Measurement of paclitaxel in biological matrices: high-throughput liquid chromatographic-tandem mass spectrometric quantification of paclitaxel and metabolites in human and dog plasma.
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.
Alexander
Michael S
MS
LC Resources Inc, 3138 NE Rivergate, Bldg 301C, McMinnville, OR 97128, USA. mike.alexander@lcresources.com
Kiser
Melissa M
MM
Culley
Travis
T
Kern
John R
JR
Dolan
John W
JW
McChesney
James D
JD
Zygmunt
Jan
J
Bannister
Steve J
SJ
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
D000972:Antineoplastic Agents, Phytogenic; D017239:Paclitaxel