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, H-1-NMR and MS)

The reversed-phase chromatography of a number of model pharmaceuticals usin g deuterium oxide (D2O) as the mobile phase at elevated temperatures, inclu ding superheated conditions (greater than 100 degreesC), is described. Foll owing elution the analytes were characterised on-line via a combination of diode array UV, H-1-NMR, FT-IR spectroscopy and mass spectrometry. This com bination of spectrometers enabled the on-flow collection of full UV, H-1-NM R, 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 phas e for chromatography with multiple spectroscopic characterisation of analyt es is discussed.