Development of a deep eutectic solvent-based ultrasound-assisted homogenous liquid-liquid microextraction method for simultaneous extraction of daclatasvir and sofosbuvir from urine samples.

An ultrasound-assisted homogenous liquid-liquid microextraction method using a new deep eutectic solvent was proposed for the extraction of daclatasvir and sofosbuvir from urine. The analytes were determined by high performance liquid chromatography-diode array detector. The deep eutectic solvent was prepared by mixing p-aminophenol with tetrabutyl ammonium chloride. It was used in the extraction procedure as an extraction solvent. The amine group in structure of the prepared deep eutectic solvent led to its various solubility in different pHs. In this method, urine sample was placed in a glass test tube and then mixed with sodium chloride and its temperature adjusted at 50 °C. Then, the deep eutectic solvent was dissolved in the solution by manually shaking. In the following, an ammonia solution was added to the solution and the mixture was sonicated for 4 min. After centrifugation, an aliquat of the sedimented phase was injected into the determination system. Low limits of detection (daclatasvir 1.0 and sofosbuvir 1.3 μg/L) and quantification (daclatasvir 3.3 and sofosbuvir 4.0 μg/L), high enrichment factor (daclatasvir 96 and sofosbuvir 90) and extraction recovery (daclatasvir 96 and sofosbuvir 90 %), and good percision (relative standard deviation ≤9.3 %) were obtained. The introduced method was successfully applied in the determination of daclatasvir and sofosbuvir concentrations in urine samples.