Abstract

Considering the importance of robust analytics in the pharmaceutical industry, an analytical quality by design (AQbD) based reversed phase liquid chromatographic method was developed for the quantification of propranolol hydrochloride (PRO) and etizolam (ETZ). The developed method was validated and compared with an analytical method of conventional approach. In AQbD approach, analytical target profile (ATP) and risk assessment were tailored to identify critical method variables (CMVs) as inputs. Accordingly, pH (X₁) % aqueous composition (X₂), flow rate (X₃) and % triethylamine (X₄) were identified as CMVs and the respective retention time (Y₁ and Y₂) and theoretical plate (Y₃ and Y₄) for PRO and ETZ were set as method responses. The central composite design (CCD) predicted the scientific relationship between CMVs (Xn) and method responses (Yn). Then, the method operable design region (MODR) was arrived from systematic simulation and contour plots. There were six candidate methods selected from MODR and they demonstrated the r² value > 0.9, thus the model proved its consistency for prediction. The robust range for the selected CMVs was derived from MODR, thus the ranges were 3–4 for pH, 30–35 for % aqueous composition, 0.9–1.1 mL min⁻¹ for flow rate and 0.1–0.2 for % TEA buffer. Finally, the target AQbD method was optimized with pH; 3.5 (X₁), 30% of aqueous composition in mobile phase with acetonitrile (X₂), 0.9 mL min⁻¹ of flow rate of (X₃) and 0. 15% trimethylamine as buffer (X₄) and further validated as per ICH Q2 guidelines. The result was compared with the conventional LC method with respect to specificity, accuracy, precision and robustness. The conventional HPLC method failed for robustness and showed relatively higher % RSD values than AQbD method.

摘要

考虑到强大的分析在制药行业中的重要性，开发了一种基于设计质量（AQbD）的反相液相色谱方法，用于定量盐酸普萘洛尔（PRO）和依替唑仑（ETZ）。验证了所开发的方法，并与常规方法的分析方法进行了比较。在AQbD方法中，量身定制了分析目标配置文件（ATP）和风险评估，以识别关键方法变量（CMV）作为输入。因此，将pH（X ₁）％水性组合物（X ₂），流速（X ₃）和三乙胺（X ₄）鉴定为CMV，并将各自的保留时间（Y ₁和Y ₂）和理论塔板数（Y将PRO和ETZ的₃和Y ₄）设置为方法响应。中央复合设计（CCD）预测了CMV（Xn）与方法响应（Yn）之间的科学关系。然后，通过系统仿真和等高线图得出了该方法的可操作设计区域（MODR）。从MODR中选择了六种候选方法，它们证明r ²值> 0.9，因此该模型证明了其预测的一致性。所选CMV的稳健范围是从MODR得出的，因此，pH范围为3–4，水性成分百分比为30–35，流速为0.9–1.1 mL min ^-1，％TEA缓冲液为0.1–0.2。最后，用pH优化目标AQbD方法。3.5（X ₁），流动相中30％的水性组合物和乙腈（X ₂），0.9 mL min ^-1的流速（X ₃）和0. 15％的三甲胺作为缓冲液（X ₄），并按照ICH Q2指南进一步验证。将结果与常规LC方法的特异性，准确性，精密度和鲁棒性进行了比较。常规HPLC方法的稳健性失败，并且显示出比AQbD方法更高的％RSD值。