Ascorbic acid is a vital vitamin for humans. It also serves as an antioxidant. Dehydroascorbic acid is an oxidation product of ascorbic acid. Both molecules are very polar, and cannot be retained by reverse-phase mechanism. Generic method for HPLC separation of ascorbic and dehydroascorbic acid was developed on a Primesep SB mixed-mode HPLC column. Both compounds are well separated and produce excellent peak shape. Method can be used for analysis of both compounds in biofluids. Proteins from biofluids will not retain due to repulsion effect of the stationary phase. Compounds can be monitored by common detection techniques like UV, ELSD, CAD, and LC/MS.
| Column | Primesep SB , 4.6×50 mm, 5 µm, 100A |
| Mobile Phase | MeCN/H2O |
| Buffer | Formic Acid |
| Flow Rate | 1.0 ml/min |
| Detection | ELSD |
| Class of Compounds |
Acid, Vitamin B₆, Hydrophobic, Ionizable |
| Analyzing Compounds | Ascorbic acid (Vitamin C), Dehydroascorbic acid |
Application Column
Primesep SB
The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.
Select optionsDehydroascorbic Acid
