Low-density solvent-based dispersive liquid–liquid microextraction followed by high performance liquid chromatography for determination of warfarin in human plasma

Abstract

Extraction and determination of warfarin, a widely used anticoagulant drug, in human plasma were performed using a new generation of dispersive liquid–liquid microextraction (DLLME) and high performance liquid chromatography (HPLC). The extraction procedure is based on extraction solvents lighter than water and performing of extraction in a specially designed extraction cell. Some important parameters, including kind and volume of extraction and disperser solvents, pH of the sample solution, salt concentration in the sample solution and extraction time were investigated and optimized. Under the optimized conditions (150μL 1-octanol as extraction solvent, 150μL methanol as disperser solvent, pHsample=2.3, extraction time of 2min, without salt addition), limit of detection (LOD) of 5ngmL−1 and extraction recovery of 91.0% were obtained. The calibration curve was linear within the range of 15–3000ngmL−1 with the square of correlation coefficient (R2) of 0.998. Repeatability and reproducibility of method based on five replicate extraction and determination were 2.8% and 6.5%, respectively. The proposed method was applied successfully for the determination of warfarin in plasma sample from a patient under treatment with this drug, and was demonstrated to be sensitive, efficient, and convenient.