A sustainable RP‐HPLC method for the comprehensive fingerprinting of pioglitazone and its related substances was developed and validated in alignment with green analytical chemistry principles. Utilizing eco‐friendly solvents and reducing waste generation were used to optimize chromatographic conditions. Mobile phase ethanol is mixed with buffer solution in a 50:50 (v/v) ratio, selected for its low environmental impact. Using a Thermo Hypersil BDS C18 column (25 cm × 4.6 mm, 5 μm), the separation was performed at 25°C with an autosampler temperature of 15°C and a flow rate of 0.7 mL/min. 269 nm was used for detection. A wide concentration range was achieved using the method, including pioglitazone concentrations of 25.67 to 77.03 μg/mL, pioglitazone related Compound I and pioglitazonerelated Compound II, as well as pioglitazone related Compound III concentrations of 0.049617 to 4.961744 μg/mL, 0.050265 to 5.026527 μg/mL, and 0.051584 to 5.158400 μg/mL. High resolution and reproducibility are achieved when using the method to separate pioglitazone from known impurities and degradation products. For evaluation of pioglitazone’s stability profile, we conducted forced degradation tests under acidic, basic, oxidative, thermal, and photolytic conditions. As indicated by the chromatograms, both intact and degradation products could be detected and quantified by this method. The chromatography resolution and peak purity were optimized to identify and quantify the analytes precisely. Greenness of the method was evaluated using multicolor assessment and Carbon Footprint Reduction Index, which demonstrated that it has minimal environmental impact. International Council for Harmonization guidelines were followed in the development of the validated procedure, which is suitable for quality control and stability testing in routine laboratories. Although sustainability is difficult to integrate into pharmaceutical impurity profiling, this study demonstrates its viability.
Alanazi et al. (2026) studied this question.