Measurement of paclitaxel and its metabolites in human plasma using liquid chromatography/ion trap mass spectrometry with a sonic spray ionization interface

A quantitative liquid chromatography/ion trap mass spectrometry method for the simultaneous determination of paclitaxel, 6 alpha-hydroxypaclitaxel and p-3'-hydroxypaclitaxel in human plasma has been developed and validated. 6 alpha-,p-3'-Dihydroxypaclitaxel was also quantified using paclitaxel as a reference and docetaxel as an internal standard. The substances were extracted from 0.500 mL plasma using solid-phase extraction. The elution was performed with acetonitrile and the samples were reconstituted in the mobile phase. Isocratic high-performance liquid chromatography analysis was performed by injecting 50 mu L of reconstituted material onto a 100 x 3.00 mm C12 column with a methanol:1% trifluoroacetic acid/ammonium trifluoroacetate in H2O 70:30 mobile phase at 350 mu L/min. The [M+H](+) ions generated in the sonic spray ionization interface were isolated and fragmented using two serial mass spectrometric methods: one for paclitaxel (transition 854 -> 569 & 551) and the dihydroxymetabolite (transition 886 -> 585 & 567) and one for the hydroxy metabolites (transition 870 -> 585 & 567; transition 870 -> 569 & 551) and docetaxel ([M+Na](+), transition 830 -> 550). Calibration curves were created ranging between 0.5 and 7500 ng/mL for paclitaxel, 0.5 and 750 ng/mL for 6 alpha-hydroxypaclitaxel, and 0.5 and 400 ng/mL for p-3'-hydroxypaclitaxel. Adduct ion formation was noted and investigated during method development and controlled by mobile phase optimization. In conclusion, a sensitive method for simultaneous quantification of paclitaxel and its metabolites suitable for analysis in clinical studies was obtained. Copyright (c) 2006 John Wiley & Sons, Ltd.