Homogenate-Ultrasound-Assisted Ionic Liquid Extraction of Total Flavonoids from Selaginella involven: Process Optimization, Composition Identification, and Antioxidant Activity

[Image: see text] In this paper, an efficient approach to extract total flavonoids (TFs) from Selaginella involvens (Sw.) Spring using homogenate-ultrasound-assisted ionic liquid (IL) extraction (HUA-ILE) was first developed. The results indicated that EPyBF(4) was selected as the suitable extractant. According to the single factor experiment and response surface methodology, the IL concentration of 0.10 mol/L, the extraction time of 160 s, the liquid/solid ratio of 13:1 mL/g, and the extraction power of 300 W were concluded as the best conditions. A yield of 8.48 ± 0.27 mg/g TF content was obtained. Compared with HUA ethanol extraction, ultrasound-assisted IL extraction, and percolation extraction, the TF content obtained by the HUA-ILE method could be increased by 2 to 4 times, and the extraction time could be reduced by 100 times. Furthermore, 16 compounds of the TF extract were finally identified through ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry, among which 11 compounds were first discovered in S. involven. The contents of six biflavonoids in S. involven were determined simultaneously adopting high-performance liquid chromatography, including amentoflavone, hinokiflavone, bilobetin, ginkgetin, isoginkgetin, and heveaflavone. The TF extract in S. involven was proved to have potent antioxidant activity through the four antioxidant experiments. In conclusion, HUA-ILE was applied for the first time to exploit a green, efficient, and novel approach to extract TFs, and the research also provided promising prospects for applications of S. involven.