Development and Evaluation of HPLC-UV Method for Astaxanthin in Health Functional Foods Code

The HFF (Health Functional Foods) Code provides protocols for the quantitative analysis of 68 types of HFF ingredients. Astaxanthin is one of the HFF with antioxidant properties, preventing cell damage and reducing eye fatigue, becoming popular in the HFF market. To improve the astaxanthin assay protocol, we changed the LC condition from a tertiary pump to a binary pump and optimized the mobile phase and gradient accordingly. Based on the differences in the enzymes, we also observed the efficiency of the conversion of astaxanthin ester to free astaxanthin. We validated our modified method based on AOAC guidelines, including linearity (R20.99), selectivity, accuracy (recovery 92~105%), and precision (%RSD 2). An applicability was evaluated on 15 health functional food products, and the recovery values were satisfied within the range of 80-120%. In this study, we modernized the analytical method of astaxanthin by modifying the LC conditions, and standardized the cholesterol esterase with the highest efficiency.