The structural identification of drug metabolites has been carried out using thermospray liquid chromatography/mass spectrometry (LC/MS). It has allowed the direct analysis of biological samples, in this case in vitro hepatocyte incubations, with the minimum of sample preparation. The technique also provided molecular weight information on several conjugates including glucuronides, a glutathione conjugate and one unidentified conjugate. A number of minor metabolites were also successfully identified using this method. The examples discussed in this paper illustrate the value of LC/MS in identifying unknown drug metabolites covering a wide polarity range in a complex biological mixture. However, this would not have been possible, if the interface had been unable to handle gradient separations.