Development and optimization of green extraction of naringin in Citrus grandis ‘Tomentosa’ using natural deep eutectic solvents (NADES) and investigation of lung cancer therapeutics mechanisms

Abstract

Natural bioactive compounds derived from traditional Chinese medicine have garnered widespread research interest. In this study, a green and efficient method for extracting naringin from Citrus grandis ‘Tomentosa’ (CGT) was developed using ultrasonic-assisted extraction (UAE) with the natural deep eutectic solvent (NADES) composed of choline chloride and levulinic acid (ChCl–Le). The extraction was carried out under the conditions of 40% (w/w) water content in NADES, a solid-to-liquid ratio of 1 : 65 (w/v), a temperature of 60 °C, an extraction time of 40 minutes, and an ultrasonic power of 300 W. Despite optimized and practical conditions, a high naringin yield of 4.7% was attained. This yield remains significantly greater than those obtained with methanol (4.1%) or ethanol (4.0%). Furthermore, the CGT extracts were evaluated for lung cancer therapeutics activity, leading to the identification of several key bioactive compounds, including poncirin, isosinensetin, sinensetin, nobiletin, didymin, neohesperidin, and naringenin. These components are known to exert therapeutic effects against lung cancer by targeting CYP17A1, CYP19A1, and AR, as well as by modulating pathways such as EGFR tyrosine kinase inhibitor resistance and prostate cancer. This study successfully establishes an eco-efficient extraction method for naringin and clarifies the multi-target mechanism of action of CGT extracts against lung cancer.