College of Pharmacy, Dankook University, Cheonan 31116, Korea.
School of Life Sciences, Gwangju Institute of Science and Technology (GIST), Gwangju 61005, Korea.
Molecules. 2020 Apr 27;25(9):2039. doi: 10.3390/molecules25092039.
This study aimed to develop and validate a sensitive liquid chromatography-coupled tandem mass spectrometry method for the quantification of LDD-2614, an indirubin derivative and novel FLT3 inhibitor, in rat plasma. In addition, the developed analytical method was applied to observe the pharmacokinetic properties of LDD-2614. Chromatographic separation was achieved on a Luna omega C column using a mixture of water and acetonitrile, both containing 0.1% formic acid. Quantitation was performed using positive electrospray ionization in a multiple reaction monitoring (MRM) mode. The MRM transitions were optimized as 426.2→113.1 for LDD-2614 and 390.2→113.1 for LDD-2633 (internal standard), and the lower limit of quantification (LLOQ) for LDD-2614 was determined as 0.1 ng/mL. Including the LLOQ, the nine-point calibration curve was linear with a correlation coefficient greater than 0.9991. Inter- and intraday accuracies (RE) ranged from -3.19% to 8.72%, and the precision was within 9.02%. All validation results (accuracy, precision, matrix effect, recovery, stability, and dilution integrity) met the acceptance criteria of the U.S. Food and Drug Administration and the Korea Ministry of Food and Drug Safety guidelines. The proposed method was validated and demonstrated to be suitable for the quantification of LDD-2614 for pharmacokinetics studies.
本研究旨在开发和验证一种灵敏的液相色谱-串联质谱法，用于定量测定 LDD-2614，这是一种靛玉红衍生物和新型 FLT3 抑制剂，在大鼠血浆中。此外，所开发的分析方法还用于观察 LDD-2614 的药代动力学特性。色谱分离在 Luna omega C 柱上实现，使用水和乙腈的混合物，两者均含有 0.1%甲酸。定量采用正电喷雾电离在多重反应监测（MRM）模式下进行。MRM 跃迁被优化为 426.2→113.1 用于 LDD-2614 和 390.2→113.1 用于 LDD-2633（内标），LDD-2614 的定量下限（LLOQ）确定为 0.1ng/mL。包括 LLOQ，九点校准曲线呈线性，相关系数大于 0.9991。日内和日间准确度（RE）范围为-3.19%至 8.72%，精密度在 9.02%以内。所有验证结果（准确度、精密度、基质效应、回收率、稳定性和稀释完整性）均符合美国食品和药物管理局和韩国食品药品安全部指南的验收标准。所提出的方法经过验证，适用于 LDD-2614 的药代动力学研究的定量测定。
