using different autoclave temperatures as pretreatment prior to enzyme hydrolysis. Evaluate and improve an existing method for the analysis of inositol phosphates using HPAEC with chemically suppressed conductivity detection. For the routine measurement of folates and folic acid in foods, accurate and robust procedures will incorporate: 1)sample digestion employing several enzymes in a specific sequence, 2)sample preparation using ion-exchange column technology, and 3)reversed-phase HPLC for separation and quantification of analytes.

Progress 07/03/02 to 02/21/04

Outputs
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? Current food analysis methods ignore the multiple isomers of folate, inositol phosphates and trans-fatty acids in foods. Precise and robust methods are being developed for the determination of nondigestible oligosaccharides, resistant starch, inositol phosphates, folates, trans- polyunsaturated fatty acid, and oxidized lipids. Lack of precise and robust methods, thus, unreliable data often lead to inconclusive results from a number of epidemiological studies involving the roles of various nutrients in health and diseases. Nutritionists and clinicians are in search of methods which will provide much needed food composition data on the above-mentioned food constituents. Furthermore, the release of Dietary Reference Intakes by the National Academy of Science based on dietary folate equivalent requires food data on folate isomers and folic acid. 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. This research will benefit the field of analytical chemistry, the medical community, and the general public. New and/or improved analytical methods will provide data on nutrients and bioactive compounds found in foods, will provide medical and nutrition researchers with data necessary to evaluate the efficacy of a variety of food components, and will expand the National Nutrient Database for Standard Reference which is on the Web and available to anyone in the U.S. or the world. 2. List the milestones (indicators of progress) from your Project Plan. There were no milestones when the original CRIS project was developed. The objectives were to: Develop precise and robust methods for the analysis of non-digestible oligosaccharides. Evaluate and improve existing methods for distinguishing various types of resistant starches. Modify, improve, and validate qualitative and quantitative methods for phytic acid and other inositol phosphates in dietary residues of selected foods. Develop an accurate, precise, and robust analytical method for the measurement of total folate content of foods. 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. All of the objectives were met. 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? None, this CRIS was terminated in September of 2003. 4. What were the most significant accomplishments this past year? A. Single most significant accomplishment during FY 2004: CRIS 1235-52000-043-00D was a bridging project for CRIS 1235-52000-036- 00D (Determination of bio-organic nutrients in foods) which expired 7/16/02. The bridging CRIS was replaced by a new CRIS 1235-52000-047-00D (Liquid chromatographic and stable isotope dilution mass spectrometric methods for the determination of water-soluble B vitamins in foods and dietary supplements). We received notification from the Office of Scientific Quality Review (OSQR) of our score (6 - minor revision) in September 2003, and the letter of certification from OSQR in December 2003. We have conducted research based on the new CRIS objectives since September 2003. Consequently, all research has been reported under the new project, CRIS 1235-52000-045-00D. B. Other significant accomplishments: None. C. Significant accomplishments/activities that support special target population: None. D. Progress Report: None. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. A summation of the research covered by CRIS 1235-52000-036-00D and CRIS 1235-52000-043-00D from 10/01/97 through 9/30/03 is listed below. A method was developed for the determination of total dietary fiber in order to estimate the amount of resistant starch in foods; such as cooked dried legumes and baked products containing high level of amylose. A comparison of this method with two other more labor-intensive methods showed that this is a simpler and more cost-effective method. A definitive mass spectrometric method was developed for the determination of folic and 5-methyl tetrahydrofolic acid in foods using 13C labeled isotopes. These methods have been used for the determination of folate vitamers in vegetables and breads. A second, less expensive, and more robust method for the determination of all the folate vitamers has been developed based on liquid chromatographic separation and fluorescence detection. This method uses only commercially available materials.

Impacts
(N/A)

Publications

  • Pawlosky, R.J., Flanagan, V.P., Doherty, R.F. A mass spectrometric validated high-performance liquid chromatography procedure for the determination of folates in foods. Journal of Agriculture and Food Chemistry. 2003. v.51. p.3726-3730.
  • Pawlosky, R.J., Hertrampf, E., Flanagan, V.P., Thomas, P.M. Mass spectral determinations of the folic acid content of fortified breads from Chile. Journal of Food Composition and Analysis. 2003. v.16. p.281-286.
  • Li, B.W., Andrews, K.W., Pehrsson, P.R. Individual sugars, soluble, and insoluble dietary fiber contents of 70 high consumption foods. Journal of Food Composition and Analysis. 2002. v.15. p.725-733.
  • Doherty, R.F., Beecher, G.R. A method for the analysis of natural and synthetic folate in foods. Journal of Agriculture and Food Chemistry. 2003. v.51. p.354-361.
  • Thomas, P.M., Flanagan, V.P., Pawlosky, R.J. Determination of 5- methyltetrahydrofolic acid and folic acid in citrus juices using stable isotope dilution-mass spectrometry. Journal of Agriculture and Food Chemistry. 2003. v.51. p.1293-1296.


