Comparison of high-performance liquid chromatography and spectrofluorimetry for vitamin C analysis of green beans ( Phaseolus vulgaris L.)

Abstract:  A spectrofluorimetric method and a chromatographic method [high-performance liquid chromatography – ultraviolet (HPLC-UV) detection] were compared for vitamin C analysis of green beans. For HPLC-UV, the determination was performed before [ascorbic acid (AA)] and after reduction with dithiothreitol [AA + dehydroascorbic acid (DHAA)]; for spectrofluorimetry, DHAA was determined after oxidation with activated charcoal. The fluorimetric determination of DHAA before oxidation was also carried out, and showed good precision but poor accuracy (very high recovery percentages). HPLC-UV showed better linearity and sensitivity, while spectrofluorimetry was more precise. Both methods were suitable for green bean samples, and the choice of which one to use depends on the particular interest of the analysis: spectrofluorimetry may be preferable when the total content of vitamin C is required, but not for individual analysis of each form. HPLC-UV is more suitable when the interest of the analysis is the ascorbic acid form as a deterioration index of vegetable products.