HPLC Analysis of Ascorbic and Dehydroascorbic acids on Coresep SB Column in reversed-Phase Anion-Exchange Modes

Application description

Ascorbic acid, or Vitamin C, is a vitamin found in plants, fruits and other food-related sources. It is one of the main dietary supplements. Vitamin C is an essential nutrient involved in repair of tissues. It is available as generic over-the-counter medication. Dehydroascorbic acid is an oxidized form of ascorbic acid. Both ascorbic and dehydroascorbic acids have similar biological activity as antiviral medications, but dehydroascorbic acid also has neuroprotective effects. Both compounds are very polar, with limited or no retention in reversed-phase chromatography. Both acids were analyzed and retained by weak reversed-phase and anion-exchange mechanisms on a core-shell mixed-mode anion-exchange column. The method and column can be used for analysis of these and other hydrophilic acids by mixed-mode chromatography. The method is compatible with LC/MS, ELSD and CAD and can be used for quantitation of ascorbic and dehydroascorbic acids in various foods, drug formulations and biofluids. Coresep SB column combines unique selectivity of mixed-mode with ultra-high efficiency of core-shell particles technology. HPLC method is reproducible and robust.

Conditions of Experiment
Column: Coresep SB
Separation Modes: reversed-phase and anion-exchange
Column Dimenstions: 3.2 x 100 mm, 2.7 um, 90A
Mobile Phase: 5% ACN with 0.05% of formic acid
Detection: UV 250 nm
Sample: 0.5 mg/ml
Injection: 1 uL
Flow rate: 0.8 ml/mim
Class of compounds: Aromatic acid, Organic acid, Supplement, Vitamin
Nature of compounds: Acidic, Hydrophilic, Polar
Compounds: Dehydroascorbic acid, Ascorbic acid