Progress 10/01/02 to 09/30/03

Outputs
1. What major problem or issue is being resolved and how are you resolving it? Current food analysis methods ignore the multiple isomers of folate, inositol phosphates and trans-fatty acids in foods. Precise and robust methods are being developed for the determination of nondigestible oligosaccharides, resistant starch, inositol phosphates, folates, trans- polyunsaturated fatty acid, and oxidized lipids. 2. How serious is the problem? Why does it matter? Lack of precise and robust methods, thus, unreliable data often lead to inconclusive results from a number of epidemiological studies involving the roles of various nutrients in health and diseases. Nutritionists and clinicians are in search of methods which will provide much needed food composition data on the above-mentioned food constituents. Furthermore, the release of Dietary Reference Intakes by the National Academy of Science based on dietary folate equivalent requires food data on folate isomers and folic acid. 3. How does it relate to the National Program(s) and National Program Component(s) to which it has been assigned? This program meets the need described under National Program 107, REE General Goal III: A Healthy and Well-Nourished Population Who Have Knowledge, Desire and Means to Make Health Promoting Choices; specifically under Objective 3.1 Nutritious Food with Strategy 3.1.2 for Food Composition and Consumption, among others to develop new methods for measuring selected nutrients and food components, and to transfer new measurement techniques and data to users. 4. What were the most significant accomplishments this past year? A. Single Most Significant Accomplishment During FY 2003: An adequate intake of folates has been directly linked to the prevention of neural tube defects and indirectly to cancer prevention because of the role folates play in the methylation of DNA. Two methods, based on high- pressure liquid chromatography-mass spectrometry (HPLC-MS) and HPLC- fluorescence spectrometry (HPLC-FS), have been developed to provide alternative analytical methods to the non-specific, microbiological assay. HPLC-MS provides definitive quantification of folic acid and 5-methyl- tetrahydrofolic acid (the common fortificant and dominant food folate, respectively), and has been used to validate the HPLC-FS method that is less specific, but more economical, and provides for the determination of folic, 5-methyl-tetrahydrofolic, tetrahydrofolic, 5,10-methenyl- tetrahydrofolicand 5-formyltetrahydrofolic acids (the latter 3 found only in the liver). These methods will provide accurate determinations of all folate vitamers in foods and better estimation of dietary intakes. B. Other Significant Accomplishments: Phytic acid, and other inositol phosphates that occur naturally in foods, have received increasing attention for their reported antioxidative and anticarcinogenic properties, which may be beneficial to human health. A high-performance anion-exchange chromatographic method was developed in the Food Composition Laboratory, Beltsville Human Nutrition Research Center (BHNRC), that is capable of separating 27 inositol phosphate isomer peaksand was used for the quantitative determination of phytic acid and inositol pentakisphosphates in 7 legumes and 6 nuts. Results indicated that the method has excellent reproducibility and will be useful for other analysts in generating food composition data on phytic acid and related inositol phosphates. This method will be used to establish more reliable databases on phytic acid and inositol phosphates and will allow for more definitive conclusions regarding the true health benefit of these food components. C. Significant Accomplishments that Support Special Target Populations: None. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. A method was developed for the determination of total dietary fiber in order to estimate the amount of resistant starch in foods; such as cooked dried legumes and baked products containing high level of amylose. A comparison of this method with two other more labor-intensive methods showed that this is a simpler and more cost-effective method. A definitive mass spectrometric method was developed for the determination of folic and 5-methyl tetrahydrofolic acid in foods using 13C-labeled isotopes. These methods have been used for the determination of folate vitamers in vegetables and breads. A second, less expensive, and more robust method for the determination of all the folate vitamers has been developed based on liquid chromatographic separation and fluorescence detection. This method uses only commercially available materials. 6. What do you expect to accomplish, year by year, over the next 3 years? This project will be replaced by a new project that is currently being peer reviewed. The new project is not yet approved by the Office of Scientific Quality Review (OSQR). 8. List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: This does not replace your peer-reviewed publications listed below). Finding the Folate in Foods. Agricultural Research. 2002. v.50(12). p.19.

Impacts
(N/A)

Publications

  • Thomas, P.M., Flanagan, V.P., Pawlosky, R.J. Determination of 5- methyltetrahydrofolic acid and folic acid in citrus juices using stable isotope dilution-mass spectrometry. Journal of Agriculture and Food Chemistry. 2003. v.51. p.1293-1296.
  • Pawlosky, R.J., Flanagan, V.P., Doherty, R.F. A mass spectrometric validated high-performance liquid chromatography procedure for the determination of folates in foods. Journal of Agriculture and Food Chemistry. 2003. v.51. p.3726-3730.
  • Pawlosky, R.J., Hertrampf, E., Flanagan, V.P., Thomas, P.M. Mass spectral determinations of the folic acid content of fortified breads from Chile. Journal of Food Composition and Analysis. 2003. v.16. p.281-286.
  • Li, B.W., Andrews, K.W., Pehrsson, P.R. Individual sugars, soluble, and insoluble dietary fiber contents of 70 high consumption foods. Journal of Food Composition and Analysis. 2002. v.15. p.725-733.
  • Doherty, R.F., Beecher, G.R. A method for the analysis of natural and synthetic folate in foods. Journal of Agriculture and Food Chemistry. 2003. v.51. p.354-361.