The analysis of 5'-mononucleotides in infant formulae by HPLC
2001
Perrin, C. | Meyer, L. | Mujahid, C. | Blake, C.J.
A method is described for the determination of four 5'-mononucleotides (cytidine 5'-monophosphate, uridine 5'-monophosphate, adenosine 5'-monophosphate and guanosine 5'-monophosphate) in infant formulae. Nucleosides which may be formed during processing can also be analysed simultaneously. This method is based on deproteinisation of samples and direct analysis by ion-pair HPLC using two Nucleosil 120-C18 columns in series, followed by diode-array detection. This method gives good recoveries of 5'-mononucleotides from spiked infant formula products. However, some chromatographic interferences were observed when analysing hypoallergenic infant formulae containing hydrolysed proteins which made peak quantification difficult. To overcome this problem a strong anion-exchange solid-phase extraction (SPE) column was used. Four SPE columns from different suppliers were evaluated, but the best recoveries of all four 5'-mononucleotides and highest reproducibility of results were obtained with Baker-bond quaternary amine columns. Nucleosides, which may occur in very low concentrations in hypoallergenic products, are not retained on the SPE columns and so cannot be analysed by this technique.
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