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Validation of a Column Liquid Chromatographic Method for Phytate

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A column liquid chromatographic method that uses a spectrophotometric detector for the analysis of phytate and other inositol phosphates in foodstuffs and seeds is described. It has been tested thoroughly and sent to 6 other laboratories where there was an interest in such an analytical method and the equipment was available for performing it. These samples were blinded to the individual collaborators. Also sent was a standard wheat bran sample available from the American Association of Cereal Chemists with a standard phytate value reported as 3.2. The postcolumn sulfosalicylic acid reaction was developed in the 1950s as an analytical method for ferric ion. All reagents are aqueous solutions and the specific pH values of each are important. The column separation is at pH 4, which dissociates all phosphate analogs. The ferric sulfosalicylate is at pH 1.8. The postcolumn reaction is then at pH 2.02.3. At this pH, the sulfosalicylate complexes one ferric ion and will yield it to a stronger complexing agent such as a phosphate ester. At higher pH values, sulfosalicylate complexes 2 ferric ions and will yield neither to these anions. The method sensitivity is 812 nmoles/injection.
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Document Type: Research Article

Affiliations: 1: Texas Tech University, 3408 88th St, Lubbock, TX 79423. 2: Howard University, Nutritional Sciences, Room 327, Washington, DC 20059.

Publication date: November 1, 2007

More about this publication?
  • The Journal of AOAC INTERNATIONAL publishes refereed papers and reviews in the fields of chemical, biological and toxicological analytical chemistry for the purpose of showcasing the most precise, accurate and sensitive methods for analysis of foods, food additives, supplements and contaminants, cosmetics, drugs, toxins, hazardous substances, pesticides, feeds, fertilizers and the environment available at that point in time. The scope of the Journal includes unpublished original research describing new analytical methods, techniques and applications; improved approaches to sampling, both in the field and the laboratory; better methods of preparing samples for analysis; collaborative studies substantiating the performance of a given method; statistical techniques for evaluating data. The Journal will also publish other articles of general interest to its audience, e.g., technical communications; cautionary notes; comments on techniques, apparatus, and reagents.
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