Implementation of White Analytical Chemistry–Assisted Analytical Quality by Design Approach to Green Liquid Chromatographic Method for Concomitant Analysis of Anti-Hypertensive Drugs in Human Plasma.

The article focuses on the development and validation of a white analytical chemistry (WAC)–assisted green reverse-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous analysis of three anti-hypertensive drugs—azilsartan medoxomil (AZL), chlorthalidone (CLT), and cilnidipine (CIL)—in human plasma and their fixed-dose combinations (FDCs). Employing an analytical quality by design (AQbD) approach combined with multivariate analysis (principal component analysis and partial least squares regression) and design of experiments (Box–Behnken design), the method optimizes critical method variables to ensure robustness, accuracy, and regulatory compliance per International Council for Harmonization (ICH) Q2(R1) and M10 guidelines. The method uses isopropyl alcohol, a Class 3 solvent considered safer and more environmentally friendly than commonly used Class 2 solvents, aligning with green analytical chemistry principles. Validation results demonstrate the method’s specificity, precision, accuracy, stability, and sensitivity, and its application to human plasma, FDCs, and laboratory mixtures confirms its suitability as an eco-friendly, economical, and effective analytical tool for pharmaceutical quality control and pharmacokinetic studies.