A High-Performance Liquid Chromatography with Electrochemical Detection Method Developed for the Sensitive Determination of Ascorbic Acid: Validation, Application, and Comparison with Titration, Spectrophotometric, and High-Performance Liquid Chromatography with Diode-Array Detection Methods

L-ascorbic acid (vitamin C, VC), an essential nutrient obtained from the diet to maintain various vital signs for the human body, is a crucial indicator of food quality and nutritional value. Herein, high-performance liquid chromatography with electrochemical detection (HPLC-ECD) was developed and validated with the advantages of higher sensitivity, simpler operation processes, and more rapid detection in measuring VC levels in honey samples when compared with the common methods (titration, spectrophotometric, and HPLC-DAD methods). The results of the HPLC-ECD methodological validation showed that the limit of detection (LOD) was 0.0043 µg mL-1; the relative standard deviations (RSDs) of the intra- and inter-day values were between 2.51% and 5.15%, and the regression coefficient was >0.999 in the linear range of 0.1 to 20 µg mL-1. The validated HPLC-ECD method was also successfully utilized to evaluate the VC levels in different varieties of honey samples with various storage durations as well as in fruit and biological samples. This study provided a perspective for the further accurate determination of VC content in food and biological samples.