HPLC Separation of Ascorbic and Dehydroascorbic Acids


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.

Condition

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

 

Description

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 options
Application Analytes:
Ascorbic Acid
Dehydroascorbic Acid

Application Detection:
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.