Department of Pharmacy, Henan Provincial People's Hospital, Zhengzhou University People's Hospital, Zhengzhou, Henan, 450003, China.
Department of Respiratory and Critical Care Medicine, Henan Provincial People's Hospital, Zhengzhou University People's Hospital, Zhengzhou, Henan, 450003, China.
J Pharm Biomed Anal. 2021 Feb 20;195:113876. doi: 10.1016/j.jpba.2020.113876. Epub 2020 Dec 30.
In this study, we developed a sensitive and efficient analytical approach combining a 96-well plate-based protein precipitation strategy with ultra-performance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC-MS/MS) in order to assess the pharmacokinetic (PK) properties of sivelestat and its metabolite XW-IMP-A in samples of plasma from ALI/ARDS patients with SIRS. The samples were separated via gradient elution with a C18 column (Phenomenex Kinetex, C18, 2.6 μm, 100 Å, 50 × 2.1 mm) using 0.1 % formic acid aqueous solution (A) and acetonitrile-methanol (1:1, V:V) (B) as a mobile phase at a 0.6 mL/min flow rate. UPLC-MS/MS spectra were generated in positive ion mode, and multiple reaction monitoring (MRM) was used to detect the following transitions: m/z 435.1 → 360.0 for sivelestat, m/z 469.0 → 394.0 for sivelestat-IS, m/z 351.0 → 276.0 for XW-IMP-A, and m/z 384.9 → 310.0 for XW-IMP-A-IS. This assay was run for 2.5 min in total, and achieved lowest limit of quantitation values of 2.0 ng/mL and 0.5 ng/mL for sivelestat and XW-IMP-A, respectively, while remaining highly linear from 2-500 ng/mL for sivelestat (r ≥ 0.9900) and from 0.5-125 ng/mL for XW-IMP-A (r ≥ 0.9900). These validated data were consistent with US Food and Drug Administration (FDA) and European Medicines Agency (EMA) acceptance criteria. In addition, this method was successfully applied to the steady-state PK evaluation of ALI/ARDS patients with SIRS.
在这项研究中，我们开发了一种灵敏高效的分析方法，将 96 孔板蛋白沉淀策略与超高效液相色谱电喷雾串联质谱（UPLC-MS/MS）相结合，以评估 SIRS 患者急性肺损伤/急性呼吸窘迫综合征（ALI/ARDS）中西维来司他及其代谢物 XW-IMP-A 的药代动力学（PK）特性。使用 Phenomenex Kinetex C18 柱（C18，2.6 μm，100 Å，50×2.1 mm），以 0.1%甲酸水溶液（A）和乙腈-甲醇（1:1，V:V）（B）为流动相，采用梯度洗脱，流速为 0.6 mL/min 对样品进行分离。UPLC-MS/MS 谱在正离子模式下生成，采用多重反应监测（MRM）检测以下离子对：m/z 435.1→360.0 用于西维来司他，m/z 469.0→394.0 用于西维来司他-IS，m/z 351.0→276.0 用于 XW-IMP-A，m/z 384.9→310.0 用于 XW-IMP-A-IS。该分析总用时 2.5 分钟，西维来司他和 XW-IMP-A 的定量下限分别为 2.0 ng/mL 和 0.5 ng/mL，而西维来司他在 2-500 ng/mL 范围内具有高度线性（r≥0.9900），XW-IMP-A 在 0.5-125 ng/mL 范围内具有高度线性（r≥0.9900）。这些验证数据符合美国食品和药物管理局（FDA）和欧洲药品管理局（EMA）的接受标准。此外，该方法成功应用于 SIRS 患者 ALI/ARDS 的稳态 PK 评价。
