存在两种对映体:(R)-和(S)-雷贝拉唑。(R)-雷贝拉唑具有特定的药代动力学优势并增强了治疗效果,保证进一步的调查和发展。这里,我们开发了一种简单快速的手性液相色谱-串联质谱(LC-MS/MS)方法来同时定量雷贝拉唑对映体及其代谢物(雷贝拉唑亚砜和去甲基雷贝拉唑对映体),并开发了一种LC-MS来定量雷贝拉唑硫醚。至于手性LC-MS/MS方法,手性AGP柱(150×4mm,使用5μm),其流动相为乙腈(流动相A)和10mmol/L乙酸铵(流动相B)(线性梯度曲线:0min,10%B;5分钟,15%B;9分钟,15%B;9.01分钟,10%B;13分钟,10%B)。m/z360.3→242.1、376.2→240.1、346.2→228.2和368.2→190.2的多个反应监测转变被用于定量雷贝拉唑对映体,雷贝拉唑亚砜,去甲基雷贝拉唑对映体和内标奥美拉唑。通过简单的液-液萃取方法制备分析物样品。至于LC-MS方法,分析物在Inertsil®ODS-3色谱柱上分离(4.6×150毫米,5μm)。流动相为乙腈-5mmol/L乙酸铵水溶液(65:35,v/v)。使用ESI+并监测m/z344.2(雷贝拉唑硫醚)和285.1(内标地西泮)的离子峰。这两种方法都经过了特异性验证,线性度精度,准确度,基体效应和提取回收率,and,特别是,分析物在各种条件下的稳定性。在静脉注射(R)-(80、20、5mg/kg/d)和外消旋(80mg/kg/d)雷贝拉唑钠后,我们成功地将这些方法应用于大鼠雷贝拉唑的13周毒代动力学研究。结果表明,雷贝拉唑及其代谢产物在大鼠体内没有积累。然而,去甲基雷贝拉唑和雷贝拉唑硫醚在最后一次给药中显示出较高的暴露量和较低的清除率。与(S)-雷贝拉唑相比,(R)-雷贝拉唑在大鼠中显示出更高的暴露量和更慢的消除速率。这些发现为(R)-雷贝拉唑的进一步临床前研究和临床应用提供了实验证据和理论基础。
There exist two enantiomers: (R)- and (S)-rabeprazole. (R)-rabeprazole offers specific pharmacokinetic advantages and enhanced therapeutic efficacy, warranting further investigation and development. Here, we developed a simple and rapid chiral liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to simultaneously quantify rabeprazole enantiomers and their metabolites (
rabeprazole sulfoxide and desmethyl
rabeprazole enantiomers) and a LC-MS to quantify rabeprazole thioether. As for the chiral LC-MS/MS method, Chiral-AGP column (150 × 4 mm, 5 μm) was used and its mobile phase was acetonitrile (mobile phase A) and 10 mmol/L ammonium acetate (mobile phase B) (linear gradient profile: 0 min, 10 % B; 5 min, 15 % B; 9 min, 15 % B; 9.01 min, 10 % B; 13 min, 10 % B). The multiple reactions monitoring transitions of m/z 360.3 → 242.1, 376.2 → 240.1, 346.2 → 228.2 and 368.2 → 190.2 were opted for quantifying rabeprazole enantiomers,
rabeprazole sulfoxide, desmethyl
rabeprazole enantiomers and internal standard omeprazole. The analyte samples were prepared by a simple liquid-liquid extraction method. As for the LC-MS method, analytes were separated on a Inertsil® ODS-3 column (4.6 × 150 mm, 5 μm). The mobile phase was acetonitrile-5 mmol/L ammonium acetate water solution (65:35, v/v). ESI+ was used and ion peaks with m/z 344.2 (rabeprazole thioether) and 285.1 (internal standard diazepam) were monitored. Both these 2 methods were validated for specificity, linearity, precision, accuracy, matrix effect and extraction recovery, and, particularly, the stability of analytes under various conditions. We successfully applied these methods to a 13-week toxicokinetic
study of
rabeprazole in rats after intravenous administration of (R)- (80, 20, 5 mg/kg/d) and racemic (80 mg/kg/d) rabeprazole sodium. The results showed that rabeprazole and its metabolites did not accumulate in rats. However, desmethyl
rabeprazole and
rabeprazole thioether showed higher exposure and lower clearance rate in the last administration than in the first one. (R)-
rabeprazole showed a higher exposure and a slower elimination rate than (S)-
rabeprazole in rats. These findings offer experimental evidence and a theoretical foundation for further preclinical investigations and clinical applications of (R)-rabeprazole.