A sensitive green HPLC-fluorescence method for simultaneous analysis of sacubitril and valsartan in pure forms, pharmaceutical dosage form and human plasma

The development of green methods of analysis has recently become a global trend in analytical chemistry, especially in routine HPLC analysis. This study presents a sensitive and economic green HPLC method with fluorescence detection for the simultaneous determination of sacubitril and valsartan in bulk, and pharmaceutical dosage forms in spiked human plasma using ibuprofen as an internal standard. Analysis was performed on a C18 column (150 mm × 4.6 mm, 5 mm) at ambient temperature with isocratic elution using 30 mM phosphate (pH 2.5) and ethanol in a ratio of (40:60 v/v) at a flow rate of 1.0 mL/min. The fluorescence detector was programmed for excitation and emission wavelengths from 0 to 3.2 min, where λ_Excitation = 250.0 nm and λ_Emission = 380.0 nm. Then the λ _Emission was changed to 320.0 nm from 3.2 to 5.2 min while the λ _Excitation was not changed. After 5.2 min, the λ _Excitation and λ _Emission were changed to 220.0 and 289.0 nm, respectively. Validation of the proposed method was performed in accordance with the ICH guidelines. The method showed good linearity for sacubitril and valsartan in the concentration ranges of 0.035 to 2.205 µg/mL and 0.035 to 4.430 µg/mL, respectively. The proposed method was proven eco-friendly and applicable for routine analysis of the studied drugs in human plasma through assessment by the Analytical Eco-Scale, AGREE, the complex GAPI, the AGSA, the CaFRI, the RGBfast and the Click Analytical Chemistry Index methods.