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An RP-HPLC method for the routine determination of supplemented 5-mononucleotides (uridine 5-monophosphate, inosine 5-monophosphate, adenosine 5-monophosphate, guanosine 5-monophosphate, and cytidine 5-monophosphate) in pediatric formulas and milk products is described. Following sample dissolution, potential interferences were removed by anion-exchange SPE. Chromatographic analyses were performed using a C18 stationary phase with gradient elution, UV detection, and quantitation by an internal standard technique. A single-laboratory validation was performed, with recoveries of 92101 and repeatability RSDs of 1.02.3. The method was optimized for the rapid, routine analysis of nucleotide-supplemented bovine milk-based infant and follow-on formulas.

A Liquid Chromatographic Method for Routine Analysis of 5-Mononucleotides in Pediatric Formulas