HPLC analysis with fluorometric detection of vitamin C in food samples

A high performance liquid chromatographic (HPLC) procedure has been developed for the analysis of ascorbic acid and dehydroascorbic acid in complex matrices. Separation is accomplished with an anion-exchange resin and fluorescent detection is achieved through post-column inline chemistry, involving oxidation of ascorbic acid to dehydroascorbic acid followed by reaction with o-phenylenediamine to form a fluorescent product. Lower limits of detection for both forms of vitamin C are well below the levels found in the usual food sources of this vitamin. The extraction procedures developed yield clean samples for analysis with minimal loss of the vitamers during the analytical procedures. Recoveries are in the range of 90-107%. The results obtained with this HPLC procedure agree well with those obtained with a modified version of the classical procedure of Deutsch and Weeks. A variety of foods including fruit juices, vegetables, and fruits were analyzed.