Cyclosporine (CsA) is extensively metabolized, with over 14 metabolites having been characterized to date. The confirmation of structure and purity is a prerequisite for studies involving CsA metabolites. Analytical techniques such as fast atom bombardment/mass spectroscopy (FAB/MS), tandem mass spectrometry (MS), 1H- and 13C-nuclear magnetic resonance (NMR) can be used for such purposes. In vitro experiments indicate that metabolites are considerably less immunosuppressive and toxic than CsA. In vivo studies have been hampered by sufficient quantities of metabolites and a suitable animal model. Preliminary results in the rat suggest that CsA metabolites are less immunosuppressive and toxic than CsA, although these results must be confirmed using a more suitable animal model. Present data indicate that the routine monitoring of metabolites is not warranted in transplant patients, although additional information is required to confirm these findings.