Rocha Ariel Lucas, Lourenço Felipe Rebello
Departamento de Farmácia, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, Av. Prof. Lineu Prestes, 580 - Bloco 15, CEP 05508-000, São Paulo, SP, Brazil.
Departamento de Farmácia, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, Av. Prof. Lineu Prestes, 580 - Bloco 15, CEP 05508-000, São Paulo, SP, Brazil.
Talanta. 2023 Jul 1;259:124481. doi: 10.1016/j.talanta.2023.124481. Epub 2023 Mar 24.
The concept of Analytical Quality by Design (AQbD) comes as a more robust, economical, and scientifically based alternative for analytical development, to the detriment of OFAT (one factor at a time). This new understanding applicable to analytical development is recommended since regulatory flexibility can be achieved and ensure more reliable results throughout the life of the product. This new approach was applied to develop an analytical procedure indicative of stability for a pharmaceutical product of association of Losartan Potassium and Hydrochlorothiazide, considered a potential first line for the treatment of hypertension. The first stage of the analytical development consisted of defining analytical target profile (ATP), follow by a bibliographic survey of the physicochemical properties of the molecules in question to define an initial method. After defining the initial analytical conditions, statistical tools for design of experiments (DoE) were used for the screening and optimization steps. In the screening stage, the Plackett-Burman design was chosen, using 11 factors and 2 levels, through which it was possible to evaluate numerous variables and determine their significance in relation to the responses. Next, optimization was carried out with the experimental design of a central composite with 4 factors and 5 levels, which allowed modeling a complex response surface and evaluating the phenomena of interactions between the factors. Thus, the optimized analytical conditions were defined, considering a 0.3% formic acid gradient as eluent A and a mixture of acetonitrile and methanol (80:20) as eluent B, X-Bridge C18 chromatographic column (150 mm × 4 .6 mm × 3.5 μm), column temperature of 40°C, flow rate of 1.3 mL/min, injection volume of 10 μL. Through this methodology, it was possible to identify an unknown degradation product of Hydrochlorothiazide, formed by the reaction with lactose (excipient present in the drug formulation), proving that the method can be applicable both to DAD detectors and to spectrometry and mass detectors. It was also proven through the forced degradation study that the method is indicative of stability, in addition to being validated and robust for its purpose.
设计分析质量（AQbD）的概念作为一种更稳健、经济且基于科学的分析方法开发替代方案，取代了传统的单因素实验法（OFAT）。推荐这种适用于分析方法开发的新认识，因为它既能实现法规灵活性，又能在产品的整个生命周期内确保更可靠的结果。这种新方法被应用于开发一种分析程序，用于指示氯沙坦钾和氢氯噻嗪复方药物产品的稳定性，该复方药物被视为治疗高血压的潜在一线药物。分析方法开发的第一阶段包括定义分析目标轮廓（ATP），随后对相关分子的物理化学性质进行文献调研以确定初始方法。在确定初始分析条件后，使用实验设计（DoE）的统计工具进行筛选和优化步骤。在筛选阶段，选择了Plackett-Burman设计，使用11个因素和2个水平，通过该设计可以评估众多变量并确定它们对响应的显著性。接下来，使用具有4个因素和5个水平的中心复合实验设计进行优化，这使得能够对复杂的响应面进行建模并评估因素之间的相互作用现象。因此，确定了优化的分析条件，即以0.3%甲酸梯度作为流动相A，乙腈和甲醇（80:20）的混合物作为流动相B，X-Bridge C18色谱柱（150 mm×4.6 mm×3.5 μm），柱温40°C，流速1.3 mL/min，进样量10 μL。通过这种方法，有可能鉴定出氢氯噻嗪的一种未知降解产物，它是由与乳糖（药物制剂中的辅料）反应形成的，证明该方法既适用于二极管阵列检测器（DAD），也适用于光谱和质谱检测器。通过强制降解研究还证明，该方法除了针对其目的进行了验证且具有稳健性外，还能指示稳定性。
