Fares Michel Y, Hegazy Maha A, El-Sayed Ghada M, Abdelrahman Maha M, Abdelwahab Nada S
Pharmaceutical Chemistry Department, Faculty of Pharmacy, Nahda University Sharq El-Nile 62511 Beni-Suef Egypt.
Analytical Chemistry Department, Faculty of Pharmacy, Cairo University Kasr El-Aini Street Cairo 11562 Egypt
RSC Adv. 2022 May 9;12(22):13896-13916. doi: 10.1039/d2ra00966h. eCollection 2022 May 5.
This work implements a combined experimental approach of analytical quality-by-design (AQbD) and green analytical chemistry (GAC) to develop an HPLC method for simultaneous determination of the two thalassemia drugs, deferasirox (DFX) and deferiprone (DFP), in biological fluid for the first time. This integration was designed to maximize efficiency and minimize environmental impacts, as well as energy and solvent consumption. To accomplish this goal, an analytical quality-by-design approach was performed, beginning with quality risk assessment and scouting analysis, followed by Placket-Burman design screening for five chromatographic parameters. Critical method parameters were thoroughly recognized and then optimized by using a two levels-three factors custom experimental design to evaluate the optimum conditions that achieved the highest resolution with acceptable peak symmetry within the shortest run time. The desirability function was used to define the optimal chromatographic conditions, and the optimal separation was achieved using an XBridge® HPLC RP-C18 (4.6 × 250 mm, 5 μm) column with ethanol : acidic water at pH 3.0 adjusted by phosphoric acid in the ratio of (70 : 30, v/v) as the mobile phase at a flow rate of 1 mL min with UV detection at 225 nm at a temperature of 25 °C. Linearity was obtained over the concentration range of 0.30-20.00 μg mL and 0.20-20.00 μg mL for DFX and DFP, respectively, using 20.00 μg mL ibuprofen (IBF) as an internal standard. The established method's greenness profile was evaluated and measured using various assessment tools, and the developed method was green. For the validation of the developed method, FDA recommendations were followed, and all the results obtained met the acceptance criteria. The suggested method was successfully used to study the pharmacokinetic parameters of DFX and DFP in rat plasma. Due to the substantial increase in bioavailability of the two iron chelating drugs, the results from this study strongly recommend their co-administration.
本研究首次采用分析质量源于设计（AQbD）与绿色分析化学（GAC）相结合的实验方法，开发了一种用于同时测定生物流体中两种地中海贫血药物地拉罗司（DFX）和去铁酮（DFP）的高效液相色谱法。这种整合旨在提高效率、减少对环境的影响以及能源和溶剂消耗。为实现这一目标，首先进行了质量风险评估和探索性分析，随后采用Plackett-Burman设计对五个色谱参数进行筛选，实施了分析质量源于设计的方法。通过使用二水平三因素的定制实验设计，全面识别关键方法参数并进行优化，以评估在最短运行时间内实现最高分离度且峰形对称可接受的最佳条件。使用合意函数定义最佳色谱条件，采用XBridge® HPLC RP-C18（4.6×250 mm，5μm）色谱柱，以乙醇∶用磷酸调节至pH 3.0的酸性水（体积比70∶30）作为流动相，流速为1 mL/min，在25℃、225 nm波长处进行紫外检测，实现了最佳分离。分别以20.00μg/mL布洛芬（IBF）为内标，DFX和DFP在0.30 - 20.00μg/mL和0.20 - 20.00μg/mL浓度范围内呈线性。使用多种评估工具对所建立方法的绿色度进行评估和测定，结果表明所开发的方法是绿色的。为验证所开发的方法，遵循了美国食品药品监督管理局（FDA）的建议，所有获得的结果均符合验收标准。所建议的方法成功用于研究DFX和DFP在大鼠血浆中的药代动力学参数。由于这两种铁螯合剂药物的生物利用度大幅提高，本研究结果强烈建议联合使用这两种药物。
