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.

由于基质效应和预期的低治疗水平，生物体液中药物的光谱测定具有挑战性。在这项工作中，我们采用盐析诱导的均相液-液微萃取制备和预浓缩人血浆中的达卡他韦，通过二阶导数分光光度法进行测量。步骤包括将 500 μL 乙腈与 500 μL 含达卡他韦的血浆涡旋，4000 rpm 离心 5 分钟，然后将 600 μL 上层转移到另一个试管中，并用碱性水定容至 1 mL (pH 7.5)。然后，加入 200 mg 醋酸铵，将管以 4000 rpm 的速度离心 5 分钟，以诱导上层乙腈的相分离，这可以在石英微量杯中进行测量。通过绘制以 ng/mL 为单位的 daclatasvir 浓度与 357 nm 处二阶导数光谱的幅度，构建校准曲线。发现所提出的方法在 500–5000 ng/mL (r²  = 0.9991)，检测限为 130 ng/mL。该方法准确且精确，回收率在 99.70% 到 101.51% 之间。与传统的液-液萃取技术相比，所开发的方法简单、快速、经济且需要最少的有机溶剂。使用绿色分析程序指数 (GAPI) 和分析生态量表评估方法绿色度。该应用揭示了在光谱生物医学应用中使用 SALLME 作为样品制备方法的潜力。