*4.5. UHPLC—Purity of Extracted Hesperidin*

The HSD purity was determined utilizing Ultra-High-Performance Liquid (UHPLC—Waters, Milford, MA, USA) with the reverse stationary phase (Zorbax Eclipse C18, length 10 cm, 5 μm, Agilent Technologies Inc., Santa Clara, CA, USA). The mobile phase was prepared with methanol and water (60:40 *v*/*v*) with the addition of phosphoric acid (0.1%). A hesperidin analytical standard (Sigma-Aldrich, >97% of purity, St. Louis, MO, USA) was used for calibration curve construction from the injection of aliquots of 1, 2, 4, 6, 8, and 10 μL of standard solution, HSD in methanol (100 ppm). Calibration curves and the analysis of the sample from the extraction were performed in triplicate. The following method was adapted from Weon et al. (2012) [59].
