Cancer Science Institute of Singapore, National University of Singapore, Singapore.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Sep 15;878(26):2409-14. doi: 10.1016/j.jchromb.2010.07.015. Epub 2010 Jul 30.
A novel, sensitive and reliable liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed and validated for the determination of belinostat (PXD101) in human plasma. Oxamflatin was used as the internal standard. Liquid-liquid extraction of the plasma sample was performed using tert-butyl methyl ether as the organic solvent. Chromatographic separation was achieved on a BDS Hypersil C18 column (2.1 mm x1 00 mm, 5 microm) using gradient elution mode using 0.05% formic acid in water and 0.05% formic acid in acetonitrile as solvents A and B, respectively, 60/40. The run time was 6 min. The mass spectrometer was operated under a positive electrospray ionization condition and a multiple reaction monitoring mode. An excellent linear calibration was achieved in the range of 0.5-1000 ng/mL. An average recovery of belinostat for four quality controls was 72.6% and the recovery of the internal standard at 1000 ng/mL was 67.8%. The intra-day and inter-day precisions for belinostat were <or=8.0 and <or=10.3%, respectively, and their accuracy ranged from 100.2 to 106.7%. No significant matrix effect was identified. In analysis of patient samples, belinostat glucuronide was identified and baseline separated from belinostat. This well-validated assay has been applied for quantification of belinostat in plasma samples within 24h after the start of infusion for Asian hepatocellular carcinoma patients in a dose escalation study.
建立并验证了一种灵敏、可靠的液相色谱-串联质谱（LC-MS/MS）法，用于测定人血浆中的贝林司他（PXD101）。奥沙氟嗪为内标。采用叔丁基甲醚作为有机溶剂，对血浆样品进行液-液萃取。采用 BDS Hypersil C18 柱（2.1mm×100mm，5μm），以水相 0.05%甲酸和乙腈相 0.05%甲酸为流动相（A 相和 B 相），采用梯度洗脱方式进行色谱分离，洗脱程序为 60/40。运行时间为 6min。质谱采用正电喷雾电离源，多反应监测模式进行检测。贝林司他在 0.5-1000ng/ml 范围内呈良好线性关系。4 个质控样品的平均回收率为 72.6%，内标在 1000ng/ml 时的回收率为 67.8%。贝林司他日内和日间精密度均<8.0%和<10.3%，准确度为 100.2%-106.7%。未观察到明显的基质效应。在分析患者样本时，鉴定出贝林司他葡萄糖醛酸结合物，并与贝林司他基线分离。该方法经过充分验证，已用于亚洲肝细胞癌患者剂量递增研究中，在输注开始后 24h 内对血浆样本中的贝林司他进行定量分析。
