A previously reported method was slightly modified for hydrolysis and high-performance liquid chromatography (HPLC) analysis (28 (link)). EE was dissolved in ethanol–hydrochloric acid–water (7/2/1, v/v/v) solution and at 75°C by ultrasound (40 kHz) for 60 min. Then, the supernatant was used in the following analysis.
HPLC was conducted by using a Shimadzu HPLC system (Shimadzu, Japan), which consisted of a pump (LC-20AT), diode array detector (DAD, SPD-M20A), C18 column (Agilent 250 × 4.6 mm), and LC-solution system manager program. The mobile phase comprised methyl alcohol–water–acetic acid in a ratio of 500/500/0.4 (v/v/v). The flow rate was 1 mL/min, and the eluate absorbance was monitored at 370 nm using a scanning range of 200–600 nm. The injection volume of the extract was 10 μL.
HPLC was conducted by using a Shimadzu HPLC system (Shimadzu, Japan), which consisted of a pump (LC-20AT), diode array detector (DAD, SPD-M20A), C18 column (Agilent 250 × 4.6 mm), and LC-solution system manager program. The mobile phase comprised methyl alcohol–water–acetic acid in a ratio of 500/500/0.4 (v/v/v). The flow rate was 1 mL/min, and the eluate absorbance was monitored at 370 nm using a scanning range of 200–600 nm. The injection volume of the extract was 10 μL.
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