Pharmaceutical Chemistry Department, Faculty of Pharmacy, Future University in Egypt, Cairo, Egypt.
Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, Cairo 11562, Egypt.
J Pharm Biomed Anal. 2023 Oct 25;235:115598. doi: 10.1016/j.jpba.2023.115598. Epub 2023 Jul 23.
This work implements a stability indicating HPLC method developed to simultaneously determine xylometazoline (XYLO) and antazoline (ANT) in their binary mixture, rabbit aqueous humor and cited drug's degradates by applying analytical quality-by-design (AQbD) combined with green analytical chemistry (GAC) experiment for the first time. This integration was designed to maximize efficiency and minimize environmental impacts, as well as energy and solvent consumption. Analytical quality-by-design was applied to achieve our aim starting with evaluation of quality risk and scouting analysis, tracked via five parameters chromatographic screening using Placket-Burman design namely: pH, temperature, organic solvent percentage, flow rate, and wavelength detection. Recognizing the critical method parameters was done followed by optimization employing central composite design and Derringer's desirability toward assess optimum conditions that attained best resolution with satisfactory peak symmetry with short run time. Optimal chromatographic separation was attained by means of an XBridge® C18 (4.6 × 250 mm, 5 µm) column through isocratic elution using a mobile phase consists of phosphate buffer (pH 3.0): ethanol (60:40, by volume) at a 1.6 mL/min flow rate and 230.0 nm UV detection. Linearity acquired over a concentration range of 1.0-100.0 µg/mL and 0.5-100.0 µg/mL for XYLO and ANT, respectively. Furthermore, imperiling cited drugs' stock solutions to stress various conditions and satisfactory peaks of degradation products were obtained indicating that cited drugs are vulnerable to oxidative degradation and basic hydrolysis. Degradates' structures were elucidated using mass spectrometry. Applying various assessment tools; namely: analytical greenness (AGREE), green analytical procedure index (GAPI), analytical eco-scale, and national environmental method index (NEMI), Greenness method's evaluation was applied and proved to be green. In fact, the developed method is established to be perceptive, accurate, and selective to assess cited drugs for routine analysis.
本工作首次应用分析质量源于设计（AQbD）结合绿色分析化学（GAC）实验，建立了一个同时测定二元混合物中盐酸羟甲唑啉（XYLO）和盐酸苯海拉明（ANT）以及两者降解产物的稳定性指示 HPLC 方法。这种整合旨在最大限度地提高效率，最大限度地减少环境影响以及能源和溶剂消耗。应用分析质量源于设计从评估质量风险和探索性分析开始，通过 Placket-Burman 设计的五个参数色谱筛选进行跟踪，这五个参数分别为：pH 值、温度、有机溶剂百分比、流速和波长检测。通过中心复合设计和 Derringer 的适宜性进行优化，确定关键方法参数，以评估最佳条件，从而实现最佳分辨率，同时获得满意的峰对称性和较短的运行时间。通过等度洗脱，使用由磷酸盐缓冲液（pH 3.0）：乙醇（60:40，体积比）组成的流动相，在 1.6mL/min 的流速和 230.0nm 的 UV 检测下，在 XBridge® C18（4.6×250mm，5μm）柱上实现最佳的色谱分离。XYLO 和 ANT 的线性范围分别为 1.0-100.0μg/mL 和 0.5-100.0μg/mL。此外，将受威胁药物的储备溶液置于各种条件下进行胁迫，得到令人满意的降解产物峰，表明受威胁药物易发生氧化降解和碱性水解。使用质谱法阐明降解产物的结构。应用各种评估工具，即：分析绿色度（AGREE）、绿色分析程序指数（GAPI）、分析生态尺度和国家环境方法指数（NEMI），对绿色方法进行评估，证明该方法是绿色的。事实上，该方法的建立是为了对受威胁药物进行常规分析，具有敏感性、准确性和选择性。
