HPLC Separation of Isomers of Ascorbic Acid and Ascorbic Acid-2-Phosphate in HILIC/Mixed-Mode on Amaze HD Column

Conditions of Experiment
Column: Amaze HA
Separation Modes: hydrophilic acidic
Column Dimenstions: 3.0x100 mm, 3 um, 100A
Mobile Phase: from 85% ACN with 30 mM Ammonium Acetate pH 4
to 50% ACN with 10 mM Ammonium Acetate pH 4 in 10 min
Detection: 280 nm
Sample: 0.2 mg/ml
Injection: 2 uL
Flow rate: 0.6 ml/min
Analytes
Class of compounds: Aromatic acid, Metabolite, Organic acid, Supplement, Vitamin
Nature of compounds: Acidic, Hydrophilic, Polar
Compounds: Isoascorbic acid, Ascorbic acid, Ascorbic acid 2-phosphate