Blueness, Whiteness, and greenness Appraisal of sensitive chromatographic approach for determination of semaglutide concurrently with Pioglitazone and metformin using green mobile Phase; alongside with pharmacokinetic study

Abstract

The replacement of hazardous solvents with greener alternatives was under consideration to achieve a sustainable and green approach. The assessment of reliable white analytical method for the determination of semaglutide as a novel member in glucagon-like peptide agonists class is still of a great importance. That is because pharmacological, clinical, and therapeutic drug monitoring purposes. This research proposes and fully validates a simple and sensitive RP-HPLC method for the determination of semaglutide simultaneously with pioglitazone and metformin. The suggested procedures utilizing a green mobile phase of 8 % glycerol in 0.01 M phosphate buffer (pH 6). Separation was achieved using a cyano (5.0 μm, 150 × 4.6 mm) column maintained at a temperature of 40 °C. The mixture was isocratically eluted under a flow rate of 1 mL/min with a total run time of less than 8 min. The UV-detector was adjusted at 215 nm. The method provides low detection (ranging from 0.003 to 0.63 µg/mL) and quantification (ranging from 0.008 to 1.9 µg/mL) limits, with good precision (%RSD ranging from 0.66 % to 0.89 %). The suggested technique has been effectively used to determine each drug in its pharmaceutical preparation and spiked human plasma. Beside the remarkable sensitivity and sustainability of the evolved approach, the relevance of therapeutic drug monitoring motivated us to perform a pharmacokinetics study for both semaglutide and metformin in real human plasma samples. Greenness, whiteness and blueness assessment of the developed method have been carried out using different tools (Green Analytical Procedure Index (GAPI), the analytical GREEnness metric approach (AGREE), RGB-12 model and BAGI). The proven green and white profile of the evolved method could open future prospective for the clinical research and regular quality control examination of the investigated mixture with the least negative environmental impacts.