HPLC Method for Naringenin Detection

Naringenin titers were detected at 290 nm using an Agilent 1260 HPLC system (Waldbronn, Germany) equipped with a diode array detector (DAD) 1260 model VL + (G7115A) and a C18 column (3 × 100 mm 2.7 µm). The column was eluted with gradient elution at 30°C and 0.3 mL min⁻¹ flow rate: 10% to 40% acetonitrile/water (vol/vol) for 5 min, 40% acetonitrile (vol/vol) for 7 min, 40% to 95% acetonitrile (vol/vol) for 3 min, and 95% to 10% acetonitrile (vol/vol) for 3 min. It was noted that 0.3% acetic acid (vol/vol) was added to above mobile phases, including the acetonitrile or water, which contributed to naringenin separation.