Ultrasound-surfactant-assisted extraction of flavonoids from Zanthoxylum bungeanum leaves: process optimization, metabolomic characterization, and bioactivity evaluation.
Abstract
This study employed an ultrasound-assisted surfactant extraction method to optimize the extraction conditions for flavonoids from Zanthoxylum bungeanum leaves (FZBL) using the response surface methodology. With fixed ultrasonic power of 300 W and frequency of 20 kHz, the maximum extraction yield of 21.05% was achieved when the optimal conditions were ultrasonic temperature of 50°C, sodium dodecyl sulfate addition of 2.9% (w/v), ultrasonic time of 50 min, ethanol concentration of 61% (v/v), and liquid/solid ratio of 26 mL/g. The resulting crude flavonoid extract was further fractionated using D-101 macroporous resin, producing three fractions eluted with 30%, 50%, and 70% ethanol, designated as FZBL-1, FZBL-2, and FZBL-3, respectively. UV-vis and fourier transform infrared characterization indicated that all three fractions possessed the structural features typical of flavonoids. Comprehensive ultra-high-performance liquid chromatography-tandem mass spectrometry based metabolomic profiling putatively annotated 444 metabolites, including 418 flavonoid compounds, highlighting the rich flavonoid composition of the extracts. Bioactivity testing revealed that FZBL-2 showed the strongest overall antioxidant, anti-lipid peroxidation, and enzyme-inhibitory activities, along with the strongest antiproliferative activity in HuH-7 cancer cell, while FZBL-3 displayed the greatest antiproliferative activities in SGC-7901 and MDA-MB-231 cancer cells. Multivariate correlation analysis identified CDD, cinchonain Id, kaempferol triglucoside, and HKDG key metabolites associated with the bioactivities of FZBL. These findings provide valuable insights into the bioactive constituents underlying FZBL's functional properties and supports further investigation into natural health-promoting ingredient.
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