Development and Characterization of Sensitive, Eco-Friendly, and Selective Microwave-Aided Fluorodensitometric Probe...

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.