Purification method for the isolation of monophosphate nucleotides from Champagne wine and their identification by mass spectrometry

Monophosphate nucleotides are difficult to identify in Champagne wine becau se they are present in small concentrations in a complex mixture. A method for the isolation, separation and identification of reference compounds, wh ich achieved on average 79% recovery (except for cytidine derivatives), was developed and applied to wine. Some monophosphate nucleotides were then is olated from a Champagne wine aged on lees for 8 years, by ultrafiltration f ollowed by a semi-preparative HPLC step using a strong anion-exchange colum n. The fraction obtained was subjected to HPLC in a reversed-phase column t o remove the salt previously introduced, before identification of compounds by HPLC coupled to a mass spectrometer, For the first time in wine, 5'-IMP , 5'-AMP, 5'-CMP, 5'-GMP, 5'-UMP and the 3'- and/or 2'-isomers of the four latter compounds were identified by comparing their HPLC and electrospray i onization mass spectrometry data with those of reference nucleotides. (C) 2 001 Elsevier Science B.V, All rights reserved.