Li Shuang-Long, Zhu Yong-Liang, Zhang Yi, Liu Shu-Han, Wang Xiang-Die, Qiu Xiang-Jun
Medical College of Henan University of Science and Technology, Luoyang 471023, China.
Int J Anal Chem. 2020 Mar 31;2020:5084127. doi: 10.1155/2020/5084127. eCollection 2020.
In our research, a straightforward UPLC-MS/MS method, with diazepam as the internal standard (IS), was proposed and acknowledged to determine the concentrations of enasidenib in rat plasma. When preparing the sample, we used acetonitrile for protein precipitation. The gradient elution method was used, and the mobile phase was acetonitrile and 0.1% formic acid. Diazepam was used as the IS. We used the Acquity UPLC BEH C18 column to separate enasidenib and IS. Under the positive ion electrospray ionization (ESI) source conditions, the mass transfer pairs of enasidenib were monitored by multiple reaction monitoring (MRM) to be 474.2 ⟶ 456.1 and 474.2 ⟶ 267.0, and the IS mass transfer pairs were 285.0 ⟶ 154.0. Enasidenib had good linearity ( = 0.9985) in the concentration range of 1.0-1000 ng/mL. Besides, the values of intraday and interday precision were 2.25-8.40% and 3.94-5.46%, respectively, and the range of the accuracy values varied from -1.44 to 2.34%. Matrix effect, extraction recovery, and stability were compliant with FDA approval guidelines in terms of bioanalytical method validation. We had established a new method that had been applied to the pharmacokinetic study of enasidenib in rats.
在我们的研究中，提出了一种以地西泮为内标（IS）的简单超高效液相色谱-串联质谱（UPLC-MS/MS）方法，并被认可用于测定大鼠血浆中恩杂鲁胺的浓度。在制备样品时，我们使用乙腈进行蛋白沉淀。采用梯度洗脱方法，流动相为乙腈和0.1%甲酸。地西泮用作内标。我们使用 Acquity UPLC BEH C18 柱分离恩杂鲁胺和内标。在正离子电喷雾电离（ESI）源条件下，通过多反应监测（MRM）监测恩杂鲁胺的质荷比为474.2⟶456.1和474.2⟶267.0，内标的质荷比为285.0⟶154.0。恩杂鲁胺在1.0 - 1000 ng/mL浓度范围内具有良好的线性（r = 0.9985）。此外，日内和日间精密度值分别为2.25 - 8.40%和3.94 - 5.46%，准确度值范围为 -1.44至2.34%。在生物分析方法验证方面，基质效应、提取回收率和稳定性均符合美国食品药品监督管理局（FDA）的批准指南。我们建立了一种新方法，并已将其应用于恩杂鲁胺在大鼠体内的药代动力学研究。
