Instrument | Arc HPLC system(Waters, Milford, MA, USA) |
Column | Zorbax eclipse XDB-C18 4.6×250 mm, 5 μm(Agilent technologies, SantaClara, CA, USA) |
Detector | PDA Detector(Ex: 210 nm, Em: 280 nm)(2998, Waters, Milford, MA, USA) |
Mobile phase | A: 0.1% formic acid in 10% AcetonitrileB: 0.1% formic acid in 90% Acetonitrile |
Gradient | 0-2 min | A : B = 100 : 0 |
2-8 min | A : B = 100 : 0 |
8-35 min | A : B = 90 : 10 |
35-40 min | A : B = 45 : 55 |
40-45 min | A : B = 50 : 50 |
45 min | A : B = 100 : 0 |
Flow rate | 1.0 mL/min |
Column temp | 35°C |
Injection volume | 10 μL |