Separation of Ascorbic Acid and Isoascorbic Acid (DE-413L)

Ascorbic acid and isoascorbic acid were separatd using a column for polymer-based reversed phase chromatography, RSpak DE-413L. The calibration curve of ascorbic acid and isoascorbic acid shows good linearity respectively in the range from 1 to 100 μg/mL.
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Sample : 10 μL
5μg/mL each (in 0.1 % metaphosphoric acid)
1. Ascorbic acid
2. Isoascorbic acid

Column       : Shodex RSpak DE-413L (4.6 mm I.D. x 250 mm)
Eluent       : 10 mM H3PO4 aq.
Flow rate    : 0.6 mL/min
Detector     : UV (254 nm)
Column temp. : 40 °C