Ultra-sensitive quantification of paclitaxel using selective solid-phase extraction in conjunction with reversed-phase capillary liquid chromatography/tandem mass spectrometry.
The ability to quantify ultra-low concentrations of biologically active compounds in biological matrices is essential for the study of pharmacological/toxicological effects occurring at low doses. Selective solid-phase extraction (SPE) was combined with highly sensitive capillary LC (microLC)-MS/MS analysis to achieve ultra-sensitive quantification of the anti-cancer drug paclitaxel in cancer cells. The optimized SPE selectively extracted paclitaxel and eliminated undesirable matrix compounds, thus enabling a high sample loading volume on the microLC column without compromising chromatographic performance and operational robustness. The validated lower limit of quantification (LOQ) was 5pg/mL, approx. 20-fold more sensitive than published LC-MS/MS methods. The calibration curve was linear over the range of 5-6250pg/mL. Accuracy was 98-109% and the variation (CV%) was 2.3-7.4%. This method was applied successfully to quantify temporal drug accumulation by A121a ovarian cancer cells treated with sub-ng/mL concentrations of paclitaxel.
The Department of Pharmaceutical Sciences, University at Buffalo, State University of New York, Amherst, NY 14260-1200, USA.
Haoying
H
Yu
Robert M
RM
Straubinger
Jin
J
Cao
Hao
H
Wang
Jun
J
Qu
D000972:Antineoplastic Agents, Phytogenic / Q000032:analysis; D002138:Calibration; D045744:Cell Line, Tumor / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods; D005260:Female; D006801:Humans; D017239:Paclitaxel / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000379:methods*; D053719:Tandem Mass Spectrometry
D000972:Antineoplastic Agents, Phytogenic; D017239:Paclitaxel