Yu Haoying, Straubinger Robert M, Cao Jin, Wang Hao, Qu Jun
The Department of Pharmaceutical Sciences, University at Buffalo, State University of New York, Amherst, NY 14260-1200, USA.
J Chromatogr A. 2008 Nov 14;1210(2):160-7. doi: 10.1016/j.chroma.2008.09.052. Epub 2008 Sep 19.
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.
对生物基质中生物活性化合物的超低浓度进行定量分析的能力，对于研究低剂量下发生的药理/毒理效应至关重要。将选择性固相萃取（SPE）与高灵敏度毛细管液相色谱（microLC）-质谱/质谱分析相结合，以实现对癌细胞中抗癌药物紫杉醇的超灵敏定量分析。优化后的SPE能选择性地萃取紫杉醇，并去除不良基质化合物，从而可在不影响色谱性能和操作稳健性的情况下，在microLC柱上实现高进样量。验证后的定量下限（LOQ）为5pg/mL，比已发表的液相色谱-质谱/质谱方法灵敏约20倍。校准曲线在5-6250pg/mL范围内呈线性。准确度为98-109%，变异系数（CV%）为2.3-7.4%。该方法成功应用于定量分析经亚纳克/毫升浓度紫杉醇处理的A121a卵巢癌细胞的时间药物积累情况。
