1290 series uplc system

The powered E. phaseoloides leaves (100 mg) was extracted by sonication with 15 mL of acetone for 60 min at 50 °C. The extracted solution was filtered and then evaporated under reduced pressure. The acetone extract was prepared in methanol to concentration 500 ppm, it was filtered twice through Millipore 0.20 μm PTFE filters (Millex-LG, Tokyo, Japan). The experiments were performed on an Agilent 1290 Series UPLC system equipped with a 1290 photodiode array detector (DAD) (Agilent Technologies, Santa Clara, CA, USA) coupled to an Agilent 6545 q-TOF hybrid mass spectrometer (MS) with a dual electrospray ionization (ESI) source for simultaneous spraying of a mass reference solution that enabled continuous calibration of detected m/z values was used for the analysis of the samples. The column was a Poroshell 120 EC-C18 (2.1 × 100 mm, 2.7 μm), Agilent Technologies, Santa Clara, CA, USA). The gradient system of 0.1% formic acid in water (solvent A) and 0.1% formic acid in acetonitrile (solvent B) as follows 0–20 min, 10–95% B; 20–25 min, 95% B. The time for re-equilibration to initial conditions was 5 min.

The Agilent 1290 series UPLC system was used for chromatographic analysis. Samples were separated with an ACQUITY HSS T3 column (2.1 mm × 100 mm, 1.8 μm) at 4 °C. The mobile phase consisted of 0.1% formic acid in water (A) and acetonitrile (B). The conditions of the mobile phase were as follows: 0–4 min, 5–10% B; 4–10 min, 10–50% B; 10–20 min, 50–80% B; 20–25 min, 80–95% B; 25–30 min, 95–95% B; 30–32 min, 95–5% B; 32–35 min, 5–5% B. The column temperature was set at 35 ℃, the injection volume was 10 µL, and the flow rate was 0.3 mL/min.
An Agilent 6550 Q-TOF-MS instrument (Agilent Technologies, Santa Clara, CA, USA) was used. The mass spectrometry analytical conditions were as follows: The electrospray capillary voltage was 3.5 kV in the negative ionization mode and 4.0 kV in the positive ionization mode. The gas temperature was 250 °C and the gas flow was 15 L/min in both negative and positive ionization modes. The nozzle voltage was 1000 V and the mass range was set from m/z 50 to 1400.