Ion-Exchange Chromatography Followed by ESI-MS for Quantitative Analysis of Sugar Monophosphates from Glucose Catabolism

The aim of this work is to establish a quantitative method to determine the ratio of [U- 13C] labeled to unlabeled hexose monophosphates isolated from yeast extracts. This is accomplished by anion exchange chromatography and mobile phase desalting followed by electrospray (ESI) mass spectrometry. We test the method with the analysis of a sample of biological origin. Previously developed analytical techniques are not adequate to accomplish mass spectrometric analysis of these and other small monosaccharide systems because of interference from salt clusters. By lowering the ionic strength of the mobile phase and using a simplified injection system to the mass spectrometer, we were able to obtain data on the relative abundance of the hexose monophosphates.