Microfluorometry Assay for Vitamin C

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Abstract
The reaction of dehydroascorbic acid with o-phenylenediamine to give a fluorescent quinoxaline has been used as the basis of an assay to determine microgram quantities of ascorbic plus dehydroascorbic acids. The development of the fluorescent derivative of the vitamin is prevented by forming a boric acid-dehydroascorbic acid complex prior to addition of the diamine solution. This provides a means of differentiating between the fluorescence from the vitamin and that from possible interfering substances. When applied to pharmaceutical preparations, beverages, and special dietary foods, the method shows a high degree of specificity. No substances tested have been found to interfere. Comparison of results with those obtained by using other methods showed excellent agreement.

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