Deep eutectic solution elution assisted ligand affinity assay: A useful tool for the active coumarins screening from Fructus cnidii

Background: Many of the ligand affinity analyses are presented in water environment, and the hydrophilic solution such as methanol is used for dissociating the bound compounds. The obtained dissociation solution needs to be concentrated for improving the sensitivity of the assay. However, it is not good for the analysis of hydrophobic and volatile compounds such as coumarins. The solubility of them in water environment is low. And they may be lost in the sample preparation process. In this work, a deep eutectic solvent elution assisted ligand affinity assay for the screening of active coumarins in plants was proposed.

Results: For the first time, deep eutectic solvent was used as a dissociation solution for dissociating the compounds that bind with acetylcholinesterase immobilized magnetic beads. Analytes could be enriched during the phase separation process of deep eutectic solvent. The concentration of the analytes in sample solution was therefore increased. It is good for the detection of coumarins. The LOD and LOQ of the proposed assay were 0.50 (μ g/mL) and 1.50 (μ g/mL), respectively. The relative recovery of the assay was 99.4 ± 7.2 %. The intra-day and inter-day precision were 7.25 % and 6.80 %. Using the assay as a tool, the active coumarins in Fructus cnidii were analyzed. And two coumarins with acetylcholinesterase inhibition activity such as imperatorin and osthole were found.

Significance: Coumarins have good solubility in the selected deep eutectic solvent, and could be enriched into hydrophobic phase during the phase separation process. The extra concentration process is not needed. Compared with traditional methods, the proposed method has the benefits of simplified operation process, high sensitivity, and is friendly to the volatile and hydrophobic compounds.