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

Ascorbic acid and isoascorbic acid were separatd using two columns of RSpak DE-413 (a column for polymer-based reversed phase chromatography). The calibration curve of ascorbic acid and isoascorbic acid shows good linearity respectively in the range from 1 to 100 μg/mL. 


Sample : 10 μL
5 μg/mL each (in 0.1 % metaphosphoric acid)
1. Ascorbic acid
2. Isoascorbic acid

Column       : Shodex RSpak DE-413 (4.6 mm I.D. x 150 mm) x 2
Eluent       : 10 mM H3PO4 aq.
Flow rate    : 0.6 mL/min
Detector     : UV (254 nm)
Column temp. : 40 ℃