Abstract

The present studies describe the systematic development and validation of a simple, rapid, sensitive and cost-effective reversed-phase high-performance liquid chromatographic bioanalytical method for the estimation of valsartan in rat plasma employing analytical quality by design (AQbD) principles quality risk management was applied for identifying the critical method parameters (CMPs) and subsequently method optimization was performed employing Box–Behnken design by selecting mobile phase pH, flow rate and % organic modifier as the CMPs and evaluated for critical analytical attributes (CAAs) such as peak area, retention time, peak tailing and number of theoretical plates. The developed method was then transferred to bioanalysis, where liquid–liquid extraction process was used for separating the drug from rat plasma. The optimization of extraction process was performed with the help of face-centered cubic design by selecting centrifugation speed and centrifugation time as the CMPs for maximizing % recovery, signal-to-noise ratio and purity threshold of the drug peak after extraction as the CAAs. Optimum chromatographic solution was chosen by mathematical and graphical search techniques, and design space was demarcated. Validation studies performed for the developed method indicated linearity ranging between 5 and 100 ng.mL⁻¹, whereas accuracy and precision study showed good percent recovery (99–102%) along with % relative standard deviation within ±2%. Sensitivity evaluation revealed limit of detection and limit of quantification were found to be 0.76 ng.mL⁻¹ and 2.29 ng.mL⁻¹, respectively. In a nutshell, the present work demonstrates significant merits of AQbD approach for holistic process understanding and analytical method development and validation with enhanced robustness and performance.

中文翻译：

---

设计和分析质量综合分析（AQbD）方法用于开发和验证用于评估缬沙坦的生物分析液相色谱方法。

摘要

本研究描述了一种简单，快速，灵敏且具有成本效益的反相高效液相色谱生物分析方法的系统开发和验证，该方法采用设计分析质量（AQbD）原理进行质量风险管理，用于估算大鼠血浆中的缬沙坦。应用于鉴定关键方法参数（CMP），随后采用Box–Behnken设计进行方法优化，方法是选择流动相pH，流速和有机改性剂百分比作为CMP，并评估关键分析属性（CAA），例如峰面积，保留时间，峰拖尾和理论塔板数。然后将已开发的方法转移到生物分析中，在该分析中使用液-液萃取过程从大鼠血浆中分离药物。通过选择离心速度和离心时间为CMP，以最大化回收率，信噪比和药物峰的纯度阈值作为CAA，选择了离心速度和离心时间，对提取工艺进行了优化。通过数学和图形搜索技术选择了最佳色谱解决方案，并划定了设计空间。对开发的方法进行的验证研究表明线性范围为5到100 ng.mL 通过数学和图形搜索技术选择了最佳色谱解决方案，并划定了设计空间。对开发的方法进行的验证研究表明线性范围为5到100 ng.mL 通过数学和图形搜索技术选择了最佳色谱解决方案，并划定了设计空间。对开发的方法进行的验证研究表明线性范围为5到100 ng.mL^-1，而准确性和精密度研究则显示出良好的回收率（99–102％），相对标准偏差百分比在±2％以内。敏感性评估表明，检测极限和定量极限分别为0.76 ng.mL ^-1和2.29 ng.mL ^-1。简而言之，当前的工作展示了AQbD方法在整体过程理解，分析方法开发和验证方面的显着优点，并具有增强的鲁棒性和性能。