High temperature reversed-phase HPLC using deuterium oxide as a mobile phase for the separation of model pharmaceuticals with multiple on-line spectroscopic analysis (UV, IR, 1H-NMR and MS)

The reversed-phase chromatography of a number of model pharmaceuticals using deuterium oxide (D2O) as the mobile phase at elevated temperatures, including superheated conditions (greater than 100 degreesC), is described. Following elution the analytes were characterised on-line via a combination of diode array UV, H-1-NMR, FT-IR spectroscopy and mass spectrometry. This combination of spectrometers enabled the on-flow collection of full UV, H-1-NMR, IR and mass spectra for a range of compounds in amounts ranging from 46 to 500 mug on-column. The advantages of the use of D2O alone as mobile phase for chromatography with multiple spectroscopic characterisation of analytes is discussed.