A simple and sensitive assay for the quantitative analysis of paclitaxel and metabolites in human plasma using liquid chromatography/tandem mass spectrometry

Abstract

A sensitive and specific LC-MS/MS assay for the determination of paclitaxel and its 3'p- and 6-alpha-hydroxy metabolites is presented. A 200 microL plasma aliquot was spiked with a 13C6-labeled paclitaxel internal standard and extracted with 1.0 mL tert-butylmethylether. Dried extracts were reconstituted in 0.1 M ammonium acetate-acetonitrile (1:1, v/v) and 25 microL volumes were injected onto the HPLC system. Separation was performed on a 150 x 2.1 mm C18 column using an alkaline eluent (10 mm ammonium hydroxide-methanol, 30:70, v/v). Detection was performed by positive ion electrospray followed by tandem mass spectrometry. The assay quantifies a range for paclitaxel from 0.25 to 1000 ng/mL and metabolites from 0.25 to 100 ng/mL using 200 microL human plasma samples. Validation results demonstrate that paclitaxel and metabolite concentrations can be accurately and precisely quantified in human plasma. This assay is now used to support clinical pharmacologic studies with paclitaxel.

Competitive mass spectrometry binding assay for characterization of three binding sites of tubulin

“…1B, the predominant product ion m/z 272 of double-charged vinblastine was consistent with Dennison. [25] For paclitaxel (m/z 854), the main fragment ions, m/z 569, 551, 509 and 286 were consistent with Liia et al [26] In subsequent experiments using MRM modes for quantitative measurement, the MRM ion pairs, m/z 400 → 310, 406 → 272 and 854 → 286, were selected to result in a highly specific and sensitive method for quantitation of colchicine, vinblastine and paclitaxel, respectively. An internal standard, an analog of SMART compounds, was added to improve the analytical accuracy of the quantitative method.…”

Multidrug Resistance Protein 2 Is an Important Determinant of Paclitaxel Pharmacokinetics

“…Amounts of paclitaxel and its hydroxylated metabolites 3 ¶-p-hydroxypaclitaxel and 6a-hydroxypaclitaxel in small plasma samples, obtained by sampling from the tail vein, were determined using a previously described sensitive and specific liquid chromatograpy-mass spectrometry/mass spectrometry assay (22). All other samples were processed using liquid-liquid and solid-phase extraction followed by reversed-phase HPLC with UV detection (23), with minor modifications.…”

Improved high-performance liquid chromatographic detection of paclitaxel in patient's plasma using solid-phase extraction, and semi-micro-bore C18 separation and UV detection

“…Numerous analytical methods for determination of taxanes in biological fluids have been proposed [9][10][11][12][13][14]. Most of them rely on HPLC separation with UV [9][10][11][12] or mass detection [13,14].…”