ACQUITY UPLC HILIC Isocratic Separation of Isoascorbic acid and ascorbic acid

Column:

ACQUITY UPLC BEH Amide,

Isocratic Mobile Phase:

80/20 MeCN/H₂O with 10 mM KH₂PO₄, pH 4.6

Flow Rate:

0.2 mL/min

Injection Volume:

5.0 μL (PLNO)

Sample Concentration:

30 μg/mL each

Sample Diluent:

75/25 MeCN/MeOH with 0.2% HCOOH

Column Temperature:

25 °C

Weak Needle Wash:

95/5 MeCN/H₂O

Detection:

UV @ 260nm

Sampling Rate:

20 points/sec

Filter Time Constant:

0.2

Instrument:

Waters ACQUITY UPLC with ACQUITY UPLC PDA Detector