Column:

HP C18 ( 5 µm, 120 Å, 4.6 x 250 mm )

Mobile phase:

A: [ H₂O : 3.12% NaH₂PO₄ : 8%TBAOH = 576 : 200 : 24, pH 5.5 by H₃PO₄] : ACN = 800 : 200, B: [ H₂O : 3.12% NaH₂PO₄ : 8%TBAOH = 126 : 200 : 24, pH 5.5 by H₃PO₄] : ACN = 350 : 650

Flow rate:

1.0 mL/min

Column temperature:

40 ℃

Detection:

UV 220 nm

Injection volume:

20 µL