2.13. The method of analysis sample using HPLC or HPLC-MS/MS

Using Agilent 1260II HPLC system for high performance liquid chromatography (HPLC) analysis(Agilent, USA). Samples were separated using an Agilent EclipsePlus C18 column (250 × 4.6 mm, 5 μm). The mobile phase was a mixture of methanol & water (95:5, v/v), and the flow rate was 1.0 mL/min. Absorbance was monitored at 227 nm.

Using Agilent Eclipse plus C18 column (4.6 mm × 250 mm, 5 μm), gradient elution was performed at a UV detection wavelength of 227 nm. The elution program was water(A): acetonitrile(B), 0-14 min: 68%A-32%B; 1 4∼27 min: 100%A-0%B; 2 7 ～28 min: 68%A-32%B, 2 9 ∼ 33 min and the flow rate of 1 mL/min, inject 20 μL of the plasma sample to be tested.

MS conditions are shown in the table below: