Department of Pharmaceutical Analysis, School of Pharmacy, Shenyang Pharmaceutical University, No. 103 Wenhua Road, Shenhe District, Shenyang 110016, PR China.
Department of Pharmaceutical Analysis, School of Pharmacy, Shenyang Pharmaceutical University, No. 103 Wenhua Road, Shenhe District, Shenyang 110016, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Aug 1;1179:122766. doi: 10.1016/j.jchromb.2021.122766. Epub 2021 May 21.
Recently, a combination of cilostazol and ambroxol has been used in the clinical treatment of stroke-associated pneumonia (SAP). However, the pharmacokinetic drug-drug interaction (DDI) of cilostazol and ambroxol has not been reported. In this paper, a rapid, reproducible and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of cilostazol and ambroxol in Sprague-Dawley (SD) rat plasma was established and validated for the first time. Domperidone was used as the internal standard (IS) and one-step liquid-liquid extraction (LLE) method was used to extract analytes and IS from plasma samples with methyl tert-butyl ether as extractant. A rapid chromatographic separation within 4.8 min was carried on an Ultimate ® XB-C column with a mobile phase consisting of methanol-acetonitrile-formic acid (0.1%) aqueous solution (90:2:8, v/v/v) at a flow rate of 500 μL/min. The quantitative detection of the analytes and IS were performed on a positive electrospray ionization mode (ESI), and scanned by multi-reaction monitoring (MRM) with the ion transitions m/z 370.3 → m/z 288.2 for cilostazol, m/z 378.8 → m/z 263.8 for ambroxol and m/z 426.2 → m/z 175.1 for domperidone (IS), respectively. It had good linearity in the range of 5.0-1000 ng/mL for cilostazol and 1.0-200 ng/mL for ambroxol in rat plasma. The methodology was fully validated with selectivity, linearity, lower limits of quantification, precision, accuracy, extraction recovery, matrix effect, stability and carry-over effect. The validated data have met the determination requirements of biological samples in FDA guideline. The method was successfully applied to the pharmacokinetics and DDI study of cilostazol and ambroxol in male SD rats. The current study found that the interaction between cilostazol and ambroxol may be caused by CYP3A4 and the pharmacological properties of cilostazol, which may be helpful for therapeutic drug monitoring, clinical dose reference and provide a valuable tool for drug-drug interactions.
最近，联合应用西洛他唑和氨溴索已用于治疗脑卒中相关性肺炎（SAP）。然而，西洛他唑和氨溴索的药代动力学药物相互作用（DDI）尚未报道。本文首次建立并验证了一种用于同时测定 SD 大鼠血浆中西洛他唑和氨溴索的快速、重现性和灵敏的液相色谱串联质谱（LC-MS/MS）方法。多潘立酮作为内标（IS），采用一步液液萃取（LLE）法，以甲基叔丁基醚为萃取剂，从血浆样品中提取分析物和 IS。采用甲醇-乙腈-甲酸（0.1%）水溶液（90:2:8，v/v/v）作为流动相，在 Ultimate ® XB-C 柱上以 500 μL/min 的流速进行快速色谱分离。在正电喷雾电离模式（ESI）下对分析物和 IS 进行定量检测，并采用多反应监测（MRM）扫描，离子转换 m/z 370.3→m/z 288.2 用于西洛他唑，m/z 378.8→m/z 263.8 用于氨溴索，m/z 426.2→m/z 175.1 用于多潘立酮（IS）。西洛他唑在大鼠血浆中的浓度范围为 5.0-1000ng/mL，氨溴索为 1.0-200ng/mL，均具有良好的线性关系。该方法具有选择性、线性、定量下限、精密度、准确度、提取回收率、基质效应、稳定性和携带效应。验证数据符合 FDA 指南中生物样品的测定要求。该方法成功应用于雄性 SD 大鼠中西洛他唑和氨溴索的药代动力学和 DDI 研究。目前的研究发现，西洛他唑和氨溴索的相互作用可能是由 CYP3A4 和西洛他唑的药理学特性引起的，这可能有助于治疗药物监测、临床剂量参考，并为药物相互作用提供有价值的工具。
