title: Measurement of paclitaxel in biological matrices: high-throughput liquid chromatographic-tandem mass spectrometric quantification of paclitaxel and metabolites in human and dog plasma.
abstract: A GLP-validated, sensitive and specific LC-MS-MS method for the quantification of paclitaxel and its 6-alpha- and 3'-p-hydroxy metabolites is presented. A 0.400 ml plasma aliquot is spiked with a (13)C(6)-labeled paclitaxel internal standard and extracted with 1 ml methyl-tert.-butyl ether. The ether is evaporated and the residue is reconstituted in 130 microl of 30% aqueous acetonitrile (ACN) containing 0.1% trifluoroacetic acid. Isocratic HPLC analysis is performed by injecting 50 microl of the reconstituted material onto a 50x2.1 mm C(18) column with an ACN-water-acetic acid (50:50:0.1) mobile phase at 200 microl/min flow. Detection is by positive ion electrospray followed by multiple reaction monitoring of the following transitions: paclitaxel (854>509 u), 6-alpha-hydroxy paclitaxel (870>525 u), 3'-p-hydroxy paclitaxel (870>509 u) and internal standard (860>509 u). Quantification is by peak area ratio against the 13C(6) internal standard. The method range is 0.117-117 nM (0.1-100 ng/ml) for paclitaxel and both metabolites using a 0.400 ml human or dog plasma sample. Analysis time per sample is less than 5 min.
authors.affiliation: LC Resources Inc, 3138 NE Rivergate, Bldg 301C, McMinnville, OR 97128, USA. mike.alexander@lcresources.com
authors.forename: Michael S
authors.initials: MS
authors.lastname: Alexander
authors.forename: Melissa M
authors.initials: MM
authors.lastname: Kiser
authors.forename: Travis
authors.initials: T
authors.lastname: Culley
authors.forename: John R
authors.initials: JR
authors.lastname: Kern
authors.forename: John W
authors.initials: JW
authors.lastname: Dolan
authors.forename: James D
authors.initials: JD
authors.lastname: McChesney
authors.forename: Jan
authors.initials: J
authors.lastname: Zygmunt
authors.forename: Steve J
authors.initials: SJ
authors.lastname: Bannister
mesh_terms: D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000097:blood*; D001711:Biotransformation; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D004355:Drug Stability; D005615:Freezing; D006801:Humans; D013058:Mass Spectrometry; D017239:Paclitaxel / Q000097:blood*; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D017239:Paclitaxel