A green homogeneous liquid-liquid microextraction method for spectrophotometric determination of daclatasvir in human plasma

Spectroscopic determination of drugs in biological fluids is challenging due to the matrix effect and the expected low therapeutic levels. In this work, we employed salting-out induced homogenous liquid-liquid microextraction for preparation and pre-concentration of daclatasvir in human plasma to be measured by second derivative spectrophotometry. The procedures included vortexing 500 μL of acetonitrile with 500 μL of the daclatasvir-containing plasma, and centrifuging for 5 min at 4000 rpm before transferring 600 μL of the upper layer to another test tube, and making up the volume to 1 mL with alkaline water (pH 7.5). Then, 200 mg of ammonium acetate were added and the tube was centrifuged for 5 min at 4000 rpm, to induce phase separation of an upper layer of acetonitrile that could be measured in a quartz microcuvette. The calibration curve was constructed by plotting the daclatasvir concentrations in ng/mL versus the amplitude of second derivative spectra at 357 nm. The proposed method was found linear in the concentration range of 500–5000 ng/mL (r2 = 0.9991), with a limit of detection of 130 ng/mL. The method was accurate and precise with a % recovery, ranging between 99.70% and 101.51%. The developed method was simple, rapid, cost-effective and required minimal amounts of organic solvents compared with conventional liquid-liquid extraction techniques. The method greenness was evaluated using the Green Analytical Procedure Index (GAPI) and the Analytical Eco-Scale. This application sheds the light on the potential of using SALLME as a sample preparation method in spectroscopic biomedical applications.