Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Suez Canal University, Ismailia, 41522, Egypt.
Pharmaceutical Chemistry Department, Egyptian Drug Authority (EDA), Cairo, Egypt.
Sci Rep. 2024 Sep 18;14(1):21758. doi: 10.1038/s41598-024-71413-3.
A green and simple UPLC method was developed and optimized, adopting a factorial design for simultaneous determination of oseltamivir phosphate and remdesivir with dexamethasone as a co-administered drug in human plasma and using daclatasvir dihydrochloride as an internal standard within 5 min. The separation was established on UPLC column BEH C 1.7 μm (2.1 100.0 mm) connected to UPLC pre-column BEH 1.7 μm (2.1 5.0 mm) at 50 °C with an injection volume of 10 μL. The photodiode array detector (PDA) was set at three wavelengths of 220, 315, and 245 nm for oseltamivir phosphate, the internal standard, and both dexamethasone and remdesivir, respectively. The mobile phase consisted of methanol and ammonium acetate solution (40 mM) adjusted to pH 4 in a ratio of 61.5:38.5 (v/v) with a flow rate of 0.25 mL min. The calibration curves were linear over 500.0-5000.0 ng mL for oseltamivir phosphate, over 10.0-500.0 ng mL and 500.0-5000.0 ng mL for dexamethasone, and over 20.0-500 ng mL and 500.0-5000.0 ng mL for remdesivir. The Gibbs free energy and Van't Hoff plots were used to investigate the effect of column oven temperatures on retention times. Fluoride-EDTA anticoagulant showed inhibition activity on the esterase enzyme in plasma. The proposed method was validated according to the M10 ICH, FDA, and EMA's bioanalytical guidelines. According to Eco-score, GAPI, and AGREE criteria, the proposed method was considered acceptable green.
建立并优化了一个绿色、简单的 UPLC 方法，采用析因设计，同时测定人血浆中奥司他韦磷酸盐和瑞德西韦，并用盐酸达拉他韦作为内标，在 5 分钟内完成分析。分离在 UPLC 柱 BEH C1.7μm（2.1×100.0mm）上进行，与 UPLC 预柱 BEH1.7μm（2.1×5.0mm）相连，柱温为 50℃，进样量为 10μL。二极管阵列检测器（PDA）分别在 220nm、315nm 和 245nm 处设置三个检测波长，用于检测奥司他韦磷酸盐、内标和地塞米松及瑞德西韦。流动相由甲醇和 40mM 乙酸铵溶液（pH4）组成，体积比为 61.5:38.5（v/v），流速为 0.25mL/min。奥司他韦磷酸盐的校准曲线在 500.0-5000.0ng/mL 范围内呈线性，地塞米松在 10.0-500.0ng/mL 和 500.0-5000.0ng/mL 范围内呈线性，瑞德西韦在 20.0-500ng/mL 和 500.0-5000.0ng/mL 范围内呈线性。采用 Gibbs 自由能和 Van't Hoff 图谱研究柱温对保留时间的影响。氟化物-EDTA 抗凝剂对血浆中的酯酶有抑制作用。该方法按照 ICH、FDA 和 EMA 的生物分析指南进行验证。根据 Eco-score、GAPI 和 AGREE 标准，该方法被认为是可接受的绿色方法。
