AOAC Official Method 2011.20: Analysis of 5′‑Mononucleotides in Infant Formula and Adult/Pediatric Nutritional Formula


The method for the analysis of nucleotide 5′‑monophosphates was successfully trialled in ten laboratories around the world as part of the MLT study; which led to its adoption as Official Final Action method 2011.20 by AOAC International and as ISO Standard 20638:2015 and was subsequently endorsed by Codex Alimentarius as a Type II Reference Method.

Method Principle: The sample is dissolved in high-salt solution to inhibit protein and fat interactions. The 5′‑mononucleotides—uridine 5′‑monophosphate (UMP), inosine 5′‑monophosphate (IMP), adenosine 5′‑monophosphate (AMP), guanosine 5′‑monophosphate (GMP), and cytidine 5′‑monophosphate (CMP) are separated from the sample matrix by strong-anion exchange solid-phase extraction (SPE), followed by chromatographic analysis using a C18 stationary phase with gradient elution, UV detection, and quantitation by an internal standard (IS) technique using thymidine 5′‑monophosphate (TMP).


Method Link
Reference Style

Journal: Journal of AOAC International
Publisher: AOAC International

AOAC International (2016) 5′‑Mononucleotides in Infant Formula and Adult/Pediatric Nutritional Formula: Gaithersburg MD, USA. Official Method 2011.20

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