초록
A homogeneous liquid-liquid microextraction (HLLME) was established based on a switchable deep eutectic solvent (DES) for the preconcentration and determination of six flavonoids with different polarity in 'Scutellariae Radix' combined with high performance liquid chromatography (HPLC). A switchable DES composed of N,N-dimethylethanolamine (DMEA) and heptanoic acid was used as an extraction solvent in the HLLME method, which was miscible thoroughly with the aqueous sample phase initially, and then underwent rapid phase transition induced by the addition of an inorganic acid. After the extraction, the upper hydrophobic layer was recovered for HPLC analysis. Different experimental parameters were optimized, and the optimal extraction conditions were as follows: the switchable DES extraction phase, 90 µL of DMEA-heptanoic acid (1:1 mole ratio); phase-switching trigger, 100 µL of 5 mol/L HCl; 10% (w/v) of salt concentration in sample phase; extraction time, 0.3 min. Furthermore, the structures of the switchable DES and the upper hydrophobic layer were characterized by Fourier transform infrared spectroscopy, proton nuclear magnetic resonance spectroscopy and differential scanning calorimetry to illustrate the phase-switching mechanism of the extraction phase during the extraction process. Under the optimized conditions, the enrichment factors for six target analytes were between 0.4 and 104. The calibration curves were linear (r≥0.9866) in the range of 0.033-8.65 mg/L for scutellarin, 0.022-5.77 mg/L for baicalin, 0.0033-0.865 mg/L for scutellarein and wogonoside, and 0.0022-0.577 mg/L for baicalein and wogonin, respectively. Low detection limits (≤8.0 × 10-3 mg/L) and quantification limits (≤2.4 × 10-2 mg/L) as well as good precisions (relative standard deviations lower than 9.2%) and acceptable accuracies (spiked recoveries 89.3-114.4%) were also obtained. The proposed method is a simple, fast, and eco-friendly sample pretreatment method.