Analytical chromatographic conditions

Column

Eclipse Plus C18 (50 × 4.6 1.8 μ)

Mobile phase

Acetonitrile (B), 10mM Ammonium bicarbonate (A)

Program

Gradient

Flow rate

0.6 mL/min

Wavelength

280 nm

Column Temperature

35˚C

Injection volume

2 μL

Diluent

Mobile phase (50:50)

Gradient program

0/3, 0.3/3, 3.2/98, 4/98, 4.1/3

(Time/%B)