A rapid and reproducible method with high selectivity was developed for simultaneous determination of a promising anti-brain tumor agent CAT3 and its two metabolites PF403 and GLU-PF403 in mouse plasma and brain. An economic deproteinization with septuple acetonitrile (v/v) was applied to pretreat the samples in this study. All analytes were well retained and separated on a CAPCELL CORE PC (2.7 μm, 2.1 mm I.D. × 150 mm, SHISEIDO Technologies) column with an eluting solvent of acetonitrile /water containing 0.1 % formic acid (v/v) at the flow rate of 0.2 mL per minute. The detection was carried out on a Q Exactive high resolution mass spectrometer equipped with a HESI ion source in parallel reaction monitoring (PRM) mode. The corresponding transitions for quantitation were 434.23→ 70.07 for CAT3, 350.17→70.07 for PF403, 526.21→70.07 for GLU-PF403, 364.19→70.07 for IS-1 and 625.18→317.07 for IS-2, respectively. A well-linear fit curve was achieved among the range of 0.1∼50 ng/mL for CAT3, 0.2∼100 ng/mL for PF403 and 2.5∼600 ng/mL for GLU-PF403 both in mouse plasma and brain homogenate. The intra-/inter-day accuracies of three analytes were within ±14.5 % and precisions were below to 13.44 %. The mean values of recovery of three compounds in mouse plasma and brain homogenate were among 98.06 ∼ 118.63 % and 81.04∼108.69 %. The analytes in NaF-treated ice cold blood of mouse was stable within tested 30 min. Plasma and brain homogenate samples had no obvious changes during all storage, sample treatment and analytic process of mouse plasma sample. The reproducible and reliable method was well employed to the research of CAT3 pharmacokinetic characteristics in mouse plasma and brain after a single intragastric administration at dose of 10 mg/kg.

中文翻译：

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通过LC-MS / MS方法同时测定小鼠血浆和脑中有希望的抗脑肿瘤药物CAT3及其两种主要代谢产物。

建立了一种快速且可重现的高选择性方法，用于同时测定小鼠血浆和脑中有希望的抗脑肿瘤药物CAT3及其两种代谢物PF403和GLU-PF403。在这项研究中，采用七元乙腈（v / v）进行经济的脱蛋白预处理样品。所有分析物均被很好地保留并在CAPCELL CORE PC（2.7μm，2.1 mm ID×150 mm，SHISEIDO Technologies）色谱柱上，用乙腈/水的洗脱溶剂以0.1％的甲酸（v / v）洗脱每分钟0.2 mL。该检测在配备有HESI离子源的Q Exactive高分辨率质谱仪上以并行反应监控（PRM）模式进行。CAT3的定量转换为434.23→70.07，PF403的定量为350.17→70.07，GLU-PF403的为526.21→70.07，对于IS-1，分别为364.19→70.07，对于IS-2为625.18→317.07。在小鼠血浆和脑匀浆中，CAT3在0.1〜50 ng / mL，PF403在0.2〜100 ng / mL和GLU-PF403在2.5〜600 ng / mL的范围内获得了良好的线性拟合曲线。三种分析物的日内/日间准确度在±14.5％之内，而精度低于13.44％。小鼠血浆和脑匀浆中三种化合物的回收率平均值分别为98.06〜118.63％和81.04〜108.69％。经NaF处理的小鼠冰冷血液中的分析物在测试的30分钟内稳定。在小鼠血浆样品的所有储存，样品处理和分析过程中，血浆和脑匀浆样品均无明显变化。