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

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 92-101% and repeatability RSDs of 1.0-2.3%. The method was optimized for the rapid, routine analysis of nucleotide-supplemented bovine milk-based infant and follow-on formulas.