Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia.
Bioavailability Laboratory, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia.
PLoS One. 2019 Mar 14;14(3):e0213786. doi: 10.1371/journal.pone.0213786. eCollection 2019.
A fixed dose combination of lesinurad and allopurinol has been recently approved by USFDA and EMA for treatment of gout-associated hyperuricemia in patients who have not achieved target serum uric acid levels with allopurinol alone. In this study, an ultra-performance hydrophilic interaction liquid chromatography (UPHILIC) coupled with tandem mass spectrometry method was developed and validated for simultaneous determination of allopurinol, oxypurinol and lesinurad in rat plasma. Liquid liquid extraction using ethyl acetate as extracting agent was used for samples extraction procedure. Acquity UPLC HILIC column (100 mm x 2.1, 1.7μm) was used for separation of allopurinol, oxypurinol, lesinurad and internal standard (5-Florouracil). The mobile phase consisting of acetonitrile, water and formic acid (95:5:0.1, v/v/v), were eluted at 0.3 mL/min flow rate having total chromatographic run time of 3 min per sample. The analytes were detected on Acquity triple quadrupole mass spectrometer equipped with a Z-Spray electrospray ionization (ESI). The ESI source was operated in negative mode and multiple reaction monitoring was used for ion transition for all compounds. The precursor to product ion transition of m/z 134.94 > 64.07 for allopurinol, 150.89 > 41.91 for oxypurinol, 401.90 > 176.79 for lesinurad and 128.85 >41.92 for internal standard were used for identification and quantification. The calibration curves for all analytes were found to be linear with weighing factor of 1/x2 using regression analysis. The developed assay was successfully applied in an oral pharmacokinetic study of allopurinol, oxypurinol and lesinurad in rats.
一种来昔布和别嘌醇的固定剂量复方制剂最近已被美国食品药品监督管理局和欧洲药品管理局批准用于治疗单独使用别嘌醇未能达到目标血清尿酸水平的痛风相关性高尿酸血症患者。在这项研究中，建立并验证了一种超高效亲水相互作用液相色谱（UPLC）-串联质谱法，用于同时测定大鼠血浆中的别嘌醇、氧嘌呤醇和来昔布。采用乙酸乙酯作为提取剂的液液萃取法用于样品提取程序。 Acquity UPLC HILIC 柱（100mm×2.1mm，1.7μm）用于分离别嘌醇、氧嘌呤醇、来昔布和内标（5-氟尿嘧啶）。以乙腈、水和甲酸（95:5:0.1，v/v/v）为流动相，流速为 0.3mL/min，每个样品的总色谱运行时间为 3min。采用配备 Z-喷雾电喷雾电离（ESI）源的 Acquity 三重四极杆质谱仪对分析物进行检测。ESI 源在负离子模式下工作，所有化合物均采用多反应监测进行离子转换。m/z 134.94 > 64.07 的 134.94 > 64.07（别嘌醇）、150.89 > 41.91（氧嘌呤醇）、401.90 > 176.79（来昔布）和 128.85 > 41.92（内标）的前体到产物离子转换用于鉴定和定量。所有分析物的校准曲线均采用回归分析，以权重因子 1/x2 呈线性。该方法成功应用于大鼠体内别嘌醇、氧嘌呤醇和来昔布的口服药代动力学研究。
