Department of Pharmacy, Luzhou People's Hospital, Luzhou, China.
School of Pharmacy, Southwest Medical University, Luzhou, China.
Biomed Chromatogr. 2021 Feb;35(2):e4968. doi: 10.1002/bmc.4968. Epub 2020 Sep 16.
In this study, a simple and sensitive UHPLC-ESI-MS/MS method was established for the determination of LXH254 in rat plasma. The developed method was validated according to the Food and Drug administration guidelines. After extraction using ethyl acetate, the sample was separated on an ACQUITY BEH C column. The mobile phase consisted of 2 mM ammonium acetate containing 0.1% formic acid and acetonitrile as the mobile phase with gradient elution. The flow rate was 0.3 mL/min. A TSQ triple quadrupole mass spectrometer operated in positive-ion mode was used for mass detection, with multiple reaction monitoring transitions of m/z 503.3 > 459.1 and m/z 435.3 > 367.1 for LXH254 and olaparib (internal standard), respectively. An excellent linearity was achieved in the concentration range of 0.1-1000 ng/mL, with correlation coefficient >0.998. The mean recovery was more than 78.55%. Inter- and intra-day precision (percentage of relative standard deviation) did not exceed 12.87%, and accuracy was in the range of -2.50 to 13.50%. LXH254 was demonstrated to be stable under the tested storage conditions. The validated UHPLC-MS/MS method was further applied to the pharmacokinetic study of LXH254 in rat plasma after oral (2, 5, and 15 mg/kg) and intravenous (2 mg/kg) administrations. The pharmacokinetic study revealed that LXH254 showed low clearance, moderate bioavailability (~30%), and linear pharmacokinetic profile over the oral dose range of 2-15 mg/kg. To the best of our knowledge, this is the first report on the method development and validation of the determination of LXH254 and its application to pharmacokinetic study.
在这项研究中，建立了一种简单灵敏的 UHPLC-ESI-MS/MS 方法，用于测定大鼠血浆中的 LXH254。该方法按照美国食品和药物管理局的指南进行了验证。样品经乙酸乙酯提取后，在 ACQUITY BEH C 柱上进行分离。流动相由 2 mM 乙酸铵和 0.1%甲酸的水溶液与乙腈组成，采用梯度洗脱。流速为 0.3 mL/min。采用 TSQ 三重四极杆质谱仪，以正离子模式进行质谱检测，分别对 m/z 503.3 > 459.1 和 m/z 435.3 > 367.1 进行多重反应监测转换，用于 LXH254 和奥拉帕利（内标）的质量检测。在 0.1-1000 ng/mL 的浓度范围内，线性关系良好，相关系数>0.998。平均回收率大于 78.55%。日内和日间精密度（相对标准偏差的百分比）不超过 12.87%，准确度在-2.50 至 13.50%范围内。LXH254 在测试的储存条件下表现出稳定性。经验证的 UHPLC-MS/MS 方法进一步应用于大鼠口服（2、5 和 15 mg/kg）和静脉（2 mg/kg）给予 LXH254 后的药代动力学研究。药代动力学研究表明，LXH254 清除率低，生物利用度适中（约 30%），口服剂量范围为 2-15 mg/kg 时呈线性药代动力学特征。据我们所知，这是首次报道 LXH254 的测定方法开发和验证及其在药代动力学研究中的应用。
