Eco-Scale, Blueness, ComplexMoGAPI, and AGREEprep comparison of developed UPLC-fluorescence method with a UPLC-PDA method for remdesivir determination in human plasma

Abstract

A sensitive ultra-performance liquid chromatographic method connected to fluorescence detector (UPLC-fluorescence) was developed for remdesivir determination in human plasma using a simple plasma extraction method. After factorial optimization, the analysis was validated on column BEH C₁₈ (2.1 × 100.0 mm, 1.7 μm) connected with pre column BEH C₁₈ (2.1 × 5.0 mm, 1.7 μm) at 50 °C with 10 μL injection volume and 0.25 mL min⁻¹ flow rate. The fluorescence detector was set at an ex./em. of 278/397 nm. The mobile phase was composed of a mixture of methanol and 40 mM ammonium acetate solution at pH 4 (61.5:38.5 (v/v)). The retention time of the internal standard (daclatasvir dihydrochloride) was 3.510 ± 0.009 min, while that of remdesivir was 3.939 ± 0.010 min. Two linear calibration curves were constructed (5–500 ng mL⁻¹ & 500–5000 ng mL⁻¹). The two linearity ranges had coefficients of determination (R²) of 0.9930 and 0.9994, respectively. The new method was validated according to the guidelines of the international council for harmonisation (ICH) for bioanalytical method validation. Eco-Scale, blue applicability grade index (BAGI), complex modified green analytical procedure index (ComplexMoGAPI), and analytical greenness metric for sample preparation (AGREEprep) assessments indicated that the developed method was acceptable in terms of environmental impact. The developed method's greenness was compared with that of a reference method that used a photodiode array (PDA) detector instead of a fluorescence detector.