In food analysis, a method for determination of vitamin C should enable measuring of total content of
ascorbic acid (AA) and dehydroascorbic acid (DHAA) because both chemical forms exhibit biological
activity. The aim of the work was to confirm applicability of HPLC–DAD method for analysis of total
content of vitamin C (TC) and ascorbic acid in various types of food by determination of validation parameters
such as: selectivity, precision, accuracy, linearity and limits of detection and quantitation. The
results showed that the method applied for determination of TC and AA was selective, linear and precise.
Precision of DHAA determination by the subtraction method was also evaluated. It was revealed that the
results of DHAA determination obtained by the subtraction method were not precise which resulted
directly from the assumption of this method and the principles of uncertainty propagation. The proposed
chromatographic method should be recommended for routine determinations of total vitamin C in
various food.