A NON-ION-FAIRING HPLC METHOD FOR MEASURING NEW FORMS OF ASCORBATE AND ASCORBIC-ACID

A new high-performance liquid-chromatography (HPLC) method separates v itamins L-ascorbic acid (C-1) and L-ascorbyl-2-sulfate (C-2) Previous HPLC reverse phase methods utilized ion-pairing reagents and resulted in co-elution with other substances. Vitamin C-1 preceded C-2 in reten tion time. In this new method, which deletes the ion-pairing reagent, vitamin C-2 precedes the elution of vitamin C-1. This new direct metho d allows for a better separation from other conflicting metabolites an d constituents of extraction mixtures. In addition, this method provid es a way of identifying ascorbyl-2-monophosphate (C-3), a form of vita min C that may be found in animal diets. Examples will be shown of HPL C profiles of extractives from tissue and diets known to contain these new forms of vitamin C.