title: Measurement of paclitaxel and its metabolites in human plasma using liquid chromatography/ion trap mass spectrometry with a sonic spray ionization interface.
abstract: A quantitative liquid chromatography/ion trap mass spectrometry method for the simultaneous determination of paclitaxel, 6alpha-hydroxypaclitaxel and p-3'-hydroxypaclitaxel in human plasma has been developed and validated. 6alpha-,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 microL of reconstituted material onto a 100 x 3.00 mm C12 column with a methanol:1% trifluoroacetic acid/ammonium trifluoroacetate in H(2)O 70:30 mobile phase at 350 microL/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 6alpha-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.
authors.affiliation: Division of Clinical Pharmacology, Department of Medicine and Care, Faculty of Health Sciences, Linköping University, SE-581 85 Linköping, Sweden. henrik.green@imv.liu.se
authors.forename: Henrik
authors.initials: H
authors.lastname: Gréen
authors.forename: Karin
authors.initials: K
authors.lastname: Vretenbrant
authors.forename: Björn
authors.initials: B
authors.lastname: Norlander
authors.forename: Curt
authors.initials: C
authors.lastname: Peterson
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000097:blood* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D017239:Paclitaxel / Q000031:analogs & derivatives / Q000097:blood* / Q000378:metabolism; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods; D043823:Taxoids / Q000097:blood*
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D043823:Taxoids; C102050:3'-p-hydroxypaclitaxel; C086219:6-hydroxytaxol; D017239:Paclitaxel