Abstract
Theabrownins (TBs) in dark tea have received increasing attention for their multiple health benefits. In this study, ultrasound assisted extraction with deep eutectic solvent (UAE-DES) was developed for the extraction of TBs from Kangzhuan dark tea (KZDT). The highest yield (12.59%) of TBs was obtained using UAE-choline (ChCl)/malic acid (MA) with a liquid to solid ratio of 20:1 (v/w), ultrasonic power of 577 W, ultrasonic time of 25 min, and water content of 30%. TBs were further eluded by silica gel to obtain six theabrownine fractions (TBFs), namely, TBFs1, TBFs2, TBFs3, TBFs4, TBFs5, and TBFs6. LC-MS/MS revealed that flavonoids, terpenes, phenolic acids, alkaloids, lipids, and amino acids are the leading components of TBFs. The TBFs4, with the DPPH, ABTS, and FRAP values of 45.08 ± 0.42 μM Ascorbic acid/g DW, 178.52 ± 0.29 μM Trolox/g DW, and 370.85 ± 6.00 μM Fe(II)/g DW, respectively, showed the highest antioxidant activity among all the TBFs. Overall, this study first provided the evidence that UAE-ChCl/MA combining with silica gel was effective to extract TBs from KZDT, and the 6,7-dihydroxycoumarin-6-glucoside and neohesperidin were found as the leading compounds in the TBFs, providing a guidance for the chemical research and further utilization of dark tea and its TBs.