A design of experiments (DoE)-driven RP-HPLC method conditions was employed to analyze simultaneously chloroquine (CQ) phosphate and flavopiridol (FLAP) in emulsions and solution. After subjecting the various critical method attributes to preliminary risk assessment and screening by Pareto-chart-based fractional factorial design, the 17 runs were produced in Box-Behnken design for optimization. Analysis of variance, lack of fit, prediction equations, 3D response surface plots and contour plots were used to evaluate the critical analytical attributes such as retention time, tailing factor and theoretical plate count. The optimized RP-HPLC method conditions include 262 nm as detection wavelength, 37°C temperature for column, 20-μl injection volume, 1-ml/min flow rate and mobile phase mixture [70:30 ratio of 0.4% triethylamine in methanol&sodium phosphate buffer (11 mM, pH 3.0)]. The studied validation parameters were found within the ICH-prescribed limits. Exposing the combined drug solution at oxidative stress condition resulted to diminish the FLAP recovery value (53.39 ± 0.86) and arrival of an extra chromatographic peak. However, the % drug entrapment efficiency values of 96.22 ± 2.47 and 85.86 ± 3.66, respectively, were noticed for CQ phosphate and FLAP in emulsions. Thus, DoE-driven approach could be helpful for systematically optimizing RP-HPLC method conditions.

中文翻译：

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基于分析质量设计的 RP-HPLC 方法条件的系统优化，用于同时分析应力诱导的组合药物溶液和药物乳剂中的磷酸氯喹和黄酮吡醇。

采用实验设计 (DoE) 驱动的 RP-HPLC 方法条件同时分析乳液和溶液中的磷酸氯喹 (CQ) 和黄吡醇 (FLAP)。通过基于帕累托图的部分因子设计对各种关键方法属性进行初步风险评估和筛选后，在 Box-Behnken 设计中进行了 17 次运行以进行优化。使用方差分析、失拟分析、预测方程、3D 响应曲面图和等高线图来评估关键的分析属性，例如保留时间、拖尾因子和理论塔板数。优化后的RP-HPLC方法条件为检测波长262 nm、柱温37℃、进样量20μl、流速1ml/min、流动相混合液[0.4%三乙胺的甲醇溶液和磷酸钠比例70:30]缓冲液（11 mM，pH 3.0）]。研究的验证参数在 ICH 规定的限度内。将组合药物溶液暴露在氧化应激条件下会导致 FLAP 恢复值 (53.39 ± 0.86) 降低并出现额外的色谱峰。然而，乳液中 CQ 磷酸盐和 FLAP 的药物包封率百分比值分别为 96.22 ± 2.47 和 85.86 ± 3.66。因此，DoE 驱动的方法可能有助于系统地优化 RP-HPLC 方法条件。