Simultaneous analysis of ascorbic and dehydroascorbic acid in fruit juice using HILIC chromatography coupled with mass spectrometry

A HILIC-MS/MS method was developed for the simultaneous analysis of ascorbic acid (AA) and dehydroascorbic acid (DHA). Ammonium acetate buffer and acetonitrile were used as mobile phases under gradient elution mode at a flow rate of 0.3 mL/min. A SCIEX Triple QUAD 5500+ system was operated under MRM mode to acquire data with 175 → 115 and 173→113 as the quantifier ions for AA and DHA, respectively. Samples were prepared with a simple dilute and shoot method. A calibration and accuracy study was performed as part of method validation in fruit juices. The accuracy profiles were estimated from the measurement uncertainty determined from the validation data according to the ISO/DTS 21748. The recoveries of the analytes were between 92 -130%. The LOQs achieved in the current study for AA and DHA were 4.5 µg/L and 18 µg/L, respectively. These LOQs were substantially lower than what has been previously reported in the literature. The study concludes with a straightforward analytical method for the determination of total vitamin C content that does not involve timing consuming steps i.e., reduction or derivatization reaction. The method can be implemented in quality control labs for routine analysis of food samples.