Development and validation of a highly sensitive liquid chromatography/mass spectrometry method for simultaneous quantification of lenalidomide and flavopiridol in human plasma
Development and validation of a highly sensitive liquid chromatography/mass spectrometry method for simultaneous quantification of lenalidomide and flavopiridol in human plasma
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DOI：
10.1097/ftd.0b013e318185813d
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发表时间：
2008-10-01
影响因子：
2.5
通讯作者：
Phelps, Mitch A.
中科院分区：
文献类型：
--
作者：
Liu, Qing;Farley, Katherine L.;Phelps, Mitch A.
来那度胺（一种免疫调节剂）和黄吡醇（一种广泛的细胞周期蛋白依赖性激酶抑制剂）是临床用于基因组高危慢性淋巴细胞白血病的积极疗法。开发了一种具有串联质谱检测功能的高效液相色谱测定法，可同时定量人和小鼠血浆中的来那度胺和黄吡醇，以促进其联合临床开发。使用含有乙腈 (ACN) 的内标染料木黄酮进行液-液萃取，然后蒸发溶剂并在 95/5 H2O/ACN 中复溶来制备样品。通过反相液相色谱法在 C-18 柱上使用 H2O 和 ACN（各含 0.1% 甲酸）梯度分离来那度胺和内标。采用正离子模式的大气压化学电离，并在三重四极杆质谱仪上进行单反应监测，用于检测来那度胺 (260.06 > 149. 10) 和黄吡醇 (402.09 > 341.02) 的转变。来那度胺和黄酮吡多的定量下限分别为 1 和 0.3 nM。在整个线性范围内，来那度胺和黄吡醇从人血浆中的回收率为 99% 至 116%。重复样品的运行内和运行间精密度和准确度均低于 15%。这是迄今为止报道的来那度胺和黄酮吡多最灵敏的分析方法。这种灵敏度将使在慢性淋巴细胞白血病患者中使用来那度胺和黄吡醇进行的计划临床试验中进行晚期终末期浓度测量和准确的药代动力学参数估计成为可能。
Lenalidomide, an immunomodulatory agent, and flavopiridol, a broad cyclin-dependent kinase inhibitor, are active therapies for clinical use in genomic high-risk chronic lymphocytic leukemia. A high-performance liquid chromatographic assay with tandem mass spectrometric detection has been developed to simultaneously quantify lenalidomide and flavopiridol in human and mouse plasma to facilitate their combined clinical development. Samples were prepared by liquid-liquid extraction with acetonitrile (ACN)-containing internal standard, genistein, followed by evaporation of solvent and reconstitution in 95/5 H2O/ACN. Lenalidomide and internal standard were separated by reversed-phase liquid chromatography on a C-18 column using a gradient of H2O and ACN, each with 0.1% formic acid. Atmospheric pressure chemical ionization in positive ion mode with single reaction monitoring on a triple quadrupole mass spectrometer was applied to detect transitions of lenalidomide (260.06 > 149. 10) and flavopiridol (402.09 > 341.02). Lower limits of quantification of lenalidomide and flavopiridol were I and 0.3 nM, respectively. Recoveries of lenalidomide and flavopiridol from human plasma ranged from 99% to 116% throughout their linear ranges. Within- and between-run precision and accuracy of replicate samples were all less than 15%. This is the most sensitive analytical method reported to date for both lenalidomide and flavopiridol. This sensitivity will enable late terminal phase concentration measurements and accurate pharmacokinetic parameter estimation in a planned clinical trial with lenalidomide and flavopiridol in patients with chronic lymphocytic leukemia.