For the analysis of ascorbic acid, amino columns or silica columns are usually used. Analysis using amino columns is particularly wide spread because isoascorbic acid (erythorbic acid) as well as ascorbic acid can be analyzed simultaneously. However, conventional silica-based amino columns are not chemically stable, therefore, they have certain disadvantage such as a decrease in retention and short column life. Since Asahipak NH2P-50 4E is a polymer-based column, it does not suffer from such disadvantages. Moreover, since NH2P-50 column can be washed with alkaline solvents, the column life can be extended.
Sample : 5micro-g/mL each, 10micro-L
1. Erythorbic acid
2. L-Ascorbic acid
Column : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm)
Eluent : 20mM NaH2PO4+30mM H3PO4 aq./CH3CN=20/80
Flow rate : 1.0mL/min
Detector : UV(254nm)
Column temp. : 35deg-C
Equilibration of NH2P-50 4E column : After passing 60mM Phosphoric acid in the column by 0.5 mL/min for 2 hours, it replaces by eluent.