Directly coupled HPLC–NMR and HPLC–NMR–MS in pharmaceutical research and development

The methodology for the direct coupling of HPLC with NMR spectroscopy and the simultaneous double coupling of HPLC with NMR and mass spectrometry (MS) is described. Indications of the necessary technical developments to achieve this are given, and the applications of these new techniques to studies of pharmaceutical relevance are reviewed. These include studies of combinatorial chemistry libraries, synthetic chemical impurities, characterisation of drug mixtures, identification of natural products of possible pharmaceutical interest and identification of xenobiotic metabolites in human, animal and in vitro systems. In addition, HPLC–NMR has been used to investigate xenobiotic metabolite reactivity. Finally, the potential future directions of the techniques are discussed.