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Analysis of Nucleotide 5′-Monophosphates in Infant Formulas by HPLC-UV: Collaborative Study, Final Action 2011.20

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A collaborative study was conducted on AOAC First Action Method 2011.20: 5′-Mononucleotides in Infant Formula and Adult/Pediatric Nutritional Formula. After the successful analysis of National Institute of Standards and Technology (NIST) 1849a Standard Reference Material (SRM) as a practice sample, 12 laboratories participated in the analysis of duplicate samples of six different infant formula products. The samples were dissolved in high-salt solution to inhibit protein and fat interactions, with the nucleotides [uridine 5′-monophosphate (UMP), inosine 5′-monophosphate (IMP), adenosine 5′-monophosphate (AMP), guanosine 5′-monophosphate (GMP), and cytidine 5′-monophosphate (CMP)] separated from the sample matrix by strong-anion exchange SPE, followed by chromatographic analysis using a C18 stationary phase with gradient elution, UV detection, and quantitation by an internal standard technique using thymidine 5′-monophosphate. For nucleotide-supplemented products, precision is within the Standard Method Performance Requirements SM (SMPR) 2011.008 target reproducibility limit of ≤11%, with the reproducibility RSD (RSDR) estimated at 7.1–8.7% for CMP, 7.9–9.0% for UMP, 2.8–7.7% for GMP, 5.5–10.3% for IMP, and 2.7–6.2% for AMP, and Horwitz ratio (HorRat) values of 0.9–1.0 for CMP, 0.9–1.0 for UMP, 0.3–0.7 for GMP, 0.6–1.0 for IMP, and 0.3–0.7 for AMP.
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Document Type: Research Article

Affiliations: Fonterra Co-operative Group Ltd, PO Box 7, Waitoa 3341, New Zealand

Publication date: July 1, 2015

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 The scope of the Journal includes unpublished original research describing new analytical methods, techniques and applications; improved approaches to sampling, both in the field and the laboratory; better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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