A multimodal HPLC stability indicating approach for the estimation of Semaglutide and Tirzepatide in bulk, pharmaceutical dosage forms, and rat plasma: a six-edged sustainability appraisal

Over the recent years, there has been a notable surge in consumer demand for rapid and effective weight-loss pharmaceuticals that are also capable of managing type 2 diabetes. Owing to their exceptional efficacy, GLP-1 receptor agonists, including Tirzepatide (TIR) and Semaglutide (SEM), have had phenomenal outcomes and confidence among consumers. A rapid, straightforward, and thorough approach for quantifying SEM and TIR is essential for quality control purposes given the rising use of these drugs in the pharmaceutical industry. This work presents the first stability-indicating HPLC method for quantifying TIR and SEM under various stress conditions (acidic and basic hydrolysis, oxidative, and photolytic degradation) without interference from degradants. In addition, the proposed methods are capable of accurately quantifying each drug in bulk, pharmaceutical dosage forms, and spiked plasma. The analysis of TIR and SEM was performed using an Inertsil ODS-3 (4.6 × 250 mm, 5 μm particle size) C18 column and the elution of the drugs was achieved isocratically using 0.1% formic acid (pH 2.5) and ACN in the ratio 30:70 with a flow rate 1 mL/min using DAD detector at 220 nm with SEM and TIR eluting at 1.42 and 1.68 min respectively. The proposed method was validated in line with the International Conference of Harmonization (ICH) guidelines and has demonstrated excellent accuracy, linearity, and superior sensitivity with LOD values of 10 and 16 ng/mL for TIR and for SEM, respectively. The obtained linearity range for both TIR and SEM was 1-500 µg/mL with correlation coefficients > 0.9999. An in-depth six- edged sustainability assessment of the proposed method was conducted using greenness, whiteness, blueness and violet innovation metrics was performed using Analytical Greenness Metric (AGREE), Modified Green Analytical Procedure Index (MoGAPI), Analytical Eco-scale, Analytical Green Star Area (AGSA), Carbon Footprint Reduction Index (CaFRI), Whiteness using RGB algorithm, Blue Applicability Grade Index (BAGI), Click Analytical Chemistry Index (CACI), Violet Innovation Grade Index (VIGI) tools and Stability Toolkit for the Appraisal of Bio/Pharmaceuticals' Level of Endurance (STABLE).