Development and Characterization of Sensitive, Eco‐Friendly, and Selective Microwave‐Aided Fluorodensitometric Probe for Estimation of Alfuzosin Using Comprehensive Approach of Multicolored Analytical Chemistry and Design of Experiments

Existing methods for AFZ estimation lacked sensitivity for picoscale detection in biological samples, faced matrix interferences, and used environmentally harmful solvents. A sensitive, selective, and eco-friendly microwave-assisted fluorodensitometric probe was developed for picoscale estimation of AFZ in pharmaceutical formulations and human plasma. The method utilized a microwave-assisted microlevel Hantzsch reaction for the generation of a fluorodensitometric probe for AFZ derivatization in the microenvironment of a silica plate. A design of experiments (DoE) approach was implemented using a Plackett-Burman design for risk assessment and a minimum run resolution IV design for response surface analysis. HPTLC-MS was used for the characterization of the fluorodensitometric probe generated by microwave-aided microlevel condensation of organic liquids and AFZ. The method demonstrated linearity in the range of 10-50 pg/band with high sensitivity (LOD: 3.0 pg/band, LOQ: 10.0 pg/band). Precision and accuracy were within acceptable limits. The method was successfully applied to pharmaceutical formulations and different plasma samples. The proposed method offers a sensitive, selective, and environmentally friendly approach for AFZ estimation at picoscale levels, adhering to white analytical chemistry principles and the QbD approach.